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Volumn 295, Issue 1, 2008, Pages

In vivo administration of calpeptin attenuates calpain activation and cardiomyocyte loss in pressure-overloaded feline myocardium

Author keywords

Cardiomyopathy; Caspases; Cytoskeleton; Heart failure

Indexed keywords

CALPAIN; CALPASTATIN; CALPEPTIN; CASPASE 3; CASPASE INHIBITOR; GELSOLIN; HISTONE H2B; N BENZOYLCARBONYLVALYLALANYLASPARTYLFLUOROMETHYLKETONE; UNCLASSIFIED DRUG; BENZYLOXYCARBONYL VALYL ASPARTIC ACID FLUOROMETHYL KETONE; BENZYLOXYCARBONYL-VALYL-ASPARTIC ACID FLUOROMETHYL KETONE; CASP3 PROTEIN, RAT; CYSTEINE PROTEINASE INHIBITOR; DIPEPTIDE; HISTONE; PEPTIDE CHLOROMETHYL KETONE;

EID: 49849088828     PISSN: 03636135     EISSN: 15221539     Source Type: Journal    
DOI: 10.1152/ajpheart.00085.2008     Document Type: Article
Times cited : (47)

References (68)
  • 2
    • 41249092847 scopus 로고    scopus 로고
    • Ultrastructural definition of apoptosis in heart failure
    • Arbustini E, Brega A, Narula J. Ultrastructural definition of apoptosis in heart failure. Heart Fail Rev 13: 121-135, 2008.
    • (2008) Heart Fail Rev , vol.13 , pp. 121-135
    • Arbustini, E.1    Brega, A.2    Narula, J.3
  • 3
    • 0034635549 scopus 로고    scopus 로고
    • Gelsolin in complex with phosphatidylinositol 4,5-bisphosphate inhibits caspase-3 and -9 to retard apoptotic progression
    • Azuma T, Koths K, Flanagan L, Kwiatkowski D. Gelsolin in complex with phosphatidylinositol 4,5-bisphosphate inhibits caspase-3 and -9 to retard apoptotic progression. J Biol Chem 275: 3761-3766, 2000.
    • (2000) J Biol Chem , vol.275 , pp. 3761-3766
    • Azuma, T.1    Koths, K.2    Flanagan, L.3    Kwiatkowski, D.4
  • 5
    • 0031049642 scopus 로고    scopus 로고
    • A new mechanism of methylprednisolone and other corticosteroids action demonstrated in vitro: Inhibition of a proteinase (calpain) prevents myelin and cytoskeletal protein degradation
    • Banik NL, Matzelle D, Terry E, Hogan EL. A new mechanism of methylprednisolone and other corticosteroids action demonstrated in vitro: inhibition of a proteinase (calpain) prevents myelin and cytoskeletal protein degradation. Brain Res 748: 205-210, 1997.
    • (1997) Brain Res , vol.748 , pp. 205-210
    • Banik, N.L.1    Matzelle, D.2    Terry, E.3    Hogan, E.L.4
  • 6
    • 0030851194 scopus 로고    scopus 로고
    • Calpain and calpastatin in myoblast differentiation and fusion: Effects of inhibitors
    • Barnoy S, Glaser T, Kosower NS. Calpain and calpastatin in myoblast differentiation and fusion: effects of inhibitors. Biochim Biophys Acta 1358: 181-188, 1997.
    • (1997) Biochim Biophys Acta , vol.1358 , pp. 181-188
    • Barnoy, S.1    Glaser, T.2    Kosower, N.S.3
  • 8
    • 3543012019 scopus 로고    scopus 로고
    • Structure of the N-terminal half of gelsolin bound to actin: Roles in severing, apoptosis and FAF
    • Burtnick LD, Urosev D, Irobi E, Narayan K, Robinson RC. Structure of the N-terminal half of gelsolin bound to actin: roles in severing, apoptosis and FAF. EMBO J 23: 2713-2722, 2004.
    • (2004) EMBO J , vol.23 , pp. 2713-2722
    • Burtnick, L.D.1    Urosev, D.2    Irobi, E.3    Narayan, K.4    Robinson, R.C.5
  • 10
    • 0037453582 scopus 로고    scopus 로고
    • Death hath a thousand doors to let out life
    • Chandrashekhar Y, Narula J. Death hath a thousand doors to let out life. Circ Res 92: 710-714, 2003.
    • (2003) Circ Res , vol.92 , pp. 710-714
    • Chandrashekhar, Y.1    Narula, J.2
  • 11
    • 0037047326 scopus 로고    scopus 로고
    • Calpain and mitochondria in ischemia/reperfusion injury
    • Chen M, Won DJ, Krajewski S, Gottlieb RA. Calpain and mitochondria in ischemia/reperfusion injury. J Biol Chem 277: 29181-29186, 2002.
    • (2002) J Biol Chem , vol.277 , pp. 29181-29186
    • Chen, M.1    Won, D.J.2    Krajewski, S.3    Gottlieb, R.A.4
  • 13
    • 23844464107 scopus 로고    scopus 로고
    • The N-terminal fragment of gelsolin inhibits the interaction of DNase I with isolated actin, but not with the cofilin-actin complex
    • Chhabra D, Nosworthy NJ, dos Remedios CG. The N-terminal fragment of gelsolin inhibits the interaction of DNase I with isolated actin, but not with the cofilin-actin complex. Proteomics 5: 3131-3136, 2005.
    • (2005) Proteomics , vol.5 , pp. 3131-3136
    • Chhabra, D.1    Nosworthy, N.J.2    dos Remedios, C.G.3
  • 14
    • 29144456760 scopus 로고    scopus 로고
    • Chohan PK, Singh RB, Dhalla NS, Netticadan T. L-Arginine administration recovers sarcoplasmic reticulum function in ischemic reperfused hearts by preventing calpain activation. Cardiovasc Res 69: 152-163, 2006.
    • Chohan PK, Singh RB, Dhalla NS, Netticadan T. L-Arginine administration recovers sarcoplasmic reticulum function in ischemic reperfused hearts by preventing calpain activation. Cardiovasc Res 69: 152-163, 2006.
  • 15
    • 57049085795 scopus 로고    scopus 로고
    • Colucci WS, Braunwald E. Pathophysiology of heart failure. In: Heart Disease (6th ed.), edited by Braunwald E, Zippes DP, Libby P. Philadelphia: Saunders, 2001, p. 503-533.
    • Colucci WS, Braunwald E. Pathophysiology of heart failure. In: Heart Disease (6th ed.), edited by Braunwald E, Zippes DP, Libby P. Philadelphia: Saunders, 2001, p. 503-533.
  • 16
    • 0033564419 scopus 로고    scopus 로고
    • Increased cardiomyocyte apoptosis and changes in proapoptotic and antiapoptotic genes bax and bcl-2 during left ventricular adaptations to chronic pressure overload in the rat
    • Condorelli G, Morisco C, Stassi G, Notte A, Farina F, Sgaramella G, de Rienzo A, Roncarati R, Trimarco B, Lembo G. Increased cardiomyocyte apoptosis and changes in proapoptotic and antiapoptotic genes bax and bcl-2 during left ventricular adaptations to chronic pressure overload in the rat. Circulation 99: 3071-3078, 1999.
    • (1999) Circulation , vol.99 , pp. 3071-3078
    • Condorelli, G.1    Morisco, C.2    Stassi, G.3    Notte, A.4    Farina, F.5    Sgaramella, G.6    de Rienzo, A.7    Roncarati, R.8    Trimarco, B.9    Lembo, G.10
  • 17
    • 0031058676 scopus 로고    scopus 로고
    • Basic determinants of myocardial hypertrophy: A review of molecular mechanisms
    • Cooper GT. Basic determinants of myocardial hypertrophy: a review of molecular mechanisms. Annu Rev Med 48: 13-23, 1997.
    • (1997) Annu Rev Med , vol.48 , pp. 13-23
    • Cooper, G.T.1
  • 18
    • 0015693837 scopus 로고
    • Mechanisms for the abnormal energetics of pressure-induced hypertrophy of cat myocardium
    • Cooper GT, Satava RM Jr, Harrison CE, Coleman HN. Mechanisms for the abnormal energetics of pressure-induced hypertrophy of cat myocardium. Circ Res 33: 213-223, 1973.
    • (1973) Circ Res , vol.33 , pp. 213-223
    • Cooper, G.T.1    Satava Jr, R.M.2    Harrison, C.E.3    Coleman, H.N.4
  • 19
    • 33846906024 scopus 로고    scopus 로고
    • The calpain system as a modulator of stress/damage response
    • Demarchi F, Schneider C. The calpain system as a modulator of stress/damage response. Cell Cycle 6: 136-138, 2007.
    • (2007) Cell Cycle , vol.6 , pp. 136-138
    • Demarchi, F.1    Schneider, C.2
  • 21
    • 0027444694 scopus 로고
    • On the role of the platelet membrane skeleton in mediating signal transduction. Association of GP IIb-IIIa, pp60c-src, pp62c-yes, and the p21ras GTPase-activating protein with the membrane skeleton
    • Fox JE, Lipfert L, Clark EA, Reynolds CC, Austin CD, Brugge JS. On the role of the platelet membrane skeleton in mediating signal transduction. Association of GP IIb-IIIa, pp60c-src, pp62c-yes, and the p21ras GTPase-activating protein with the membrane skeleton. J Biol Chem 268: 25973-25984, 1993.
    • (1993) J Biol Chem , vol.268 , pp. 25973-25984
    • Fox, J.E.1    Lipfert, L.2    Clark, E.A.3    Reynolds, C.C.4    Austin, C.D.5    Brugge, J.S.6
  • 22
    • 0032426430 scopus 로고    scopus 로고
    • Caspase-3-induced gelsolin fragmentation contributes to actin cytoskeletal collapse, nucleolysis, and apoptosis of vascular smooth muscle cells exposed to proinflammatory cytokines
    • Geng YJ, Azuma T, Tang JX, Hartwig JH, Muszynski M, Wu Q, Libby P, Kwiatkowski DJ. Caspase-3-induced gelsolin fragmentation contributes to actin cytoskeletal collapse, nucleolysis, and apoptosis of vascular smooth muscle cells exposed to proinflammatory cytokines. Eur J Cell Biol 77: 294-302, 1998.
    • (1998) Eur J Cell Biol , vol.77 , pp. 294-302
    • Geng, Y.J.1    Azuma, T.2    Tang, J.X.3    Hartwig, J.H.4    Muszynski, M.5    Wu, Q.6    Libby, P.7    Kwiatkowski, D.J.8
  • 24
    • 0029123733 scopus 로고
    • Exogenous gelsolin binds to sarcomeric thin filaments without severing
    • Gonsior S, Hinssen H. Exogenous gelsolin binds to sarcomeric thin filaments without severing. Cell Motil Cytoskeleton 31: 196-206, 1995.
    • (1995) Cell Motil Cytoskeleton , vol.31 , pp. 196-206
    • Gonsior, S.1    Hinssen, H.2
  • 26
    • 37549040627 scopus 로고    scopus 로고
    • Calpain inhibition attenuates right ventricular contractile dysfunction after acute pressure overload
    • Greyson CR, Schwartz GG, Lu L, Ye S, Helmke S, Xu Y, Ahmad H. Calpain inhibition attenuates right ventricular contractile dysfunction after acute pressure overload. J Mol Cell Cardiol 44: 59-68, 2008.
    • (2008) J Mol Cell Cardiol , vol.44 , pp. 59-68
    • Greyson, C.R.1    Schwartz, G.G.2    Lu, L.3    Ye, S.4    Helmke, S.5    Xu, Y.6    Ahmad, H.7
  • 27
    • 25844513258 scopus 로고    scopus 로고
    • Left ventricular dysfunction after acute intracranial hypertension is associated with increased hydroxyl free radical production, cardiac ryanodine hyperphosphorylation, and troponin I degradation
    • Hall SR, Wang L, Milne B, Hong M. Left ventricular dysfunction after acute intracranial hypertension is associated with increased hydroxyl free radical production, cardiac ryanodine hyperphosphorylation, and troponin I degradation. J Heart Lung Transplant 24: 1639-1649, 2005.
    • (2005) J Heart Lung Transplant , vol.24 , pp. 1639-1649
    • Hall, S.R.1    Wang, L.2    Milne, B.3    Hong, M.4
  • 29
    • 0034625535 scopus 로고    scopus 로고
    • Apoptosis and heart failure: A critical review of the literature
    • Kang PM, Izumo S. Apoptosis and heart failure: a critical review of the literature. Circ Res 86: 1107-1113, 2000.
    • (2000) Circ Res , vol.86 , pp. 1107-1113
    • Kang, P.M.1    Izumo, S.2
  • 30
    • 34547093646 scopus 로고    scopus 로고
    • Calpain-dependent calpastatin cleavage regulates caspase-3 activation during apoptosis of Jurkat T cells induced by Entamoeba histolytica
    • Kim KA, Lee YA, Shin MH. Calpain-dependent calpastatin cleavage regulates caspase-3 activation during apoptosis of Jurkat T cells induced by Entamoeba histolytica. Int J Parasitol 37: 1209-1219, 2007.
    • (2007) Int J Parasitol , vol.37 , pp. 1209-1219
    • Kim, K.A.1    Lee, Y.A.2    Shin, M.H.3
  • 32
    • 0034685808 scopus 로고    scopus 로고
    • Gelsolin inhibits apoptosis by blocking mitochondrial membrane potential loss and cytochrome c release
    • Koya RC, Fujita H, Shimizu S, Ohtsu M, Takimoto M, Tsujimoto Y, Kuzumaki N. Gelsolin inhibits apoptosis by blocking mitochondrial membrane potential loss and cytochrome c release. J Biol Chem 275: 15343-15349, 2000.
    • (2000) J Biol Chem , vol.275 , pp. 15343-15349
    • Koya, R.C.1    Fujita, H.2    Shimizu, S.3    Ohtsu, M.4    Takimoto, M.5    Tsujimoto, Y.6    Kuzumaki, N.7
  • 33
    • 33744525885 scopus 로고    scopus 로고
    • How do cardiomyocytes die? Apoptosis and autophagic cell death in cardiac myocytes
    • Kunapuli S, Rosanio S, Schwarz ER. "How do cardiomyocytes die?" Apoptosis and autophagic cell death in cardiac myocytes. J Card Fail 12: 381-391, 2006.
    • (2006) J Card Fail , vol.12 , pp. 381-391
    • Kunapuli, S.1    Rosanio, S.2    Schwarz, E.R.3
  • 34
    • 0031058040 scopus 로고    scopus 로고
    • Association of tyrosine-phosphorylated c-Src with the cytoskeleton of hypertrophying myocardium
    • Kuppuswamy D, Kerr C, Narishige T, Kasi VS, Menick DR, Cooper GT. Association of tyrosine-phosphorylated c-Src with the cytoskeleton of hypertrophying myocardium. J Biol Chem 272: 4500-4508, 1997.
    • (1997) J Biol Chem , vol.272 , pp. 4500-4508
    • Kuppuswamy, D.1    Kerr, C.2    Narishige, T.3    Kasi, V.S.4    Menick, D.R.5    Cooper, G.T.6
  • 35
    • 0034609764 scopus 로고    scopus 로고
    • Human gelsolin prevents apoptosis by inhibiting apoptotic mitochondrial changes via closing VDAC
    • Kusano H, Shimizu S, Koya RC, Fujita H, Kamada S, Kuzumaki N, Tsujimoto Y. Human gelsolin prevents apoptosis by inhibiting apoptotic mitochondrial changes via closing VDAC. Oncogene 19: 4807-4814, 2000.
    • (2000) Oncogene , vol.19 , pp. 4807-4814
    • Kusano, H.1    Shimizu, S.2    Koya, R.C.3    Fujita, H.4    Kamada, S.5    Kuzumaki, N.6    Tsujimoto, Y.7
  • 36
    • 0032966363 scopus 로고    scopus 로고
    • Functions of gelsolin: Motility, signaling, apoptosis, cancer
    • Kwiatkowski DJ. Functions of gelsolin: motility, signaling, apoptosis, cancer. Curr Opin Cell Biol 11: 103-108, 1999.
    • (1999) Curr Opin Cell Biol , vol.11 , pp. 103-108
    • Kwiatkowski, D.J.1
  • 39
    • 0034698878 scopus 로고    scopus 로고
    • Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis
    • Nakagawa T, Yuan J. Cross-talk between two cysteine protease families. Activation of caspase-12 by calpain in apoptosis. J Cell Biol 150: 887-894, 2000.
    • (2000) J Cell Biol , vol.150 , pp. 887-894
    • Nakagawa, T.1    Yuan, J.2
  • 42
    • 0141728522 scopus 로고    scopus 로고
    • Differential effects of calpain inhibitors on hypertrophy of cardiomyocytes
    • Pollack JR, Witt RC, Sugimoto JT. Differential effects of calpain inhibitors on hypertrophy of cardiomyocytes. Mol Cell Biochem 251: 47-50, 2003.
    • (2003) Mol Cell Biochem , vol.251 , pp. 47-50
    • Pollack, J.R.1    Witt, R.C.2    Sugimoto, J.T.3
  • 43
    • 0041969008 scopus 로고    scopus 로고
    • Calpain and its involvement in the pathophysiology of CNS injuries and diseases: Therapeutic potential of calpain inhibitors for prevention of neurodegeneration
    • Ray SK, Banik NL. Calpain and its involvement in the pathophysiology of CNS injuries and diseases: therapeutic potential of calpain inhibitors for prevention of neurodegeneration. Curr Drug Targets CNS Neurol Disord 2: 173-189, 2003.
    • (2003) Curr Drug Targets CNS Neurol Disord , vol.2 , pp. 173-189
    • Ray, S.K.1    Banik, N.L.2
  • 44
    • 0037989744 scopus 로고    scopus 로고
    • Calpain in the pathophysiology of spinal cord injury: Neuroprotection with calpain inhibitors
    • Ray SK, Hogan EL, Banik NL. Calpain in the pathophysiology of spinal cord injury: neuroprotection with calpain inhibitors. Brain Res Brain Res Rev 42: 169-185, 2003.
    • (2003) Brain Res Brain Res Rev , vol.42 , pp. 169-185
    • Ray, S.K.1    Hogan, E.L.2    Banik, N.L.3
  • 45
    • 33645846638 scopus 로고    scopus 로고
    • Inhibition of calpain and caspase-3 prevented apoptosis and preserved electrophysiological properties of voltage-gated and ligand-gated ion channels in rat primary cortical neurons exposed to glutamate
    • Ray SK, Karmakar S, Nowak MW, Banik NL. Inhibition of calpain and caspase-3 prevented apoptosis and preserved electrophysiological properties of voltage-gated and ligand-gated ion channels in rat primary cortical neurons exposed to glutamate. Neuroscience 139: 577-595, 2006.
    • (2006) Neuroscience , vol.139 , pp. 577-595
    • Ray, S.K.1    Karmakar, S.2    Nowak, M.W.3    Banik, N.L.4
  • 47
    • 34247345833 scopus 로고    scopus 로고
    • The apoptosome: Signalling platform of cell death
    • Riedl SJ, Salvesen GS. The apoptosome: signalling platform of cell death. Nat Rev Mol Cell Biol 8: 405-413, 2007.
    • (2007) Nat Rev Mol Cell Biol , vol.8 , pp. 405-413
    • Riedl, S.J.1    Salvesen, G.S.2
  • 48
    • 0036310279 scopus 로고    scopus 로고
    • Microinjection of cathepsin D induces caspase-dependent apoptosis in fibroblasts
    • Roberg K, Kagedal K, Ollinger K. Microinjection of cathepsin D induces caspase-dependent apoptosis in fibroblasts. Am J Pathol 161: 89-96, 2002.
    • (2002) Am J Pathol , vol.161 , pp. 89-96
    • Roberg, K.1    Kagedal, K.2    Ollinger, K.3
  • 49
    • 35848940084 scopus 로고    scopus 로고
    • Reperfusion injury as a therapeutic challenge in patients with acute myocardial infarction
    • Rodriguez-Sinovas A, Abdallah Y, Piper HM, Garcia-Dorado D. Reperfusion injury as a therapeutic challenge in patients with acute myocardial infarction. Heart Fail Rev 12: 207-216, 2007.
    • (2007) Heart Fail Rev , vol.12 , pp. 207-216
    • Rodriguez-Sinovas, A.1    Abdallah, Y.2    Piper, H.M.3    Garcia-Dorado, D.4
  • 52
    • 11444263337 scopus 로고    scopus 로고
    • Preconditioning of arteriogenesis
    • Scholz D, Schaper W. Preconditioning of arteriogenesis. Cardiovasc Res 65: 513-523, 2005.
    • (2005) Cardiovasc Res , vol.65 , pp. 513-523
    • Scholz, D.1    Schaper, W.2
  • 53
    • 0032831745 scopus 로고    scopus 로고
    • Calpain activity and expression are increased in splenic inflammatory cells associated with experimental allergic encephalomyelitis
    • Shields DC, Schaecher KE, Goust JM, Banik NL. Calpain activity and expression are increased in splenic inflammatory cells associated with experimental allergic encephalomyelitis. J Neuroimmunol 99: 1-12, 1999.
    • (1999) J Neuroimmunol , vol.99 , pp. 1-12
    • Shields, D.C.1    Schaecher, K.E.2    Goust, J.M.3    Banik, N.L.4
  • 55
    • 3142654669 scopus 로고    scopus 로고
    • The sarcoplasmic reticulum proteins are targets for calpain action in the ischemic-reperfused heart
    • Singh RB, Chohan PK, Dhalla NS, Netticadan T. The sarcoplasmic reticulum proteins are targets for calpain action in the ischemic-reperfused heart. J Mol Cell Cardiol 37: 101-110, 2004.
    • (2004) J Mol Cell Cardiol , vol.37 , pp. 101-110
    • Singh, R.B.1    Chohan, P.K.2    Dhalla, N.S.3    Netticadan, T.4
  • 58
    • 0030137184 scopus 로고    scopus 로고
    • Cytoskeletal role in the contractile dysfunction of cardiocytes from hypertrophied and failing right ventricular myocardium
    • Tagawa H, Koide M, Sato H, Cooper GT. Cytoskeletal role in the contractile dysfunction of cardiocytes from hypertrophied and failing right ventricular myocardium. Proc Assoc Am Physicians 108: 218-229, 1996.
    • (1996) Proc Assoc Am Physicians , vol.108 , pp. 218-229
    • Tagawa, H.1    Koide, M.2    Sato, H.3    Cooper, G.T.4
  • 60
    • 33745727523 scopus 로고    scopus 로고
    • Possible involvement of calpain activation in pathogenesis of chronic heart failure after acute myocardial infarction
    • Takahashi M, Tanonaka K, Yoshida H, Koshimizu M, Daicho T, Oikawa R, Takeo S. Possible involvement of calpain activation in pathogenesis of chronic heart failure after acute myocardial infarction. J Cardiovasc Pharmacol 47: 413-421, 2006.
    • (2006) J Cardiovasc Pharmacol , vol.47 , pp. 413-421
    • Takahashi, M.1    Tanonaka, K.2    Yoshida, H.3    Koshimizu, M.4    Daicho, T.5    Oikawa, R.6    Takeo, S.7
  • 61
    • 33744918550 scopus 로고    scopus 로고
    • Ubiquitous calpains promote caspase-12 and JNK activation during endoplasmic reticulum stress-induced apoptosis
    • Tan Y, Dourdin N, Wu C, De Veyra T, Elce JS, Greer PA. Ubiquitous calpains promote caspase-12 and JNK activation during endoplasmic reticulum stress-induced apoptosis. J Biol Chem 281: 16016-16024, 2006.
    • (2006) J Biol Chem , vol.281 , pp. 16016-16024
    • Tan, Y.1    Dourdin, N.2    Wu, C.3    De Veyra, T.4    Elce, J.S.5    Greer, P.A.6
  • 63
    • 0035661090 scopus 로고    scopus 로고
    • Implications of CAD and DNase II in ischemic neuronal necrosis specific for the primate hippocampus
    • Tsukada T, Watanabe M, Yamashima T. Implications of CAD and DNase II in ischemic neuronal necrosis specific for the primate hippocampus. J Neurochem 79: 1196-1206, 2001.
    • (2001) J Neurochem , vol.79 , pp. 1196-1206
    • Tsukada, T.1    Watanabe, M.2    Yamashima, T.3
  • 65
    • 0033956278 scopus 로고    scopus 로고
    • Calpain and caspase: Can you tell the difference?
    • Wang KK. Calpain and caspase: can you tell the difference? Trends Neurosci 23: 59, 2000.
    • (2000) Trends Neurosci , vol.23 , pp. 59
    • Wang, K.K.1


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