메뉴 건너뛰기




Volumn 88, Issue 1, 2010, Pages 150-158

Trimetazidine inhibits pressure overload-induced cardiac fibrosis through NADPH oxidase-ROS-CTGF pathway

Author keywords

Cardiac fibrosis; Connective tissue growth factor; NADPH oxidase; Reactive oxygen species; Trimetazidine

Indexed keywords

ANGIOTENSIN II; COLLAGEN; CONNECTIVE TISSUE GROWTH FACTOR; RAC1 PROTEIN; REACTIVE OXYGEN METABOLITE; REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE PHOSPHATE OXIDASE; TRIMETAZIDINE;

EID: 77956816714     PISSN: 00086363     EISSN: 17553245     Source Type: Journal    
DOI: 10.1093/cvr/cvq181     Document Type: Article
Times cited : (76)

References (41)
  • 1
    • 85047680324 scopus 로고
    • Myocardial fibrosis: Functional significance and regulatory factors
    • Weber KT, Brilla CG, Janicki JS. Myocardial fibrosis: functional significance and regulatory factors. Cardiovasc Res 1993;27:341-348.
    • (1993) Cardiovasc Res , vol.27 , pp. 341-348
    • Weber, K.T.1    Brilla, C.G.2    Janicki, J.S.3
  • 2
    • 0036272927 scopus 로고    scopus 로고
    • Myocardial stiffness is determined by ventricular fibrosis, but not by compensatory or excessive hypertrophy in hypertensive heart
    • Yamamoto K, Masuyama T, Sakata Y, Nishikawa N, Mano T, Yoshida J et al. Myocardial stiffness is determined by ventricular fibrosis, but not by compensatory or excessive hypertrophy in hypertensive heart. Cardiovasc Res 2002;55:76-82.
    • (2002) Cardiovasc Res , vol.55 , pp. 76-82
    • Yamamoto, K.1    Masuyama, T.2    Sakata, Y.3    Nishikawa, N.4    Mano, T.5    Yoshida, J.6
  • 3
    • 0037465434 scopus 로고    scopus 로고
    • Progression from compensated hypertrophy to failure in the pressure-overloaded human heart: Structural deterioration and compensatory mechanisms
    • Hein S, Arnon E, Kostin S, Schonburg M, Elsasser A, Polyakova V et al. Progression from compensated hypertrophy to failure in the pressure-overloaded human heart: structural deterioration and compensatory mechanisms. Circulation 2003;107: 984-991.
    • (2003) Circulation , vol.107 , pp. 984-991
    • Hein, S.1    Arnon, E.2    Kostin, S.3    Schonburg, M.4    Elsasser, A.5    Polyakova, V.6
  • 4
    • 0034678082 scopus 로고    scopus 로고
    • The antianginal drug trimetazidine shifts cardiac energy metabolism from fatty acid oxidation to glucose oxidation by inhibiting mitochondrial long-chain 3-ketoacyl coenzyme A thiolase
    • Kantor PF, Lucien A, Kozak R, Lopaschuk GD. The antianginal drug trimetazidine shifts cardiac energy metabolism from fatty acid oxidation to glucose oxidation by inhibiting mitochondrial long-chain 3-ketoacyl coenzyme A thiolase. Circ Res 2000; 86:580-588.
    • (2000) Circ Res , vol.86 , pp. 580-588
    • Kantor, P.F.1    Lucien, A.2    Kozak, R.3    Lopaschuk, G.D.4
  • 5
    • 0141499890 scopus 로고    scopus 로고
    • Beneficial effects of trimetazidine in ex vivo working ischemic hearts are due to a stimulation of glucose oxidation secondary to inhibition of long-chain 3-ketoacyl coenzyme a thiolase
    • Lopaschuk GD, Barr R, Thomas PD, Dyck JR. Beneficial effects of trimetazidine in ex vivo working ischemic hearts are due to a stimulation of glucose oxidation secondary to inhibition of long-chain 3-ketoacyl coenzyme a thiolase. Circ Res 2003;93:e33-e37.
    • (2003) Circ Res , vol.93
    • Lopaschuk, G.D.1    Barr, R.2    Thomas, P.D.3    Dyck, J.R.4
  • 6
    • 0027237307 scopus 로고
    • Infarct size in rabbits: A modified method illustrated by the effects of propranolol and trimetazidine
    • Drake-Holland AJ, Belcher PR, Hynd J, Noble MI. Infarct size in rabbits: a modified method illustrated by the effects of propranolol and trimetazidine. Basic Res Cardiol 1993;88:250-258.
    • (1993) Basic Res Cardiol , vol.88 , pp. 250-258
    • Drake-Holland, A.J.1    Belcher, P.R.2    Hynd, J.3    Noble, M.I.4
  • 7
    • 0028018335 scopus 로고
    • Some biochemical aspects of the protective effect of trimetazidine on rat cardiomyocytes during hypoxia and reox-ygenation
    • Fantini E, Demaison L, Sentex E, Grynberg A, Athias P. Some biochemical aspects of the protective effect of trimetazidine on rat cardiomyocytes during hypoxia and reox-ygenation. J Mol Cell Cardiol 1994;26:949-958.
    • (1994) J Mol Cell Cardiol , vol.26 , pp. 949-958
    • Fantini, E.1    Demaison, L.2    Sentex, E.3    Grynberg, A.4    Athias, P.5
  • 8
    • 34548363767 scopus 로고    scopus 로고
    • Trimetazidine improves endothelial dysfunction in chronic heart failure: An antioxidant effect
    • Belardinelli R, Solenghi M, Volpe L, Purcaro A. Trimetazidine improves endothelial dysfunction in chronic heart failure: an antioxidant effect. Eur Heart J 2007;28: 1102-1108.
    • (2007) Eur Heart J , vol.28 , pp. 1102-1108
    • Belardinelli, R.1    Solenghi, M.2    Volpe, L.3    Purcaro, A.4
  • 9
    • 0242408768 scopus 로고    scopus 로고
    • Evidence for a protective role of trimetazidine during cold ischemia: Targeting inflammation and nephron mass
    • Faure JP, Baumert H, Han Z, Goujon JM, Favreau F, Dutheil D et al. Evidence for a protective role of trimetazidine during cold ischemia: targeting inflammation and nephron mass. Biochem Pharmacol 2003;66:2241-2250.
    • (2003) Biochem Pharmacol , vol.66 , pp. 2241-2250
    • Faure, J.P.1    Baumert, H.2    Han, Z.3    Goujon, J.M.4    Favreau, F.5    Dutheil, D.6
  • 10
    • 0029090052 scopus 로고
    • Improvement of long-term preservation of the isolated arrested rat heart by trimetazidine: Effects on the energy state and mitochondrial function
    • Kay L, Finelli C, Aussedat J, Guarnieri C, Rossi A. Improvement of long-term preservation of the isolated arrested rat heart by trimetazidine: effects on the energy state and mitochondrial function. Am J Cardiol 1995;76:45B-49B.
    • (1995) Am J Cardiol , vol.76
    • Kay, L.1    Finelli, C.2    Aussedat, J.3    Guarnieri, C.4    Rossi, A.5
  • 11
    • 0035208524 scopus 로고    scopus 로고
    • Effects of trimetazidine on the contractile response of chronically dysfunctional myocardium to low-dose dobutamine in ischaemic cardio-myopathy
    • Belardinelli R, Purcaro A. Effects of trimetazidine on the contractile response of chronically dysfunctional myocardium to low-dose dobutamine in ischaemic cardio-myopathy. Eur Heart J 2001;22:2164-2170.
    • (2001) Eur Heart J , vol.22 , pp. 2164-2170
    • Belardinelli, R.1    Purcaro, A.2
  • 12
    • 33747887613 scopus 로고    scopus 로고
    • A randomized clinical trial of trimetazidine, a partial free fatty acid oxidation inhibitor, in patients with heart failure
    • Fragasso G, Palloshi A, Puccetti P, Silipigni C, Rossodivita A, Pala M et al. A randomized clinical trial of trimetazidine, a partial free fatty acid oxidation inhibitor, in patients with heart failure. J Am Coll Cardiol 2006;48:992-998.
    • (2006) J Am Coll Cardiol , vol.48 , pp. 992-998
    • Fragasso, G.1    Palloshi, A.2    Puccetti, P.3    Silipigni, C.4    Rossodivita, A.5    Pala, M.6
  • 13
    • 33645466318 scopus 로고    scopus 로고
    • Effects of metabolic modulation by trimetazidine on left ventricular function and phospho-creatine/adenosine triphosphate ratio in patients with heart failure
    • Fragasso G, Perseghin G, De Cobelli F, Esposito A, Palloshi A, Lattuada G et al. Effects of metabolic modulation by trimetazidine on left ventricular function and phospho-creatine/adenosine triphosphate ratio in patients with heart failure. Eur Heart J 2006;27:942-948.
    • (2006) Eur Heart J , vol.27 , pp. 942-948
    • Fragasso, G.1    Perseghin, G.2    De Cobelli, F.3    Esposito, A.4    Palloshi, A.5    Lattuada, G.6
  • 14
    • 1542496389 scopus 로고    scopus 로고
    • Short-and long-term beneficial effects of trimetazidine in patients with diabetes and ischemic cardiomyopathy
    • Fragasso G, Piatti Md PM, Monti L, Palloshi A, Setola E, Puccetti P et al. Short-and long-term beneficial effects of trimetazidine in patients with diabetes and ischemic cardiomyopathy. Am Heart J 2003;146:E18.
    • (2003) Am Heart J , vol.146
    • Fragasso, G.1    Piatti Md, P.M.2    Monti, L.3    Palloshi, A.4    Setola, E.5    Puccetti, P.6
  • 15
    • 4944224833 scopus 로고    scopus 로고
    • Trimetazidine improves left ventricular function and quality of life in elderly patients with coronary artery disease
    • Vitale C, Wajngaten M, Sposato B, Gebara O, Rossini P, Fini M et al. Trimetazidine improves left ventricular function and quality of life in elderly patients with coronary artery disease. Eur Heart J 2004;25:1814-1821.
    • (2004) Eur Heart J , vol.25 , pp. 1814-1821
    • Vitale, C.1    Wajngaten, M.2    Sposato, B.3    Gebara, O.4    Rossini, P.5    Fini, M.6
  • 16
    • 53449101397 scopus 로고    scopus 로고
    • Trimetazidine, a metabolic modulator, has cardiac and extracardiac benefits in idiopathic dilated car-diomyopathy
    • Tuunanen H, Engblom E, Naum A, Nagren K, Scheinin M, Hesse B et al. Trimetazidine, a metabolic modulator, has cardiac and extracardiac benefits in idiopathic dilated car-diomyopathy. Circulation 2008;118:1250-1258.
    • (2008) Circulation , vol.118 , pp. 1250-1258
    • Tuunanen, H.1    Engblom, E.2    Naum, A.3    Nagren, K.4    Scheinin, M.5    Hesse, B.6
  • 17
    • 1542358803 scopus 로고    scopus 로고
    • Connective tissue growth factor\a novel mediator of angiotensin II-stimulated cardiac fibroblast activation in heart failure in rats
    • Ahmed MS, Oie E, Vinge LE, Yndestad A, Oystein Andersen G, Andersson Y et al. Connective tissue growth factor\a novel mediator of angiotensin II-stimulated cardiac fibroblast activation in heart failure in rats. J Mol Cell Cardiol 2004;36:393-404.
    • (2004) J Mol Cell Cardiol , vol.36 , pp. 393-404
    • Ahmed, M.S.1    Oie, E.2    Vinge, L.E.3    Yndestad, A.4    Oystein Andersen, G.5    Andersson, Y.6
  • 18
    • 18144380288 scopus 로고    scopus 로고
    • Individual domains of connective tissue growth factor regulate fibroblast proliferation and myofibroblast differentiation
    • Grotendorst GR, Duncan MR. Individual domains of connective tissue growth factor regulate fibroblast proliferation and myofibroblast differentiation. FASEB J 2005;19: 729-738.
    • (2005) FASEB J , vol.19 , pp. 729-738
    • Grotendorst, G.R.1    Duncan, M.R.2
  • 19
    • 34247892260 scopus 로고    scopus 로고
    • Increased connective tissue growth factor relative to brain natriuretic peptide as a determinant of myocardial fibrosis
    • Koitabashi N, Arai M, Kogure S, Niwano K, Watanabe A, Aoki Y et al. Increased connective tissue growth factor relative to brain natriuretic peptide as a determinant of myocardial fibrosis. Hypertension 2007;49:1120-1127.
    • (2007) Hypertension , vol.49 , pp. 1120-1127
    • Koitabashi, N.1    Arai, M.2    Kogure, S.3    Niwano, K.4    Watanabe, A.5    Aoki, Y.6
  • 20
    • 0141954211 scopus 로고    scopus 로고
    • Induction of connective tissue growth factor by angiotensin II: Integration of signaling pathways
    • Iwanciw D, Rehm M, Porst M, Goppelt-Struebe M. Induction of connective tissue growth factor by angiotensin II: integration of signaling pathways. Arterioscler Thromb Vasc Biol 2003;23:1782-1787.
    • (2003) Arterioscler Thromb Vasc Biol , vol.23 , pp. 1782-1787
    • Iwanciw, D.1    Rehm, M.2    Porst, M.3    Goppelt-Struebe, M.4
  • 21
    • 34447525522 scopus 로고    scopus 로고
    • Tissue-specific mechanisms for CCN2/CTGF persistence in fibrotic gingiva: Interactions between cAMP and MAPK signaling pathways, and prostaglandin E2-EP3 receptor mediated activation of the c-JUN N-terminal kinase
    • Black SA Jr, Palamakumbura AH, Stan M, Trackman PC. Tissue-specific mechanisms for CCN2/CTGF persistence in fibrotic gingiva: interactions between cAMP and MAPK signaling pathways, and prostaglandin E2-EP3 receptor mediated activation of the c-JUN N-terminal kinase. J Biol Chem 2007;282:15416-15429.
    • (2007) J Biol Chem , vol.282 , pp. 15416-15429
    • Black Jr., S.A.1    Palamakumbura, A.H.2    Stan, M.3    Trackman, P.C.4
  • 22
    • 18744430508 scopus 로고    scopus 로고
    • Connective tissue growth factor is a mediator of angiotensin II-induced fibrosis
    • DOI 10.1161/01.CIR.0000089129.51288.BA
    • Ruperez M, Lorenzo O, Blanco-Colio LM, Esteban V, Egido J, Ruiz-Ortega M. Connective tissue growth factor is a mediator of angiotensin II-induced fibrosis. Circulation 2003;108:1499-1505. (Pubitemid 37176458)
    • (2003) Circulation , vol.108 , Issue.12 , pp. 1499-1505
    • Ruperez, M.1    Lorenzo, O.2    Blanco-Colio, L.M.3    Esteban, V.4    Egido, J.5    Ruiz-Ortega, M.6
  • 23
    • 20144371323 scopus 로고    scopus 로고
    • Trimeta-zidine protects isolated rat hearts against ischemia-reperfusion injury in an experimental timing-dependent manner
    • Pantos C, Bescond-Jacquet A, Tzeis S, Paizis I, Mourouzis I, Moraitis P et al. Trimeta-zidine protects isolated rat hearts against ischemia-reperfusion injury in an experimental timing-dependent manner. Basic Res Cardiol 2005;100:154-160.
    • (2005) Basic Res Cardiol , vol.100 , pp. 154-160
    • Pantos, C.1    Bescond-Jacquet, A.2    Tzeis, S.3    Paizis, I.4    Mourouzis, I.5    Moraitis, P.6
  • 26
    • 17444404892 scopus 로고    scopus 로고
    • 14-3-3 proteins regulate glycogen synthase 3beta phosphorylation and inhibit cardiomyocyte hypertrophy
    • Liao W, Wang S, Han C, Zhang Y. 14-3-3 proteins regulate glycogen synthase 3beta phosphorylation and inhibit cardiomyocyte hypertrophy. FEBS J 2005;272: 1845-1854.
    • (2005) FEBS J , vol.272 , pp. 1845-1854
    • Liao, W.1    Wang, S.2    Han, C.3    Zhang, Y.4
  • 27
    • 39549087890 scopus 로고    scopus 로고
    • Inhibitory crosstalk between ERK and AMPK in the growth and proliferation of cardiac fibroblasts
    • Du J, Guan T, Zhang H, Xia Y, Liu F, Zhang Y. Inhibitory crosstalk between ERK and AMPK in the growth and proliferation of cardiac fibroblasts. Biochem Biophys Res Commun 2008;368:402-407.
    • (2008) Biochem Biophys Res Commun , vol.368 , pp. 402-407
    • Du, J.1    Guan, T.2    Zhang, H.3    Xia, Y.4    Liu, F.5    Zhang, Y.6
  • 28
    • 57649186957 scopus 로고    scopus 로고
    • A novel protein kinase A-independent, beta-arrestin-1-dependent signaling pathway for p38 mitogen-activated protein kinase activation by beta2-adrenergic receptors
    • Gong K, Li Z, Xu M, Du J, Lv Z, Zhang Y. A novel protein kinase A-independent, beta-arrestin-1-dependent signaling pathway for p38 mitogen-activated protein kinase activation by beta2-adrenergic receptors. J Biol Chem 2008;283: 29028-29036.
    • (2008) J Biol Chem , vol.283 , pp. 29028-29036
    • Gong, K.1    Li, Z.2    Xu, M.3    Du, J.4    Lv, Z.5    Zhang, Y.6
  • 29
    • 0028365310 scopus 로고
    • Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells
    • Griendling KK, Minieri CA, Ollerenshaw JD, Alexander RW. Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells. Circ Res 1994;74:1141-1148.
    • (1994) Circ Res , vol.74 , pp. 1141-1148
    • Griendling, K.K.1    Minieri, C.A.2    Ollerenshaw, J.D.3    Alexander, R.W.4
  • 30
    • 20944445197 scopus 로고    scopus 로고
    • Oxidant stress from nitric oxide synthase-3 uncoupling stimulates cardiac pathologic remodeling from chronic pressure load
    • Takimoto E, Champion HC, Li M, Ren S, Rodriguez ER, Tavazzi B et al. Oxidant stress from nitric oxide synthase-3 uncoupling stimulates cardiac pathologic remodeling from chronic pressure load. J Clin Invest 2005;115:1221-1231.
    • (2005) J Clin Invest , vol.115 , pp. 1221-1231
    • Takimoto, E.1    Champion, H.C.2    Li, M.3    Ren, S.4    Rodriguez, E.R.5    Tavazzi, B.6
  • 31
    • 33750205980 scopus 로고    scopus 로고
    • Pressure overload-induced transient oxidative stress mediates perivascular inflammation and cardiac fibrosis through angiotensin II
    • Kai H, Mori T, Tokuda K, Takayama N, Tahara N, Takemiya K et al. Pressure overload-induced transient oxidative stress mediates perivascular inflammation and cardiac fibrosis through angiotensin II. Hypertens Res 2006;29:711-718.
    • (2006) Hypertens Res , vol.29 , pp. 711-718
    • Kai, H.1    Mori, T.2    Tokuda, K.3    Takayama, N.4    Tahara, N.5    Takemiya, K.6
  • 32
    • 41049089282 scopus 로고    scopus 로고
    • Evaluation of the renin-angiotensin system in cardiac tissues of cats with pressure-overload cardiac hypertrophy
    • Uechi M, Tanaka Y, Aramaki Y, Hori Y, Ishikawa Y, Ebisawa T et al. Evaluation of the renin-angiotensin system in cardiac tissues of cats with pressure-overload cardiac hypertrophy. Am J Vet Res 2008;69:343-348.
    • (2008) Am J Vet Res , vol.69 , pp. 343-348
    • Uechi, M.1    Tanaka, Y.2    Aramaki, Y.3    Hori, Y.4    Ishikawa, Y.5    Ebisawa, T.6
  • 33
    • 41449097556 scopus 로고    scopus 로고
    • Plasma connective tissue growth factor is a novel potential biomarker of cardiac dysfunction in patients with chronic heart failure
    • Koitabashi N, Arai M, Niwano K, Watanabe A, Endoh M, Suguta M et al. Plasma connective tissue growth factor is a novel potential biomarker of cardiac dysfunction in patients with chronic heart failure. Eur J Heart Fail 2008;10:373-379.
    • (2008) Eur J Heart Fail , vol.10 , pp. 373-379
    • Koitabashi, N.1    Arai, M.2    Niwano, K.3    Watanabe, A.4    Endoh, M.5    Suguta, M.6
  • 34
    • 0029808645 scopus 로고    scopus 로고
    • A novel transforming growth factor beta response element controls the expression of the connective tissue growth factor gene
    • Grotendorst GR, Okochi H, Hayashi N. A novel transforming growth factor beta response element controls the expression of the connective tissue growth factor gene. Cell Growth Differ 1996;7:469-480.
    • (1996) Cell Growth Differ , vol.7 , pp. 469-480
    • Grotendorst, G.R.1    Okochi, H.2    Hayashi, N.3
  • 35
    • 33646814422 scopus 로고    scopus 로고
    • Attenuation of myocardial ischemia-reperfusion injury by trimetazidine derivatives functionalized with antioxidant properties
    • Kutala VK, Khan M, Mandal R, Ganesan LP, Tridandapani S, Kalai T et al. Attenuation of myocardial ischemia-reperfusion injury by trimetazidine derivatives functionalized with antioxidant properties. J Pharmacol Exp Ther 2006;317:921-928.
    • (2006) J Pharmacol Exp Ther , vol.317 , pp. 921-928
    • Kutala, V.K.1    Khan, M.2    Mandal, R.3    Ganesan, L.P.4    Tridandapani, S.5    Kalai, T.6
  • 36
    • 20544457454 scopus 로고    scopus 로고
    • Smad3 and extracellular signal-regulated kinase 1/2 coordinately mediate transforming growth factor-beta-induced expression of connective tissue growth factor in human fibroblasts
    • Leivonen SK, Hakkinen L, Liu D, Kahari VM. Smad3 and extracellular signal-regulated kinase 1/2 coordinately mediate transforming growth factor-beta-induced expression of connective tissue growth factor in human fibroblasts. J Invest Dermatol 2005;124: 1162-1169.
    • (2005) J Invest Dermatol , vol.124 , pp. 1162-1169
    • Leivonen, S.K.1    Hakkinen, L.2    Liu, D.3    Kahari, V.M.4
  • 38
    • 0027217406 scopus 로고
    • Effect of trimetazidine on mitochondrial function and oxi-dative damage during reperfusion of ischemic hypertrophied rat myocardium
    • Guarnieri C, Muscari C. Effect of trimetazidine on mitochondrial function and oxi-dative damage during reperfusion of ischemic hypertrophied rat myocardium. Pharmacology 1993;46:324-331.
    • (1993) Pharmacology , vol.46 , pp. 324-331
    • Guarnieri, C.1    Muscari, C.2
  • 39
    • 0037785315 scopus 로고    scopus 로고
    • NADPH oxidase-derived superoxide anion mediates angiotensin II-induced cardiac hypertrophy
    • Nakagami H, Takemoto M, Liao JK. NADPH oxidase-derived superoxide anion mediates angiotensin II-induced cardiac hypertrophy. J Mol Cell Cardiol 2003;35:851-859.
    • (2003) J Mol Cell Cardiol , vol.35 , pp. 851-859
    • Nakagami, H.1    Takemoto, M.2    Liao, J.K.3
  • 40
    • 0031656261 scopus 로고    scopus 로고
    • Role of NADH/NADPH oxidase-derived H2O2 in angiotensin II-induced vascular hypertrophy
    • Zafari AM, Ushio-Fukai M, Akers M, Yin Q, Shah A, Harrison DG et al. Role of NADH/NADPH oxidase-derived H2O2 in angiotensin II-induced vascular hypertrophy. Hypertension 1998;32:488-495.
    • (1998) Hypertension , vol.32 , pp. 488-495
    • Zafari, A.M.1    Ushio-Fukai, M.2    Akers, M.3    Yin, Q.4    Shah, A.5    Harrison, D.G.6
  • 41
    • 0032496326 scopus 로고    scopus 로고
    • Role of Rac1 and oxygen radicals in collagenase-1 expression induced by cell shape change
    • Kheradmand F, Werner E, Tremble P, Symons M, Werb Z. Role of Rac1 and oxygen radicals in collagenase-1 expression induced by cell shape change. Science 1998;280: 898-902.
    • (1998) Science , vol.280 , pp. 898-902
    • Kheradmand, F.1    Werner, E.2    Tremble, P.3    Symons, M.4    Werb, Z.5


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.