Circulation. 2002;106:130-135
Published online before print June 10, 2002, doi: 10.1161/01.CIR.0000020689.12472.E0
Published online before print June 10, 2002, doi: 10.1161/01.CIR.0000020689.12472.E0
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
(Circulation. 2002;106:130.)
© 2002 American Heart Association, Inc.
Basic Science Reports |
Transforming Growth Factor-ß Function Blocking Prevents Myocardial Fibrosis and Diastolic Dysfunction in Pressure-Overloaded Rats
From the Cardiovascular Research Institute and Internal Medicine III, Kurume University School of Medicine, Kurume (F.K., H.K., K.T., T.I.); the Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka (M.K.); and Cardiovascular Medicine, Graduate School of Medical Sciences, Kyushu University, Fukuoka (A.T., K.E.), Japan.
Correspondence to Hisashi Kai, MD, PhD, Cardiovascular Research Institute and Internal Medicine III, Kurume University School of Medicine, 67 Asahimachi, Kurume, Fukuoka 830-0011, Japan. E-mail naikai@ med.kurume-u.ac.jp
Background— Excessive myocardial fibrosis impairs cardiac function in hypertensive hearts. Roles of transforming growth factor (TGF)-ß in myocardial remodeling and cardiac dysfunction were examined in pressure-overloaded rats.
Methods and Results— Pressure overload was induced by a suprarenal aortic constriction in Wistar rats. Fibroblast activation (proliferation and phenotype transition to myofibroblasts) was observed after day 3 and peaked at days 3 to 7. Thereafter, myocyte hypertrophy and myocardial fibrosis developed by day 28. At day 28, echocardiography showed normal left ventricular fractional shortening, but the decreased ratio of early to late filling velocity of the transmitral Doppler velocity and hemodynamic measurement revealed left ventricular end-diastolic pressure elevation, indicating normal systolic but abnormal diastolic function. Myocardial TGF-ß mRNA expression was induced after day 3, peaked at day 7, and remained modestly increased at day 28. An anti–TGF-ß neutralizing antibody, which was administered intraperitoneally daily from 1 day before operation, inhibited fibroblast activation and subsequently prevented collagen mRNA induction and myocardial fibrosis, but not myocyte hypertrophy. Neutralizing antibody reversed diastolic dysfunction without affecting blood pressure and systolic function.
Conclusions— TGF-ß plays a causal role in myocardial fibrosis and diastolic dysfunction through fibroblast activation in pressure-overloaded hearts. Our findings may provide an insight into a new therapeutic strategy to prevent myocardial fibrosis and diastolic dysfunction in pressure-overloaded hearts.
Key Words: growth substances • fibrosis • hypertension • diastole
This article has been cited by other articles:
 |
|
 |
|
  M. M. Shafiq and A. B. Miller Blocking aldosterone in heart failure Therapeutic Advances in Cardiovascular Disease, October 1, 2009; 3(5): 379 - 385. [Abstract] [PDF]
|
|
|
|
 |
|
 H. Kudo, H. Kai, H. Kajimoto, M. Koga, N. Takayama, T. Mori, A. Ikeda, S. Yasuoka, T. Anegawa, H. Mifune, et al. Exaggerated Blood Pressure Variability Superimposed on Hypertension Aggravates Cardiac Remodeling in Rats via Angiotensin II System-Mediated Chronic Inflammation Hypertension, October 1, 2009; 54(4): 832 - 838. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Berni, M. Savi, L. Bocchi, F. Delucchi, E. Musso, C. Chaponnier, G. Gabbiani, S. Clement, and D. Stilli Modulation of actin isoform expression before the transition from experimental compensated pressure-overload cardiac hypertrophy to decompensation Am J Physiol Heart Circ Physiol, May 1, 2009; 296(5): H1625 - H1632. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Frantz, J. Bauersachs, and G. Ertl Post-infarct remodelling: contribution of wound healing and inflammation Cardiovasc Res, February 15, 2009; 81(3): 474 - 481. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-Y. Liu, W.-F. Cai, H.-Z. Yang, B. Cui, Z.-R. Chen, H.-Z. Liu, J. Yan, W. Jin, H.-M. Yan, B.-M. Xin, et al. Bacillus Calmette-Guerin and TLR4 Agonist Prevent Cardiovascular Hypertrophy and Fibrosis by Regulating Immune Microenvironment J. Immunol., June 1, 2008; 180(11): 7349 - 7357. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Cihakova, J. G. Barin, M. Afanasyeva, M. Kimura, D. Fairweather, M. Berg, M. V. Talor, G. C. Baldeviano, S. Frisancho, K. Gabrielson, et al. Interleukin-13 Protects Against Experimental Autoimmune Myocarditis by Regulating Macrophage Differentiation Am. J. Pathol., May 1, 2008; 172(5): 1195 - 1208. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. L.O. Buxton and D. Duan Cyclic GMP/Protein Kinase G Phosphorylation of Smad3 Blocks Transforming Growth Factor-{beta}-Induced Nuclear Smad Translocation: A Key Antifibrogenic Mechanism of Atrial Natriuretic Peptide Circ. Res., February 1, 2008; 102(2): 151 - 153. [Full Text] [PDF]
|
|
|
|
|
|
B. P. Shapiro, T. E. Owan, S. Mohammed, M. Kruger, W. A. Linke, J. C. Burnett Jr, and M. M. Redfield Mineralocorticoid Signaling in Transition to Heart Failure With Normal Ejection Fraction Hypertension, February 1, 2008; 51(2): 289 - 295. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Kanemitsu, S. Takai, H. Tsuneyoshi, E. Yoshikawa, T. Nishina, M. Miyazaki, T. Ikeda, and M. Komeda Chronic chymase inhibition preserves cardiac function after left ventricular repair in rats Eur. J. Cardiothorac. Surg., January 1, 2008; 33(1): 25 - 31. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Li, X. Li, H. Zheng, B. Xie, K. R. Bidasee, and G. J. Rozanski Pro-oxidant effect of transforming growth factor-{beta}1 mediates contractile dysfunction in rat ventricular myocytes Cardiovasc Res, January 1, 2008; 77(1): 107 - 117. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Belmadani, J. Bernal, C.-C. Wei, M. A. Pallero, L. Dell'Italia, J. E. Murphy-Ullrich, and K. H. Berecek A Thrombospondin-1 Antagonist of Transforming Growth Factor-{beta} Activation Blocks Cardiomyopathy in Rats with Diabetes and Elevated Angiotensin II Am. J. Pathol., September 1, 2007; 171(3): 777 - 789. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. P. Shapiro, C. S.P. Lam, J. B. Patel, S. F. Mohammed, M. Kruger, D. M. Meyer, W. A. Linke, and M. M. Redfield Acute and Chronic Ventricular-Arterial Coupling in Systole and Diastole: Insights From an Elderly Hypertensive Model Hypertension, September 1, 2007; 50(3): 503 - 511. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Koga, H. Kai, H. Yasukawa, T. Yamamoto, Y. Kawai, S. Kato, K. Kusaba, M. Kai, K. Egashira, Y. Kataoka, et al. Inhibition of Progression and Stabilization of Plaques by Postnatal Interferon-{gamma} Function Blocking in ApoE-Knockout Mice Circ. Res., August 17, 2007; 101(4): 348 - 356. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Yamamoto, T. Shirayama, T. Sakatani, T. Takahashi, H. Tanaka, T. Takamatsu, K. W. Spitzer, and H. Matsubara Enhanced activity of ventricular Na+-HCO3 cotransport in pressure overload hypertrophy Am J Physiol Heart Circ Physiol, August 1, 2007; 293(2): H1254 - H1264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Wang, A. Omar, T. Angelovska, V. Drobic, S. G. Rattan, S. C. Jones, and I. M. C. Dixon Regulation of collagen synthesis by inhibitory Smad7 in cardiac myofibroblasts Am J Physiol Heart Circ Physiol, August 1, 2007; 293(2): H1282 - H1290. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. D Metcalfe and M. W.J Ferguson Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration J R Soc Interface, June 22, 2007; 4(14): 413 - 437. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Shingu, N. Shiiya, T. Mikami, K. Matsuzaki, T. Kunihara, and Y. Matsui Left Ventricular Diastolic Dysfunction in Chronic Aortic Dissection Ann. Thorac. Surg., April 1, 2007; 83(4): 1356 - 1360. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Kusaba, H. Kai, M. Koga, N. Takayama, A. Ikeda, H. Yasukawa, Y. Seki, K. Egashira, and T. Imaizumi Inhibition of Intrinsic Interferon-{gamma} Function Prevents Neointima Formation After Balloon Injury Hypertension, April 1, 2007; 49(4): 909 - 915. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Poobalarahi, C. F. Baicu, and A. D. Bradshaw Cardiac myofibroblasts differentiated in 3D culture exhibit distinct changes in collagen I production, processing, and matrix deposition Am J Physiol Heart Circ Physiol, December 1, 2006; 291(6): H2924 - H2932. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. A. Baudino, W. Carver, W. Giles, and T. K. Borg Cardiac fibroblasts: friend or foe? Am J Physiol Heart Circ Physiol, September 1, 2006; 291(3): H1015 - H1026. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Milting, A. Kassner, L. Arusoglu, H. E. Meyer, M. Morshuis, R. Brendel, B. Klauke, A. El Banayosy, and R. Korfer Influence of ACE-inhibition and mechanical unloading on the regulation of extracellular matrix proteins in the myocardium of heart transplantation candidates bridged by ventricular assist devices Eur J Heart Fail, May 1, 2006; 8(3): 278 - 283. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Matsumoto-Ida, Y. Takimoto, T. Aoyama, M. Akao, T. Takeda, and T. Kita Activation of TGF-{beta}1-TAK1-p38 MAPK pathway in spared cardiomyocytes is involved in left ventricular remodeling after myocardial infarction in rats Am J Physiol Heart Circ Physiol, February 1, 2006; 290(2): H709 - H715. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Misao, G. Takemura, M. Arai, S. Sato, K. Suzuki, S. Miyata, K.-i. Kosai, S. Minatoguchi, T. Fujiwara, and H. Fujiwara Bone marrow-derived myocyte-like cells and regulation of repair-related cytokines after bone marrow cell transplantation Cardiovasc Res, February 1, 2006; 69(2): 476 - 490. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Naugle, E. R. Olson, X. Zhang, S. E. Mase, C. F. Pilati, M. B. Maron, H. G. Folkesson, W. I. Horne, K. J. Doane, and J. G. Meszaros Type VI collagen induces cardiac myofibroblast differentiation: implications for postinfarction remodeling Am J Physiol Heart Circ Physiol, January 1, 2006; 290(1): H323 - H330. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Cucoranu, R. Clempus, A. Dikalova, P. J. Phelan, S. Ariyan, S. Dikalov, and D. Sorescu NAD(P)H Oxidase 4 Mediates Transforming Growth Factor-{beta}1-Induced Differentiation of Cardiac Fibroblasts Into Myofibroblasts Circ. Res., October 28, 2005; 97(9): 900 - 907. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Peng, O. A. Carretero, N. Vuljaj, T.-D. Liao, A. Motivala, E. L. Peterson, and N.-E. Rhaleb Angiotensin-Converting Enzyme Inhibitors: A New Mechanism of Action Circulation, October 18, 2005; 112(16): 2436 - 2445. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Wang, N. Xu, X. Feng, N. Hou, J. Zhang, X. Cheng, Y. Chen, Y. Zhang, and X. Yang Targeted Disruption of Smad4 in Cardiomyocytes Results in Cardiac Hypertrophy and Heart Failure Circ. Res., October 14, 2005; 97(8): 821 - 828. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Vellaichamy, M. L. Khurana, J. Fink, and K. N. Pandey Involvement of the NF-{kappa}B/Matrix Metalloproteinase Pathway in Cardiac Fibrosis of Mice Lacking Guanylyl Cyclase/Natriuretic Peptide Receptor A J. Biol. Chem., May 13, 2005; 280(19): 19230 - 19242. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Boynton-Jarrett, J. Rich-Edwards, S. Malspeis, S. A. Missmer, and R. Wright A Prospective Study of Hypertension and Risk of Uterine Leiomyomata Am. J. Epidemiol., April 1, 2005; 161(7): 628 - 638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Saito, N. Ishizaka, T. Aizawa, M. Sata, N. Iso-o, E. Noiri, I. Mori, M. Ohno, and R. Nagai Iron chelation and a free radical scavenger suppress angiotensin II-induced upregulation of TGF-{beta}1 in the heart Am J Physiol Heart Circ Physiol, April 1, 2005; 288(4): H1836 - H1843. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Tsuruda, J. Kato, K. Hatakeyama, H. Masuyama, Y.-N. Cao, T. Imamura, K. Kitamura, Y. Asada, and T. Eto Antifibrotic effect of adrenomedullin on coronary adventitia in angiotensin II-induced hypertensive rats Cardiovasc Res, March 1, 2005; 65(4): 921 - 929. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. R. Olson, J. E. Naugle, X. Zhang, J. A. Bomser, and J. G. Meszaros Inhibition of cardiac fibroblast proliferation and myofibroblast differentiation by resveratrol Am J Physiol Heart Circ Physiol, March 1, 2005; 288(3): H1131 - H1138. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Motomura, H. Kanbayashi, W. I. Khan, Y. Deng, P. A. Blennerhassett, P. J. Margetts, J. Gauldie, K. Egashira, and S. M. Collins The gene transfer of soluble VEGF type I receptor (Flt-1) attenuates peritoneal fibrosis formation in mice but not soluble TGF-{beta} type II receptor gene transfer Am J Physiol Gastrointest Liver Physiol, January 1, 2005; 288(1): G143 - G150. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Chen, J. L. Mehta, D. Li, L. Joseph, and J. Joseph Transforming Growth Factor {beta} Receptor Endoglin Is Expressed in Cardiac Fibroblasts and Modulates Profibrogenic Actions of Angiotensin II Circ. Res., December 10, 2004; 95(12): 1167 - 1173. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Fairweather, S. Frisancho-Kiss, S. A. Yusung, M. A. Barrett, S. E. Davis, S. J.L. Gatewood, D. B. Njoku, and N. R. Rose Interferon-{gamma} Protects against Chronic Viral Myocarditis by Reducing Mast Cell Degranulation, Fibrosis, and the Profibrotic Cytokines Transforming Growth Factor-{beta}1, Interleukin-1{beta}, and Interleukin-4 in the Heart Am. J. Pathol., December 1, 2004; 165(6): 1883 - 1894. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Ikeuchi, H. Tsutsui, T. Shiomi, H. Matsusaka, S. Matsushima, J. Wen, T. Kubota, and A. Takeshita Inhibition of TGF-{beta} signaling exacerbates early cardiac dysfunction but prevents late remodeling after infarction Cardiovasc Res, December 1, 2004; 64(3): 526 - 535. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Pokharel, P. P. van Geel, U. C. Sharma, J. P.M. Cleutjens, H. Bohnemeier, X.-L. Tian, H. Schunkert, H. J.G.M. Crijns, M. Paul, and Y. M. Pinto Increased Myocardial Collagen Content in Transgenic Rats Overexpressing Cardiac Angiotensin-Converting Enzyme Is Related to Enhanced Breakdown of N-Acetyl-Ser-Asp-Lys-Pro and Increased Phosphorylation of Smad2/3 Circulation, November 9, 2004; 110(19): 3129 - 3135. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Philip-Couderc, F. Smih, J. E. Hall, A. Pathak, J. Roncalli, R. Harmancey, P. Massabuau, M. Galinier, P. Verwaerde, J.-M. Senard, et al. Kinetic analysis of cardiac transcriptome regulation during chronic high-fat diet in dogs Physiol Genomics, September 16, 2004; 19(1): 32 - 40. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Rosenkranz TGF-{beta}1 and angiotensin networking in cardiac remodeling Cardiovasc Res, August 15, 2004; 63(3): 423 - 432. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Stawowy, C. Margeta, H. Kallisch, N. G Seidah, M. Chretien, E. Fleck, and K. Graf Regulation of matrix metalloproteinase MT1-MMP/MMP-2 in cardiac fibroblasts by TGF-{beta}1 involves furin-convertase Cardiovasc Res, July 1, 2004; 63(1): 87 - 97. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. A. Kass, J. G.F. Bronzwaer, and W. J. Paulus What Mechanisms Underlie Diastolic Dysfunction in Heart Failure? Circ. Res., June 25, 2004; 94(12): 1533 - 1542. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. G. Petrich, B. C. Eloff, D. L. Lerner, A. Kovacs, J. E. Saffitz, D. S. Rosenbaum, and Y. Wang Targeted Activation of c-Jun N-terminal Kinase in Vivo Induces Restrictive Cardiomyopathy and Conduction Defects J. Biol. Chem., April 9, 2004; 279(15): 15330 - 15338. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Kuwahara, H. Kai, K. Tokuda, M. Takeya, A. Takeshita, K. Egashira, and T. Imaizumi Hypertensive Myocardial Fibrosis and Diastolic Dysfunction: Another Model of Inflammation? Hypertension, April 1, 2004; 43(4): 739 - 745. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Kapoun, F. Liang, G. O'Young, D. L. Damm, D. Quon, R. T. White, K. Munson, A. Lam, G. F. Schreiner, and A. A. Protter B-Type Natriuretic Peptide Exerts Broad Functional Opposition to Transforming Growth Factor-{beta} in Primary Human Cardiac Fibroblasts: Fibrosis, Myofibroblast Conversion, Proliferation, and Inflammation Circ. Res., March 5, 2004; 94(4): 453 - 461. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yoshida, K. Yamamoto, T. Mano, Y. Sakata, N. Nishikawa, M. Nishio, T. Ohtani, T. Miwa, M. Hori, and T. Masuyama AT1 Receptor Blocker Added to ACE Inhibitor Provides Benefits at Advanced Stage of Hypertensive Diastolic Heart Failure Hypertension, March 1, 2004; 43(3): 686 - 691. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Tokuda, H. Kai, F. Kuwahara, H. Yasukawa, N. Tahara, H. Kudo, K. Takemiya, M. Koga, T. Yamamoto, and T. Imaizumi Pressure-Independent Effects of Angiotensin II on Hypertensive Myocardial Fibrosis Hypertension, February 1, 2004; 43(2): 499 - 503. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. P. Czubryt and E. N. Olson Balancing Contractility and Energy Production: The Role of Myocyte Enhancer Factor 2 (MEF2) in Cardiac Hypertrophy Recent Prog. Horm. Res., January 1, 2004; 59(1): 105 - 124. [Abstract] [Full Text]
|
|
|
|
 |
|
 H. Huang, S. B. Petkova, A. W. Cohen, B. Bouzahzah, J. Chan, J.-n. Zhou, S. M. Factor, L. M. Weiss, M. Krishnamachary, S. Mukherjee, et al. Activation of Transcription Factors AP-1 and NF-{kappa}B in Murine Chagasic Myocarditis Infect. Immun., May 1, 2003; 71(5): 2859 - 2867. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Takai, D. Jin, M. Sakaguchi, S. Katayama, M. Muramatsu, M. Sakaguchi, E. Matsumura, S. Kim, and M. Miyazaki A Novel Chymase Inhibitor, 4-[1-{[bis-(4-Methyl-phenyl)-methyl]-carbamoyl}-3-(2-ethoxy-benzyl)-4-oxo-azetidine-2-yloxy]-benzoic acid (BCEAB), Suppressed Cardiac Fibrosis in Cardiomyopathic Hamsters J. Pharmacol. Exp. Ther., April 1, 2003; 305(1): 17 - 23. [Abstract] [Full Text]
|
|
|
|
 |
|
 T. Nagata, H. Kai, R. Shibata, M. Koga, A. Yoshimura, and T. Imaizumi Oncostatin M, an Interleukin-6 Family Cytokine, Upregulates Matrix Metalloproteinase-9 Through the Mitogen-Activated Protein Kinase Kinase-Extracellular Signal-Regulated Kinase Pathway in Cultured Smooth Muscle Cells Arterioscler Thromb Vasc Biol, April 1, 2003; 23(4): 588 - 593. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Kuwahara, H. Kai, K. Tokuda, H. Niiyama, N. Tahara, K. Kusaba, K. Takemiya, A. Jalalidin, M. Koga, T. Nagata, et al. Roles of Intercellular Adhesion Molecule-1 in Hypertensive Cardiac Remodeling Hypertension, March 1, 2003; 41(3): 819 - 823. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.F. Cordeiro, F. Kuwahara, H. Kai, K. Tokuda, T. Imaizumi, M. Kai, A. Takeshita, and K. Egashira Transforming Growth Factor-{beta} Function Blocking Already Effective as Therapeutic Strategy * Response Circulation, February 11, 2003; 107 (5): e37 - e37. [Full Text] [PDF]
|
|
|
|
|
|
I. Manabe, T. Shindo, and R. Nagai Gene Expression in Fibroblasts and Fibrosis: Involvement in Cardiac Hypertrophy Circ. Res., December 13, 2002; 91(12): 1103 - 1113. [Abstract] [Full Text] [PDF]
|
|