Activation of host tissue trophic factors through JAK-STAT3 signaling: A mechanism of mesenchymal stem cell-mediated cardiac repair

American Journal of Physiology - Heart and Circulatory Physiology

Volumn 299, Issue 5, 2010, Pages

  Shabbir, Arsalan ^a   Zisa, David ^a   Lin, Huey ^a   Mastri, Michalis ^a   Roloff, Gregory ^a   Suzuki, Gen ^a   Lee, Techung ^a  

^a UNIVERSITY AT BUFFALO   (United States)

Author keywords

IL 6 type cytokine; Signal transducer and activator of transcription 3; Skeletal muscle

Indexed keywords

AGENTS INTERACTING WITH TRANSMITTER, HORMONE OR DRUG RECEPTORS; INTERLEUKIN 6; JANUS KINASE; SCATTER FACTOR; STAT3 PROTEIN; UNCLASSIFIED DRUG; VASCULOTROPIN; WP 1066;

ANIMAL EXPERIMENT; ARTICLE; CELL PERMEABILIZATION; CYTOKINE PRODUCTION; DRUG INHIBITION; ENZYME LINKED IMMUNOSORBENT ASSAY; HEART; HUMAN; HUMAN CELL; IN VITRO STUDY; MALE; MESENCHYMAL STEM CELL; NONHUMAN; PRIORITY JOURNAL; SKELETAL MUSCLE; TISSUE REPAIR;

ANIMALS; CELLS, CULTURED; CRICETINAE; CYTOKINES; HEPATOCYTE GROWTH FACTOR; JANUS KINASES; MALE; MESENCHYMAL STEM CELL TRANSPLANTATION; MESENCHYMAL STEM CELLS; MODELS, ANIMAL; MUSCLE FIBERS, SKELETAL; MYOCARDIUM; PYRIDINES; SIGNAL TRANSDUCTION; STAT3 TRANSCRIPTION FACTOR; TYRPHOSTINS; VASCULAR ENDOTHELIAL GROWTH FACTOR A;

EID: 78249278186     PISSN: 03636135     EISSN: 15221539     Source Type: Journal    
DOI: 10.1152/ajpheart.00488.2010     Document Type: Article

References (77)

1. Abraham D, Hofbauer R, Schafer R, Blumer R, Paulus P, Miksovsky A, Traxler H, Kocher A, Aharinejad S. Selective downregulation of VEGF-A(165), VEGF-R(1), and decreased capillary density in patients with dilative but not ischemic cardiomyopathy. Circ Res 87: 644-647, 2000.
2. Boengler K, Buechert A, Heinen Y, Roeskes C, Hilfiker-Kleiner D, Heusch G, Schulz R. Cardioprotection by ischemic postconditioning is lost in aged and STAT3-deficient mice. Circ Res 102: 131-135, 2008.
3. Brunswig-Spickenheier B, Boche J, Westenfelder C, Peimann F, Gruber AD, Jaquet K, Krause K, Zustin J, Zander AR, Lange C. Limited immune-modulating activity of porcine mesenchymal stromal cells abolishes their protective efficacy in acute kidney injury. Stem Cells Dev 19: 719-729, 2010.
4. Caplan A. Why are MSCs therapeutic? New data: new insight. J Pathol 217: 318-324, 2009.
5. Chae CH, Jung SL, An SH, Park BY, Wang SW, Cho IH, Cho JY, Kim HT. Treadmill exercise improves cognitive function and facilitates nerve growth factor signaling by activating mitogen-activated protein kinase/extracellular signal-regulated kinase1/2 in the streptozotocin-induced diabetic rat hippocampus. Neuroscience 164: 1665-1673, 2009.
6. Cho HJ, Lee N, Lee JY, Choi YJ, Ii M, Wecker A, Jeong JO, Curry C, Qin G, Yoon YS. Role of host tissues for sustained humoral effects after endothelial progenitor cell transplantation into the ischemic heart. J Exp Med 204: 3257-3269, 2007.
7. Darnell JE Jr. STATs and gene regulation. Science 277: 1630-1635, 1997.
8. Dixon IM, Ju H, Reid NL, Scammell-La Fleur T, Werner JP, Jasmin G. Cardiac collagen remodeling in the cardiomyopathic Syrian hamster and the effect of losartan. J Mol Cell Cardiol 29: 1837-1850, 1997.
9. Djouad F, Charbonnier LM, Bouffi C, Louis-Plence P, Bony C, Apparailly F, Cantos C, Jorgensen C, Noel D. Mesenchymal stem cells inhibit the differentiation of dendritic cells through an interleukin-6- dependent mechanism. Stem Cells 25: 2025-2032, 2007.
10. Ernst M, Jenkins BJ. Acquiring signalling specificity from the cytokine receptor gp130. Trends Genet 20: 23-32, 2004.
11. Funamoto M, Fujio Y, Kunisada K, Negoro S, Tone E, Osugi T, Hirota H, Izumi M, Yoshizaki K, Walsh K, Kishimoto T, Yamauchi- Takihara K. Signal transducer and activator of transcription 3 is required for glycoprotein 130-mediated induction of vascular endothelial growth factor in cardiac myocytes. J Biol Chem 275: 10561-10566, 2000.
12. Gnecchi M, Zhang Z, Ni A, Dzau VJ. Paracrine mechanisms in adult stem cell signaling and therapy. Circ Res 103: 1204-1219, 2008.
13. Goineau S, Pape D, Guillo P, Ramee MP, Bellissant E. Hemodynamic and histomorphometric characteristics of dilated cardiomyopathy of Syrian hamsters (Bio TO-2 strain). Can J Physiol Pharmacol 79: 329-337, 2001.
14. Hendriks JJ, Slaets H, Carmans S, de Vries HE, Dijkstra CD, Stinissen P, Hellings N. Leukemia inhibitory factor modulates production of inflammatory mediators and myelin phagocytosis by macrophages. J Neuroimmunol 204: 52-57, 2008.
15. Hilfiker-Kleiner D, Hilfiker A, Drexler H. Many good reasons to have STAT3 in the heart. Pharmacol Ther 107: 131-137, 2005.
16. Hirota H, Chen J, Betz UA, Rajewsky K, Gu Y, Ross J Jr, Muller W, Chien KR. Loss of a gp130 cardiac muscle cell survival pathway is a critical event in the onset of heart failure during biomechanical stress. Cell 97: 189-198, 1999.
17. Hughes RA, Sendtner M, Goldfarb M, Lindholm D, Thoenen H. Evidence that fibroblast growth factor 5 is a major muscle-derived survival factor for cultured spinal motoneurons. Neuron 10: 369-377, 1993. (Pubitemid 23111730)
18. Iwamaru A, Szymanski S, Iwado E, Aoki H, Yokoyama T, Fokt I, Hess K, Conrad C, Madden T, Sawaya R, Kondo S, Priebe W, Kondo Y. A novel inhibitor of the STAT3 pathway induces apoptosis in malignant glioma cells both in vitro and in vivo. Oncogene 26: 2435-2444, 2007. (Pubitemid 46597261)
19. Jacoby JJ, Kalinowski A, Liu MG, Zhang SS, Gao Q, Chai GX, Ji L, Iwamoto Y, Li E, Schneider M, Russell KS, Fu XY. Cardiomyocyte-restricted knockout of STAT3 results in higher sensitivity to inflammation, cardiac fibrosis, and heart failure with advanced age. Proc Natl Acad Sci USA 100: 12929-12934, 2003. (Pubitemid 37340002)
20. Kato Y, Iwase M, Takagi K, Nishizawa T, Kanazawa H, Matsushita A, Umeda H, Izawa H, Noda A, Koike Y, Nagata K, Yokota M. Differential myolysis of myocardium and skeletal muscle in hamsters with dilated cardiomyopathy. Circ J 70: 1497-1502, 2006.
21. Kaye DM, Vaddadi G, Gruskin SL, Du XJ, Esler MD. Reduced myocardial nerve growth factor expression in human and experimental heart failure. Circ Res 86: E80-E84, 2000.
22. Khalife WI, Tang YD, Kuzman JA, Thomas TA, Anderson BE, Said S, Tille P, Schlenker EH, Gerdes AM. Treatment of subclinical hypothyroidism reverses ischemia and prevents myocyte loss and progressive LV dysfunction in hamsters with dilated cardiomyopathy. Am J Physiol Heart Circ Physiol 289: H2409-H2415, 2005. (Pubitemid 41698550)
23. Kilroy GE, Foster SJ, Wu X, Ruiz J, Sherwood S, Heifetz A, Ludlow JW, Stricker DM, Potiny S, Green P, Halvorsen YD, Cheatham B, Storms RW, Gimble JM. Cytokine profile of human adipose-derived stem cells: expression of angiogenic, hematopoietic, and pro-inflammatory factors. J Cell Physiol 212: 702-709, 2007. (Pubitemid 47271882)
24. Komamura K, Tatsumi R, Miyazaki J, Matsumoto K, Yamato E, Nakamura T, Shimizu Y, Nakatani T, Kitamura S, Tomoike H, Kitakaze M, Kangawa K, Miyatake K. Treatment of dilated cardiomyopathy with electroporation of hepatocyte growth factor gene into skeletal muscle. Hypertension 44: 365-371, 2004. (Pubitemid 39167237)
25. Kositprapa C, Zhang B, Berger S, Canty JM, Lee TC. Calpainmediated proteolytic cleavage of troponin I induced by hypoxia or metabolic inhibition in cultured neonatal cardiomyocytes. Mol Cell Biochem 214: 47-55, 2000. (Pubitemid 32010325)
26. Kunisada K, Negoro S, Tone E, Funamoto M, Osugi T, Yamada S, Okabe M, Kishimoto T, Yamauchi-Takihara K. Signal transducer and activator of transcription 3 in the heart transduces not only a hypertrophic signal but a protective signal against doxorubicin-induced cardiomyopathy. Proc Natl Acad Sci USA 97: 315-319, 2000. (Pubitemid 30055828)
27. Kurdi M, Booz GW. Can the protective actions of JAK-STAT in the heart be exploited therapeutically? Parsing the regulation of interleukin-6-type cytokine signaling. J Cardiovasc Pharmacol 50: 126-141, 2007.
28. Lehrke S, Mazhari R, Durand DJ, Zheng M, Bedja D, Zimmet JM, Schuleri KH, Chi AS, Gabrielson KL, Hare JM. Aging impairs the beneficial effect of granulocyte colony-stimulating factor and stem cell factor on post-myocardial infarction remodeling. Circ Res 99: 553-560, 2006.
29. Lin H, Shabbir A, Molnar M, Yang J, Marion S, Canty JM, Lee T. Adenoviral expression of vascular endothelial growth factor splice variants differentially regulate bone marrow-derived mesenchymal stem cells. J Cell Physiol 216: 458-468, 2008.
30. Lipskaia L, Pinet C, Fromes Y, Hatem S, Cantaloube I, Coulombe A, Lompre AM. Mutation of delta-sarcoglycan is associated with Ca(+) -dependent vascular remodeling in the Syrian hamster. Am J Pathol 171: 162-171, 2007.
31. Lista I, Sorrentino G. Biological mechanisms of physical activity in preventing cognitive decline. Cell Mol Neurobiol 30: 493-503, 2010.
32. MacDougald OA, Burant CF. The rapidly expanding family of adipokines. Cell Metab 6: 159-161, 2007.
33. Madden T, Kazerooni R, Myer J, Culotta K, Donato N, Johansen MJ, Kondo Y, Mack D, Priebe W. The preclinical pharmacology of WP 1066, a potent small molecule inhibitor of the JAK2/STAT3 pathway. In: AACR Meeting, Experimental and Molecular Therapeutics 40, Abstract #4853, 2006.
34. Minieri M, Fiaccavento R, Carosella L, Peruzzi G, Di Nardo P. The cardiomyopathic hamster as model of early myocardial aging. Mol Cell Biochem 198: 1-6, 1999.
35. Missihoun C, Zisa D, Shabbir A, Lin H, Lee T. Myocardial oxidative stress, osteogenic phenotype, and energy metabolism are differentially involved in the initiation and early progression of delta-sarcoglycan-null cardiomyopathy. Mol Cell Biochem 321: 45-52, 2008.
36. Miyata S, Takemura G, Kawase Y, Li Y, Okada H, Maruyama R, Ushikoshi H, Esaki M, Kanamori H, Li L, Misao Y, Tezuka A, Toyo-Oka T, Minatoguchi S, Fujiwara T, Fujiwara H. Autophagic cardiomyocyte death in cardiomyopathic hamsters and its prevention by granulocyte colony-stimulating factor. Am J Pathol 168: 386-397, 2006.
37. Mudd JO, Kass DA. Tackling heart failure in the twenty-first century. Nature 451: 919-928, 2008.
38. Nakamura T, Matsumoto K, Mizuno S, Sawa Y, Matsuda H, Nakamura T. Hepatocyte growth factor prevents tissue fibrosis, remodeling, and dysfunction in cardiomyopathic hamster hearts. Am J Physiol Heart Circ Physiol 288: H2131-H2139, 2005.
39. Negoro S, Kunisada K, Fujio Y, Funamoto M, Darville MI, Eizirik DL, Osugi T, Izumi M, Oshima Y, Nakaoka Y, Hirota H, Kishimoto T, Yamauchi-Takihara K. Activation of signal transducer and activator of transcription 3 protects cardiomyocytes from hypoxia/reoxygenation-induced oxidative stress through the upregulation of manganese superoxide dismutase. Circulation 104: 979-981, 2001.
40. Novotny NM, Markel TA, Crisostomo PR, Meldrum DR. Differential IL-6 and VEGF secretion in adult and neonatal mesenchymal stem cells: role of NFkB. Cytokine 43: 215-219, 2008.
41. O'Reilly C, McKay B, Phillips S, Tarnopolsky M, Parise G. Hepatocyte growth factor (HGF) and the satellite cell response following muscle lengthening contractions in humans. Muscle Nerve 38: 1434-1442, 2008.
42. Ortiz LA, Dutreil M, Fattman C, Pandey AC, Torres G, Go K, Phinney DG. Interleukin 1 receptor antagonist mediates the antiinflammatory and antifibrotic effect of mesenchymal stem cells during lung injury. Proc Natl Acad Sci USA 104: 11002-11007, 2007.
43. Osugi T, Oshima Y, Fujio Y, Funamoto M, Yamashita A, Negoro S, Kunisada K, Izumi M, Nakaoka Y, Hirota H, Okabe M, Yamauchi- Takihara K, Kawase I, Kishimoto T. Cardiac-specific activation of signal transducer and activator of transcription 3 promotes vascular formation in the heart. J Biol Chem 277: 6676-6681, 2002.
44. Park KS, Kim YS, Kim JH, Choi B, Kim SH, Tan AH, Lee MS, Lee MK, Kwon CH, Joh JW, Kim SJ, Kim KW. Trophic molecules derived from human mesenchymal stem cells enhance survival, function, and angiogenesis of isolated islets after transplantation. Transplantation 89: 509-517, 2010.
45. Pedersen BK, Febbraio MA. Muscle as an endocrine organ: focus on muscle-derived interleukin-6. Physiol Rev 88: 1379-1406, 2008.
46. Podewski EK, Hilfiker-Kleiner D, Hilfiker A, Morawietz H, Lichtenberg A, Wollert KC, Drexler H. Alterations in Janus kinase (JAK)-signal transducers and activators of transcription (STAT) signaling in patients with end-stage dilated cardiomyopathy. Circulation 107: 798-802, 2003.
47. Prockop DJ. "Stemness" does not explain the repair of many tissues by mesenchymal stem/multipotent stromal cells (MSCs). Clin Pharmacol Ther 82: 241-243, 2007.
48. Rissanen TT, Vajanto I, Hiltunen MO, Rutanen J, Kettunen MI, Niemi M, Leppanen P, Turunen MP, Markkanen JE, Arve K, Alhava E, Kauppinen RA, Yla-Herttuala S. Expression of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 (KDR/ Flk-1) in ischemic skeletal muscle and its regeneration. Am J Pathol 160: 1393-1403, 2002.
49. Ryoke T, Gu Y, Ikeda Y, Martone ME, Oh SS, Jeon ES, Knowlton KU, Ross J Jr. Apoptosis and oncosis in the early progression of left ventricular dysfunction in the cardiomyopathic hamster. Basic Res Cardiol 97: 65-75, 2002.
50. Sakamoto A, Ono K, Abe M, Jasmin G, Eki T, Murakami Y, Masaki T, Toyo-Oka T, Hanaoka F. Both hypertrophic and dilated cardiomyopathies are caused by mutation of the same gene, delta-sarcoglycan, in hamster: an animal model of disrupted dystrophin-associated glycoprotein complex. Proc Natl Acad Sci USA 94: 13873-13878, 1997.
51. Serose A, Prudhon B, Salmon A, Doyennette MA, Fiszman MY, Fromes Y. Administration of insulin-like growth factor-1 (IGF-1) improves both structure and function of delta-sarcoglycan deficient cardiac muscle in the hamster. Basic Res Cardiol 100: 161-170, 2005.
52. Serrano AL, Baeza-Raja B, Perdiguero E, Jardi M, Munoz-Canoves P. Interleukin-6 is an essential regulator of satellite cell-mediated skeletal muscle hypertrophy. Cell Metab 7: 33-44, 2008.
53. Shabbir A, Zisa D, Leiker M, Johnston C, Lin H, Lee T. Muscular dystrophy therapy by non-autologous mesenchymal stem cells: muscle regeneration without immunosuppression and inflammation. Transplantation 87: 1275-1282, 2009.
54. Shabbir A, Zisa D, Suzuki G, Lee T. Heart failure therapy mediated by the trophic activities of bone marrow mesenchymal stem cells: a noninvasive therapeutic regimen. Am J Physiol Heart Circ Physiol 296: H1888-H1897, 2009.
55. Shimizu T, Okamoto H, Watanabe M, Kumamoto H, Chiba S, Matsui Y, Sugawara T, Onozuka H, Mikami T, Kitabatake A. Altered microvasculature is involved in remodeling processes in cardiomyopathic hamsters. Jpn Heart J 44: 111-126, 2003.
56. Slomiany MG, Black LA, Kibbey MM, Day TA, Rosenzweig SA. IGF-1 induced vascular endothelial growth factor secretion in head and neck squamous cell carcinoma. Biochem Biophys Res Commun 342: 851-858, 2006.
57. Spangenburg EE, Booth FW. Multiple signaling pathways mediate LIF-induced skeletal muscle satellite cell proliferation. Am J Physiol Cell Physiol 283: C204-C211, 2002.
58. Suetta C, Clemmensen C, Andersen JL, Magnusson SP, Schjerling P, Kjaer M. Coordinated increase in skeletal muscle fiber area and expression of IGF-I with resistance exercise in elderly post-operative patients. Growth Horm IGF Res 20: 134-140, 2010.
59. Suzuki G, Lee T, Fallavollita JA, Canty JM Jr. Adenoviral gene transfer of FGF-5 to hibernating myocardium improves function and stimulates myocytes to hypertrophy and reenter the cell cycle. Circ Res 96: 767-775, 2005.
60. Tatsumi R, Anderson JE, Nevoret CJ, Halevy O, Allen RE. HGF/SF is present in normal adult skeletal muscle and is capable of activating satellite cells. Dev Biol 194: 114-128, 1998. (Pubitemid 28115263)
61. Testa U, Pannitteri G, Condorelli GL. Vascular endothelial growth factors in cardiovascular medicine. J Cardiovasc Med (Hagerstown) 9: 1190-1221, 2008.
62. Thompson PD, Buchner D, Pina IL, Balady GJ, Williams MA, Marcus BH, Berra K, Blair SN, Costa F, Franklin B, Fletcher GF, Gordon NF, Pate RR, Rodriguez BL, Yancey AK, Wenger NK. American Heart Association Council on Clinical Cardiology Subcommittee on Exercise, Rehabilitation, and Prevention; American Heart Association Council on Nutrition, Physical Activity, and Metabolism Subcommittee on Physical Activity. Exercise and physical activity in the prevention and treatment of atherosclerotic cardiovascular disease: a statement from the Council on Clinical Cardiology (Subcommittee on Exercise, Rehabilitation, and Prevention) and the Council on Nutrition, Physical Activity, and Metabolism (Subcommittee on Physical Activity). Circulation 107: 3109-3116, 2003.
63. Trenerry MK, Carey KA, Ward AC, Cameron-Smith D. STAT3 signaling is activated in human skeletal muscle following acute resistance exercise. J Appl Physiol 102: 1483-1489, 2007. (Pubitemid 46571044)
64. Tyndall A, Walker UA, Cope A, Dazzi F, De Bari C, Fibbe W, Guiducci S, Jones S, Jorgensen C, Le Blanc K, Luyten F, McGonagle D, Martin I, Bocelli-Tyndall C, Pennesi G, Pistoia V, Pitzalis C, Uccelli A, Wulffraat N, Feldmann M. Immunomodulatory properties of mesenchymal stem cells: a review based on an interdisciplinary meeting held at the Kennedy Institute of Rheumatology Division, London, UK, 31 October 2005. Arthritis Res Ther 9: 301, 2007.
65. Vacanti V, Kong E, Suzuki G, Sato K, Canty JM Jr, Lee T. Phenotypic changes of adult porcine mesenchymal stem cells induced by prolonged passaging in culture. J Cell Physiol 205: 194-201, 2005. (Pubitemid 41428901)
66. Van Belle E, Witzenbichler B, Chen D, Silver M, Chang L, Schwall R, Isner JM. Potentiated angiogenic effect of scatter factor/hepatocyte growth factor via induction of vascular endothelial growth factor: the case for paracrine amplification of angiogenesis. Circulation 97: 381-390, 1998. (Pubitemid 28074339)
67. Van Berendoncks AM, Beckers P, Hoymans VY, Possemiers N, Wuytss FL, Vrints CJ, Conraads VM. Exercise training reduces circulating adiponectin levels in patients with chronic heart failure. Clin Sci (Lond) 118: 281-289, 2010.
68. Van Berendoncks AM, Garnier A, Beckers P, Hoymans VY, Possemiers N, Fortin D, Martinet W, Van Hoof V, Vrints CJ, Ventura- Clapier R, Conraads VM. Functional adiponectin resistance at the level of the skeletal muscle in mild to moderate chronic heart failure. Circ Heart Fail 3: 185-194, 2010.
69. Walsh K. Adipokines, myokines and cardiovascular disease. Circ J 73: 13-18, 2009.
70. Wang K, Wang C, Xiao F, Wang H, Wu Z. JAK2/STAT2/STAT3 are required for myogenic differentiation. J Biol Chem 283: 34029-34036, 2008.
71. Wehner R, Wehrum D, Bornhauser M, Zhao S, Schakel K, Bachmann MP, Platzbecker U, Ehninger G, Rieber EP, Schmitz M. Mesenchymal stem cells efficiently inhibit the proinflammatory properties of 6-sulfo LacNAc dendritic cells. Haematologica 94: 1151-1156, 2009.
72. Wojcik EJ, Sharifpoor S, Miller NA, Wright TG, Watering R, Tremblay EA, Swan K, Mueller CR, Elliott BE. A novel activating function of c-Src and Stat3 on HGF transcription in mammary carcinoma cells. Oncogene 25: 2773-2784, 2006.
73. Wojta J, Kaun C, Breuss JM, Koshelnick Y, Beckmann R, Hattey E, Mildner M, Weninger W, Nakamura T, Tschachler E, Binder BR. Hepatocyte growth factor increases expression of vascular endothelial growth factor and plasminogen activator inhibitor-1 in human keratinocytes and the vascular endothelial growth factor receptor flk-1 in human endothelial cells. Lab Invest 79: 427-438, 1999. (Pubitemid 29190432)
74. Wu G, Rana JS, Wykrzykowska J, Du Z, Ke Q, Kang P, Li J, Laham RJ. Exercise-induced expression of VEGF and salvation of myocardium in the early stage of myocardial infarction. Am J Physiol Heart Circ Physiol 296: H389-H395, 2009.
75. Xin X, Yang S, Ingle G, Zlot C, Rangell L, Kowalski J, Schwall R, Ferrara N, Gerritsen ME. Hepatocyte growth factor enhances vascular endothelial growth factor-induced angiogenesis in vitro and in vivo. Am J Pathol 158: 1111-1120, 2001. (Pubitemid 32221803)
76. Zisa D, Shabbir A, Mastri M, Suzuki G, Lee T. Intramuscular VEGF repairs the failing heart: role of host-derived growth factors and mobilization of progenitor cells. Am J Physiol Regul Integr Comp Physiol 297: R1503-R1515, 2009.
77. Zisa D, Shabbir A, Suzuki G, Lee T. Vascular endothelial growth factor (VEGF) as a key therapeutic trophic factor in bone marrow mesenchymal stem cell-mediated cardiac repair. Biochem Biophys Res Commun 390: 834-838, 2009.