Versican in inflammation and tissue remodeling: The impact on lung disorders

Glycobiology

Volumn 25, Issue 3, 2014, Pages 243-251

  Andersson Sjöland, Annika ^a   Hallgren, Oskar ^a,b   Rolandsson, Sara ^a   Weitoft, Maria ^a   Tykesson, Emil ^a   Larsson Callerfelt, Anna Karin ^a   Rydell Törmänen, Kristina ^a   Bjermer, Leif ^b   Malmström, Anders ^b   Karlsson, Jenny C ^a   Westergren Thorsson, Gunilla ^a  

^a LUND UNIVERSITY   (Sweden)

^b LUND UNIVERSITY HOSPITAL   (Sweden)

Author keywords

extracellular matrix; lung disorders; remodeling; versican

Indexed keywords

TRANSFORMING GROWTH FACTOR BETA; VERSICAN;

ASTHMA; BRONCHIOLITIS OBLITERANS; CHRONIC OBSTRUCTIVE LUNG DISEASE; HUMAN; INFLAMMATION; LUNG DISEASE; NONHUMAN; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN FUNCTION; PROTEIN STRUCTURE; REGULATORY MECHANISM; REVIEW; TISSUE REMODELING; TISSUES; ANIMAL; CHEMISTRY; EXTRACELLULAR MATRIX; GENETICS; METABOLISM;

ANIMALS; EXTRACELLULAR MATRIX; HUMANS; LUNG DISEASES; VERSICANS;

EID: 84943261780     PISSN: 09596658     EISSN: 14602423     Source Type: Journal    
DOI: 10.1093/glycob/cwu120     Document Type: Review

References (93)

1. Akhmetshina A, Palumbo K, Dees C, Bergmann C, Venalis P, Zerr P, Horn A, Kireva T, Beyer C, Zwerina J, et al. 2012. Activation of canonical Wnt signalling is required for TGF-beta-mediated fibrosis. Nat Commun. 3:735.
2. Alber A, Howie SE, Wallace WA, Hirani N. 2012. The role of macrophages in healing the wounded lung. Int J Exp Pathol. 93:243-251.
3. Andersson-Sjoland A, Erjefalt JS, Bjermer L, Eriksson L, Westergren-Thorsson G. 2009. Fibrocytes are associated with vascular and parenchymal remodelling in patients with obliterative bronchiolitis. Respir Res. 10:103.
4. Andersson-Sjoland A, Nihlberg K, Eriksson L, Bjermer L, Westergren- Thorsson G. 2011. Fibrocytes and the tissue niche in lung repair. Respir Res. 12:76.
5. Andersson-Sjoland A, Thiman L, Nihlberg K, Hallgren O, Rolandsson S, Skog I, Mared L, Hansson L, Eriksson L, Bjermer L, et al. 2011. Fibroblast phenotypes and their activity are changed in the wound healing process after lung transplantation. J Heart Lung Transpl. 30:945-954.
6. Annoni R, Lancas T, Yukimatsu Tanigawa R, de Medeiros Matsushita M, de Morais Fernezlian S, Bruno A, Fernando Ferraz da Silva L, Roughley PJ, Battaglia S, Dolhnikoff M, et al. 2012. Extracellular matrix composition in COPD. Eur Respir J. 40:1362-1373.
7. Araujo BB, Dolhnikoff M, Silva LF, Elliot J, Lindeman JH, Ferreira DS, Mulder A, Gomes HA, Fernezlian SM, James A, et al. 2008. Extracellular matrix components and regulators in the airway smooth muscle in asthma. Eur Respir J. 32:61-69.
8. Bartolini B, Thelin MA, Svensson L, Ghiselli G, van Kuppevelt TH, Malmstrom A, Maccarana M. 2013. Iduronic acid in chondroitin/dermatan sulfate affects directional migration of aortic smooth muscle cells. PLoS ONE. 8:e66704.
9. Berdiaki A, Zafiropoulos A, Fthenou E, Katonis P, Tsatsakis A, Karamanos NK, Tzanakakis GN. 2008. Regulation of hyaluronan and versican deposition by growth factors in fibrosarcoma cell lines. Biochim Biophys Acta. 1780:194-202.
10. Beyer C, Schramm A, Akhmetshina A, Dees C, Kireva T, Gelse K, Sonnylal S, de Crombrugghe B, Taketo MM, Distler O, et al. 2012. Beta-catenin is a central mediator of pro-fibrotic Wnt signaling in systemic sclerosis. Ann Rheum Dis. 71:761-767.
11. Bianchetti L, Barczyk M, Cardoso J, Schmidt M, Bellini A, Mattoli S. 2012. Extracellular matrix remodelling properties of human fibrocytes. J Cell Mol Med. 16:483-495.
12. Black PN, Ching PS, Beaumont B, Ranasinghe S, Taylor G, Merrilees MJ. 2008. Changes in elastic fibres in the small airways and alveoli in COPD. Eur Respir J. 31:998-1004.
13. Boue S, De Leon H, Schlage WK, Peck MJ, Weiler H, Berges A, Vuillaume G, Martin F, Friedrichs B, Lebrun S, et al. 2013. Cigarette smoke induces molecular responses in respiratory tissues of ApoE(-/-) mice that are progressively deactivated upon cessation. Toxicology. 314:112-124.
14. Chang MY, Chan CK, Braun KR, Green PS, O'Brien KD, Chait A, Day AJ, Wight TN. 2012. Monocyte-to-macrophage differentiation: Synthesis and secretion of a complex extracellular matrix. J Biol Chem. 287: 14122-14135.
15. Chiappara G, Gagliardo R, Siena A, Bonsignore MR, Bousquet J, Bonsignore G, Vignola AM. 2001. Airway remodelling in the pathogenesis of asthma. Curr Opin Allergy Clin Immunol. 1:85-93.
16. Colombat M, Mal H, Groussard O, Capron F, Thabut G, Jebrak G, Brugiere O, Dauriat G, Castier Y, Leseche G, et al. 2007. Pulmonary vascular lesions in end-stage idiopathic pulmonary fibrosis: Histopathologic study on lung explant specimens and correlations with pulmonary hemodynamics. Hum Pathol. 38:60-65.
17. de Medeiros Matsushita M, da Silva LF, dos Santos MA, Fernezlian S, Schrumpf JA, Roughley P, Hiemstra PS, Saldiva PH, Mauad T, Dolhnikoff M. 2005. Airway proteoglycans are differentially altered in fatal asthma. J Pathol. 207:102-110.
18. Distler A, Deloch L, Huang J, Dees C, Lin NY, Palumbo-Zerr K, Beyer C, Weidemann A, Distler O, Schett G, et al. 2012. Inactivation of tankyrases reduces experimental fibrosis by inhibiting canonical Wnt signalling. Ann Rheum Dis. 72:1575-1580.
19. Dournes G, Laurent F. 2012. Airway remodelling in asthma and COPD: Findings, similarities, and differences using quantitative CT. Pulm Med. 2012:670414.
20. Du WW, Yang W, Yee AJ. 2013. Roles of versican in cancer biology - Tumorigenesis, progression and metastasis. Histol Histopathol. 28:701-713.
21. Duvernelle C, Freund V, Frossard N. 2003. Transforming growth factor-beta and its role in asthma. Pulm Pharmacol Therapeu. 16:181-196.
22. Evanko SP, Potter-Perigo S, Bollyky PL, Nepom GT, Wight TN. 2012. Hyaluronan and versican in the control of human T-lymphocyte adhesion and migration. Matrix Biol. 31:90-100.
23. Evanko SP, Potter-Perigo S, Johnson PY, Wight TN. 2009. Organization of hyaluronan and versican in the extracellular matrix of human fibroblasts treated with the viral mimetic poly I:C. J Histochem Cytochem. 57: 1041-1060.
24. Foster LJ, Zeemann PA, Li C, Mann M, Jensen ON, Kassem M. 2005. Differential expression profiling of membrane proteins by quantitative proteomics in a human mesenchymal stem cell line undergoing osteoblast differentiation. Stem Cells. 23:1367-1377.
25. Gill S, Wight TN, Frevert CW. 2010. Proteoglycans: Key regulators of pulmonary inflammation and the innate immune response to lung infection. Anat Rec (Hoboken). 293:968-981.
26. Gorowiec MR, Borthwick LA, Parker SM, Kirby JA, Saretzki GC, Fisher AJ. 2012. Free radical generation induces epithelial-to-mesenchymal transition in lung epithelium via a TGF-beta1-dependent mechanism. Free Radical Biol Med. 52:1024-1032.
27. Hallgren O, Nihlberg K, Dahlback M, Bjermer L, Eriksson LT, Erjefalt JS, Lofdahl CG, Westergren-Thorsson G. 2010. Altered fibroblast proteoglycan production in COPD. Respir Res. 11:55.
28. Hallgren O, Rolandsson S, Andersson-Sjoland A, Nihlberg K, Wieslander E, Kvist-Reimer M, Dahlback M, Eriksson L, Bjermer L, Erjefalt J, et al. 2012. Enhanced ROCK1 dependent contractility in fibroblast from chronic obstructive pulmonary disease patients. J Transl Med. 10:171.
29. Hirose J, Kawashima H, Yoshie O, Tashiro K, Miyasaka M. 2001. Versican interacts with chemokines and modulates cellular responses. J Biol Chem. 276:5228-5234.
30. Hitchcock AM, Costello CE, Zaia J. 2006. Glycoform quantification of chondroitin/ dermatan sulfate using a liquid chromatography-tandem mass spectrometry platform. Biochemistry. 45:2350-2361.
31. Huang J, Olivenstein R, Taha R, Hamid Q, Ludwig M. 1999. Enhanced proteoglycan deposition in the airway wall of atopic asthmatics. Am J Respirand Crit Care Med. 160:725-729.
32. Huang R, Merrilees MJ, Braun K, Beaumont B, Lemire J, Clowes AW, Hinek A, Wight TN. 2006. Inhibition of versican synthesis by antisense alters smooth muscle cell phenotype and induces elastic fiber formation in vitro and in neointima after vessel injury. Circ Res. 98:370-377.
33. Hwang JY, Johnson PY, Braun KR, Hinek A, Fischer JW, O'Brien KD, Starcher B, Clowes AW, Merrilees MJ, Wight TN. 2008. Retrovirally mediated overexpression of glycosaminoglycan-deficient biglycan in arterial smooth muscle cells induces tropoelastin synthesis and elastic fiber formation in vitro and in neointimae after vascular injury. Am J Pathol. 173:1919-1928.
34. Kamato D, Burch ML, Piva TJ, Rezaei HB, Rostam MA, Xu S, Zheng W, Little PJ, Osman N. 2013. Transforming growth factor-beta signalling: Role and consequences of Smad linker region phosphorylation. Cell Signal. 25:2017-2024.
35. Kawashima H, Atarashi K, Hirose M, Hirose J, Yamada S, Sugahara K, Miyasaka M. 2002. Oversulfated chondroitin/dermatan sulfates containing GlcAbeta1/IdoAalpha1-3GalNAc(4, 6-O-disulfate) interact with L- and P-selectin and chemokines. J Biol Chem. 277:12921-12930.
36. Khalil N, Greenberg AH. 1991. The role of TGF-beta in pulmonary fibrosis. Ciba Found Symp. 157:194-207; discussion 207-111.
37. Kim S, Takahashi H, Lin WW, Descargues P, Grivennikov S, Kim Y, Luo JL, Karin M. 2009. Carcinoma-produced factors activate myeloid cells through TLR2 to stimulate metastasis. Nature. 457:102-106.
38. Kischel P, Waltregny D, Dumont B, Turtoi A, Greffe Y, Kirsch S, De Pauw E, Castronovo V. 2010. Versican overexpression in human breast cancer lesions: Known and new isoforms for stromal tumor targeting. Int J Cancer. 126:640-650.
39. Konigshoff M, Kneidinger N, Eickelberg O. 2009. TGF-beta signaling in COPD: Deciphering genetic and cellular susceptibilities for future therapeutic regimen. Swiss Med Weekly. 139:554-563.
40. Korinek V, Barker N, Morin PJ, van Wichen D, de Weger R, Kinzler KW, Vogelstein B, Clevers H. 1997. Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC-/- colon carcinoma. Science. 275: 1784-1787.
41. Krause DS. 2008. Bone marrow-derived cells and stem cells in lung repair. Proc Am Thoracic Soc. 5:323-327.
42. Kunz LI, Strebus J, Budulac SE, Lapperre TS, Sterk PJ, Postma DS, Mauad T, Timens W, Hiemstra PS. 2013. Inhaled steroids modulate extracellular matrix composition in bronchial biopsies of COPD patients: A randomized, controlled trial. PLoS ONE. 8:e63430.
43. Kuznetsova SA, Issa P, Perruccio EM, Zeng B, Sipes JM, Ward Y, Seyfried NT, Fielder HL, Day AJ, Wight TN, et al. 2006. Versican-thrombospondin-1 binding in vitro and colocalization in microfibrils induced by inflammation on vascular smooth muscle cells. J Cell Sci. 119:4499-4509.
44. Lam AP, Flozak AS, Russell S, Wei J, Jain M, Mutlu GM, Budinger GR, Feghali-Bostwick CA, Varga J, Gottardi CJ. 2011. Nuclear beta-catenin is increased in systemic sclerosis pulmonary fibrosis and promotes lung fibroblast migration and proliferation. Am J Respir Cell Mol Biol. 45:915-922.
45. Lange P. 2013. Persistent airway obstruction in asthma. Am J Respir Crit Care Med. 187:1-2.
46. Lee DY, Jeyapalan Z, Fang L, Yang J, Zhang Y, Yee AY, Li M, Du WW, Shatseva T, Yang BB. 2010. Expression of versican 3'-untranslated region modulates endogenous microRNA functions. PLoS ONE. 5:e13599.
47. Lemire JM, Chan CK, Bressler S, Miller J, LeBaron RG, Wight TN. 2007. Interleukin-1beta selectively decreases the synthesis of versican by arterial smooth muscle cells. J Cell Biochem. 101:753-766.
48. Logan CY, Nusse R. 2004. The Wnt signaling pathway in development and disease. Annu Rev Cell Dev Biol. 20:781-810.
49. Ludwig MS, Ftouhi-Paquin N, Huang W, Page N, Chakir J, Hamid Q. 2004. Mechanical strain enhances proteoglycan message in fibroblasts from asthmatic subjects. Clin Exp Allergy. 34:926-930.
50. Malmstrom A, Bartolini B, Thelin MA, Pacheco B, Maccarana M. 2012. Iduronic acid in chondroitin/dermatan sulfate: Biosynthesis and biological function. J Histochem Cytochem. 60:916-925.
51. Margolis RU, Margolis RK. 1994. Aggrecan-versican-neurocan family proteoglycans. Methods Enzymol. 245:105-126.
52. Masuda A, Yasuoka H, Satoh T, Okazaki Y, Yamaguchi Y, Kuwana M. 2013. Versican is upregulated in circulating monocytes in patients with systemic sclerosis and amplifies a CCL2-mediated pathogenic loop. Arthritis Res Ther. 15:R74.
53. Maydan M, McDonald PC, Sanghera J, Yan J, Rallis C, Pinchin S, Hannigan GE, Foster LJ, Ish-Horowicz D, Walsh MP, et al. 2010. Integrin-linked kinase is a functional Mn2+-dependent protein kinase that regulates glycogen synthase kinase-3beta (GSK-3beta) phosphorylation. PLoS ONE. 5:e12356.
54. Merrilees MJ, Ching PS, Beaumont B, Hinek A, Wight TN, Black PN. 2008. Changes in elastin, elastin binding protein and versican in alveoli in chronic obstructive pulmonary disease. Respir Res. 9:41.
55. Merrilees MJ, Lemire JM, Fischer JW, Kinsella MG, Braun KR, Clowes AW, Wight TN. 2002. Retrovirally mediated overexpression of versican v3 by arterial smooth muscle cells induces tropoelastin synthesis and elastic fiber formation in vitro and in neointima after vascular injury. Circ Res. 90:481-487.
56. Nihlberg K, Andersson-Sjoland A, Tufvesson E, Erjefalt JS, Bjermer L, Westergren-Thorsson G. 2010. Altered matrix production in the distal airways of individuals with asthma. Thorax. 65:670-676.
57. Nihlberg K, Lundström M, Larsson A-K, Tufvesson E, Bjermer L, Westergren-Thorsson G. 2011. Matrix production in parenchymal fibroblasts from uncontrolled asthmatics negatively correlate to nitric oxide in these patients. Am J Respir Crit Care Med. 183:A3603.
58. Norian JM, Malik M, Parker CY, Joseph D, Leppert PC, Segars JH, Catherino WH. 2009. Transforming growth factor beta3 regulates the versican variants in the extracellular matrix-rich uterine leiomyomas. Reprod Sci. 16:1153-1164.
59. Phipps S, Lam CE, Foster PS, Matthaei KI. 2007. The contribution of toll-like receptors to the pathogenesis of asthma. Immunol Cell Biol. 85:463-470.
60. Piccinini AM, Midwood KS. 2010. DAMPening inflammation by modulating TLR signalling. Mediators Inflamm. 2010:1-21.
61. Pini L, Hamid Q, Shannon J, Lemelin L, Olivenstein R, Ernst P, Lemiere C, Martin JG, Ludwig MS. 2007. Differences in proteoglycan deposition in the airways of moderate and severe asthmatics. Eur Respir J. 29:71-77.
62. Plotnikov SV, Waterman CM. 2013. Guiding cell migration by tugging. Curr Opin Cell Biol. 25:619-626.
63. Potter-Perigo S, Johnson PY, Evanko SP, Chan CK, Braun KR, Wilkinson TS, Altman LC, Wight TN. 2010. Polyinosine-polycytidylic acid stimulates versican accumulation in the extracellular matrix promoting monocyte adhesion. Am J Respir Cell Mol Biol. 43:109-120.
64. Rahmani M, Carthy JM, McManus BM. 2012. Mapping of the Wnt/betacatenin/ TCF response elements in the human versican promoter. Methods Mol Biol. 836:35-52.
65. Rahmani M, Read JT, Carthy JM, McDonald PC, Wong BW, Esfandiarei M, Si X, Luo Z, Luo H, Rennie PS, et al. 2005. Regulation of the versican promoter by the beta-catenin-T-cell factor complex in vascular smooth muscle cells. J Biol Chem. 280:13019-13028.
66. Roberts CR. 1995. Is asthma a fibrotic disease? Chest. 107:111S-117S.
67. Rutnam ZJ, Wight TN, Yang BB. 2013. miRNAs regulate expression and function of extracellular matrix molecules. Matrix Biol. 32:74-85.
68. Rydell-Tormanen K, Andreasson K, Hesselstrand R, Risteli J, Heinegard D, Saxne T, Westergren-Thorsson G. 2012. Extracellular matrix alterations and acute inflammation; developing in parallel during early induction of pulmonary fibrosis. Lab Investig. 92:917-925.
69. Schaefer L, Babelova A, Kiss E, Hausser HJ, Baliova M, Krzyzankova M, Marsche G, Young MF, Mihalik D, Gotte M, et al. 2005. The matrix component biglycan is proinflammatory and signals through Toll-like receptors 4 and 2 in macrophages. J Clin Invest. 115:2223-2233.
70. Schonherr E, Kinsella MG, Wight TN. 1997. Genistein selectively inhibits platelet-derived growth factor-stimulated versican biosynthesis in monkey arterial smooth muscle cells. Arch Biochem Biophys. 339:353-361.
71. Sheng W, Wang G, La Pierre DP, Wen J, Deng Z, Wong CK, Lee DY, Yang BB. 2006. Versican mediates mesenchymal-epithelial transition. Mol Biol Cell. 17:2009-2020.
72. Sheng W, Wang G, Wang Y, Liang J, Wen J, Zheng PS, Wu Y, Lee V, Slingerland J, Dumont D, et al. 2005. The roles of versican V1 and V2 isoforms in cell proliferation and apoptosis. Mol Biol Cell. 16:1330-1340.
73. Shimizu K, Hasegawa M, Makita H, Nasuhara Y, Konno S, Nishimura M. 2011. Comparison of airway remodelling assessed by computed tomography in asthma and COPD. Respir Med. 105:1275-1283.
74. Smits NC, Shworak NW, Dekhuijzen PN, van Kuppevelt TH. 2010. Heparan sulfates in the lung: Structure, diversity, and role in pulmonary emphysema. Anat Rec (Hoboken). 293:955-967.
75. Sotoodehnejadnematalahi F, Burke B. 2013. Structure, function and regulation of versican: The most abundant type of proteoglycan in the extracellular matrix. Acta Medica Iranica. 51:740-750.
76. Takahashi A, Majumdar A, Parameswaran H, Bartolak-Suki E, Suki B. 2014. Proteoglycans maintain lung stability in an elastase-treated mouse model of emphysema. Am J Respir Cell Mol Biol. 51:26-33.
77. Tiedemann K, Malmstrom A, Westergren-Thorsson G. 1997. Cytokine regulation of proteoglycan production in fibroblasts: Separate and synergistic effects. Matrix Biol. 15:469-478.
78. Tiedemann K, Olander B, Eklund E, Todorova L, Bengtsson M, Maccarana M, Westergren-Thorsson G, Malmstrom A. 2005. Regulation of the chondroitin/dermatan fine structure by transforming growth factor-beta1 through effects on polymer-modifying enzymes. Glycobiology. 15: 1277-1285.
79. Toeda K, Nakamura K, Hirohata S, Hatipoglu OF, Demircan K, Yamawaki H, Ogawa H, Kusachi S, Shiratori Y, Ninomiya Y. 2005. Versican is induced in infiltrating monocytes in myocardial infarction. Mol and Cell Biochem. 280:47-56.
80. Tufvesson E, Westergren-Thorsson G. 2000. Alteration of proteoglycan synthesis in human lung fibroblasts induced by interleukin-1beta and tumor necrosis factor-alpha. J Cell Biochem. 77:298-309.
81. Tufvesson E, Westergren-Thorsson G. 2003. Biglycan and decorin induce morphological and cytoskeletal changes involving signalling by the small GTPases RhoA and Rac1 resulting in lung fibroblast migration. J Cell Sci. 116:4857-4864.
82. van Amerongen R, Nusse R. 2009. Towards an integrated view of Wnt signaling in development. Development. 136:3205-3214.
83. Velleman SG, Sporer KR, Ernst CW, Reed KM, Strasburg GM. 2012. Versican, matrix Gla protein, and death-associated protein expression affect muscle satellite cell proliferation and differentiation. Poultry Sci. 91:1964-1973.
84. Wang W, Xu GL, Jia WD, Ma JL, Li JS, Ge YS, Ren WH, Yu JH, Liu WB. 2009. Ligation of TLR2 by versican: A link between inflammation and metastasis. Arch Med Res. 40:321-323.
85. Wang X, Hu G, Zhou J. 2010. Repression of versican expression by microRNA-143. J Biol Chem. 285:23241-23250.
86. Westergren-Thorsson G, Antonsson P, Malmstrom A, Heinegard D, Oldberg A. 1991. The synthesis of a family of structurally related proteoglycans in fibroblasts is differently regulated by TFG-beta. Matrix. 11:177-183.
87. Westergren-Thorsson G, Chakir J, Lafreniere-Allard MJ, Boulet LP, Tremblay GM. 2002. Correlation between airway responsiveness and proteoglycan production by bronchial fibroblasts from normal and asthmatic subjects. Int J Biochem Cell Biol. 34:1256-1267.
88. Weitoft M, Andersson C, Andersson-Sjöland A, Tufvesson E, Bjermer L, Erjefält J, Westergren Thorsson G. 2014. Controlled and uncontrolled asthma display distinct alveolar tissue matrix compositions. Respir Res. 15:67. doi:10.1186/1465-9921-15-67
89. Wu YJ, La Pierre DP, Wu J, Yee AJ, Yang BB. 2005. The interaction of versican with its binding partners. Cell Res. 15:483-494.
90. Yang YC, Zhang N, Van Crombruggen K, Hu GH, Hong SL, Bachert C. 2012. Transforming growth factor-beta1 in inflammatory airway disease: A key for understanding inflammation and remodeling. Allergy. 67:1193-1202.
91. Zanini A, Chetta A, Imperatori AS, Spanevello A, Olivieri D. 2010. The role of the bronchial microvasculature in the airway remodelling in asthma and COPD. Respir Res. 11:132.
92. Zimmermann DR, Dours-Zimmermann MT, Schubert M, Bruckner- Tuderman L. 1994. Versican is expressed in the proliferating zone in the epidermis and in association with the elastic network of the dermis. J Cell Biol. 124:817-825.
93. Zimmermann DR, Ruoslahti E. 1989. Multiple domains of the large fibroblast proteoglycan, versican. EMBO J. 8:2975-2981.