Blocking type TSH receptor antibodies

Autoimmunity Highlights

Volumn 4, Issue 1, 2013, Pages 11-26

  Furmaniak, Jadwiga ^a   Sanders, Jane ^a   Rees Smith, Bernard ^a  

^a RSR LTD   (United Kingdom)

Author keywords

Autoimmunity; Graves' disease; Thyroid blocking autoantibody; Thyroid stimulating autoantibody; TSH receptor

Indexed keywords

MONOCLONAL ANTIBODY; MONOCLONAL ANTIBODY 1H7; MONOCLONAL ANTIBODY 5C9; MONOCLONAL ANTIBODY B2; MONOCLONAL ANTIBODY CS 17; MONOCLONAL ANTIBODY K1 70; MONOCLONAL ANTIBODY M22; THYROID HORMONE; THYROTROPIN RECEPTOR ANTIBODY; UNCLASSIFIED DRUG;

AMINO ACID SUBSTITUTION; ANTIBODY BLOOD LEVEL; BINDING AFFINITY; COMPLEX FORMATION; CONCENTRATION RESPONSE; CRYSTAL STRUCTURE; DOSE RESPONSE; DRUG DOSE COMPARISON; DRUG MECHANISM; DRUG RECEPTOR BINDING; DRUG SYNTHESIS; GENE MUTATION; GRAVES DISEASE; HUMAN; IMMUNOHISTOCHEMISTRY; INHIBITION KINETICS; INSULIN DEPENDENT DIABETES MELLITUS; MOLECULAR INTERACTION; NONHUMAN; PATHOGENESIS; PRIORITY JOURNAL; PROTEIN EXPRESSION; RECEPTOR UPREGULATION; REVIEW; THYROID FUNCTION; THYROID HORMONE RELEASE; THYROID HORMONE SYNTHESIS;

EID: 84877331608     PISSN: 20380305     EISSN: 20383274     Source Type: Journal    
DOI: 10.1007/s13317-012-0028-1     Document Type: Review

References (78)

1. Rees Smith B, McLachlan SM, Furmaniak J (1988) Autoantibodies to the thyrotropin receptor. Endocr Rev 9: 106-121. doi: 10. 1210/edrv-9-1-106.
2. Rapoport B, Chazenbalk GD, Jaume JC, McLachlan SM (1998) The thyrotropin (TSH) receptor: interaction with TSH and autoantibodies. Endocr Rev 19: 673-716. doi: 10. 1210/er. 19. 6. 673.
3. Rees Smith B, Bolton J, Young S, Collyer A, Weeden A, Bradbury J, Weightman D, Perros P, Sanders J, Furmaniak J (2004) A new assay for thyrotropin receptor autoantibodies. Thyroid 14: 830-835. doi: 10. 1089/thy. 2004. 14. 830.
4. Rees Smith B, Sanders J, Furmaniak J (2007) TSH receptor antibodies. Thyroid 17: 923-938. doi: 10. 1089/thy. 2007. 0239.
5. McLachlan SM, Rapoport B (1996) Monoclonal, human autoantibodies to the TSH receptor-the holy grail and why are we looking for it? J Clin Endocrinol Metab 81: 3152-3154.
6. McLachlan SM, Rapoport B (2004) Thyroid stimulating monoclonal antibodies: overcoming the road blocks and the way forward. Clin Endocrinol 61: 10-18. doi: 10. 1111/j. 1365-2265. 2004. 02028. x.
7. Sanders J, Jeffreys J, Depraetere H, Richards T, Evans M, Kiddie A, Brereton K, Groenen M, Oda Y, Furmaniak J, Rees Smith B (2002) Thyroid stimulating monoclonal antibodies. Thyroid 12: 1043-1050. doi: 10. 1089/105072502321085135.
8. Ando T, Latif R, Pritsker A, Moran T, Nagayama Y, Davies TF (2002) A monoclonal thyroid-stimulating antibody. J Clin Investig 110: 1667-1674. doi: 10. 1172/JCI16991.
9. Costagliola S, Franssen JDF, Bonomi M, Urizar E, Willnich M, Bergmann A, Vassart G (2002) Generation of a mouse monoclonal TSH receptor antibody with stimulating activity. Biochem Biophys Res Commun 299: 891-896. doi: 10. 1016/S0006-291X(02)02762-6.
10. Costagliola S, Bonomi M, Morganthaler NG, van Durme J, Panneels V, Refetoff S, Vassart G (2004) Delineation of the discontinuous-conformational epitope of a monoclonal antibody displaying full in vitro and in vivo thyrotropin activity. Mol Endocrinol 18: 3020-3034. doi: 10. 1210/me. 2004-0231.
11. Gilbert J, Gianoukakis A, Salehi S, Moorhead J, Rao P, Kahn MZ, McGregor A, Smith T, Banga JP (2006) Monoclonal pathogenic antibodies to the thyroid stimulating hormone receptor in Graves' disease with potent thyroid-stimulating activity but differential blocking activity activate multiple signalling pathways. J Immunol 176: 5084-5092.
12. Sanders J, Allen F, Jeffreys J, Bolton J, Richards T, Depraetere H, Nakatake N, Evans M, Kiddie A, Premawardhana LDKE, Chirgadze DY, Miguel RN, Blundell TL, Furmaniak J, Rees Smith B (2005) Characteristics of a monoclonal antibody to the thyrotropin receptor that acts as a powerful thyroid-stimulating autoantibody antagonist. Thyroid 15: 672-682. doi: 10. 1089/thy. 2005. 15. 672.
13. Shepherd PS, Da Costa CR, Cridland JC, Gilmore KS, Johnstone AP (1999) Identification of an important thyrotrophin binding site on the human thyrotrophin receptor using monoclonal antibodies. Mol Cell Endocrinol 149: 197-206.
14. Jeffreys J, Depraetere H, Sanders J, Oda Y, Evans M, Kiddie A, Richards T, Furmaniak J, Rees Smith B (2002) Characterization of the thyrotropin binding pocket. Thyroid 12: 1051-1061. doi: 10. 1089/105072502321085144.
15. Costagliola S, Panneels V, Bonomi M, Koch J, Many MC, Smits G, Vassart G (2002) Tyrosine sulfation is required for agonist recognition by glycoprotein hormone receptors. EMBO J 21: 504-513. doi: 10. 1093/emboj/21. 4. 504.
16. Davies TF, Bobovnikova Y, Weiss M, Vlase H, Moran T, Graves PN (1998) Development and characterization of monoclonal antibodies specific for the murine thyrotropin receptor. Thyroid 8: 693-701.
17. Lenzner C, Morgenthaler NG (2003) The effect of thyrotropin-receptor blocking antibodies on stimulating autoantibodies from patients with Graves' disease. Thyroid 13: 1153-1161. doi: 10. 1089/10507250360731569.
18. Minich WB, Lenzner C, Morgenthaler NG (2004) Antibodies to TSH-receptor in thyroid autoimmune disease interact with monoclonal antibodies whose epitopes are broadly distributed on the receptor. Clin Exp Immunol 136: 129-136. doi: 10. 1111/j. 1365-2249. 2004. 02417. x.
19. Oda Y, Sanders J, Roberts S, Maruyama M, Kato R, Perez M, Petersen VB, Wedlock N, Furmaniak J, Rees Smith B (1998) Binding characteristics of antibodies to the TSH receptor. J Mol Endocrinol 20: 233-244. doi: 10. 1677/jme. 0. 0200233.
20. Oda Y, Sanders J, Evans M, Kiddie A, Munkley A, James C, Richards T, Wills J, Furmaniak J, Rees Smith B (2000) Epitope analysis of the human thyrotropin (TSH) receptor using monoclonal antibodies. Thyroid 10: 1051-1059. doi: 10. 1089/thy. 2000. 10. 1051.
21. Vlase H, Weiss M, Graves PN, Davies TF (1998) Characterization of the murine immune response to the murine TSH receptor ectodomain: induction of hypothyroidism and TSH receptor antibodies. Clin Exp Immunol 113: 111-118. doi: 10. 1046/j. 1365-2249. 1998. 00622. x.
22. Sanders J, Evans M, Premawardhana LDKE, Depraetere H, Jeffreys J, Richards T, Furmaniak J, Rees Smith B (2003) Human monoclonal thyroid stimulating autoantibody. Lancet 362: 126-128. doi: 10. 1016/S0140-6736(03)13866-4.
23. Sanders J, Jeffreys J, Depraetere H, Evans M, Richards T, Kiddie A, Brereton K, Premawardhana LD, Chirgadze DY, Núñez Miguel R, Blundell TL, Furmaniak J, Rees Smith B (2004) Characteristics of a human monoclonal autoantibody to the thyrotropin receptor: sequence structure and function. Thyroid 14: 560-570. doi: 10. 1089/1050725041692918.
24. Sanders J, Chirgadze DY, Sanders P, Baker S, Sullivan A, Bhardwaja A, Bolton J, Reeve M, Nakatake N, Evans M, Richards T, Powell M, Núñez Miguel R, Blundell TL, Furmaniak J, Rees Smith B (2007) Crystal structure of the TSH receptor in complex with a thyroid-stimulating autoantibody. Thyroid 17: 395-410. doi: 10. 1089/thy. 2007. 0034.
25. Sanders J, Evans M, Betterle C, Sanders P, Bhardwaja A, Young S, Roberts E, Wilmot J, Richards T, Kiddie A, Small K, Platt H, Summerhayes S, Harris R, Reeve M, Coco G, Zanchetta R, Chen S, Furmaniak J, Rees Smith B (2008) A human monoclonal autoantibody to the thyrotropin receptor with thyroid-stimulating blocking activity. Thyroid 18: 735-746. doi: 10. 1089/thy. 2007. 0327.
26. Rees Smith B, Sanders J, Evans M, Tagami T, Furmaniak J (2009) TSH receptor-autoantibody interactions. Hormon Metab Res 41: 448-455. doi: 10. 1055/s-0029-1220913.
27. Evans M, Sanders J, Tagami T, Sanders P, Young S, Roberts E, Wilmot J, Hu X, Kabelis K, Clark J, Holl S, Richards T, Collyer A, Furmaniak J, Rees Smith B (2010) Monoclonal autoantibodies to the TSH receptor, one with stimulating activity and one with blocking activity, obtained from the same blood sample. Clin Endocrinol 73: 404-412. doi: 10. 1111/j. 1365-2265. 2010. 03831. x.
28. Sanders P, Young S, Sanders J, Kabelis K, Baker S, Sullivan A, Evans M, Clark J, Wilmot J, Hu X, Roberts E, Powell M, Núñez Miguel R, Furmaniak J, Rees Smith B (2011) Crystal structure of the TSH receptor (TSHR) bound to a blocking-type TSHR autoantibody. J Mol Endocrinol 46: 81-99. doi: 10. 1530/JME-10-0127.
29. Sanders J, Núñez Miguel R, Furmaniak J, Rees Smith B (2010) TSH receptor monoclonal antibodies with agonist, antagonist and inverse agonist activities. Methods Enzymol 485: 393-420. doi: 10. 1016/B978-0-12-381296-4. 00022-1.
30. Chen CR, McLachlan SM, Rapoport B (2007) Suppression of thyrotropin receptor constitutive activity by a monoclonal antibody with inverse agonist activity. Endocrinology 148: 2375-2382. doi: 10. 1210/en. 2006-1754.
31. Chen CR, McLachlan SM, Rapoport B (2008) Identification of key amino acid residues in a thyrotropin receptor monoclonal antibody epitope provides insight into its inverse agonist and antagonist properties. Endocrinology 149: 3427-3434. doi: 10. 1210/en. 2008-0207.
32. Zhang M, Phuong K, Tong T, Fremont V, Chen J, Narayan P, Puett D, Weintraub BD, Szkudlinski MW (2000) The extracellular domain suppresses constitutive activity of the transmembrane domain of the human TSH receptor: implications for hormone-receptor interaction and antagonist design. Endocrinology 141: 3514-3517. doi: 10. 1210/en. 141. 9. 3514.
33. Vlaeminck-Guillem VK, Ho SC, Rodien P, Vassart G, Costagliola S (2002) Activation of the cAMP pathway by the TSH receptor involves switching of the ectodomain from a tethered inverse agonist to an agonist. Mol Endocrinol 16: 736-746. doi: 10. 1210/me. 16. 4. 736.
34. Sanders J, Oda Y, Roberts S, Maruyama M, Furmaniak J, Rees Smith B (1997) Understanding the thyrotropin receptor function-structure relationship. Baillière's Clin Endocr Metab 11: 451-479. doi: 10. 1016/S0950-351X(97)80693-3.
35. Krohn K, Führer D, Bayer Y, Eszlinger M, Brauer V, Neumann S, Paschke R (2005) Molecular pathogenesis of euthyroid and toxic multinodulargoiter. Endocr Rev 26: 504-524. doi: 10. 1210/er. 2004-0005.
36. Kopp P, Muirhead S, Jourdain N, Gu W-X, Jameson JL, Rodd C (1997) Congenital hyperthyroidism caused by solitary toxic adenoma harboring a novel somatic mutation (serine281 to isoleucine) in the extracellular domain of the thyrotropin receptor. J Clin Investig 100: 1634-1639. doi: 10. 1172/JCI119687.
37. Parma J, van Sande J, Swillens S, Tonacchera M, Dumont J, Vassart G (1995) Somatic mutations causing constitutive activity of the thyrotropin receptor are the major cause of hyperfunctioning thyroid adenomas: identification of additional mutations activating both the cyclic adenosine 3′, 5′-monophosphate and inositol phosphate-Ca2 + cascades. Mol Endocrinol 9: 725-733. doi: 10. 1210/me. 9. 6. 725.
38. Parma J, Duprez L, van Sande J, Cochaux P, Gervy C, Mockel J, Dumont J, Vassart G (1993) Somatic mutations in the thyrotropin receptor gene cause hyperfunctioning thyroid adenomas. Nature 365: 649-651. doi: 10. 1038/365649a0.
39. Sanders J, Bolton J, Sanders P, Jeffreys J, Nakatake N, Richards T, Evans M, Kiddie A, Summerhayes S, Roberts E, Núñez Miguel R, Furmaniak J, Rees Smith B (2006) Effects of TSH receptor mutations on binding and biological activity of monoclonal antibodies and TSH. Thyroid 16: 1195-1206. doi: 10. 1089/thy. 2006. 16. 1195.
40. Kosugi S, Ban T, Akamizu T, Kohn LD (1991) Site-directed mutagenesis of a portion of the extracellular domain of the rat thyrotropin receptor important in autoimmune thyroid disease and nonhomologous with gonadotropin receptors: relationship of functional and immunogenic domains. J Biol Chem 266: 19413-19418.
41. Mueller S, Kleinau G, Jaeschke H, Paschke R, Krause G (2008) Extended hormone binding site of the human thyroid stimulating hormone receptor: distinctive acidic residues in the hinge region are involved in bovine thyroid stimulating hormone binding and receptor activation. J Biol Chem 283: 18048-18055. doi: 10. 1074/jbc. M800449200.
42. Mueller S, Kleinau G, Szkudlinski MW, Jaeschke H, Krause G, Paschke R (2009) The superagonistic activity of bovine thyroid-stimulating hormone (TSH) and the human TR1401 TSH analog is determined by specific amino acids in the hinge region of the human TSH receptor. J Biol Chem 284: 16317-16324. doi: 10. 1074/jbc. M109. 005710.
43. Sanders J, Núñez Miguel R, Bolton J, Bhardwaja A, Sanders P, Nakatake N, Evans M, Furmaniak J, Rees Smith B (2007) Molecular interactions between the TSH receptor and a thyroid-stimulating monoclonal autoantibody. Thyroid 17: 699-706. doi: 10. 1089/thy. 2007. 0041.
44. Fan QR, Hendrickson WA (2005) Structure of human follicle-stimulating hormone in complex with its receptor. Nature 433: 269-277. doi: 10. 1038/nature03206.
45. Chen C-R, Tanaka K, Chazenbalk GD, McLachlan SM, Rapoport B (2001) A full biological response to autoantibodies in Graves' disease requires a disulfide-bond loop in the thyrotropin N-terminus homologous to a laminin EGF-like domain. J Biol Chem 276: 14767-14772. doi: 10. 1074/jbc. M008001200.
46. Chazenbalk GD, Latrofa F, McLachlan SM, Rapoport B (2004) Thyroid stimulation does not require antibodies with identical epitopes but does involve recognition of a critical conformation at the N terminus of the thyrotropin receptor A-subunit. J Clin Endocrinol Metab 89: 1788-1793. doi: 10. 1210/jc. 2003-031554.
47. Hamidi S, Chen C-R, McLachlan SM, Rapoport B (2011) Insight into thyroid-stimulating autoantibody interaction with the thyrotropin receptor N-terminus based on mutagenesis and re-evaluation of ambiguity in this region of the receptor crystal structure. Thyroid 21: 1013-1020. doi: 10. 1089/thy. 2011. 0147.
48. Kobe B, Deisenhofer J (1994) The leucine-rich repeat: a versatile binding motif. Trends Biochem Sci 19: 415-421. doi: 10. 1016/0968-0004(94)90090-6.
49. Buchanan SG, Gay NJ (1996) Structural and functional diversity in the leucine rich repeat family of proteins. Prog Biophys Mol Biol 65: 1-44. doi: 10. 1016/S0079-6107(96)00003-X.
50. Núñez Miguel R, Sanders J, Jeffreys J, Depraetere H, Evans M, Richards T, Blundell TL, Rees Smith B, Furmaniak J (2004) Analysis of the thyrotropin receptor-thyrotropin interaction by comparative modeling. Thyroid 14: 991-1011. doi: 10. 1089/thy. 2004. 14. 991.
51. Núñez Miguel R, Sanders J, Blundell TL, Rees Smith B, Furmaniak J (2005) Comparative modeling of the thyrotropin receptor. Thyroid 15: 746-747. doi: 10. 1089/thy. 2005. 15. 746.
52. Núñez Miguel R, Sanders J, Chirgadze DY, Blundell TL, Furmaniak J, Rees Smith B (2008) FSH and TSH binding to their respective receptors: similarities, differences and implication for glycoprotein hormone specificity. J Mol Endocrinol 41: 145-164. doi: 10. 1677/JME-08-0040.
53. Núñez Miguel R, Sanders J, Chirgadze DY, Furmaniak J, Rees Smith B (2009) Thyroid stimulating autoantibody M22 mimics TSH binding to the TSH receptor leucine rich domain: a comparative structural study of protein-protein interactions. J Mol Endocrinol 42: 381-395. doi: 10. 1677/JME-08-0152.
54. Horimoto M, Petersen VB, Pegg CAS, Fukuma N, Wabayashi N, Kiso Y, Furmaniak J, Rees Smith B (1993) Production and characterization of a human monoclonal thyroid peroxidase autoantibody. Autoimmunity 14: 1-7. doi: 10. 3109/08916939309077350.
55. Costagliola S, Rodien P, Many MC, Ludgate M, Vassart G (1998) Genetic immunisation against the human thyrotropin receptor causes thyroiditis and allows production of monoclonal antibodies recognising the native receptor. J Immunol 160: 1458-1465.
56. Sequeira M, Jasani B, Fuhrer D, Wheeler M, Ludgate M (2002) Demonstration of reduced in vivo surface expression of activating mutant thyrotropin receptors in thyroid sections. Eur J Endocrinol 146: 163-171. doi: 10. 1530/eje. 0. 1460163.
57. Boschi A, Daumerie Ch, Spiritus M, Beguin C, Senou M, Yuksel D, Duplicy M, Costagliola S, Ludgate M, Many MC (2005) Quantification of cells expressing the thyrotropin receptor in extraocular muscles in thyroid associated orbitopathy. Br J Ophthalmol 89: 724-729. doi: 10. 1136/bjo. 2004. 050807.
58. de Lloyd A, Bursell J, Gregory JW, Rees DA, Ludgate M (2010) TSH receptor activation and body composition. J Endocrinol 204: 13-20. doi: 10. 1677/JOE-09-0262.
59. Williams GR (2011) Extrathyroidal expression of TSH receptor. Ann Endocrinol 72: 68-73. doi: 10. 1016/j. ando. 2011. 03. 006.
60. Cooper DS (2005) Antithyroid drugs. N Engl J Med 352: 905-917. doi: 10. 1056/NEJMra042972.
61. Pearce EN (2006) Diagnosis and management of thyrotoxicosis. Br Med J 332: 1369-1373. doi: 10. 1136/bmj. 332. 7554. 1369.
62. Brent GA (2008) Graves' disease. N Engl J Med 358: 2594-2605. doi: 10. 1056/NEJMcp0801880.
63. Astwood EB (1943) Treatment of hyperthyroidism with thiourea and thiouracil. J Am Med Assoc 122: 78-86. doi: 10. 1001/jama. 1943. 02840190008003.
64. Volpé R (1978) The pathogenesis of Graves' disease: an overview. Clin Endocr Metab 7: 3-29. doi: 10. 1016/S0300-595X(78)80033-4.
65. Laurberg P (2006) Remission of Graves' disease during anti-thyroid drug therapy: time to reconsider the mechanism? Eur J Endocrinol 155: 783-786. doi: 10. 1530/eje. 1. 02295.
66. Prabhakar BS, Bahn RS, Smith TJ (2003) Current perspective on the pathogenesis of Graves' disease and ophthalmopathy. Endocr Rev 24: 802-835. doi: 10. 1210/er. 2002-0020.
67. Kumar S, Schiefer R, Coenen MJ, Bahn RS (2010) A stimulatory thyrotropin receptor antibody (M22) and thyrotropin increase interleukin-6 expression and secretion in Graves' orbital preadipocyte fibroblasts. Thyroid 20: 59-65. doi: 10. 1089/thy. 2009. 0278.
68. El Fassi D, Banga JP, Gilbert JA, Padoa C, Hegedüs L, Nielsen CH (2009) Treatment of Graves' disease with rituximab specifically reduces the production of thyroid stimulating antibodies. Clin Immunol 130: 252-258. doi: 10. 1016/j. clim. 2008. 09. 007.
69. Ueki I, Abiru N, Kobayashi M, Nakahara M, Ichikawa T, Egchi K, Nagayama Y (2011) B cell-targeted therapy with anti-CD20 monoclonal antibody in a mouse model of Graves' hyperthyroidism. Clin Exp Immunol 163: 309-317. doi: 10. 1111/j. 1365-2249. 2010. 04301. x.
70. Rapoport B, Alsabeh R, Aftergood D, McLachlan SM (2000) Elephantiasic pretibial myedema: insight into and a hypothesis regarding the pathogenesis of the extrathyroidal manifestations of Graves' disease. Thyroid 10: 685-692. doi: 10. 1089/10507250050137761.
71. Beck-Peccoz P, Brucker-Davis F, Persani L, Smallridge RC, Weintraub BD (1996) Thyrotropin-secreting pituitary tumors. Endocr Rev 17: 610-638. doi: 10. 1210/edrv-17-6-610.
72. Brekke OH, Sandlie I (2003) Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 2: 52-62. doi: 10. 1038/nrd984.
73. Roskos LK, Davis CG, Schwab GM (2004) The clinical pharmacology of therapeutic monoclonal antibodies. Drug Dev Res 61: 108-120. doi: 10. 1002/ddr. 10346.
74. Luster M (2006) Present status of the use of recombinant human TSH in thyroid cancer management. Acta Oncol 45: 1018-1030. doi: 10. 1080/02841860600979013.
75. Alley SC, Okeley NM, Senter PD (2010) Antibody-drug conjugates: targeted drug delivery for cancer. Curr Opin Chem Biol 14: 529-537. doi: 10. 1016/j. cbpa. 2010. 06. 170.
76. Hayakawa N, Premawardhana LDKE, Powell M, Masuda M, Arnold C, Sanders J, Evans M, Chen S, Jaume JC, Baekkeskov S, Rees Smith B, Furmaniak J (2002) Isolation and characterization of human monoclonal autoantibodies to glutamic acid decarboxylase. Autoimmunity 35: 343-355. doi: 10. 1080/0891693021000003206.
77. Sanders J, Evans M, Betterle C, Sanders P, Bhardwaja A, Young S, Roberts E, Wilmot J, Richards T, Kiddie A, Small K, Platt H, Summerhayes S, Harris R, Reeve M, Coco G, Zanchetta R, Chen S, Furmaniak J, Rees Smith B (2008) A human monoclonal autoantibody to the thyrotropin receptor with thyroid-stimulating blocking activity. Thyroid 18(7): 735-746. doi: 10. 1089/thy. 2007. 0327.
78. Furmaniak J, Sanders J, Young S, Kabelis K, Sanders P, Evans M et al (2012) In vivo effects of a human thyroid timulating monoclonal autoantibody (M22) and a human thyroid blocking autoantibody (K1-70). Autoimmun Highlights (in press) doi: 10. 1007/s13317-011-0025-9.