Relaxin as a natural agent for vascular health

Vascular Health and Risk Management

Volumn 4, Issue 3, 2008, Pages 515-524

  Bani, Daniele ^a,b  

^a UNIVERSITY OF FLORENCE   (Italy)

^b NONE   (Italy)

Author keywords

Blood vessels; Cardiovascular disease; Endothelial cells; Nitric oxide; Relaxin; Vascular smooth muscle

Indexed keywords

ACETYLCHOLINE; GLYCEROL NITRATE; NITRIC OXIDE; NITROPRUSSIDE SODIUM; PLACEBO; RECOMBINANT RELAXIN; RELAXIN; RELEASIN; SEX HORMONE; STEROID;

ANGIOGENESIS; ANTIINFLAMMATORY ACTIVITY; BLOOD VESSEL WALL; CARDIOVASCULAR DISEASE; CAUSE OF DEATH; CLINICAL EFFECTIVENESS; CLINICAL TRIAL; DEVELOPING COUNTRY; DISABILITY; DISEASE SEVERITY; DRUG MECHANISM; ENDOTHELIAL DYSFUNCTION; HEART PROTECTION; HORMONE ACTION; HORMONE RELEASE; HORMONE SYNTHESIS; HUMAN; IN VITRO STUDY; INCIDENCE; LEUKOCYTE ACTIVATION; MORTALITY; NONHUMAN; OVARY; PATHOPHYSIOLOGY; PERIPHERAL VASCULAR DISEASE; POSTMENOPAUSE; PREECLAMPSIA; RAYNAUD PHENOMENON; REVIEW; RISK REDUCTION; SEX DIFFERENCE; THROMBOCYTE ACTIVATION; VASCULAR ENDOTHELIUM; VASODILATATION;

ANIMALS; CARDIOVASCULAR DISEASES; ENDOTHELIUM, VASCULAR; FEMALE; HUMANS; MENOPAUSE; NEOVASCULARIZATION, PHYSIOLOGIC; NITRIC OXIDE; OXIDATIVE STRESS; PRE-ECLAMPSIA; PREGNANCY; RELAXIN; VASODILATION;

EID: 47849084197     PISSN: 11766344     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Review

References (118)

1. Ambrose JA, Barua RS. 2004. The pathophysiology of cigarette smoking and cardiovascular disease. J Am Coll Cardiol, 43: 1731-7.
2. Bani D, Bigazzi M, Masini E, et al. 1995. Relaxin depresses platelet aggregation: in vitro studies on isolated human and rabbit platelets. Lab Invest, 73:709-16.
3. Bani D. 1997. Relaxin, a pleiotropic hormone. Gen Pharmacol, 28:13-22.
4. Bani D, Ballati L, Masini E, et al. 1997. Relaxin counteracts asthma-like reaction induced by inhaled antigen in sensitized guinea pigs. Endocrinology, 138:1909-15.
5. Bani D, Failli P, Bello MG, et al. 1998a. Relaxin activates the L-arginine-nitric oxide pathway in vascular smooth muscle cells in culture. Hypertension, 31:1240-7.
6. Bani D, Masini E, Bello MG, et al. 1998b. Relaxin protects against myocardial injury caused by ischemia and reperfusion in rat heart. Am J Pathol, 152:1367-76.
7. Bani D, Nistri S, Quattrone S, et al. 2001. The vasorelaxant hormone relaxin induces changes in liver sinusoid microcirculation: a morphologic study in the rat. J Endocrinol, 171:541-9.
8. Bani D, Bigazzi M. 2005. Clinical aspects and therapeutic perspectives of relaxin. Curr Med Chem - Immunology, Endocrine and Metabolic Agents (CMC-IEMA) 5:403-10.
9. Bani D, Nistri S, Bani Sacchi T, et al. 2005. Basic progress and future therapeutic perspectives of relaxin in ischemic heart disease. Ann N Y Acad Sci, 1041:423-30.
10. Bani D, Nistri S, Cinci L, et al. 2007. A novel, simple bioactivity assay for relaxin based on inhibition of platelet aggregation. Regul Peptides, in press.
11. Bani G, Bani Sacchi T, Bigazzi M, et al. 1988. Effects of relaxin on the microvasculature of mouse mammary gland. Histol Histopath, 3:337-43.
12. Bani G, Maurizi M, Bigazzi M, et al. 1995. Effects of relaxin on the endometrial stroma. Studies in mice. Biol Reprod, 53:253-62.
13. Bani Sacchi T, Bigazzi M, Bani D, et al. 1995. Relaxin-induced increased coronary flow through stimulation of nitric oxide production. Brit J Pharmacol, 116:1589-94.
14. Barua RS, Ambrose JA, Eales-Reynolds LJ, et al. 2001. Dysfunctional endothelial nitric oxide biosynthesis in healthy smokers with impaired endothelium-dependent vasodilatation. Circulation, 104:1905-10.
15. Barua RS, Ambrose JA, Saba DC, et al. 2002. Smoking is associated with altered endothelial-derived fibrinolytic and antithrombotic factors: an in vitro demonstration. Circulation, 106:905-8.
16. Bathgate RA, Samuel CS, Burazin TC, et al. 2003. Relaxin: new peptides, receptors and novel actions. Trends Endocrinol Metab, 14:207-13.
17. Bathgate RA, Ivell R, Sanborn BM, et al. 2006. International Union of Pharmacology LVII: Recommendations for the nomenclature of receptors for relaxin family peptides. Pharmacol Rev, 58:7-31.
18. Bigazzi M, Del Mese A, Petrucci F, et al. 1986. The local administration of relaxin induces changes in the microcirculation ofthe rat mesocaecum. Acta Endocrinol, 112:296-9.
19. Boehnert MU, Hilbig H, Armbruster FP. 2005. Relaxin as an additional protective substance in preserving and reperfusion solution for liver transplantation, shown in a model of isolated perfused rat liver. Ann N Y Acad Sci, 1041:434-40.
20. Bonnar J. 1987. Coagulation effects of oral contraception. Am J Obstet Gynecol, 157:1042-8.
21. Casten GG, Boucek RJ. 1958. Use of relaxin in the treatment of scleroderma. JAMA, 166:319-24.
22. Casten GG, Gilmore HR, Houghton FE, et al. 1960. A new approach to the management of obliterative peripheral arterial disease. Angiology, 11:408-14.
23. Conrad KP, Gandley RE, Ogawa T, et al. 1999. Endothelin mediates renal vasodilation and hyperfiltration during pregnancy in chronically instrumented conscious rats. Am J Physiol Renal Physiol, 276:F767-76.
24. Conrad KP, Novak J. 2004. Emerging role of relaxin in renal and cardiovascular function. Am J Physiol Regul Integr Comp Physiol, 287:250-61.
25. Cosen-Binker LI, Binker MG, Cosen R, et al. 2006. Relaxin prevents the development of severe acute pancreatitis. World J Gastroenterol, 12:1558-68.
26. Dallenbach-Hellweg G, Dawson AB, Hisaw FL. 1966. The effect of relaxin on the endometrium of monkeys-histological and histochemical studies. Am J Anat, 119:61-78.
27. Danielson LA, Sherwood OD, Conrad KP. 1999. Relaxin is a potent renal vasodilator in conscious rats. J Clin Invest, 103:525-33.
28. Danielson LA, Kercher LJ, Conrad KP. 2000. Impact of gender and endothelin on renal vasodilation and hyperfiltration induced by relaxin in conscious rats. Am J Physiol Regal Integr Comp Physiol, 279: R1298-304.
29. Davison JM, Homuth V, Jeyabalan A, et al. 2004. New aspects in the pathophysiology of preeclampsia. J Am Soc Nephrol, 15:2440-8.
30. Dschietzig T, Ritcher C, Bartsch C, et al. 2001. The pregnancy hormone relaxin is a player in human heart failure. FASEB J, 165:2187-95.
31. Dschietzig T, Bartsch C, Stangl V, et al. 2004. Identification of the pregnancy hormone relaxin as glucocorticoid receptor agonist. FASEB J, 13: 1536-8.
32. Dschietzig T, Bartsch C, Baumann G, et al. 2006. Relaxin - a pleiotropic hormone and its emerging role for experimental and clinical therapeutics. Pharm Ther, 112:38-56.
33. Eddie LW, Bell RJ, Lester A, et al. 1986. Radioimmunoassay of relaxin in pregnancy with an analogue of human relaxin. Lancet, 8494:1344-6.
34. Failli P, Nistri S, Quattrone S, et al. 2001. Relaxin up-regulates inducible nitric oxide synthase expression and nitric oxide generation in rat coronary endothelial cells. FASEB J, 16:252-4.
35. Fevold HL, Hisaw FL, Meyer RK. 1930. The relaxative hormone of the corpus luteum. Its purification and concentration. J Am Chem Soc, 52:3340-8.
36. Formigli L, Perna AM, Meacci E, et al. 2007. Paracrine effects of transplanted myoblasts and relaxin on post-infarction heart remodeling. J Cell Mol Med, 11: 1101-4.
37. Forte P, Kneale BJ, Milne E, et al. 1998. Evidence for a difference in nitric oxide biosynthesis between healthy women and men. Hypertension, 32:730-4.
38. Garber SL, Mirochnik Y, Brecklin CS, et al. 2001. Relaxin decreases renal interstitial fibrosis and slows progression of renal disease. Kidney Int, 59:876-82.
39. Garber SL, Mirochnik Y, Brecklin C, et al. 2003. Effect of relaxin in two models of renal mass reduction. Am J Nephrol, 23:8-12.
40. Grimm S, Baeuerle PA. 1993. Review article: the inducible transcription factor NF-κB: structure-function relationship of its protein subunits. Biochem J, 290:297-308.
41. Grodstein F, Stampfer MJ, Manson JE, et al. 1996. Postmenopausal estrogen and progestin use and the risk of cardiovascular disease. N Engl J Med, 335:453-61.
42. Hayashi S, Yamada K, Esaki T et al. 1995. estrogen increases endothelial nitric oxide by a receptor-mediated system. Biochim Biophys Res Commun, 214:847-55.
43. Herrington DM, Reboussin DM, Brosnihan KB, et al. 2000. Effects of estrogen replacement on the progression of coronary-artery atherosclerosis. N Engl J Med, 343:522-9.
44. Hisaw FL. 1926. Experimental relaxation of the pubic ligament of the guinea pig. Proc Soc Exp Biol Med, 23:661-3.
45. Hisaw FL, Hisaw FL Jr, Dawson AB. 1967. Effect of relaxin on the endothelium of endometrial blood vessels in monkey (Macaca mulatta). Endocrinology, 81:375-85.
46. Hsu SY, Nakabayashi K, Nishi S, et al. 2002. Activation of orphan receptors by the hormone relaxin. Science, 295:671-4.
47. Hu FB, Stampfer MJ, Manson JE, et al. 2000. Trends in the incidence of coronary heart disease and changes in diet and lifestyle in women. N Engl J Med, 343:530-7.
48. Hudson P, John M, Crawford R, et al. 1984. Relaxin gene expression in human ovaries and the predicted structure of a human preprorelaxin by analysis of cDNA clones. EMBO J, 3:2333-9.
49. Hulley S, Grady D, Bush T, et al. 1998. Randomized trial of estrogen plus progestin for secondary prevention of coronary heart disease in postmenopausal women. Heart and Estrogen/progestin Replacement Study (HERS) Research Group. JAMA, 280:605-13.
50. Igarashi J, Michel T. 2001. Sphingosine 1-phosphate and isoform-specific activation of phosphoinositide 3-kinase beta. Evidence for divergence and convergence of receptor-regulated endothelial nitric-oxide synthase signaling pathways. J Biol Chem, 276:36281-8.
51. Ignarro LJ. 1991. Signal transduction mechanisms involving nitric oxide. Biochem Pharmacol, 41:485-90.
52. Jonas MA, Oates JA, Ockene JK, et al. 1992. Statement on smoking and cardiovascular disease for health care professionals. American Heart Association. Circulation, 86:1664-9.
53. Kalinka J, Hanke W, Sobala W. 2005. Impact of prenatal tobacco smoke exposure, as measured by midgestation serum cotinine levels, on fetal biometry and umbilical flow velocity waveforms. Am J Perinatol, 22:41-7.
54. Khalil RA. 2005. Sex hormones as potential modulators of vascular function in hypertension. Hypertension, 46:249-54.
55. Khan-Dawood FS, Goldsmith LT, Weiss G, et al. 1989. Human corpus luteum secretion of relaxin, oxytocin and progesterone. J Clin Endocrinol Metab, 68:627-31.
56. Kim NY, Pae HO, Kim YC, et al. 2002. Pentoxifylline potentiates nitric oxide production in interleukin-1beta-stimulated vascular smooth muscle cells through cyclic AMP-dependent protein kinase A pathway. Gen Pharmacol, 35:205-11.
57. Kohsaka T, Min G, Lukas G, et al. 1998. Identification of specific relaxin-binding cells in the human female. Biol Reprod, 59:991-9.
58. Kubes P, Suzuki M, Granger DN. 1991. Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA, 88:4651-5.
59. Lefer AM, Tsao PS, Lefer DJ, et al. 1991. Role of endothelial dysfunction in the pathogenesis of reperfusion injury after myocardial ischemia. FASEB J, 5:2029-34.
60. Lewis M, Deshpande U, Guzman L, et al. 2001. Systemic relaxin administration stimulates angiogenic cytokine expression and vessel formation in a rat myocardial infarct model. In: Tregear GW, Ivell R, Bathgate RA, Wade DJ eds. Relaxin 2001: Proceedings of the 3rd international conference on relaxin and related peptides. Dordrecht, The Netherlands: Kluwer, pp 159-67.
61. Liu J, Conklin BR, Blin N, et al. 1995. Identification of a receptor/ G-protein contact site critical for signaling specificity and G-protein activation. Proc Natl Acad Sci USA, 92:11642-6.
62. Liu C, Chen J, Sutton S, et al. 2003a. Identification of relaxin-3/INSL7 as a ligand for GPCR142. J Biol Chem, 278:50765-70.
63. Liu C, Eriste E, Sutton S, et al. 2003b. Identification of relaxin-3/ INSL7 as an endogenous ligand for the orphan G-protein-coupled receptor GPCR135. J Biol Chem, 278:50754-64.
64. Liu C, Kuei C, Sutton S, et al. 2005. INSL5 is a high affinity specific agonist for GPCR142 (GPR100). J Biol Chem, 280:292-300.
65. Manson JE, Hsia J, Johnson KC, et al. 2003. Women's Health Initiative Investigators. Estrogen plus progestin and the risk of coronary heart disease. N Engl J Med, 349:523-34.
66. Masini E, Bani D, Bello MG, et al. 1997. Relaxin counteracts myocardial damage induced by ischemia-reperfusion in isolated guinea pig hearts: evidence for an involvement of nitric oxide. Endocrinology, 138:4713-20.
67. Masini E, Nistri S, Vannacci A, et al. 2004. Relaxin inhibits the activation of human neutrophils: involvement of the nitric oxide pathway. Endocrinology, 145:1106-12.
68. Masini E, Cuzzocrea S, Mazzon E, et al. 2006. Protective effects of relaxin in ischemia/reperfusion-induced intestinal injury due to splanchnic artery occlusion. Brit J Pharmacol, 148:1124-32.
69. McDonald GA, Sarkar P, Rennke H, et al. 2003. Relaxin increases ubiquitin-dependent degradation of fibronectin in vitro and ameliorates renal fibrosis in vivo. Am J Physiol Renal Physiol, 285:F59-F67.
70. Mendelsohn ME, Karas RH. 2005. Molecular and cellular basis of cardiovascular gender differences. Science, 308:1583-7.
71. Mensah GA. 2007. Healthy endothelium: the scientific basis for cardiovascular health promotion and chronic disease prevention. Vasc Pharmacol, 46:310-4.
72. Min G, Sherwood OD. 1996. Identification of specific relaxin-binding sites in the cervix, mammary glands, nipples, small intestine and skin of pregnant pigs. Biol Reprod, 55:1243-52.
73. Moncada S, Palmer RMJ, Higgs EA. 1991. Nitric oxide physiology, pathophysiology, and pharmacology. Pharmacol Rev, 43:109-42.
74. Napoli C, de Nigris F, Williams-Ignarro S, et al. 2006. Nitric oxide and atherosclerosis: an update. Nitric Oxide, 15:265-79.
75. Nathan C. 1992. Nitric oxide as a secretory product of mammalian cells. FASEB J, 6:3051-64.
76. Nedeljkovic ZS, Gokce N, Loscalzo J. 2003. Mechanisms of oxidative stress and vascular dysfunction. Postgrad Med, J 79:195-9.
77. Nistri S, Bani D. 2003. Relaxin receptors and nitric oxide synthases. Search for the missing link. Reprod Biol Endocrinol, 1:5.
78. Nistri S, Chiappini L, Sassoli C, et al. 2003. Relaxin inhibits lipopolysaccharide-induced adhesion of neutrophils to coronary endothelial cells by a nitric oxide-mediated mechanism. FASEB J, 17:2109-11.
79. Nistri S, Bigazzi M, Bani D. 2007. Relaxin as a cardiovascular hormone. Physiology, pathophysiology and therapeutic promises. Cardiovasc Hematol Agents Med Chem (CHA-MC), 5:101-8.
80. Novak J, Danielson LA, Kerchner LJ, et al. 2001. Relaxin is essential for renal vasodilation during pregnancy in conscious rats. J Clin Invest, 107:1469-75.
81. O'Byrne EM, Carriere BT, Sorensen L, et al. 1978. Plasma immunoreactive relaxin levels in pregnant and nonpregnant women. J Clin Endocrinol Metab, 47:1106-10.
82. Orshal JM, Khalil RA. 2004. Gender, sex hormones, and vascular tone. Am J Physiol Regul Integr Comp Physiol, 286:R233-49.
83. Osto E, Coppolino G, Volpe M, et al. 2007. Restoring the dysfunctional endothelium. Curr Pharm Des, 13:1053-68.
84. Palmer RMJ, Ferrige A, Moncada S. 1987. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor Nature, 327:524-6.
85. Perna AM, Masini E, Nistri S, et al. 2005. Novel drug development opportunity for relaxin in acute myocardial infarction. evidences from a swine model. FASEB J, 19:1525-7
86. Price JF, Mowbray PI, Lee AJ, et al. 1999. Relationship between smoking and cardiovascular risk factors in the development of peripheral arterial disease and coronary artery disease: Edinburgh Artery Study. Eur Heart J, 20:344-53.
87. Quattrone S, Chiappini L, Scapagnini G, et al. 2004. Relaxin potentiates the expression of inducible nitric oxide synthase by endothelial cells from human umbilical vein in in vitro culture. Mol Hum Reprod, 10:325-30.
88. Radomski MV, Palmer RMJ, Moncada S. 1987. The role of nitric oxide and cGMP in platelet adhesion to vascular endothelium. Biochem Biophys Res Commun, 148:1482-9.
89. Rosano GM, Vitale C, Fini M. 2006. hormone replacement therapy and cardioprotection: what is good and what is bad for the cardiovascular system? Ann N Y Acad Sci, 1092:341-8.
90. Sader MA, Celermajer DS. Endothelial function, vascular reactivity and gender differences in the cardiovascular system. Cardiovasc Res, 53:597-604.
91. Samuel CS, Unemori EN, Mookerjee I, et al. 2003. Relaxin modulates cardiac fibroblast proliferation, differentiation and collagen production and reverses cardiac fibrosis in vivo. Endocrinology, 145:4125-33.
92. Samuel CS, Du XJ, Bathgate RAD, et al. 2006. Relaxin' the stiffened heart and arteries: the therapeutic potential for relaxin in the treatment of cardiovascular disease. Pharm Ther, 112:529-52.
93. Schwabe C, Billesbach EE. 1994. Relaxin: structure, functions, promises and nonevolution. FASEB J, 8:1152-60.
94. Sherwood OD. 2004. Relaxin's physiological roles and other diverse actions. Endocr Rev, 25:205-34.
95. Siow RCM, Li FYL, Rowlands DJ et al. 2007. Cardiovascular targets for estrogens and phytoestrogens: transcriptional regulation of nitric oxide synthase and antioxidant defense genes. Free Rad Bio Med, 42:909-25.
96. Sluiter W, Pietersma A, Lamers JMJ, et al. 1993. Leukocyte adhesion molecules on the vascular endothelium: their role in the pathogenesis of cardiovascular disease and the mechanism underlying their expression. J Cardiovasc Pharmacol, 22(Suppl 4):S37-44.
97. Smith GC, Pell JP, Walsh D. 2001. Pregnancy complications and maternal risk of ischaemic heart disease: A retrospective cohort study of 129,290 births. Lancet, 357:2002-6.
98. Stewart DR, Celniker AC, Taylor CA Jr, et al. 1990, Relaxin in the peri-implantation period. J Clin Endocrinol Metab, 70: 1771-3.
99. St-Louis J, Massicotte G. 1985. Chronic decrease of blood pressure by rat relaxin in spontaneously hypertensive rats. Life Sci, 37:1351-7.
100. Sudo S, Kumagai J, Nishi S, et al. 2003. H3 Relaxin is a specific ligand for LGR7 and activates the receptor by interacting with both the ectodomain and the exoloop 2. J Biol Chem, 278:7855-62.
101. Taylor MJ, Clark CL. 1994. Evidence for a novel source of relaxin: atrial cardiocytes. J Endocrinol, 143:R5-8.
102. Tunstall PH, Kuulasmaa K, P. Amouyel P, et al. 1994. Myocardial infarction and coronary deaths in the World Health Organization MONICA Project. Registration procedures, event rates, and case-fatality rates in 38 populations from 21 countries in four continents. Circulation, 90:583-612.
103. Unemori EN, Amento EP. 1990. Relaxin modulates synthesis and secretion of procollagenase and collagen by human dermal fibroblasts. J Biol Chem, 265:10681-5.
104. Unemori EN, Bauer EA, Amento EP. 1992. Relaxin alone and in conjunction with interferon-γ decreases collagen synthesis by cultured human scleroderma fibroblasts. J Invest Dermatol, 99:337-42.
105. Unemori EN, Beck LS, Lee WP, et al. 1993. Human relaxin decreases collagen accumulation in vivo in two rodent models of fibrosis. J Invest Dermatol, 101:290-5.
106. Unemori EN, Pickford LB, Salles AL, et al.1996. Relaxin induces an extracellular matrix-degrading phenotype in human lung fibroblasts in vitro and inhibits lung fibrosis in a routine model in vivo. J Clin Invest, 98:2739-45.
107. Unemori EN, Erikson ME, Rocco SE, et al. 1999. Relaxin stimulates expression on vascular endothelial growth factor in normal human endometrial cells in vitro and is associated with menometrorrhagia in women. Hum Reprod, 14:800-6.
108. Vanhoutte PM. 1997. Endothelial dysfunction and atherosclerosis. Eur Heart J, 18:E19-29.
109. Vasilenko P, Mead JP, Weidmann JE. 1986 Uterine growth-promoting effects of relaxin: A morphometric and histological analysis. Biol Reprod, 35:987-95.
110. Wei JY. 1992. Age and the cardiovascular system. N Engl J Med, 327:1735-9.
111. Wenger NK, Speroff L, Packard B. 1993. Cardiovascular health and disease in women. N Engl J Med, 329:247-56.
112. Williams EJ, Benyon RC, Trim N, et al. 2001. Relaxin inhibits effective collagen deposition by cultured hepatic stellate cells and decreases rat liver fibrosis in vivo. Gut, 49:577-83.
113. Wilson BC, Connell B, Saleh TM. 2006, Relaxin-induced reduction of infarct size in male rats receiving MCAO is dependent on nitric oxide synthesis and not estrogenic mechanisms. Neurosci Lett, 393:160-4.
114. Winslow JW, Shih A, Bourell JH, et al. 1992. Human seminal relaxin is a product of the same gene as human luteal relaxin. Endocrinology 130:2660-88.
115. Wyckoff MH, Chambliss KL, Mineo C, et al. 2001. Plasma membrane estrogen receptors are coupled to endothelial nitric oxide synthase through Ga, J Biol Chem, 276:27071-6.
116. Zdravkovic T, Genbacev O, McMaster MT, et al. 2005. The adverse effects of maternal smoking on the human placenta: a review. Placenta, 26(Suppl A):S81-6.
117. Zhang J, Qi YF, Geng B, et al. 2005. Effect of relaxin on myocardial ischemia injury induced by isoproterenol. Peptides, 26:1632-9.
118. Ziche M, Morbidelli L, Masini E, et al. 1994 Nitric oxide mediates angiogenesis in vivo and endothelial cell growth and migration in vitro promoted by substance P. J Clin Invest, 94:2036-44.