Two types of TNF-α exist in teleost fish: Phylogeny, expression, and bioactivity analysis of type-II TNF-α3 in rainbow trout Oncorhynchus mykiss

Journal of Immunology

Volumn 191, Issue 12, 2013, Pages 5959-5972

  Hong, Suhee ^a,b   Li, Ronggai ^a   Xu, Qiaoqing ^a,c   Secombes, Chris J ^a   Wang, Tiehui ^a  

^a UNIVERSITY OF ABERDEEN   (United Kingdom)

^b Gangneung Wonju National University   (South Korea)

^c YANGTZE UNIVERSITY   (China)

Author keywords

[No Author keywords available]

Indexed keywords

CATHELICIDIN; CYTOKINE; HEPCIDIN; INTERLEUKIN 17; INTERLEUKIN 1BETA; INTERLEUKIN 34; INTERLEUKIN 6; INTERLEUKIN 8; LIPOPOLYSACCHARIDE; MEMBRANE PROTEIN; PEPTIDOGLYCAN; POLYINOSINIC POLYCYTIDYLIC ACID; RECOMBINANT GAMMA INTERFERON; SUPPRESSOR OF CYTOKINE SIGNALING 1; SUPPRESSOR OF CYTOKINE SIGNALING 2; SUPPRESSOR OF CYTOKINE SIGNALING 3; TRANSFORMING GROWTH FACTOR BETA1; TUMOR NECROSIS FACTOR ALPHA; TUMOR NECROSIS FACTOR ALPHA1; TUMOR NECROSIS FACTOR ALPHA2; TUMOR NECROSIS FACTOR ALPHA3; UNCLASSIFIED DRUG;

AMINO ACID SEQUENCE; ANIMAL CELL; ARTICLE; BACTERIAL INFECTION; BIOLOGICAL ACTIVITY; CONTROLLED STUDY; ENZYME SYNTHESIS; GENE; IN VITRO STUDY; IN VIVO STUDY; MACROPHAGE; MOLECULAR CLONING; NONHUMAN; PHYLOGENY; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN EXPRESSION; PROTEIN FUNCTION; RAINBOW TROUT; SEQUENCE ANALYSIS; SYNTENY; TNF ALPHA GENE; VIRUS INFECTION;

EID: 84890402747     PISSN: 00221767     EISSN: 15506606     Source Type: Journal    
DOI: 10.4049/jimmunol.1301584     Document Type: Article

References (86)

1. Chu, W. M. 2013. Tumor necrosis factor. Cancer Lett. 328: 222-225.
2. Wollenberg, G. K., L. E. DeForge, G. Bolgos, and D. G. Remick. 1993. Differential expression of tumor necrosis factor and interleukin-6 by peritoneal macrophages in vivo and in culture. Am. J. Pathol. 143: 1121-1130.
3. Kayama, F., T. Yoshida, Y. Kodama, T. Matsui, J. M. Matheson, and M. I. Luster. 1997. Pro-inflammatory cytokines and interleukin 6 in the renal response to bacterial endotoxin. Cytokine 9: 688-695.
4. Amiot, F., C. Fitting, K. J. Tracey, J. M. Cavaillon, and F. Dautry. 1997. Lipopolysaccharide-induced cytokine cascade and lethality in LT alpha/TNF alpha-deficient mice. Mol. Med. 3: 864-875.
5. Horiuchi, T., H. Mitoma, S. Harashima, H. Tsukamoto, and T. Shimoda. 2010. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford) 49: 1215-1228.
6. Rahman, M. M., and G. McFadden. 2006. Modulation of tumor necrosis factor by microbial pathogens. PLoS Pathog. 2: e4.
7. Yarilina, A., K. H. Park-Min, T. Antoniv, X. Hu, and L. B. Ivashkiv. 2008. TNF activates an IRF1-dependent autocrine loop leading to sustained expression of chemokines and STAT1-dependent type I interferon-response genes. Nat. Immunol. 9: 378-387.
8. Parameswaran, N., and S. Patial. 2010. Tumor necrosis factor-a signaling in macrophages. Crit. Rev. Eukaryot. Gene Expr. 20: 87-103.
9. Lo, S. Z., J. H. Steer, and D. A. Joyce. 2011. TNF-α renders macrophages resistant to a range of cancer chemotherapeutic agents through NF-κB-mediated antagonism of apoptosis signalling. Cancer Lett. 307: 80-92.
10. Aggarwal, B. B., S. C. Gupta, and J. H. Kim. 2012. Historical perspectives on tumor necrosis factor and its superfamily: 25 years later, a golden journey. Blood 119: 651-665.
11. Glenney, G. W., and G. D. Wiens. 2007. Early diversification of the TNF superfamily in teleosts: genomic characterization and expression analysis. J. Immunol. 178: 7955-7973.
12. Wiens, G. D., and G. W. Glenney. 2011. Origin and evolution of TNF and TNF receptor superfamilies. Dev. Comp. Immunol. 35: 1324-1335.
13. Bodmer, J. L., P. Schneider, and J. Tschopp. 2002. The molecular architecture of the TNF superfamily. Trends Biochem. Sci. 27: 19-26.
14. Ware, C. F. 2005. Network communications: lymphotoxins, LIGHT, and TNF. Annu. Rev. Immunol. 23: 787-819.
15. Hirono, I., B. H. Nam, T. Kurobe, and T. Aoki. 2000. Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus. J. Immunol. 165: 4423-4427.
16. Laing, K. J., T. Wang, J. Zou, J. Holland, S. Hong, N. Bols, I. Hirono, T. Aoki, and C. J. Secombes. 2001. Cloning and expression analysis of rainbow trout Oncorhynchus mykiss tumour necrosis factor-alpha. Eur. J. Biochem. 268: 1315-1322.
17. García-Castillo, J., P. Pelegrín, V. Mulero, and J. Meseguer. 2002. Molecular cloning and expression analysis of tumor necrosis factor alpha from a marine fish reveal its constitutive expression and ubiquitous nature. Immunogenetics 54: 200-207.
18. Saeij, J. P., R. J. Stet, B. J. de Vries, W. B. van Muiswinkel, and G. F. Wiegertjes. 2003. Molecular and functional characterization of carp TNF: a link between TNF polymorphism and trypanotolerance? Dev. Comp. Immunol. 27: 29-41.
19. Zou, J., C. J. Secombes, S. Long, N. Miller, L. W. Clem, and V. G. Chinchar. 2003. Molecular identification and expression analysis of tumor necrosis factor in channel catfish (Ictalurus punctatus). Dev. Comp. Immunol. 27: 845-858.
20. Savan, R., T. Kono, D. Igawa, and M. Sakai. 2005. A novel tumor necrosis factor (TNF) gene present in tandem with theTNF-alpha gene on the same chromosome in teleosts. Immunogenetics 57: 140-150.
21. Nascimento, D. S., P. J. Pereira, M. I. Reis, A. do Vale, J. Zou, M. T. Silva, C. J. Secombes, and N. M. dos Santos. 2007. Molecular cloning and expression analysis of sea bass (Dicentrarchus labrax L.) tumor necrosis factor-alpha (TNF-alpha). Fish Shellfish Immunol. 23: 701-710.
22. Zhang, A., D. Chen, H. Wei, L. Du, T. Zhao, X. Wang, and H. Zhou. 2012. Functional characterization of TNF-α in grass carp head kidney leukocytes: induction and involvement in the regulation of NF-kB signaling. Fish Shellfish Immunol. 33: 1123-1132.
23. Cai, Z. H., L. S. Song, C. P. Gao, L. T. Wu, L. H. Qiu, and J. H. Xiang. 2003. [Cloning and expression of tumor necrosis factor (TNFalpha) cDNA from red seabream Pagrus major.]. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 35: 1111-1116 (Shanghai).
24. Kim, M. S., Y. J. Hwang, K. J. Yoon, K. Zenke, Y. K. Nam, S. K. Kim, and K. H. Kim. 2009. Molecular cloning of rock bream (Oplegnathus fasciatus) tumor necrosis factor-alpha and its effect on the respiratory burst activity of phagocytes. Fish Shellfish Immunol. 27: 618-624.
25. Ordás, M. C., M. M. Costa, F. J. Roca, G. López- Castejón, V. Mulero, J. Meseguer, A. Figueras, and B. Novoa. 2007. Turbot TNFalpha gene: molecular characterization and biological activity of the recombinant protein. Mol. Immunol. 44: 389-400.
26. Uenobe, M., C. Kohchi, N. Yoshioka, A. Yuasa, H. Inagawa, K. Morii, T. Nishizawa, Y. Takahashi, and G. Soma. 2007. Cloning and characterization of a TNF-like protein of Plecoglossus altivelis (ayu fish). Mol. Immunol. 44: 1115-1122.
27. Savan, R., and M. Sakai. 2004. Presence of multiple isoforms of TNF alpha in carp (Cyprinus carpio L.): genomic and expression analysis. Fish Shellfish Immunol. 17: 87-94.
28. Zou, J., T. Wang, I. Hirono, T. Aoki, H. Inagawa, T. Honda, G. I. Soma, M. Ototake, T. Nakanishi, A. E. Ellis, and C. J. Secombes. 2002. Differential expression of two tumor necrosis factor genes in rainbow trout, Oncorhynchus mykiss. Dev. Comp. Immunol. 26: 161-172.
29. Kadowaki, T., H. Harada, Y. Sawada, C. Kohchi, G. Soma, Y. Takahashi, and H. Inagawa. 2009. Two types of tumor necrosis factor-alpha in bluefin tuna (Thunnus orientalis) genes: Molecular cloning and expression profile in response to several immunological stimulants. Fish Shellfish Immunol. 27: 585-594.
30. Lam, F. W., S. Y. Wu, S. J. Lin, C. C. Lin, Y. M. Chen, H. C. Wang, T. Y. Chen, H. T. Lin, and J. H. Lin. 2011. The expression of two novel orange-spotted grouper (Epinephelus coioides) TNF genes in peripheral blood leukocytes, various organs, and fish larvae. Fish Shellfish Immunol. 30: 618-629.
31. Meyer, A., and Y. Van de Peer. 2005. From 2R to 3R: evidence for a fish-specific genome duplication (FSGD). Bioessays 27: 937-945.
32. Wang, T., P. C. Hanington, M. Belosevic, and C. J. Secombes. 2008. Two macrophage colony-stimulating factor genes exist in fish that differ in gene organization and are differentially expressed. J. Immunol. 181: 3310-3322.
33. Husain, M., S. Bird, R. van Zwieten, C. J. Secombes, and T. Wang. 2012. Cloning of the IL-1β3 gene and IL-1β4 pseudogene in salmonids uncovers a second type of IL-1β gene in teleost fish. Dev. Comp. Immunol. 38: 431-446.
34. He, X., and J. Zhang. 2005. Rapid subfunctionalization accompanied by prolonged and substantial neofunctionalization in duplicate gene evolution. Genetics 169: 1157-1164.
35. Zou, J., S. Peddie, G. Scapigliati, Y. Zhang, N. C. Bols, A. E. Ellis, and C. J. Secombes. 2003. Functional characterisation of the recombinant tumor necrosis factors in rainbow trout, Oncorhynchus mykiss. Dev. Comp. Immunol. 27: 813-822.
36. Grayfer, L., J. G. Walsh, and M. Belosevic. 2008. Characterization and functional analysis of goldfish (Carassius auratus L.) tumor necrosis factor-alpha. Dev. Comp. Immunol. 32: 532-543.
37. Roca, F. J., I. Mulero, A. López-Muñoz, M. P. Sepulcre, S. A. Renshaw, J. Meseguer, and V. Mulero. 2008. Evolution of the inflammatory response in vertebrates: fish TNF-alpha is a powerful activator of endothelial cells but hardly activates phagocytes. J. Immunol. 181: 5071-5081.
38. Forlenza, M., S. Magez, J. P. Scharsack, A. Westphal, H. F. Savelkoul, and G. F. Wiegertjes. 2009. Receptor-mediated and lectin-like activities of carp (Cyprinus carpio) TNF-alpha. J. Immunol. 183: 5319-5332.
39. Altschul, S. F., T. L. Madden, A. A. Schäffer, J. Zhang, Z. Zhang, W. Miller, and D. J. Lipman. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25: 3389-3402.
40. Wang, T., and C. J. Secombes. 2003. Complete sequencing and expression of three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss. Immunogenetics 55: 615-628.
41. Ganassin, R. C., and N. C. Bols. 1998. Development of a monocyte/macrophage-like cell line, RTS11, from rainbow trout spleen. Fish Shellfish Immunol. 8: 457-476.
42. Wang, T., P. Diaz-Rosales, M. M. Costa, S. Campbell, M. Snow, B. Collet, S. A. Martin, and C. J. Secombes. 2011a. Functional characterization of a non-mammalian IL-21: rainbow trout Oncorhynchus mykiss IL-21 upregulates the expression of the Th cell signature cytokines IFN-gamma, IL-10, and IL-22. J. Immunol. 186: 708-721.
43. Chenna, R., H. Sugawara, T. Koike, R. Lopez, T. J. Gibson, D. G. Higgins, and J. D. Thompson. 2003. Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res. 31: 3497-3500.
44. Campanella, J. J., L. Bitincka, and J. Smalley. 2003. MatGAT: an application that generates similarity/identity matrices using protein or DNA sequences. BMC Bioinformatics 4: 29.
45. Hofmann, K., and W. Stoffel. 1993. TMbase - A database of membrane spanning protein segments. Biol. Chem. Hoppe Seyler 373: 166.
46. Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 2731-2739.
47. Muffato, M., A. Louis, C. E. Poisnel, and H. Roest Crollius. 2010. Genomicus: a database and a browser to study gene synteny in modern and ancestral genomes. Bioinformatics 26: 1119-1121.
48. Wang, T., W. Huang, M. M. Costa, S. A. Martin, and C. J. Secombes. 2011b. Two copies of the genes encoding the subunits of putative interleukin (IL)-4/IL-13 receptors, IL-4Rα, IL-13Rα1 and IL-13Rα2, have been identified in rainbow trout (Oncorhynchus mykiss) and have complex patterns of expression and modulation. Immunogenetics 63: 235-253.
49. Costa, M. M., T. Maehr, P. Diaz-Rosales, C. J. Secombes, and T. Wang. 2011. Bioactivity studies of rainbow trout (Oncorhynchus mykiss) interleukin-6: effects on macrophage growth and antimicrobial peptide gene expression. Mol. Immunol. 48: 1903-1916.
50. Hong, S., J. Zou, M. Crampe, S. Peddie, G. Scapigliati, N. Bols, C. Cunningham, and C. J. Secombes. 2001. The production and bioactivity of rainbow trout (Oncorhynchus mykiss) recombinant IL-1 beta. Vet. Immunol. Immunopathol. 81: 1-14.
51. Lee, L. E., J. H. Clemons, D. G. Bechtel, S. J. Caldwell, K. B. Han, M. Pasitschniak-Arts, D. D. Mosser, and N. C. Bols. 1993. Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity. Cell Biol. Toxicol. 9: 279-294.
52. Wolf, K., and M. C. Quimby. 1962. Established eurythermic line of fish cells in vitro. Science 135: 1065-1066.
53. Schirmer, K., A. G. Chan, B. M. Greenberg, D. G. Dixon, and N. C. Bols. 1998. Ability of 16 priority PAHs to be photocytotoxic to a cell line from the rainbow trout gill. Toxicology 127: 143-155.
54. Wang, T., M. M. Monte, W. Huang, P. Boudinot, S. A. Martin, and C. J. Secombes. 2010. Identification of two FoxP3 genes in rainbow trout (Oncorhynchus mykiss) with differential induction patterns. Mol. Immunol. 47: 2563-2574.
55. Campbell, S., B. Collet, K. Einer-Jensen, C. J. Secombes, and M. Snow. 2009. Identifying potential virulence determinants in viral haemorrhagic septicaemia virus (VHSV) for rainbow trout. Dis. Aquat. Organ. 86: 205-212.
56. Wang, T., S. Bird, A. Koussounadis, J. W. Holland, A. Carrington, J. Zou, and C. J. Secombes. 2009. Identification of a novel IL-1 cytokine family member in teleost fish. J. Immunol. 183: 962-974.
57. Kou, T. C., L. Fan, Y. Zhou, Z. Y. Ye, X. P. Liu, L. Zhao, and W. S. Tan. 2011. Detailed understanding of enhanced specific productivity in Chinese hamster ovary cells at low culture temperature. J. Biosci. Bioeng. 111: 365-369.
58. Wang, T., J. W. Holland, N. Bols, and C. J. Secombes. 2005. Cloning and expression of the first nonmammalian interleukin-11 gene in rainbow trout Oncorhynchus mykiss. FEBS J. 272: 1136-1147.
59. Wang, T., J. W. Holland, A. Carrington, J. Zou, and C. J. Secombes. 2007. Molecular and functional characterization of IL-15 in rainbow trout Oncorhynchus mykiss: a potent inducer of IFN-gamma expression in spleen leukocytes. J. Immunol. 179: 1475-1488.
60. Wang, T., S. A. Martin, and C. J. Secombes. 2010. Two interleukin-17C-like genes exist in rainbow trout Oncorhynchus mykiss that are differentially expressed and modulated. Dev. Comp. Immunol. 34: 491-500.
61. Wang, T., T. Kono, M. M. Monte, H. Kuse, M. M. Costa, H. Korenaga, T. Maehr, M. Husain, M. Sakai, and C. J. Secombes. 2013. Identification of IL-34 in teleost fish: differential expression of rainbow trout IL-34, MCSF1 and MCSF2, ligands of the MCSF receptor. Mol. Immunol. 53: 398-409.
62. Chang, C. I., Y. A. Zhang, J. Zou, P. Nie, and C. J. Secombes. 2006. Two cathelicidin genes are present in both rainbow trout (Oncorhynchus mykiss) and atlantic salmon (Salmo salar) . Antimicrob. Agents Chemother. 50: 185-195.
63. Douglas, S. E., J. W. Gallant, R. S. Liebscher, A. Dacanay, and S. C. Tsoi. 2003. Identification and expression analysis of hepcidin-like antimicrobial peptides in bony fish. Dev. Comp. Immunol. 27: 589-601.
64. Wang, T., and C. J. Secombes. 2008. Rainbow trout suppressor of cytokine signalling (SOCS)-1, 2 and 3: molecular identification, expression and modulation. Mol. Immunol. 45: 1449-1457.
65. Harun, N. O., M. M. Costa, C. J. Secombes, and T. Wang. 2011. Sequencing of a second interleukin-10 gene in rainbow trout Oncorhynchus mykiss and comparative investigation of the expression and modulation of the paralogues in vitro and in vivo. Fish Shellfish Immunol. 31: 107-117.
66. Maehr, T., M. M. Costa, J. L. Vecino, S. Wadsworth, S. A. Martin, T. Wang, and C. J. Secombes. 2013. Transforming growth factor-b1b: a second TGF-β1 paralogue in the rainbow trout (Oncorhynchus mykiss) that has a lower constitutive expression but is more responsive to immune stimulation. Fish Shellfish Immunol. 34: 420-432.
67. Jones, E. Y., D. I. Stuart, and N. P. Walker. 1989. Structure of tumour necrosis factor. Nature 338: 225-228.
68. Narachi, M. A., J. M. Davis, Y. R. Hsu, and T. Arakawa. 1987. Role of single disulfide in recombinant human tumor necrosis factor-alpha. J. Biol. Chem. 262: 13107-13110.
69. MacKenzie, S. A., N. Roher, S. Boltaña, and F. W. Goetz. 2010. Peptidoglycan, not endotoxin, is the key mediator of cytokine gene expression induced in rainbow trout macrophages by crude LPS. Mol. Immunol. 47: 1450-1457.
70. Brenchley, J. M., D. C. Douek, D. R. Ambrozak, M. Chatterji, M. R. Betts, L. S. Davis, and R. A. Koup. 2002. Expansion of activated human naïve T-cells precedes effector function. Clin. Exp. Immunol. 130: 432-440.
71. Shalaby, M. R., T. Espevik, G. C. Rice, A. J. Ammann, I. S. Figari, G. E. Ranges, and M. A. Palladino, Jr. 1988. The involvement of human tumor necrosis factors-alpha and -beta in the mixed lymphocyte reaction. J. Immunol. 141: 499-503.
72. Semenzato, G. 1990. Tumour necrosis factor: a cytokine with multiple biological activities. Br. J. Cancer 61: 354-361.
73. Flajnik, M. F., and M. Kasahara. 2010. Origin and evolution of the adaptive immune system: genetic events and selective pressures. Nat. Rev. Genet. 11: 47-59.
74. Ravi, V., and B. Venkatesh. 2008. Rapidly evolving fish genomes and teleost diversity. Curr. Opin. Genet. Dev. 18: 544-550.
75. Brunet, F. G., H. Roest Crollius, M. Paris, J. M. Aury, P. Gibert, O. Jaillon, V. Laudet, and M. Robinson-Rechavi. 2006. Gene loss and evolutionary rates following whole-genome duplication in teleost fishes. Mol. Biol. Evol. 23: 1808-1816.
76. Eissner, G., W. Kolch, and P. Scheurich. 2004. Ligands working as receptors: reverse signaling by members of the TNF superfamily enhance the plasticity of the immune system. Cytokine Growth Factor Rev. 15: 353-366.
77. Calmon-Hamaty, F., B. Combe, M. Hahne, and J. Morel. 2011. Lymphotoxin α revisited: general features and implications in rheumatoid arthritis. Arthritis Res. Ther. 13: 232.
78. Beil, E. J., G. A. Heavner, S. J. Wu, and J. F. Nemeth. 2012. Probing the solution structure of tumor necrosis factor-a homotrimer and heterotrimer after complex perturbation using electrospray ionization mass spectrometry. J. Mol. Recognit. 25: 174-183.
79. Schottelius, A. J., L. L. Moldawer, C. A. Dinarello, K. Asadullah, W. Sterry, and C. K. Edwards, III. 2004. Biology of tumor necrosis factor-alpha- implications for psoriasis. Exp. Dermatol. 13: 193-222.
80. Praveen, K., D. L. Evans, and L. Jaso-Friedmann. 2006. Constitutive expression of tumor necrosis factor-alpha in cytotoxic cells of teleosts and its role in regulation of cell-mediated cytotoxicity. Mol. Immunol. 43: 279-291.
81. DeForge, L. E., J. S. Kenney, M. L. Jones, J. S. Warren, and D. G. Remick. 1992. Biphasic production of IL-8 in lipopolysaccharide (LPS)-stimulated human whole blood. Separation of LPS- and cytokine-stimulated components using anti-tumor necrosis factor and anti-IL-1 antibodies. J. Immunol. 148: 2133-2141.
82. Li, H., and X. Lin. 2008. Positive and negative signaling components involved in TNFalpha-induced NF-kappaB activation. Cytokine 41: 1-8.
83. Ehlting, C., W. S. Lai, F. Schaper, E. D. Brenndörfer, R. J. Matthes, P. C. Heinrich, S. Ludwig, P. J. Blackshear, M. Gaestel, D. Häussinger, and J. G. Bode. 2007. Regulation of suppressor of cytokine signaling 3 (SOCS3) mRNA stability by TNF-alpha involves activation of the MKK6/p38MAPK/MK2 cascade. J. Immunol. 178: 2813-2826.
84. Pappu, R., S. Rutz, and W. Ouyang. 2012. Regulation of epithelial immunity by IL-17 family cytokines. Trends Immunol. 33: 343-349.
85. Secombes, C. J., T. Wang, and S. Bird. 2011. The interleukins of fish. Dev. Comp. Immunol. 35: 1336-1345.
86. Nemeth, E., S. Rivera, V. Gabayan, C. Keller, S. Taudorf, B. K. Pedersen, and T. Ganz. 2004. IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin. J. Clin. Invest. 113: 1271-1276.