Association of endothelial nitric oxide synthase (NOS3) gene polymorphisms with primary open-angle glaucoma in a Saudi cohort

Autoři: Altaf A. Kondkar aff001;  Taif A. Azad aff001;  Tahira Sultan aff001;  Essam A. Osman aff001;  Faisal A. Almobarak aff001;  Saleh A. Al-Obeidan aff001
Působiště autorů: Department of Ophthalmology, College of Medicine, King Saud University, Riyadh, Saudi Arabia aff001;  Glaucoma Research Chair in Ophthalmology, College of Medicine, King Saud University, Riyadh, Saudi Arabia aff002
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article



To investigate the association of endothelial nitric oxide synthase (NOS3) gene polymorphisms in patients with primary open-angle glaucoma (POAG) of Saudi origin.


This case-control study included 173 patients with POAG (94 men and 79 women) and 171 controls (98 men and 73 women). Genotyping of rs2070744 (T-786C) and rs1799983 (G894T) variants of the NOS3 gene was performed using TaqMan® assay.


Rs1799983 genotypes showed a significant association with POAG but did not survive Bonferroni correction (pcorrection = 0.01). The minor ‘T’ allele was significantly associated with the risk of POAG among men (p = 0.025, odds ratio (OR) = 1.77, 95% confidence interval (CI) = 1.07–2.94). Likewise, the genotypes were significantly associated with POAG among men in dominant (p = 0.030, OR = 1.92, 95% CI = 1.06–3.48) and log-additive models (p = 0.022, OR = 1.82, 95% CI = 1.08–3.07), and after adjustment for age and smoking. Genotype and allele frequencies of rs2070744 were not significantly different between POAG cases and controls, and after sex stratification. CG haplotype was significantly protective (p = 0.011, OR = 0.52, 95% CI = 0.32–0.87) and CT haplotype conferred significantly increased risk of POAG (p = 0.016, OR = 2.60, 95% CI = 1.16–5.82) among men. Rs1799983 showed trend (p = 0.054) towards risk of POAG independent of age, gender, smoking, and rs2070744 polymorphism in logistic regression analysis. Both the polymorphisms showed no association with POAG phenotypes such as intraocular pressure and cup/disc ratio.


Our results suggest that the polymorphism rs1799983 and the haplotypes of rs20707440 and rs1799983 in the NOS3 gene may significantly modulate the risk of POAG in Saudi’s, particularly among men. Further larger studies are needed to confirm these findings.

Klíčová slova:

Alleles – Genetics of disease – Glaucoma – Haplotypes – Human genetics – Molecular genetics – Nitric oxide – Variant genotypes


1. Kwon YH, Fingert JH, Kuehn MH, Alward WL. Primary open-angle glaucoma. N Engl J Med. 2009;360(11):1113–24. Epub 2009/03/13. doi: 10.1056/NEJMra0804630 360/11/1113 [pii]. 19279343; PubMed Central PMCID: PMC3700399.

2. Al Obeidan SA, Dewedar A, Osman EA, Mousa A. The profile of glaucoma in a Tertiary Ophthalmic University Center in Riyadh, Saudi Arabia. Saudi J Ophthalmol. 2011;25(4):373–9. Epub 2011/10/01. doi: 10.1016/j.sjopt.2011.09.001 SJOPT118 [pii]. 23960951; PubMed Central PMCID: PMC3729326.

3. Abu-Amero K, Kondkar AA, Chalam KV. An Updated Review on the Genetics of Primary Open Angle Glaucoma. Int J Mol Sci. 2015;16(12):28886–911. Epub 2015/12/23. doi: 10.3390/ijms161226135 ijms161226135 [pii]. 26690118; PubMed Central PMCID: PMC4691082.

4. Topouzis F, Wilson MR, Harris A, Founti P, Yu F, Anastasopoulos E, et al. Association of open-angle glaucoma with perfusion pressure status in the Thessaloniki Eye Study. Am J Ophthalmol. 2013;155(5):843–51. Epub 2013/02/12. doi: 10.1016/j.ajo.2012.12.007 S0002-9394(12)00861-6 [pii]. 23394905.

5. Orzalesi N, Rossetti L, Omboni S. Vascular risk factors in glaucoma: the results of a national survey. Graefes Arch Clin Exp Ophthalmol. 2007;245(6):795–802. Epub 2006/11/23. doi: 10.1007/s00417-006-0457-5 17120006.

6. Su WW, Cheng ST, Ho WJ, Tsay PK, Wu SC, Chang SH. Glaucoma is associated with peripheral vascular endothelial dysfunction. Ophthalmology. 2008;115(7):1173–8 e1. Epub 2007/12/14. S0161-6420(07)01178-5 [pii] doi: 10.1016/j.ophtha.2007.10.026 18076992.

7. Janssen SF, Gorgels TG, Ramdas WD, Klaver CC, van Duijn CM, Jansonius NM, et al. The vast complexity of primary open angle glaucoma: disease genes, risks, molecular mechanisms and pathobiology. Prog Retin Eye Res. 2013;37:31–67. Epub 2013/09/24. doi: 10.1016/j.preteyeres.2013.09.001 S1350-9462(13)00057-8 [pii]. 24055863.

8. Charlesworth J, Kramer PL, Dyer T, Diego V, Samples JR, Craig JE, et al. The path to open-angle glaucoma gene discovery: endophenotypic status of intraocular pressure, cup-to-disc ratio, and central corneal thickness. Invest Ophthalmol Vis Sci. 2010;51(7):3509–14. Epub 2010/03/20. doi: 10.1167/iovs.09-4786 iovs.09-4786 [pii]. 20237253; PubMed Central PMCID: PMC2904007.

9. Bian K, Murad F. What is next in nitric oxide research? From cardiovascular system to cancer biology. Nitric Oxide. 2014;43:3–7. Epub 2014/08/26. doi: 10.1016/j.niox.2014.08.006 S1089-8603(14)00280-8 [pii]. 25153032.

10. Stamer WD, Lei Y, Boussommier-Calleja A, Overby DR, Ethier CR. eNOS, a pressure-dependent regulator of intraocular pressure. Invest Ophthalmol Vis Sci. 2011;52(13):9438–44. Epub 2011/11/01. doi: 10.1167/iovs.11-7839 iovs.11-7839 [pii]. 22039240; PubMed Central PMCID: PMC3293415.

11. Lei Y, Zhang X, Song M, Wu J, Sun X. Aqueous Humor Outflow Physiology in NOS3 Knockout Mice. Invest Ophthalmol Vis Sci. 2015;56(8):4891–8. Epub 2015/08/01. doi: 10.1167/iovs.15-16564 2422124 [pii]. 26225628.

12. Nathanson JA, McKee M. Identification of an extensive system of nitric oxide-producing cells in the ciliary muscle and outflow pathway of the human eye. Invest Ophthalmol Vis Sci. 1995;36(9):1765–73. Epub 1995/08/01. 7543462.

13. Chang JY, Stamer WD, Bertrand J, Read AT, Marando CM, Ethier CR, et al. Role of nitric oxide in murine conventional outflow physiology. Am J Physiol Cell Physiol. 2015;309(4):C205–14. Epub 2015/06/05. doi: 10.1152/ajpcell.00347.2014 ajpcell.00347.2014 [pii]. 26040898; PubMed Central PMCID: PMC4537932.

14. Galassi F, Renieri G, Sodi A, Ucci F, Vannozzi L, Masini E. Nitric oxide proxies and ocular perfusion pressure in primary open angle glaucoma. Br J Ophthalmol. 2004;88(6):757–60. Epub 2004/05/19. doi: 10.1136/bjo.2003.028357 15148207; PubMed Central PMCID: PMC1772173.

15. Cavet ME, Vittitow JL, Impagnatiello F, Ongini E, Bastia E. Nitric oxide (NO): an emerging target for the treatment of glaucoma. Invest Ophthalmol Vis Sci. 2014;55(8):5005–15. Epub 2014/08/16. doi: 10.1167/iovs.14-14515 55/8/5005 [pii]. 25125670.

16. Toda N, Nakanishi-Toda M. Nitric oxide: ocular blood flow, glaucoma, and diabetic retinopathy. Prog Retin Eye Res. 2007;26(3):205–38. Epub 2007/03/06. S1350-9462(07)00005-5 [pii] doi: 10.1016/j.preteyeres.2007.01.004 17337232.

17. Wink DA, Miranda KM, Espey MG, Pluta RM, Hewett SJ, Colton C, et al. Mechanisms of the antioxidant effects of nitric oxide. Antioxid Redox Signal. 2001;3(2):203–13. Epub 2001/06/09. doi: 10.1089/152308601300185179 11396476.

18. Ster AM, Popp RA, Petrisor FM, Stan C, Pop VI. The Role of Oxidative Stress and Vascular Insufficiency in Primary Open Angle Glaucoma. Clujul Med. 2014;87(3):143–6. Epub 2014/01/01. doi: 10.15386/cjmed-295 cm8703p143 [pii]. 26528013; PubMed Central PMCID: PMC4508597.

19. Lin HJ, Tsai CH, Tsai FJ, Chen WC, Tsai SW, Fan SS. Distribution of oxidation enzyme eNOS and myeloperoxidase in primary open angle glaucoma. J Clin Lab Anal. 2005;19(2):87–92. Epub 2005/03/10. doi: 10.1002/jcla.20057 15756709.

20. Emam WA, Zidan HE, Abdulhalim BE, Dabour SA, Ghali MA, Kamal AT. Endothelial nitric oxide synthase polymorphisms and susceptibility to high-tension primary open-angle glaucoma in an Egyptian cohort. Mol Vis. 2014;20:804–11. Epub 2014/06/19. 24940036; PubMed Central PMCID: PMC4057245.

21. Magalhaes da Silva T, Rocha AV, Lacchini R, Marques CR, Silva ES, Tanus-Santos JE, et al. Association of polymorphisms of endothelial nitric oxide synthase (eNOS) gene with the risk of primary open angle glaucoma in a Brazilian population. Gene. 2012;502(2):142–6. Epub 2012/05/09. doi: 10.1016/j.gene.2012.04.047 S0378-1119(12)00473-8 [pii]. 22561696.

22. Miyamoto Y, Saito Y, Nakayama M, Shimasaki Y, Yoshimura T, Yoshimura M, et al. Replication protein A1 reduces transcription of the endothelial nitric oxide synthase gene containing a -786T—>C mutation associated with coronary spastic angina. Hum Mol Genet. 2000;9(18):2629–37. Epub 2000/11/07. doi: 10.1093/hmg/9.18.2629 11063722.

23. Tesauro M, Thompson WC, Rogliani P, Qi L, Chaudhary PP, Moss J. Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs. glutamate at position 298. Proc Natl Acad Sci U S A. 2000;97(6):2832–5. Epub 2000/03/16. 97/6/2832 [pii] doi: 10.1073/pnas.97.6.2832 10717002; PubMed Central PMCID: PMC16015.

24. Xiang Y, Dong Y, Li X, Tang X. Association of Common Variants in eNOS Gene with Primary Open Angle Glaucoma: A Meta-Analysis. J Ophthalmol. 2016;2016:1348347. Epub 2016/06/01. doi: 10.1155/2016/1348347 27242919; PubMed Central PMCID: PMC4875980.

25. Kondkar AA, Azad TA, Almobarak FA, Abu-Amero KK, Al-Obeidan SA. Polymorphism rs7961953 in TMTC2 gene is not associated with primary open-angle glaucoma in a Saudi cohort. Ophthalmic Genet. 2019;40(1):74–6. Epub 2019/02/08. doi: 10.1080/13816810.2019.1576210 30729851.

26. Kondkar AA, Sultan T, Almobarak FA, Kalantan H, Abu-Amero KK, Al-Obeidan SA. Plexin domain containing 2 (PLXDC2) gene polymorphism rs7081455 may not influence POAG risk in a Saudi cohort. BMC Res Notes. 2018;11(1):733. Epub 2018/10/18. doi: 10.1186/s13104-018-3848-x [pii]. 30326957; PubMed Central PMCID: PMC6192173.

27. Kondkar AA, Azad TA, Almobarak FA, Bahabri IM, Kalantan H, Abu-Amero KK, et al. Lack of Association between Variant rs7916697 in ATOH7 and Primary Open Angle Glaucoma in a Saudi Cohort. Genet Res Int. 2018;2018:2148056. Epub 2018/12/07. doi: 10.1155/2018/2148056 30519491; PubMed Central PMCID: PMC6241241.

28. Kondkar AA, Azad TA, Sultan T, Al-Mobarak FA, Kalantan H, Al-Obeidan SA. Polymorphisms rs693421 and rs2499601 at locus 1q43 and their haplotypes are not associated with primary open-angle glaucoma: a case-control study. BMC Res Notes. 2019;12(1):453. Epub 2019/07/25. doi: 10.1186/s13104-019-4491-x [pii]. 31337432; PubMed Central PMCID: PMC6651941.

29. Ahmad SS. Controversies in the vascular theory of glaucomatous optic nerve degeneration. Taiwan J Ophthalmol. 2016;6(4):182–6. Epub 2017/10/12. doi: 10.1016/j.tjo.2016.05.009 TJO-6-182 [pii]. 29018738; PubMed Central PMCID: PMC5525630.

30. Yoshimura M, Yasue H, Nakayama M, Shimasaki Y, Sumida H, Sugiyama S, et al. A missense Glu298Asp variant in the endothelial nitric oxide synthase gene is associated with coronary spasm in the Japanese. Hum Genet. 1998;103(1):65–9. Epub 1998/09/16. doi: 10.1007/s004390050785 9737779.

31. Tunny TJ, Richardson KA, Clark CV. Association study of the 5' flanking regions of endothelial-nitric oxide synthase and endothelin-1 genes in familial primary open-angle glaucoma. Clin Exp Pharmacol Physiol. 1998;25(1):26–9. Epub 1998/03/11. doi: 10.1111/j.1440-1681.1998.tb02138.x 9493554.

32. Fan BJ, Liu K, Wang DY, Tham CC, Tam PO, Lam DS, et al. Association of polymorphisms of tumor necrosis factor and tumor protein p53 with primary open-angle glaucoma. Invest Ophthalmol Vis Sci. 2010;51(8):4110–6. Epub 2010/04/02. doi: 10.1167/iovs.09-4974 iovs.09-4974 [pii]. 20357201.

33. Kang JH, Wiggs JL, Rosner BA, Hankinson SE, Abdrabou W, Fan BJ, et al. Endothelial nitric oxide synthase gene variants and primary open-angle glaucoma: interactions with sex and postmenopausal hormone use. Invest Ophthalmol Vis Sci. 2010;51(2):971–9. Epub 2009/10/10. doi: 10.1167/iovs.09-4266 iovs.09-4266 [pii]. 19815736; PubMed Central PMCID: PMC3094851.

34. Kang JH, Wiggs JL, Rosner BA, Haines J, Abdrabou W, Pasquale LR. Endothelial nitric oxide synthase gene variants and primary open-angle glaucoma: interactions with hypertension, alcohol intake, and cigarette smoking. Arch Ophthalmol. 2011;129(6):773–80. Epub 2011/06/15. doi: 10.1001/archophthalmol.2011.118 129/6/773 [pii]. 21670344; PubMed Central PMCID: PMC3337676.

35. Logan JF, Chakravarthy U, Hughes AE, Patterson CC, Jackson JA, Rankin SJ. Evidence for association of endothelial nitric oxide synthase gene in subjects with glaucoma and a history of migraine. Invest Ophthalmol Vis Sci. 2005;46(9):3221–6. Epub 2005/08/27. 46/9/3221 [pii] doi: 10.1167/iovs.05-0368 16123422.

36. Kosior-Jarecka E, Lukasik U, Wrobel-Dudzinska D, Kocki J, Bartosinska J, Witczak A, et al. Risk Factors for Normal and High-Tension Glaucoma in Poland in Connection with Polymorphisms of the Endothelial Nitric Oxide Synthase Gene. PLoS One. 2016;11(1):e0147540. Epub 2016/01/26. doi: 10.1371/journal.pone.0147540 PONE-D-15-33145 [pii]. 26807726; PubMed Central PMCID: PMC4726562.

37. Jeoung JW, Kim DM, Oh S, Lee JS, Park SS, Kim JY. The Relation Between Endothelial Nitric Oxide Synthase Polymorphisms and Normal Tension Glaucoma. J Glaucoma. 2017;26(11):1030–5. Epub 2017/08/05. doi: 10.1097/IJG.0000000000000751 28777225.

38. Weiss J, Frankl SA, Flammer J, Grieshaber MC, Hollo G, Teuchner B, et al. No difference in genotype frequencies of polymorphisms of the nitric oxide pathway between Caucasian normal and high tension glaucoma patients. Mol Vis. 2012;18:2174–81. Epub 2012/08/25. 22919264; PubMed Central PMCID: PMC3425578.

39. Sakai T, Shikishima K, Matsushima M, Kitahara K. Endothelial nitric oxide synthase gene polymorphisms in non-arteritic anterior ischemic optic neuropathy. Graefes Arch Clin Exp Ophthalmol. 2007;245(2):288–92. Epub 2006/04/25. doi: 10.1007/s00417-005-0245-7 16633797.

40. Lay CL, Broner SW. Migraine in women. Neurol Clin. 2009;27(2):503–11. Epub 2009/03/18. doi: 10.1016/j.ncl.2009.01.002 S0733-8619(09)00003-6 [pii]. 19289228.

41. Liao Q, Wang DH, Sun HJ. Association of genetic polymorphisms of eNOS with glaucoma. Mol Vis. 2011;17:153–8. Epub 2011/01/20. 19 [pii]. 21245953; PubMed Central PMCID: PMC3021569.

42. Colombo MG, Paradossi U, Andreassi MG, Botto N, Manfredi S, Masetti S, et al. Endothelial nitric oxide synthase gene polymorphisms and risk of coronary artery disease. Clin Chem. 2003;49(3):389–95. Epub 2003/02/26. doi: 10.1373/49.3.389 12600950.

43. Matsa LS, Rangaraju A, Vengaldas V, Latifi M, Jahromi HM, Ananthapur V, et al. Haplotypes of NOS3 gene polymorphisms in dilated cardiomyopathy. PLoS One. 2013;8(7):e70523. Epub 2013/08/08. doi: 10.1371/journal.pone.0070523 PONE-D-12-31516 [pii]. 23923002; PubMed Central PMCID: PMC3726655.

44. Sandrim VC, Coelho EB, Nobre F, Arado GM, Lanchote VL, Tanus-Santos JE. Susceptible and protective eNOS haplotypes in hypertensive black and white subjects. Atherosclerosis. 2006;186(2):428–32. Epub 2005/09/20. S0021-9150(05)00521-6 [pii] doi: 10.1016/j.atherosclerosis.2005.08.003 16168996.

45. Kondkar AA, Azad TA, Almobarak FA, Kalantan H, Sultan T, Alsabaani NA, et al. Polymorphism rs10483727 in the SIX1/SIX6 Gene Locus Is a Risk Factor for Primary Open Angle Glaucoma in a Saudi Cohort. Genet Test Mol Biomarkers. 2018;22(1):74–8. Epub 2017/12/01. doi: 10.1089/gtmb.2017.0159 29190129.

46. Abu-Amero KK, Kondkar AA, Mousa A, Almobarak FA, Alawad A, Altuwaijri S, et al. Analysis of Cyclin-Dependent Kinase Inhibitor-2B rs1063192 Polymorphism in Saudi Patients with Primary Open-Angle Glaucoma. Genet Test Mol Biomarkers. 2016;20(10):637–41. Epub 2016/10/18. doi: 10.1089/gtmb.2016.0140 27541204.

47. Abu-Amero KK, Kondkar AA, Mousa A, Osman EA, Al-Obeidan SA. Lack of association of SNP rs4236601 near CAV1 and CAV2 with POAG in a Saudi cohort. Mol Vis. 2012;18:1960–5. Epub 2012/08/10. 22876122; PubMed Central PMCID: PMC3413415.

48. Kondkar AA, Azad TA, Almobarak FA, Kalantan H, Sultan T, Al-Obeidan SA, et al. Polymorphism rs11656696 in GAS7 Is Not Associated with Primary Open Angle Glaucoma in a Saudi Cohort. Genet Test Mol Biomarkers. 2017;21(12):754–8. Epub 2017/10/13. doi: 10.1089/gtmb.2017.0147 29022762.

49. Kondkar AA, Mousa A, Azad TA, Sultan T, Alawad A, Altuwaijri S, et al. Polymorphism rs7555523 in transmembrane and coiled-coil domain 1 (TMCO1) is not a risk factor for primary open angle glaucoma in a Saudi cohort. J Negat Results Biomed. 2016;15(1):17. Epub 2016/10/01. doi: 10.1186/s12952-016-0060-1 [pii]. 27687253; PubMed Central PMCID: PMC5043619.

50. Polak K, Luksch A, Berisha F, Fuchsjaeger-Mayrl G, Dallinger S, Schmetterer L. Altered nitric oxide system in patients with open-angle glaucoma. Arch Ophthalmol. 2007;125(4):494–8. Epub 2007/04/11. 125/4/494 [pii] doi: 10.1001/archopht.125.4.494 17420369.

51. Wang XL, Sim AS, Wang MX, Murrell GA, Trudinger B, Wang J. Genotype dependent and cigarette specific effects on endothelial nitric oxide synthase gene expression and enzyme activity. FEBS Lett. 2000;471(1):45–50. Epub 2000/04/13. S0014-5793(00)01356-9 [pii] doi: 10.1016/s0014-5793(00)01356-9 10760510.

52. Naber Ch K, Frey UH, Oldenburg O, Brauck K, Eggebrecht H, Schmermund A, et al. Relevance of the NOS3 T-786C and G894T variants for cholinergic and adrenergic coronary vasomotor responses in man. Basic Res Cardiol. 2005;100(5):453–60. Epub 2005/07/21. doi: 10.1007/s00395-005-0530-y 16032374.

53. Fischmann TO, Hruza A, Niu XD, Fossetta JD, Lunn CA, Dolphin E, et al. Structural characterization of nitric oxide synthase isoforms reveals striking active-site conservation. Nat Struct Biol. 1999;6(3):233–42. Epub 1999/03/13. doi: 10.1038/6675 10074942.

54. Joshi MS, Bauer JA. Preliminary computational modeling of nitric oxide synthase 3 interactions with caveolin-1: influence of exon 7 Glu298Asp polymorphism. Acta Biochim Biophys Sin (Shanghai). 2008;40(1):47–54. Epub 2008/01/09. doi: 10.1111/j.1745-7270.2008.00369.x 18180853.

55. Godfrey V, Chan SL, Cassidy A, Butler R, Choy A, Fardon T, et al. The functional consequence of the Glu298Asp polymorphism of the endothelial nitric oxide synthase gene in young healthy volunteers. Cardiovasc Drug Rev. 2007;25(3):280–8. Epub 2007/10/09. CDR017 [pii] doi: 10.1111/j.1527-3466.2007.00017.x 17919260.

56. Abu-Amero KK, Kondkar AA, Mousa A, Osman EA, Al-Obeidan SA. Decreased total antioxidants in patients with primary open angle glaucoma. Curr Eye Res. 2013;38(9):959–64. Epub 2013/05/09. doi: 10.3109/02713683.2013.794246 23651069.

57. Fairchild TA, Fulton D, Fontana JT, Gratton JP, McCabe TJ, Sessa WC. Acidic hydrolysis as a mechanism for the cleavage of the Glu(298)—>Asp variant of human endothelial nitric-oxide synthase. J Biol Chem. 2001;276(28):26674–9. Epub 2001/05/02. doi: 10.1074/jbc.M103647200 M103647200 [pii]. 11331296.

58. Hingorani AD. Polymorphisms in endothelial nitric oxide synthase and atherogenesis: John French Lecture 2000. Atherosclerosis. 2001;154(3):521–7. Epub 2001/03/21. S0021-9150(00)00699-7 [pii]. doi: 10.1016/s0021-9150(00)00699-7 11257252.

59. Golser R, Gorren AC, Mayer B, Schmidt K. Functional characterization of Glu298Asp mutant human endothelial nitric oxide synthase purified from a yeast expression system. Nitric Oxide. 2003;8(1):7–14. Epub 2003/02/15. S1089860302001313 [pii]. doi: 10.1016/s1089-8603(02)00131-3 12586536.

60. Wang J, Dudley D, Wang XL. Haplotype-specific effects on endothelial NO synthase promoter efficiency: modifiable by cigarette smoking. Arterioscler Thromb Vasc Biol. 2002;22(5):e1–4. Epub 2002/05/15. doi: 10.1161/01.ATV.0000016248.51577.1F 12006409; PubMed Central PMCID: PMC1350055.

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