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系統識別號 U0026-1708201118392300
論文名稱(中文) 腸病毒71型結構蛋白突變對病毒感染之影響
論文名稱(英文) The effect of capsid protein mutations on enterovirus 71 infection
校院名稱 成功大學
系所名稱(中) 醫學檢驗生物技術學系碩博士班
系所名稱(英) Department of Medical Laboratory Science and Biotechnology
學年度 99
學期 2
出版年 100
研究生(中文) 劉欣芸
研究生(英文) Shin-Yun Liu
學號 t36984035
學位類別 碩士
語文別 中文
論文頁數 81頁
口試委員 指導教授-王貞仁
口試委員-楊孔嘉
口試委員-劉校生
口試委員-林貴香
中文關鍵字 腸病毒71型  結構蛋白  突變  感染 
英文關鍵字 enterovirus 71  capsid protein  mutation  infection 
學科別分類
中文摘要 腸病毒71型在分類上屬於小RNA病毒科中的腸病毒屬,其外殼不具套膜,由大小約30 奈米的20面體外殼內含一條正股RNA所形成。腸病毒71型主要感染5歲以內的嬰幼兒,引起的症狀包括較輕微的手足口症、咽峽炎,重症則會侵犯中樞神經造成無菌性腦膜炎、腦幹腦炎、急性無力四肢麻痺、小兒麻痺肢體症候群甚至造成死亡。然而目前對於腸病毒71型是如何感染神經系統造成病變其原因並不清楚。2004年,王等人利用人類病毒株4643在老鼠體內培養數代後得到老鼠適應株MP4,研究指出MP4相較於4643可更容易的感染老鼠並引起神經性病變。經由本實驗室黃等人利用重組病毒及序列分析比對後,發現當4643病毒外殼蛋白VP1上145位點由Q變為E及VP2上149位點由K變為M時,此2個突變點可提升4643病毒感染老鼠神經細胞瘤 ( Neuro-2a ) 之能力或是增加小鼠死亡率。進一步研究發現當4643帶有VP1145E突變可增加病毒黏附到Neuro-2a之能力,然而VP2149M則沒有影響。因此本篇研究的目的即為探討VP1145E及VP2149M是藉由哪些機制影響了4643病毒的感染力。在腸病毒71型生活史中,病毒殼蛋白除了參與黏附宿主細胞表面受器外,也影響了病毒脫殼作用、RNA複製及包被現象。我們利用中性紅製作光敏感病毒並利用中性紅試驗結合感染率溶斑試驗來探討殼蛋白突變對於脫殼作用之影響,實驗結果發現當4643病毒帶有VP2149M突變可提升病毒脫殼效率增加RNA的釋出。另外,我們也使用Neuro-2a細胞建立了胞外轉譯/複製系統來研究殼蛋白突變對於病毒RNA複製的影響,從研究結果得知當4643帶有VP1145E及VP2149M突變時,不管是單點突變或是雙點突變都有提升RNA複製的能力。本研究顯示VP1145E及VP2149M突變可藉由影響腸病毒71型生活史中的不同階段來提升病毒感染力:VP2149M殼蛋白突變可提升病毒脫殼及RNA複製,VP1145E則影響了RNA複製,而當VP1145E及VP2149M突變同時存在時對於病毒RNA複製也有加成的效果。
英文摘要 Enterovirus 71 (EV71) is a non-enveloped positive single-stranded RNA virus which belongs to genus enterovirus in the family Picornaviridae. EV71 causes several diseases, such as hand, foot and mouth disease, herpangina, encephalitis, meningitis, and acute flaccid paralysis. However, it remains unknown how EV71 causes neurological diseases. In 2004, Wang et al. established a mouse adapted EV71 strain (MP4) from the parental EV71 strain (4643) in mice, and they found that the MP4 strain was more cytotoxic in vitro than 4643 strain. By constructing recombinant viruses of MP4 and 4643, Huang et al. found that the capsid protein region determined the infectivity in mouse neuronal Neuro-2a cells. After sequence analysis and constructing the recombinant mutant viruses, they discovered that the VP2149M and VP1145E mutations could enhance viral infection in Neuro-2a cells. Furthermore, by using the binding assay, they also discovered that only the VP1145E could increase the attachment of EV71 on Neuro-2a cells. Therefore, the mechanisms of these two mutations which affect EV71 infection deserve to be explored. It has been reported that the capsid protein may involve in several steps of virus infection, including virus binding, uncoating, replication and encapsidation. First, we performed neutral red assay and infectious center assay (NRIC assay) for viral uncoating by using NR-containing viruses. The results showed that the mutation of VP2149M could affect the viral uncoating and enhance RNA release. In order to determine whether capsid protein mutations might affect viral RNA replication, an in vitro translation/replication system was established by using mouse Neuro-2a cell extracts. The in vitro translation/replication system which can translate protein in tube was performed by adding manually prepared cell extract, initiation factor, buffer, RNA and enzymes. By using EV71 RNA with luciferase reporter gene, we found the VP2149M and/or VP1145E mutation might enhance EV71 RNA replication in in vitro translation/ replication reaction. In conclusion, the VP1145E and VP2 149M mutations could enhance EV71 infection in various steps; VP2149M enhanced viral uncoating, and VP2149M and/or VP1145E mutation increased EV71 RNA replication.
論文目次 中文摘要 I
Abstract III
致謝 V
目錄 VI
表目錄 VIII
圖目錄 IX
第一章、緒論 1
一、 腸病毒的分型及構造 1
二、 腸病毒71型的臨床感染症狀及流行病學 5
三、 腸病毒71型的病毒蛋白和宿主蛋白之關連 7
四、 腸病毒71型感染宿主細胞時引起之複製複合物 9
五、 腸病毒71型的殼蛋白研究 10
六、 病毒RNA胞外轉譯/複製系統 12
七、 中性紅於病毒學的應用 14
八、 感染性克隆及複製子系統 16
九、 研究動機及目標 17
第二章、材料與方法 21
一、細胞與病毒 21
二、 質體 ( plasmid ) 之操作 25
三、RNA之製備及RNA轉染 26
四、中性紅試驗結合感染率溶斑試驗 ( Neutral red and infectious center assay,NRIC assay ) 29
五、 胞外RNA轉譯/複製系統之建立 31
第三章、結果 35
一、 腸病毒71型殼蛋白VP2突變增加病毒感染能力 35
二、 腸病毒71型殼蛋白VP2突變增加病毒脫殼能力 37
三、 腸病毒71型殼蛋白VP2及VP1突變增加病毒RNA複製能力 40
第四章、討論 45
結語 51
參考文獻 53
表圖 60
附錄 72
自述 81

參考文獻 1. Andino, R., G. E.Rieckhof, P. L.Achacoso, and D. Baltimore. 1993. Poliovirus RNA synthesis utilizes an RNP complex formed around the 5' -end of viral RNA. The EMBO Journal 12:3587-3598.
2. Arita, M., Y. Ami, T. Wakita, and H. Shimizu. 2007. Cooperative Effect of the Attenuation Determinants Derived from Poliovirus Sabin 1 Strain Is Essential for Attenuation of Enterovirus 71 in the NOD/SCID Mouse Infection Model. Journal of Virology 82:1787-1797.
3. Barton, D. J., E. P. Black, and J. B. Flanegan. 1995. Complete replication of poliovirus in vitro: preinitiation RNA replication complexes require soluble cellular factors for the synthesis of VPg-linked RNA. Journal of Virology 69:5516-27.
4. Barton, D. J., and J. B. Flannegan. 1993. Coupled translation and replication of poliovirus RNA in vitro synthesis of functional 3D polymerase and infectious virus. Journal of Virology 67:822-831.
5. Bible, J. M., P. Pantelidis, P. K. S. Chan, and C. Y. W. Tong. 2007. Genetic evolution of enterovirus 71: epidemiological and pathological implications. Reviews in Medical Virology 17:371-379.
6. Bienz, K., D. Egger, Y. Rasser, and W. Bossart. 1980. Kinetics and location of poliovirus macromolecular synthesis in correlation to virus-induced cytopathology. Virology 100:390-399.
7. Bienz, K., D. Egger, M. Troxler, and L. Psamontes. 1990. Structrual organization of poliovirus RNA replication is mediated by viral proteins of the P2 genomic region. Journal of Virology 64:1156-1163.
8. Brandenburg, B., L. Y. Lee, M. Lakadamyali, M. J. Rust, X. Zhuang, and J. M. Hogle. 2007. Imaging poliovirus entry in live cells. PLoS Biology 5:e183.
9. Brown, B. A., M. S. Oberste, J. P. Alexander, JR., M. L. Kennett, and M. A. Pallansch. 1999. Molecular epidemiology and evolution of enterovirus 71 strains isolated from 1970 to 1998. Journal of Virology 73:9969-9975.
10. Campanella, M. 2004. The coxsackievirus 2B protein suppresses apoptotic host cell responses by manipulating intracellular Ca2+ homeostasis. Journal of Biological Chemistry 279:18440-18450.
11. Choe, S., D. Dodd, and K. Kirkegaard. 2005. Inhibition of cellular protein secretion by picornaviral 3A proteins. Virology 337:18-29.
12. Chua, B. H., P. Phuektes, S. A. Sanders, P. K. Nicholls, and P. C. McMinn. 2008. The molecular basis of mouse adaptation by human enterovirus 71. Journal of General Virology 89:1622-1632.
13. Colston, E., and V. R. Racaniello. 1994. Soluble receptor-resistant poliovirus mutants identify surface and internal capsid residues that control interaction with the cell-receptor. Embo Journal 13:5855-5862.
14. Couderc, T., F. Delpeyroux, H. LeBlay, and B. Blondel. 1996. Mouse adaptation determinants of poliovirus type 1 enhance viral uncoating. Journal of Virology 70:305-312.
15. Couderc, T., J. Hogle, H. Leblay, F. Horaud, and B. Blondel. 1993. Molecular characterization of mouse-virulent poliovirus type-1 mahoney mutants - involvement of residues of polypeptides VP1 and VP2 located on the inner surface of the capsid protein shell. Journal of Virology 67:3808-3817.
16. Crowther, D., and J. L. Melnick. 1961. The incorporation of neutral red and acridine orange into developing poliovirus particles making them photosensitive. Virology 14:11-21.
17. Dodd, D. A., T. H. Giddings, and K. Kirkegaard. 2001. Poliovirus 3A protein limits interleukin-6 (IL-6), IL-8, and beta interferon secretion during viral infection. Journal of Virology 75:8158-8165.
18. Dorner, A. J., B. L. Semler, R. J. Jackson, R. Hanecak, E. Duprey, and E. Wimmer. 1984. In vitro translation of poliovirus RNA: utilization of internal initiation sites in reticulocyte lysate. Journal of Virology 50:507-514.
19. Egger, D., L. Pasamontes, R. Bolten, V. Boyko, and K. Bienz. 1996. Reversible dissociation of the poliovirus replication complex functions and interactions of its components in viral RNA synthesis. Journal of Virology 70:8675-8683.
20. Elghonemy, S., W. G. Davis, and M. A. Brinton. 2005. The majority of the nucleotides in the top loop of the genomic 3 ' terminal stem loop structure are cis-acting in a West Nile virus infectious clone. Virology 331:238-246.
21. Franco, D., H. B. Pathak, C. E. Cameron, B. Rombaut, E. Wimmer, and A. V. Paul. 2005. Stimulation of poliovirus RNA synthesis and virus maturation in a HeLa cell-free in vitro translation-RNA replication system by viral protein 3CDpro. Virology Journal 2:86.
22. Franco, D., H. B. Pathak, C. E. Cameron, B. Rombaut, E. Wimmer, and A. V. Paul. 2005. Stimulation of poliovirus synthesis in a HeLa cell-free in vitro translation-RNA replication system by viral protein 3CDpro. Journal of Virology 79:6358-6367.
23. Giachetti, C., and B. L. Semler. 1991. Role of a viral membrane polypeptide in strand-specific initiation of poliovirus RNA synthesis. Journal of Virology 65:2647-2654.
24. Harris, K. S., S. R. Reddigari, M. J. H. Nicklin, T. Hammerle, and E. Wimmer. 1992. Purification and characterization of poliovirus polypeptide 3CD, a proteinase and a precursor for RNA polymerase. Journal of Virology 66:7481-7489.
25. Hogle, J. M. 2002. Poliovirus cell entry: common structural themes in viral cell entry pathways. Annual Review of Microbiology 56:677-702.
26. Hong, Y. L., K. Levay, J. F. Murphy, P. G. Klein, J. G. Shaw, and A. G. Hunt. 1995. A potyvirus polymerase interacts with the viral coat protein and Vpg in yeast-cells. Virology 214:159-166.
27. Huanga, Y.-P., T.-L. Lina, C.-Y. Kuoa, M.-W. Lina, C.-Y. Yaoa, H.-W. Liaoa, L.-C. Hsua, C.-F. Yanga, J.-Y. Yanga, P.-J. Chenb, and Ho-ShengWua. 2008. The circulation of subgenogroups B5 and C5 of enterovirus 71 in Taiwan from 2006 to 2007. Virus Research 137:206-212.
28. Hussain, K. M., K. L. J. Leong, M. M. L. Ng, and J. J. H. Chu. 2010. The essential role of clathrin-mediated endocytosis in the infectious entry of human enterovirus 71. Journal of Biological Chemistry 286:309-321.
29. Ikeda, M., M. K. Yi, K. Li, and S. A. Lemon. 2002. Selectable subgenomic and genome-length dicistronic RNAs derived from an infectious molecular clone of the HCV-N strain of hepatitis C virus replicate efficiently in cultured Huh7 cells. Journal of Virology 76:2997-3006.
30. Ishimaru, Y., S. Nakano, K. Yamaoka, and S. Takami. 1980. Outbreak of hand, foot, and mouth disease by enterovirus 71. Archives of Disease in Childhood 55:583-588.
31. Kirkegaard, K. 1990. Mutations in VP1 of poliovirus specifically affect both encapsidation and release of viral-RNA. Journal of Virology 64:195-206.
32. Kung, Y.-H., S.-W. Huang, P.-H. Kuo, D. Kiang, M.-S. Ho, C.-C. Liu, C.-K. Yu, I.-J. Su, and J.-R. Wang. 2010. Introduction of a strong temperature-sensitive phenotype into enterovirus 71 by altering an amino acid of virus 3D polymerase. Virology 396:1-9.
33. Kuo, R.-L., S.-H. Kung, Y.-Y. Hsu, and W.-T. Liu. 2002. Infection with enterovirus 71 or expression of its 2A protease induces apoptotic cell death. Journal of General Virology 83:1367-1376.
34. Lama, J., M. A. Sanz, and P. L. Rodríguez. 1995. A role for 3AB protein in poliovirus genome replication. The Journal of Biological Chemistry 270:14430-14438.
35. Li, M. 2002. The 3C protease activity of enterovirus 71 induces human neural cell apoptosis. Virology 293:386-395.
36. Lin, J. Y., M. L. Li, P. N. Huang, K. Y. Chien, J. T. Horng, and S. R. Shih. 2008. Heterogeneous nuclear ribonuclear protein K interacts with the enterovirus 71 5' untranslated region and participates in virus replication. Journal of General Virology 89:2540-2549.
37. Lin, J. Y., M. L. Li, and S. R. Shih. 2008. Far upstream element binding protein 2 interacts with enterovirus 71 internal ribosomal entry site and negatively regulates viral translation. Nucleic Acids Research 37:47-59.
38. Lin, K.-H., K.-P. Hwang, G.-M. Ke, C.-F. Wang, L.-Y. Ke, Y.-T. Hsu, Y.-C. Tung, P.-Y. Chu, B.-H. Chen, H.-L. Chen, C.-L. Kao, J.-R. Wang, H.-L. Eng, S.-Y. Wang, L.-C. Hsu, and H.-Y. Chen. 2006. Evolution of EV71 genogroup in Taiwan from 1998 to 2005: An emerging of subgenogroup C4 of EV71. Journal of Medical Virology 78:254-262.
39. Lin, T.-Y., S.-H. Hsia, Y.-C. Huang, C.-T. Wu, and L.-Y. Chang. 2003. Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system. Clinical Infectious Diseases 36:269-274.
40. Lin, T.-Y., S.-J. Twu, M.-S. Ho, L.-Y. Chang, and C.-Y. Lee. 2003. Enterovirus 71 outbreaks, Taiwan: Occurrence and recognition. Emerging Infectious Diseases 9:291-293.
41. Liu, Y., C. L. Wang, S. Mueller, A. V. Paul, E. Wimmer, and P. Jiang. 2010. Direct interaction between two viral proteins, the nonstructural protein 2C(ATPase) and the capsid protein VP3, is required for enterovirus morphogenesis. PLoS Pathogens 6:1-14.
42. Lodish, H. F. 1968. Bacteriophage f2 RNA: control of translation and gene order. Nature 220:345-350.
43. M, H., C. ER, H. KH, T. SJ, C. KT, T. SF, W. JR, and S. SR. 1999. An epidemic of enterovirus 71 infection in Taiwan. The New England Journal of Medicine 341:929-935.
44. Madshus, I. H., S. Olsnes, and K. Sandvig. 1984. Mechanism of entry into the cytosol of poliovirus type-1 - requirement for low pH. Journal of Cell Biology 98:1194-1200.
45. Mandel, B. 1967. The relationshop between penatration and uncoating of poliovirus in HeLa cells. Virology 31:702-712.
46. Maraver, A., R. Clemente, J. F. Rodriguez, and E. Lombardo. 2003. Identification and molecular characterization of the RNA polymerase-binding motif of infectious bursal disease virus inner capsid protein VP3. Journal of Virology 77:2459-2468.
47. McKnight, K. L., and S. M. Lemon. 1996. Capsid coding sequence is required for efficient replication of human rhinovirus 14 RNA. Journal of Virology 70:1941-1952.
48. McKnight, K. L., and S. M. Lemon. 1998. The rhinovirus type 14 genome contains an internally located RNA structure that is required for viral replication. RNA-a Publication of the RNA Society 4:1569-1584.
49. Miyamura, K., Y. Nishimura, M. Abo, T. Wakita, and H. Shimizu. 2010. Adaptive mutations in the genomes of enterovirus 71 strains following infection of mouse cells expressing human P-selectin glycoprotein ligand-1. Journal of General Virology 92:287-291.
50. Molla, A., A. V. Paul, and E. Wimmer. 1991. Cell-free, de novo synthesis of poliovirus. Science 254:1647-1651.
51. Molla, A., A. V. Paul, and E. Wimmer. 1993. Effects of temperature and lipophilic agents on poliovirus formation and RNA synthesis in a cell-free system. Journal of Virology 67:5932-5938.
52. Nayak, A., I. G. Goodfellow, and G. J. Belsham. 2005. Factors required for the uridylylation of the foot-and-mouth disease virus 3B1, 3B2, and 3B3 peptides by the RNA-dependent RNA polymerase (3Dpol) in vitro. Journal of Virology 79:7698-7706.
53. Nishimura, Y., M. Shimojima, Y. Tano, T. Miyamura, T. Wakita, and H. Shimizu. 2009. Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71. Nature Medicine 15:794-797.
54. Oberg, B. F., and A. J. Shatkin. 1972. Initiation of picornavirus protein-synthesis in ascites cell extracts. Proceedings of the National Academy of Sciences of the United States of America 69:3589-3593.
55. Omata, T., M. Kohara, Y. Sakai, A. Kameda, N. Imura, and A. Nomoto. 1984. Cloned infectious complementary-DNA of the poliovirus sabin-1 genome - biochemical and biological properties of the recovered Virus. Gene 32:1-10.
56. Pfister, T., and E. Wimmer. 1999. Characterization of the nucleoside triphosphatase activity of poliovirus protein 2C reveals a mechanism by which guanidine inhibits poliovirus replication. Journal of Biological Chemistry 274:6992-7001.
57. Pincus, S. E., D. C. Diamond, E. A. Emini, and E. Wimmer. 1986. Guanidine-selected mutants of poliovirus: mapping of point mutations to polypeptide 2C. Journal of Virology 57:638-46.
58. Pincus, S. E., and E. Wimmer. 1986. Production of guanidine-resistant and -dependent poliovirus mutants from cloned cDNA: mutations in polypeptide 2C are directly responsible for altered guanidine sensitivity. Journal of Virology 60:739-796.
59. Pringle, C. R. 1964. Inhibition of multiplication of foot-and-mouth disease virus by guanidine hydrochloride. Nature 204:1012-3.
60. Rekosh, D. 1972. Gene order of the poliovirus capsid proteins. Journal of Virology 9:479-487.
61. Repetto, G., A. del Peso, and J. L. Zurita. 2008. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nature Protocols 3:1125-1131.
62. Rust, R. C., L. Landmann, R. Gosert, B. L. Tang, W. Hong, H. P. Hauri, D. Egger, and K. Bienz. 2001. Cellular COPII proteins are involved in production of the vesicles that form the poliovirus replication complex. Journal of Virology 75:9808-9818.
63. Taber, R., D. Rekosh, and D. Baltimore. 1971. Effect of pactamycin on synthesis of poliovirus proteins: a method for genetic mapping. Journal of Virology 8:395-401.
64. Tacken, M. G. J., B. P. H. Peeters, A. A. M. Thomas, P. J. M. Rottier, and H. J. Boot. 2002. Infectious bursal disease virus capsid protein VP3 interacts both with VP1, the RNA-dependent RNA polymerase, and with viral double-stranded RNA. Journal of Virology 76:11301-11311.
65. Tacken, M. G. J., P. J. M. Rottier, A. L. J. Gielkens, and B. P. H. Peeters. 2000. Interactions in vivo between the proteins of infectious bursal disease virus: capsid protein VP3 interacts with the RNA-dependent RNA polymerase, VP1. Journal of General Virology 81:209-218.
66. Tang, W. F., S. Y. Yang, B. W. Wu, J. R. Jheng, Y. L. Chen, C. H. Shih, K. H. Lin, H. C. Lai, P. Tang, and J. T. Horng. 2006. Reticulon 3 binds the 2C protein of enterovirus 71 and is required for viral replication. Journal of Biological Chemistry 282:5888-5898.
67. Thomas, J. V., W. A. Dunlap, and A. M. Rich. 1973. Photodynamic inactivation in treatment of experimental herpes-simplex keratitis. British Journal of Ophthalmology 57:336-338.
68. Vance, L. M., N. Moscufo, M. Chow, and B. A. Heinz. 1997. Poliovirus 2C region functions during encapsidation of viral RNA. Journal of Virology 71:8759-8765.
69. Varma, J. P. 1968. Inhibition of tobacco necrosis virus by guanidine carbonate. Virology 36:305-8.
70. Wang, S.-M., H.-Y. Lei, K.-J. Huang, J.-M. Wu, J.-R. Wang, C.-K. Yu, I.-J. Su, and C.-C. Liu. 2003. Pathogenesis of enteroviurs 71 brainstem encephalitis in pediatric patients: Roles of cytokines and cellular immune activation in patients with pulmonary edema. Journal of infectious diseases 188:564-570.
71. Wang, Y. F., C. T. Chou, H. Y. Lei, C. C. Liu, S. M. Wang, J. J. Yan, I. J. Su, J. R. Wang, T. M. Yeh, S. H. Chen, and C. K. Yu. 2004. A mouse-adapted enterovirus 71 strain causes neurological disease in mice after oral infection. Journal of Virology 78:7916-7924.
72. Wilson, J. N., and P. D. Cooper. 1963. Aspects of the growth of poliovirus as revealed by the photodynamic effects of neutral red and acridine orange. Virology 21:135-145.
73. Xiang, W., K. S. Harris, L. Alexander, and E. Wimmer. 1995. Interaction between the 5'-terminal cloverleaf and 3AB/3CDpro of poliovirus is essential for RNA replication. Journal of Virology 69:3658-3667.
74. Yamayoshi, S., Y. Yamashita, J. Li, N. Hanagata, T. Minowa, T. Takemura, and S. Koike. 2009. Scavenger receptor B2 is a cellular receptor for enterovirus 71. Nature Medicine 15:798-801.
75. Zamora, M., W. E. Marissen, and R. E. Lloyd. 2002. Multiple eIF4GI-specific protease activities present in uninfected and poliovirus-infected cells. Journal of Virology 76:165-177.
76. Zhao, Z. J., T. Date, Y. H. Li, T. Kato, M. Miyamoto, K. Yasui, and T. Wakita. 2005. Characterization of the E-138 (Glu/Lys) mutation in Japanese encephalitis virus by using a stable, full-length, infectious cDNA clone. Journal of General Virology 86:2209-2220.
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