研究業績

これまでの研究業績

原索動物ホヤ類におけるアロ認識機構の解明

＜研究代表者＞澤田 均（名古屋大学 大学院理学研究科・教授）

Yamada, L., Saito, T., Taniguchi, H., Sawada, H., and Harada, Y. (2009). Comprehensive egg coat proteome of the ascidian Ciona intestinalis reveals gamete recognition molecules involved in self-sterility. J. Biol. Chem. 284 (14), 9402-9410.

*Harada, Y., Takagaki, Y., Sunagawa, M., Saito, T., Yamada, L., Taniguchi, H., Shoguchi, E., and *Sawada, H. (2008). Mechanism of self-sterility in a hermaphroditic chordate. Science 320 (5875), 548-550.

Harada, Y., and * Sawada, H. (2008). Allorecognition mechanisms during ascidian fertilization . Int. J. Dev. Biol. 52 (5-6), 637-645.

Urayama, S., Harada, Y., Nakagawa, Y., Ban, S., Akasaka, M., Kawasaki, N., and *Sawada, H. (2008). Ascidian sperm GPI-anchored CRISP-like protein as a binding partner for an allorecognizable sperm receptor on the vitelline coat. J. Biol. Chem. 283 (31), 21725-21733.

*Harada, Y., and Sawada, H. (2007). Proteins interacting with the ascidian vitelline-coat sperm receptor HrVC70 as revealed by yeast two-hybrid screening. Mol. Reprod. Dev. 74 (9), 1478-1487.

Ban, S., Harada, Y., Yokosawa, H., and *Sawada, H. (2005). Highly polymorphic vitelline-coat protein HaVC80 from the ascidian, Halocynthia aurantium: Structural analysis and involvement in self/nonself recognition during fertilization. Dev. Biol. 286 (2), 440-451.

*Sawada, H., Tanaka, E., Ban, S., Yamasaki, C., Fujino, J., Ooura, K., Matsumoto, K., and Yokosawa, H. (2004). Self/nonself recognition in ascidian fertilization: vitelline coat protein HrVC70 is a candidate allorecognition molecule. Proc. Natl. Acad. Sci. USA 101 (44), 15615-15620.

Sakai, N., *Sawada, H., and Yokosawa, H. (2003). Extracellular ubiquitin system implicated in fertilization of the ascidian, Halocynthia roretzi: Isolation and characterization. Dev. Biol. 264 (1), 299-307.

*Sawada, H., Sakai, N., Abe, Y., Tanaka, E., Takahashi, Y., Fujino, J., Takizawa, S., and Yokosawa, H. (2002). Extracellular ubiquitination and proteasome-mediated degradation of the ascidian sperm receptor. Proc. Natl. Acad. Sci. USA 99 (3), 1223-1228.

Kodama, E., Baba, T., Yokosawa, H., and *Sawada, H. (2001). cDNA cloning and functional analysis of ascidian sperm proacrosin. J. Biol. Chem. 276 (27), 24594-24600.

*Sawada, H., and Yokosawa, H. (2001). Self-nonself recognition and lysin-system in fertilization of the ascidian Halocynthia roretzi. In “The Biology of Ascidians (H. Sawada, H. Yokosawa, and C. C. Lambert eds.)” , pp. 18-23, Springer-Verlag, Tokyo.

アブラナ科植物におけるアロ認識機構の解明

＜研究代表者＞ 岩野 恵（奈良先端科学技術大学院大学 バイオサイエンス研究科・助教）

Iwano M., Entani T., Shiba H., Kakita M., Nagai T., Mizuno H., Miyawaki A., Shoji T., Kubo K., Isogai A., Takayama S. (2009) Fine-tuning of the cytoplasmic Ca2+ concentration is essential for pollen tube growth. Plant Physiol. 150(3), 1322-34.

*Iwano, M., Shiba, H., Matoba, K., Miwa, T., Funato, M., Entani, T., Nakayama, P., Shimosato, H., Takaoka, A., Isogai, A., and Takayama, S. (2007). Actin dynamics in papilla cells of Brassica rapa during self-and cross-pollination. Plant Physiol. 144 (1), 72-81.

Kakita, M., Murase, K., Iwano, M., Matsumoto, T., Isogai, A., and *Takayama, S. (2007). Two distinct forms of M-Locus protein kinase localize to the plasma membrane and interact directly with S-Locus receptor kinase to transduce self-incompatibility signaling in Brassica rapa. Plant Cell 19 (12), 39613973.

Shimosato, H., Yokota, N., Shiba, H., Iwano, M., Entani, T., Che, F. S., Watanabe, M., Isogai, A., and *Takayama, S. (2007). Characterization of the SP11/SCR high-affinity binding site involved in self/nonself recognition in Brassica self-incompatibility. Plant Cell 19 (1), 107-117.

Shiba, H., Kakizaki, T., Iwano, M., Tarutani, Y., Watanabe, M., Isogai, A., and *Takayama, S. (2006). Dominance relationships between self-incompatibility alleles controlled by DNA methylation. Nature Genet. 38 (3), 297-299.

*Iwano, M., Shiba, H., Miwa, T., Che, F. S., Takayama, S., Nagai, T., Miyawaki, A., and Isogai, A. (2004). Ca²⁺dynamics in a pollen grain and papilla cell during pollination of Arabidopsis. Plant Physiol. 136 (3), 3562-3571.

Murase, K., Shiba, H., Iwano, M., Che, F. S., Watanabe, M., Isogai, A., and *Takayama, S. (2004). A membrane-anchored protein kinase involved in Brassica self-incompatibility signaling. Science 303 (5663), 1516-1519.

Iwano, M., Shiba, H., Funato, M., Shimosato, H., Takayama, S., and *Isogai, A. (2003). Immunohistochemical studies on translocation of pollen S-haplotype determinant in self-incompatibility of Brassica rapa. Plant Cell Physiol. 44 (1), 428-436.

Shiba, H., Iwano, M., Entani, T., Ishimoto, K., Shimosato, H., Che, F. S., Satta, Y., Ito, A., Takada, Y., Watanabe, M., Isogai, A., and *Takayama, S. (2002). The dominance of alleles controlling self-incompatibility in Brassica pollen is regulated at the RNA level. Plant Cell 14 (2), 491-504.

Takayama, S., Shimosato, H., Shiba, H., Funato, M., Che, F. S., Watanabe, M., Iwano, M., and *Isogai, A. (2001). Direct ligand-receptor complex interaction controls Brassica self-incompatibility. Nature 413 (6855), 534-538.

Takayama, S., Shiba, H., Iwano, M., Shimosato, H., Che, F. S., Kai, N., Watanabe, M., Suzuki, G., Hinata, K., and *Isogai, A. (2000). The pollen determinant of self-incompatibility in Brassica campestris. Proc. Natl. Acad. Sci. USA 97 (4), 1920-1925.

アロ認証研究のための次世代技術の開発と活用

＜研究代表者＞ 稲葉 一男（筑波大学 大学院生命環境科学研究科・教授）

Nomura, M., Nakajima, A., and *Inaba, K. (2008). Proteomic profiles of embryonic development in the ascidian Ciona intestinalis. Dev. Biol. 325: 468-481.

Mizuno, K., Padma, P., Konno, A., Satouh, Y., Ogawa, K., and *Inaba, K. (2008). A novel neuronal calcium sensor family protein, calaxin, is a potential Ca²⁺-dependent regulator for outer arm dynein of metazoan cilia and flagella. Biol. Cell 101: 91-103.

*Satou, Y., Mineta, K., Ogasawara, M., and et al. (16 authors), and Inaba, K. (2008). Improved genome assembly and evidence-based global gene model set for the chordate Ciona intestinalis: new insight into intron and operon populations. Genome Biol. 9 (10), R152.

*Inaba, K. (2007). Molecular basis of sperm flagellar axonemes: structural and evolutional aspects. Ann. NY Acad. Sci. 1101, 506-526.

*Inaba, K., Nomura, M., Nakajima, A., and Hozumi, A. (2007). Functional proteomics in Ciona intestinalis -A breakthrough in the exploration of the molecular and cellular mechanism of ascidian development. Dev. Dyn. 236 (7), 1782-1789.

Satouh, Y., Padma, P., Toda, T., Satoh, N., Ide, H., and *Inaba, K. (2005). Molecular characterization of radial spoke subcomplex containing radial spoke protein 3 and heat shock protein 40 in sperm flagella of the ascidian Ciona intestinalis. Mol. Biol. Cell 16 (2), 626-636.

Murata, Y., Iwasaki, H., Sasaki, M., Inaba, K., and *Okamura, Y. (2005). Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. Nature 435 (7046),1239-1243.

Padma, P., Satouh, Y., Wakabayashi, K., Hozumi, A., Ushimaru, Y., Kamiya, R., and *Inaba. K. (2003). Identification of a novel leucine-rich repeat protein as a component of flagellar radial spoke in the ascidian Ciona intestinalis. Mol. Biol. Cell 14 (2), 774-485.

Casey, D. M., Inaba, K., Pazour, G. J., Takada, S., Wakabayashi, K., Wilkerson, C. G., Kamiya, R., and *Witman, G. B. (2003). DC3, the 21-kD subunit of the outer dynein arm-docking complex (ODA-DC), is a novel EF-hand protein important for assembly of both the outer arm and the ODA-DC. Mol. Biol. Cell 14 (9), 3650-3663.

Dehal, P., et al. (Inaba, K., 42th from 85 authors), and *Rokhsar, D. S. (2002). The draft genome of Ciona intestinalis. Insights into chordate and vertebrate origins. Science 298 (5601), 2157-2167.

＜研究分担者＞ 笹倉 靖徳（筑波大学 大学院生命環境科学研究科・講師）

*Sasakura, Y., Konno, A., Mizuno, K., Satoh, N., and Inaba, K. (2008). Enhancer detection in the ascidian Ciona intestinalis with transposase-expressing lines of Minos. Dev Dyn. 237 (1), 39-50.

*Holland, L. Z., et al. (Sasakura, Y. 42th in 62 authors). (2008). The amphioxus genome illuminates vertebrate origins and cephalochordate biology. Genome Res 18 (7), 1100-1111.

*Awazu, S., Matsuoka, T., Inaba, K., Satoh, N., and Sasakura, Y. (2007). High-throughput enhancer trap by re-mobilization of transposon Minos in Ciona intestinalis. Genesis 45 (5), 307-317.

Sasakura, Y., Nakashima, K., Awazu, S., Matsuoka, T., Nakayama, A., Azuma, J., and *Satoh, N. (2005). Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian, Ciona intestinalis. Proc. Natl. Acad. Sci. USA 102 (42), 15134-15139.

Sasakura, Y., Awazu, S., Chiba, S., and *Satoh, N. (2003). Germ-line transgenesis of the Tc1/mariner superfamily transposon Minos in Ciona intestinalis. Proc. Natl. Acad. Sci. USA 100 (13), 7726-7730.

哺乳類の雌性殖器における精子認証機構の解明

＜研究代表者＞ 馬場 忠（筑波大学 大学院生命環境科学研究科・教授）

Kim, E., Yamashita, M., Kimura, M., Honda, A., Kashiwabara, S., and *Baba, T. (2008). Sperm penetration through cumulus mass and zona pellucida. Int. J. Dev. Biol. 52 (5-6), 677-682.

Yamashita, M., Honda, A., Ogura, A., Kashiwabra, S., Fukami, K., and *Baba, T. (2008). Reduced fertility of mouse epididymal sperm lacking Prss21/Tesp5 is rescued by sperm exposure to uterine microenvironment. Genes Cells 13 (10), 1001-1013.

Yamazaki, T., Yamagata, K., and *Baba, T. (2007). Time-lapse and retrospective analysis of DNA methylation in mouse preimplantation embryos by live cell imaging. Dev. Biol. 304 (1), 409-419.

Kim, E., Yamashita, M., Nakanishi, T., Park, K. E., Kimura, M., Kashiwabara, S., and *Baba, T. (2006). Mouse sperm lacking ADAM1b/ADAM2 fertilin can fuse with the egg plasma membrane and effect fertilization. J. Biol. Chem. 281 (9), 5634-5639.

Kim, E., Baba, D., Kimura, M., Yamashita, M., Kashiwabara, S., and *Baba, T. (2005). Identification of a hyaluronidase, Hyal5, involved in penetration of mouse sperm through cumulus mass. Proc. Acad. Natl. Sci. USA 102 (50), 18028-18033.

Nishimura, H., Kim, E., Nakanishi, T., Okabe, M., and *Baba, T. (2004). Possible function of the ADAM1a/ADAM2 fertilin complex in the appearance of ADAM3 on the sperm surface. J. Biol. Chem. 279 (33), 34957-34962.

Baba, D., Kashiwabara, S., Honda, A., Yamagata, K., Wu, Q., Ikawa, M., Okabe, M., and *Baba, T. (2002). Mouse sperm lacking cell surface hyaluronidase PH-20 can pass through the layer of cumulus cells and fertilize the egg. J. Biol. Chem. 277 (33), 30310-30314.

Honda, A., Yamagata, K., Sugiura, S., Watanabe, K., and *Baba, T. (2002). A mouse serine protease TESP5 is selectively included into lipid rafts of sperm membrane presumably as a glycosylphosphatidylinositol-anchored protein. J. Biol. Chem. 277 (19), 16976-16984.

Kashiwabara, S., Noguchi, J., Zhuang, T., Ohmura, K., Honda, A., Sugiura, S., Miyamoto, K., Takahashi, S., Inoue, K., Ogura, A., and *Baba, T. (2002). Regulation of spermatogenesis by testis-specific, cytoplasmic poly(A) polymerase TPAP. Science 298 (5600), 1999-2002.

Honda, A., Sirundawineti, J., and *Baba, T. (2002). Role of acrosomal matrix proteases in sperm-zona pellucida interactions. Hum. Reprod. Update 8 (5), 405-412.

哺乳類における配偶子融合の分子認証機構の解明

＜研究代表者＞ 宮戸 健二（国立成育医療センター研究所 生殖・細胞医療研究部・室長）

*Miyado, K., Yoshida, K., Yamagata, K., Sakakibara, K., Okabe, M., Wang, X., Miyamoto, K., Akutsu, H., Kondo, T., Takahashi, Y., Ban, T., Ito, C., Toshimori, K., Nakamura, A., Ito, M., Miyado, M., Mekada, E., and Umezawa, A. (2008). The fusing ability of sperm is bestowed by CD9-containing vesicles released from eggs in mice. Proc. Natl. Acad. Sci. USA 105 (35), 12921-12926.

*Hamatani, T., Yamada, M., Akutsu, H., Kuji, N., Mochimaru, Y., Takano, M., Toyoda, M., Miyado, K., Umezawa, A., and Yoshimura, Y. 　 (2008). What can we learn from gene expression profiling of mouse oocytes? 　 Reproduction 135 (5), 581.592.

Tanigawa, M., Miyamoto, K., Kobayashi, S., Sato, M., Akutsu, H., Okabe, M., Mekada, E., Umezawa, A., and *Miyado, K. (2008). Possible involvement of CD81 in acrosome reaction of sperm in mice. Mol. Repro. Dev. 75 (1), 150-155.

Nishiyama, N., *Miyoshi, S., Hida, N., Uyama, T., Okamoto, K., Ikegami, Y., Miyado, K., Segawa, K., Terai, M., Sakamoto, M., Ogawa, S., and Umezawa, A. (2007). The significant cardiomyogenic potential of human umbilical cord blood-derived mesenchymal stem cells in vitro. Stem Cells 25 (8), 2017-2024.

Cui, C. H., Uyama, T., Miyado, K., Terai, M., Kyo, S., Kiyono, T., and *Umezawa, A. (2007). Menstrual blood-derived cells confer human dystrophin expression in the murine model of Duchenne muscular dystrophy via cell fusion and myogenic transdifferentiation. Mol. Biol. Cell 18 (5), 1586-1594.

Nakanishi, T., Kubota, H., Ishibashi, N., Kumagai, S., Watanabe, H., Yamashita, M., Kashiwabara, S., Miyado, K., and *Baba, T. (2006). Possible role of mouse poly(A) polymerase mGLD-2 during oocyte maturation. Dev. Biol. 289 (1), 115-126.

Ishibashi, T., Ding, L., Ikenaka, K., Inoue, Y., Miyado, K., Mekada, E., and *Baba, H. (2004). Tetraspanin protein CD9 is a novel paranodal component regulating paranodal junctional formation. J. Neurosci. 24 (1), 96-102.

Iwamoto, R., Yamazaki, S., Asakura, M., Takashima, S., Hasuwa, H., Miyado, K., Adachi, S., Kitakaze, M., Hashimoto, K., Raab, G., Namba, D., Higashiyama, S., Kori, M., Klagsbrun, M., and *Mekada, E. (2003). Heparin-binding EGF-like growth factor and ErbB signaling is essential for heart function. Proc. Natl. Acad. Sci. USA 100 (6), 3221-3226.

Takeda, Y., *Tachibana, I., Miyado, K., Kobayashi, M., Miyazaki, T., Funakoshi, T., Kimura, H., Yamane, H., Saito, Y., Goto, H., Yoneda, T., Yoshida, M., Kumagai, T., Osaki, T., Hayashi, S., Kawase, I., and Mekada, E. (2003). Tetraspanins CD9 and CD81 function to prevent the fusion of blood monocytes/alveolar macrophages. J. Cell Biol. 161 (5), 945-956.

Miyado, K., Yamada, G., Yamada, S., Hasuwa, H., Nakamura, Y., Ryu, F., Suzuki, K., Kosai, K., Inoue, K., Ogura, A., Okabe, M., and *Mekada, E. (2000). Requirement of CD9 on the egg plasma membrane for fertilization. Science 287 (5451), 321-324.

＜研究分担者＞ 井上 直和（大阪大学 微生物病研究所・助教）

Inoue, N., Ikawa, M., and *Okabe, M. (2008). Putative sperm fusion protein IZUMO and the role of N-glycosylation. Biochem. Biophys. Res. Commun. 377 (3), 910-914.

*Shingai, M., *Inoue, N., Okuno, T., Okabe, M., Akazawa, T., Miyamoto, Y., Ayata, M., Honda, K., Kurita-Taniguchi, M., Matsumoto, M., et al. (2005). Wild-type measles virus infection in human CD46/CD150-transgenic mice: CD11c-positive dendritic cells establish systemic viral infection. J. Immunol. 175 (5), 3252-3261.

Inoue, N., Ikawa, M., Isotani, A., and *Okabe, M. (2005). The immunoglobulin superfamily protein Izumo is required for sperm to fuse with eggs. Nature 434 (7030), 234-238.

Inoue, N., Ikawa, M., Nakanishi, T., Matsumoto, M., Nomura, M., Seya, T., and *Okabe, M. (2003). Disruption of mouse CD46 causes an accelerated spontaneous acrosome reaction in sperm. Mol. Cell. Biol. 23 (7), 2614-2622.

Inoue, N., Fukui, A., Nomura, M., Matsumoto, M., Nishizawa, Y., Toyoshima, K., and *Seya, T. (2001). A novel chicken membrane-associated complement regulatory protein: molecular cloning and functional characterization. J. Immunol. 166 (1), 424-431.

被子植物における配偶子融合の分子認証機構の解明

＜研究代表者＞ 岡本 龍史（首都大学東京 大学院理工学研究科・准教授）

*Kranz, E., Hoshino, Y., and Okamoto, T. (2008). In vitro fertilization with isolated higher plant gametes. Methods Mol. Biol. 427, 51-69.

Uchiumi, T., Uemura, I., and *Okamoto, T. (2007). Establishment of an in vitro fertilization system in rice (Oryza sativa L.). Planta 226 (3), 581-589.

Uchiumi, T., Shinkawa, T., Isobe, T., and *Okamoto, T. (2007). Identification of the major protein components of rice egg cells. J. Plant Res. 120 (4), 575-579.

Uchiumi, T., Komatsu, S., Koshiba, T., and *Okamoto, T. (2006). Isolation of gametes and central cells from Oryza sativa L. Sex. Plant Reprod. 19 (1), 37-45.

Okamoto, T., and *Kranz, E. (2005). In vitro fertilization -a tool to dissect cell specification from a higher plant zygote. Embryology of Seed Plants Then and Now. Curr. Sci. 89 (11), 1861-1869.

*Okamoto, T., Scholten, S., Lorz, H., and Kranz, E. (2005). Identification of genes that are up-or down-regulated in the apical or basal cell of maize two-celled embryos and monitoring their expressions during zygote development by a cell manipulation-and PCR-based approach. Plant Cell Physiol. 46 (2), 332-338.

*Okamoto, T., Higuchi, K., Shinkawa, T., Isobe, T., Lorz, H., Koshiba, T., and Kranz, E. (2004). Identification of major proteins in maize egg cells. Plant Cell Physiol. 45 (10), 1206-1212.

*Okamoto, T., Toyooka, K., and Minamikawa, T. (2001). Identification of a membrane-associated cysteine protease (MCP) with possible dual roles in the endoplasmic reticulum and protein storage vacuole. J. Biol. Chem. 276 (1), 742-751.

Toyooka, K., *Okamoto, T., and Minamikawa, T. (2001). Cotyledon cells of Vigna mungo seedlings utilize at least two distinct autophagic machineries for degradation of starch granules and cellular components. Cotyledon cells of Vigna mungo seedlings use at least two distinct autophagic machineries for degradation of starch granules and cellular components. J. Cell Biol. 154 (5), 973-982.

Toyooka, K., *Okamoto, T., and Minamikawa, T. (2000). Mass transport of a KDEL-tailed cysteine proteinase (SH-EP) to protein storage vacuole by ER-derived vesicle is involved in protein mobilization in germinating seeds. J. Cell Biol. 148 (3), 453-463.

植物と動物の配偶子融合における中核機構の解明

＜研究代表者＞ 森 稔幸（理化学研究所 宮城島独立主幹研究ユニット・基礎科学特別研究員）

*Hirai, M., #Arai, M., #Mori, T., Miyagishima, S., Kawai, S., Kita, K., Kuroiwa, T., Terenius, O., and Matsuoka, H. (2008). Male fertility of malaria parasites is determined by GCS1, a plant-type reproduction factor. Curr. Biol. 18 (8), 607-613.

*Mori, T., Kuroiwa, H., Higashiyama, T., and Kuroiwa, T. (2006). GENERATIVE CELL SPECIFIC 1 is essential for angiosperm fertilization. Nature Cell Biol. 8 (1), 64-71.

*Nozaki, H., Mori, T., Misumi O., Matsunaga, S., and Kuroiwa, T. (2006). Males evolved from the dominant isogametic mating type. Curr. Biol. 16 (24), R1018-R1020.

*Higashiyama, T., Inatsugi, R., Sakamoto, S., Sasaki, N., Mori, T., Kuroiwa, H., Nakada, T., Nozaki, H., Kuroiwa, T., and Nakano, A. (2006). Species preferentiality of the pollen tube attractant derived from the synergid cell of Torenia fournieri. Plant Physiol. 142 (2), 481-491.

*Misumi, O., Matsuzaki, M., Nozaki, H., Miyagishima, S., Mori, T., Nishida, K., Yagisawa, F., Yoshida, Y., Kuroiwa, H., and Kuroiwa, T. (2005). Cyanidioschyzon merolae genome. A tool for facilitating comparable studies on organelle biogenesis in photosynthetic eukaryotes. Plant Physiol. 137 (2), 567-585.

*Matsuzaki, M., Misumi, O., Shin-i, T., et al. （ほか 39 名） (2004). Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D. Nature 428 (6983), 653-657.

*Mori, T., Kuroiwa, H., Higashiyama, T., and Kuroiwa, T. (2003). Identification of higher plant GlsA, a putative morphogenesis factor of gametic cells. Biochem. Biophys. Res. Commun. 306 (2), 564-569.

*Momoyama, Y., Miyazawa, Y., Miyagishima, S., Mori, T., Misumi, O., Kuroiwa, H., and Kuroiwa, T. (2003). The division of pleomorphic plastids with multiple FtsZ rings in tobacco BY-2 cells. Eur. J. Cell Biol. 82 (6), 323-332.

*Miyagishima, S., Nishida, K., Mori, T., Kuroiwa, H., Higashiyama, T., and Kuroiwa, T. (2003). A plant-specific dynamin-related protein forms a ring at the chloroplast division site. Plant Cell 15 (3), 655-665.

*Yagisawa, F., Mori, T., Higashiyama, T., Kuroiwa, H., and Kuroiwa, T. (2003). Regulation of Brassica rapa chloroplast proliferation in vivo and in cultured leaf disks. Protoplasma 222 (3-4), 139-148.

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