記述言語：英語   掲載種別：研究論文（学術雑誌）  

In vertebrates, estrogen receptors are essential for estrogen-associated early gonadal sex development. Our previous studies revealed sexual dimorphic expression of estrogen receptor β2 (ERβ2) during embryogenesis of medaka, and here we investigated the functional importance of ERβ2 in female gonad development and maintenance using a transgenerational ERβ2-knockdown (ERβ2-KD) line and ERβ2-null mutants. We found that ERβ2 reduction favored male-biased gene transcription, suppressed female-responsive gene expression, and affected SDF1a and CXCR4b co-assisted chemotactic primordial germ cell (PGC) migration. Co-overexpression of SDF1a and CXXR4b restored the ERβ2-KD/KO associated PGC mismigration. Further analysis confirmed that curtailment of ERβ2 increased intracellular Ca²⁺ concentration, disrupted intra- and extracellular calcium homeostasis, and instigated autophagic germ cell degradation and germ cell loss, which in some cases ultimately affected the XX female sexual development. This study is expected improve our understanding of germ cell maintenance and sex spectrum, and hence open new avenues for reproductive disorder management. In this article, Chakraborty et al. find that estrogen receptor β2 (ERβ2) plays multifaceted crucial roles in medaka gonadal development and sexuality. ERβ2 predominantly expresses in germ cells, influences the SDF1/CXCR4 chemotaxis, PGC migration, and death, and regulates germ cell proliferation and various sex-biased gene transcriptions that are essential for gonadal sex assignment and maintenance.

DOI： 10.1016/j.stemcr.2019.07.013

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1038/s41598-017-03314-7

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1038/srep19480

担当区分：最終著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Environmental Pollution  

Benzalkonium chloride (BAC), a commonly used quaternary ammonium compound in various products like antiseptics, cosmetics, and disinfectants, has raised concerns due to its potential to contaminate aquatic environments and subsequently affect the reproductive performance of the organisms within those ecosystems. The article underscores a critical concern regarding the impact of BAC on aquatic ecosystems, particularly its effect on fish reproductive quality, using medaka (Oryzias latipes) as a model organism. Firstly, while measuring lethal dose of BAC in adult medaka, we observed a dose dependent mortality in BAC treated fish (100 and 200 ppm: 100%; 60 ppm: 51.7%; 30 ppm or less: no mortality at 24 h post treatment (hpt)) and calculated the LD50 at 96 hpt as 39.291 ppm (95% confidence interval: 28.817–53.570 ppm). Further, we assessed the molecular, cellular and histological changes through long-term exposure. Enlarged sperm pockets and reduced spermatocyte were seen in BAC exposed testis while no significant structural changes were observed in the ovaries. Following BAC exposure, drastic alterations in the gene expression and cellular localization related to sex, estrogen signaling, and autophagy were also noted from gonads and liver. Subsequently, using a short-term exposure analysis, we confirmed the sex and time responsive transcriptional kinetics and found that BAC sequentially affected the gonadal somatic cells followed by germ cell differentiation. Finally, using reproductively competent male and female medaka, we conducted progeny production and performance analysis and depicted a drastic reduction in fecundity, and fertilization and hatching rate, indicating adverse effects of BAC on reproductive success. Cumulatively, these findings emphasize the consequences of widespread use of BAC on reproductive security of aquatic animals and highlights the need for further research to comprehend the potential harm posed by such compounds to aquatic animal health and ecosystem integrity.

DOI： 10.1016/j.envpol.2024.124778

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記述言語：英語   出版者・発行元：Comparative Biochemistry and Physiology Part - B: Biochemistry and Molecular Biology  

Steroidogenic acute regulatory protein (Star) plays an essential role in the biosynthesis of corticosteroids and sex steroids by mediating the transport of cholesterol from the outer to the inner membrane of mitochondria. Two duplicated Star genes, namely star1 and star2, have been identified in non-mammalian vertebrates. To investigate the roles of star genes in fish steriodogenesis, we generated two mutation lines of star1−/− and star1−/−/star2−/− in Nile tilapia (Oreochromis niloticus). Previous studies revealed that deficiency of star2 gene caused delayed spermatogenesis, sperm apoptosis and sterility in male tilapia. Our present data revealed that mutation of star genes impaired male fertility. Disordered seminiferous lobules and spermatic duct obstruction were found in the testis of both types of mutants. Moreover, significant decline in semen volume, sperm abnormality and impaired fertility were also detected in star1−/− and star1−/−/star2−/− males. In star1−/− male fish, lipid accumulation, up-regulation of steroidogenic enzymes, and significant decline of androgens were found. Additionally, hyperplasic interrenal cells, elevated steroidogenic gene expression level and decline of serum glucocorticoids were detected in star1 mutants. Intriguingly, either 11-KT or cortisol supplementation successfully rescued the impaired fertility of the star1−/− mutants. Taken together, these results further indicate that Star1 might play critical roles in the production of both 11-KT and glucocorticoids, which are indispensable for the maintenance of male fertility in fish.

DOI： 10.1016/j.cbpb.2024.110985

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担当区分：責任著者   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Marine Science  

Sexual development and reproduction are largely linked to epigenetic changes in many fish species. However, understanding of epigenetic regulation in scombrid species, such as tunas and mackerels, is limited. This study investigates DNA methylation’s impact on cyp19a expression, crucial for estrogen synthesis, focusing on chub mackerel. Given the commercial significance of scombrids and susceptibility of marine fish to environmental changes, elucidating epigenetic mechanisms, particularly in the context of global warming, becomes imperative. We aimed to generalize observations from chub mackerel to other scombrids. Additionally, we studied DNA methylation patterns across fish with different sexual systems to understand aromatase regulation’s phenotypic plasticity. Our in silico analysis revealed highly conserved promoter sequences within scombrids, sharing TFBS like Foxl2, FOS::JUN, ESRR, and Sox3, while CpG content varies. This indicates a conserved regulatory network controlling gene expression. We found sexual dimorphism in DNA methylation, with males hypermethylated and aromatase expression downregulated. Despite similar dnmt1 expression, tet1, tet2, and tet3 were higher expressed in females, suggesting that the observed DNA methylation patterns are maintained through active demethylation rather than differential methylation. Gonochoristic Japanese anchovy and protogynous bamboo leaf wrasse displayed similar trends, but species-specific methylation patterns highlight DNA methylation’s complex role in gonadal changes. In vitro assays confirmed methylation’s regulatory role and identified an SF-1 binding site relevant for promoter activation in chub mackerel. Another studied SF-1 site, present in both chub mackerel and bamboo leaf wrasse, showed regulatory effects, indicating potential similar regulatory mechanisms for cyp19a expression. Overall, our findings suggest that while global methylation affects cyp19a transcription, the variation in CpG density and location could be introducing nuances in its epigenetic regulation. This study contributes to our understanding of the cyp19a regulation in fish gonad maturation.

DOI： 10.3389/fmars.2024.1408561

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記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：General and Comparative Endocrinology  

Primordial germ cells (PGCs) are pivotal for gonadal development and reproductive success. Though artificial induction of sterility by targeting PGCs are gaining popularity due to its advantages in fish surrogacy and biodiversity management, it is often skill and time intensive. In this study, we have focused on understanding the role of PGCs and the chemotactic SDF-1/CXCR4 signaling on gonad development of Japanese anchovy (JA, Engraulis japonicus), an upcoming marine model organism with eco-commercial values, with an aim to develop a novel, easy, and versatile gonad sterilization method. Our data showed that PGC migration related genes, i.e., sdf-1a, sdf-1b, cxcr4a, cxcr4b and vasa, are phylogenetically closer relatives of respective herring (Clupea harengus) and zebrafish (Danio rerio) homolog. Subsequently, PGC marking and live tracing experiments confirmed that PGC migration in JA initiates from 16 hours post fertilization (hpf) followed by PGC settlement in the gonadal ridge at 44 hpf. We found that overexpression of zebrafish sdf-1a mRNA in the germ cell suppresses cxcr4a and increases cxcr4b transcription at 8 hpf, dose dependently disrupts PGC migration at 24–48 hpf, induces PGC death and upregulates sdf-1b at 5 days post hatching. 48 h of immersion treatment with CXCR4 antagonist (AMD3100, Abcam) also accelerated PGC mismigration and pushed the PGC away from gonadal ridge in a dose responsive manner, and further when grown to adulthood caused germ cell less gonad formation in some individuals. Cumulatively, our data, for the first time, suggests that JA PGC migration is largely regulated by SDF1/CXCR4 signaling, and modulation of this signaling has strong potential for sterile, germ cell less gonad preparation at a mass scale. However, further in-depth analysis is pertinent to apply this methodology in marine fish species to successfully catapult Japanese anchovy into a true marine fish model.

DOI： 10.1016/j.ygcen.2024.114476

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担当区分：筆頭著者,　責任著者   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Marine Science  

The gonadal sexual fate of vertebrates is either defined by genetics or environment, or a combination of both factors. Interestingly, in sequential hermaphroditism, the animal can undergo natural sex changes from female-to-male, male-to-female, and bidirectional way throughout their lives. This change exhibits the process which shifts between oogenesis and spermatogenesis and is regarded as an ideal instance of sexual plasticity. To develop the experimental model for studying the sexual plasticity of protogynous fish, the social conditions that induce sex changes were defined in wrasse, Pseudolabrus sieboldi. When six females were kept together in a tank, the largest female became a male, whereas a similar conversion did not occur when only two females were present in a tank. A semi-gonadectomy analysis developed in the present study verified the direct relationship between gonadal sex and body coloration. In P. sieboldi, the sex change is controlled by the relative body size of an individual within a group, rather than by absolute body size. When six females were kept in smaller sized tanks, delayed sex change or unchanged individuals was observed. Overall, more than 90% of the largest females demonstrated sex change after being housed with five smaller females in different sizes of tanks ranging from 80 to 500 L. Furthermore, the experiment using a transparent barrier suggested that visual stimuli are one of the major cues to initiate sex change. Our findings on the laboratory conditions leading to the initiation of sex change in wrasse suggest the usefulness of this species as a model organism for comparative studies in molecular, cellular, and physiological mechanisms of sexual plasticity, as well as on social and reproductive behaviors.

DOI： 10.3389/fmars.2022.1048506

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担当区分：筆頭著者,　責任著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：International Journal of Molecular Sciences  

Germ cells are pivotal for gonadal sexuality maintenance and reproduction. Sex lethal (sxl), the somatic sex determining gene of Drosophila, is the known regulator and initiator of germ cell femininity in invertebrates. However, the role of the Sxl homologue has rarely been investigated in vertebrates. So, we used medaka to clarify the role of sxl in vertebrate gonadogenesis and sexuality and identified two Sxl homologues, i.e., Sxl1a and Sxl1b. We found that sxl1a specifically expresses in the primordial germ cells (PGC), ovary, (early gonia and oocytes), while sxl1b distributions are ubiquitous. An mRNA overexpression of sxl1a accelerated germ cell numbers in 10 DAH XY fish, and sxl1a knockdown (KD), on the other hand, induced PGC mis-migration, aberrant PGC structuring and ultimately caused significant germ cell reduction in XX fish. Using an in vitro promoter analysis and in vivo steroid treatment, we found a strong link between sxl1a and estrogenic germ cell-population maintenance. Further, using sxl1a-KD and erβ2-knockout fish, we determined that sxl1 acts through erβ2 and controls PGC sexuality. Cumulatively, our study highlights the novel role of sxl1a in germ cell maintenance and sexual identity assignment and thus might become a steppingstone to understanding the commonalities of animal sexual development.

DOI： 10.3390/ijms232415496

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記述言語：英語   出版者・発行元：Frontiers in Endocrinology  

Background: Steroid hormones play an essential role in many reproductive processes of vertebrates. Previous studies revealed that teleost-specific Cyp17a2 (cytochrome P450 family 17 subfamily a 2) might be required for the production of cortisol in the head-kidney and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) in ovary during oocyte maturation. However, the role of Cyp17a2 in male reproduction remains to be largely unknown. The aim of this study was to explore the essentiality of cyp17a2 gene in male steroidogenesis, spermatogenesis, and male fertility. Methods: A homozygous mutation line of cyp17a2 gene was constructed in tilapia by CRISPR/Cas9 gene editing technology. The expression level of germ cell and meiosis-related genes and steroidogenic enzymes were detected by qRT-PCR, IHC, and Western blotting. EIA and LC-MS/MS assays were used to measure the steroid production levels. And sperm quality was examined by Sperm Quality Analyzer software. Results: In this study, cyp17a2 gene mutation resulted in the significant decline of serum DHP and cortisol levels. On the contrary, significant increases in intermediate products of cortisol and DHP were found in cyp17a2-/- male fish. The deficiency of cyp17a2 led to the arrest of meiotic initiation in male fish revealing as the reduction of the expression of germ cell-related genes (vasa, piwil, oct4) and meiosis-related genes (spo11 and sycp3) by 90 dah. Afterwards, spermatogenesis was gradually recovered with the development of testis in cyp17a2-/- males, but it showed a lower sperm motility and reduced fertility compared to cyp17a2+/+ XY fish. Deletion of cyp17a2 led to the abnormal upregulation of steroidogenic enzymes for cortisol production in the head-kidney. Moreover, unaltered serum androgens and estrogens, as well as unchanged related steroidogenic enzymes were found in the testis of cyp17a2-/- male fish. Conclusion: This study proved that, for the fist time, Cyp17a2 is indispensable for cortisol and DHP production, and cyp17a2 deficiency associated curtailed meiotic initiation and subfertility suggesting the essentiality of DHP and cortisol in the male fertility of fish.

DOI： 10.3389/fendo.2022.1074921

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担当区分：責任著者   掲載種別：研究論文（学術雑誌）   出版者・発行元：Frontiers in Marine Science  

Fisheries and aquaculture are the fastest-growing food-producing sector and rapidly becoming an important element for the global food security since they are the primary source of seafood and high animal protein in the human diet. Genome editing offers new possibilities such as the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein (Cas9) technology, which has the potential to accelerate the sustainable genetic improvement in fisheries and aquaculture. The CRISPR/Cas9 system has four key components, namely, target DNA, Cas9, the protospacer adjacent motif sequence, and the guide RNA or single-guide RNA. CRISPR/Cas is cheaper, easier, and more precise than the other genome editing technologies and can be used as a new breeding technology in fisheries and aquaculture to solve the far-reaching challenges. The attributes like high fecundity, external fertilization, short generation interval, the established method of breeding, and the larval rearing of most aquaculture species have advantages for CRISPR/Cas9 genome editing applications. CRISPR/Cas9 has recently been applied to the traits valued in some aquaculture species (almost >20 species), targeting the main traits of traditional genetic improvement initiatives like growth, disease resistance, reproduction, sterility, and pigmentation. Genome editing can fast forward the breeding process with precision where changes occur in the targeted genes. The probability of desired changes occurring and passing the trait in the next generation is high, so it takes 1-3 generations to establish a breed. Moreover, CRISPR/Cas genome editing rapidly introduces favorable changes by disrupting genes with targeted minor changes, in contrast to transgenesis, which introduces foreign genes into the host genome and thereby alleviates major public concerns on safety. Although the CRISPR/Cas technology has a tremendous potential, there are several technical challenges and regulatory and public issues concerning the applications in fisheries and the aquaculture breeding sector. Nonetheless, the exciting point in the CRISPR/Cas9 genome editing is that two CRISPR-edited fish, namely, red sea bream and tiger puffer developed by the Kyoto-based startup got approval and are now on the market for sale, and another fish, FLT-01 Nile tilapia developed by the AquaBounty, is not classified under genetically modified organism regulatory. However, there is still a way to go before it revolutionizes and becomes viable in commercial aquaculture as the new breeding technology for aquaculture-important traits and species.

DOI： 10.3389/fmars.2022.924475

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記述言語：英語   掲載種別：研究論文（学術雑誌）  

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： https://doi.org/10.1016/j.jsbmb.2021.105974

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.aquatox.2019.105289

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.fsi.2018.08.027

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.ygcen.2018.03.005

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.cbpb.2017.01.009

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.bbrc.2016.03.045

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1038/srep19543

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.cbpa.2015.07.003

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1371/journal.pone.0123900

記述言語：英語   掲載種別：研究論文（学術雑誌）  

Interferon gamma (. IFNγ) is an active player in estrogen dependent immuno-regulation of fish. The present work was aimed to characterize the alternatively spliced isoforms of IFNγ2 in the gonadal sex development in medaka. Phylogenetic analysis demonstrated that IFNγ2a and 2b were clustered with fish specific interferon gamma. Our in vitro promoter and mini-genome analysis data confirmed that alternative splicing of IFNγ2 is regulated by estrogens and androgens. Tissue distribution, quantitative PCR and ISH data demonstrated ubiquitous expression of IFNγ2a, while IFNγ2b was only expressed predominantly in female germ cells than males. This was further confirmed by germ cell specific GFP signals in the IFNγ2b-GFP over-expressed embryos and specific induction of IFNγ2b expression in the BrdU positive cells. All together our data suggest that steroid responsive alternatively spliced IFNγ2b isoforms might have some indirect roles in germ cell proliferation and thus can be an important candidate for immuno-reproductive interaction studies.

DOI： 10.1016/j.mce.2014.10.012

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/jwas.12144

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1371/journal.pone.0100929

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.mce.2014.05.013

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/j.1439-0396.2012.01301.x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1186/1471-213X-12-36

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.pestbp.2012.06.006

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1111/j.1365-2095.2011.00866.x

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.jsbmb.2010.11.015

記述言語：英語   掲載種別：研究論文（学術雑誌）  

DOI： 10.1016/j.mce.2010.12.002

記述言語：英語   掲載種別：研究論文（学術雑誌）  

開催年月日： 2023年5月

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国名：ブラジル連邦共和国  

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記述言語：日本語   会議種別：口頭発表（一般）  

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記述言語：日本語   会議種別：口頭発表（一般）  

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会議種別：口頭発表（一般）  

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記述言語：英語   会議種別：口頭発表（一般）  

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会議種別：口頭発表（一般）  

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記述言語：英語   会議種別：口頭発表（一般）  

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記述言語：英語   会議種別：口頭発表（招待・特別）  

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J-GLOBAL

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J-GLOBAL

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記述言語：英語  

DOI： https://doi.org/10.1007/978-981-16-3215-0_7

記述言語：英語  

DOI： https://doi.org/10.1080/15548627.2020.1797280

記述言語：英語   掲載種別：記事・総説・解説・論説等（学術雑誌）  

種別：大会・シンポジウム等 

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種別：学会・研究会等 

種別：査読等 

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種別：査読等 

外国語雑誌　査読論文数：15

種別：学会・研究会等 

種別：査読等 

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種別：学会・研究会等 

種別：査読等 

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種別：査読等 

外国語雑誌　査読論文数：25

種別：査読等 

外国語雑誌　査読論文数：10

日本語雑誌　査読論文数：0

国際会議録　査読論文数：0

国内会議録　査読論文数：0

種別：査読等 

外国語雑誌　査読論文数：15

日本語雑誌　査読論文数：0

国際会議録　査読論文数：0

国内会議録　査読論文数：0

種別：学術調査 

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種別：学術調査 

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種別：査読等 

外国語雑誌　査読論文数：5

日本語雑誌　査読論文数：0

国際会議録　査読論文数：0

国内会議録　査読論文数：0

種別：その他

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Foreign co-ordinator of COFLAA, India.

種別：その他

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対象： 幼稚園以下,　小学生,　中学生,　高校生

種別：セミナー・ワークショップ

種別：セミナー・ワークショップ

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対象： 社会人・一般,　学術団体,　企業,　市民団体,　行政機関

種別：その他

種別：その他

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organized training (volunteer) for Primary and high school teachers

種別：その他

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対象： 幼稚園以下,　小学生,　中学生,　高校生

種別：セミナー・ワークショップ

種別：セミナー・ワークショップ

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種別：研究指導

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