476 results on '"Takeshi Noda"'
Search Results
2. Compact wide-field femtoliter-chamber imaging system for high-speed and accurate digital bioanalysis
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Tatsuya Iida, Jun Ando, Hajime Shinoda, Asami Makino, Mami Yoshimura, Kazue Murai, Makiko Mori, Hiroaki Takeuchi, Takeshi Noda, Hiroshi Nishimasu, and Rikiya Watanabe
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Biomedical Engineering ,Bioengineering ,General Chemistry ,Biochemistry - Abstract
The femtoliter-chamber array is a bioanalytical platform that enables highly sensitive and quantitative analysis of biological reactions at the single-molecule level. This feature has been considered a key technology for "digital bioanalysis" in the biomedical field; however, its versatility is limited by the need for a large and expensive setup such as a fluorescence microscope, which requires a long time to acquire the entire image of a femtoliter-chamber array. To address these issues, we developed a compact and inexpensive wide-field imaging system (COWFISH) that can acquire fluorescence images with a large field of view (11.8 mm × 7.9 mm) and a high spatial resolution of ∼ 3 μm, enabling high-speed analysis of sub-million femtoliter chambers in 20 s. Using COWFISH, we demonstrated a CRISPR-Cas13a-based digital detection of viral RNA of SARS-CoV-2 with an equivalent detection sensitivity (limit of detection: 480 aM) and a 10-fold reduction in total imaging time, as compared to confocal fluorescence microscopy. In addition, we demonstrated the feasibility of COWFISH to discriminate between SARS-CoV-2-positive and -negative clinical specimens with 95% accuracy, showing its application in COVID-19 diagnosis. Therefore, COWFISH can serve as a compact and inexpensive imaging system for high-speed and accurate digital bioanalysis, paving a way for various biomedical applications, such as diagnosis of viral infections.
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- 2023
3. VEGF (vascular endothelial growth factor) provides antimicrobial effects via autophagy and lysosomal empowerment in endothelial cells
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Shiou-Ling Lu and Takeshi Noda
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- 2022
4. Structure of the bile acid transporter and HBV receptor NTCP
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Jinta Asami, Kanako Terakado Kimura, Yoko Fujita-Fujiharu, Hanako Ishida, Zhikuan Zhang, Yayoi Nomura, Kehong Liu, Tomoko Uemura, Yumi Sato, Masatsugu Ono, Masaki Yamamoto, Takeshi Noda, Hideki Shigematsu, David Drew, So Iwata, Toshiyuki Shimizu, Norimichi Nomura, and Umeharu Ohto
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Hepatitis B virus ,Multidisciplinary ,Symporters ,Cryoelectron Microscopy ,Sodium ,Organic Anion Transporters, Sodium-Dependent ,Rats ,Mutation ,Hepatocytes ,Animals ,Humans ,Receptors, Virus ,Cattle ,Apoproteins - Abstract
Chronic infection with hepatitis B virus (HBV) affects more than 290 million people worldwide, is a major cause of cirrhosis and hepatocellular carcinoma, and results in an estimated 820,000 deaths annually
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- 2022
5. Cryoelectron microscopic structure of the nucleoprotein–RNA complex of the European filovirus, Lloviu virus
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Shangfan Hu, Yoko Fujita-Fujiharu, Yukihiko Sugita, Lisa Wendt, Yukiko Muramoto, Masahiro Nakano, Thomas Hoenen, and Takeshi Noda
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Lloviu virus ,nucleocapsid ,cryo-EM ,Filovirus - Abstract
Lloviu virus (LLOV) is a novel filovirus detected in Schreiber’s bats in Europe. The isolation of the infectious LLOV from bats has raised public health concerns. However, the virological and molecular characteristics of LLOV remain largely unknown. The nucleoprotein (NP) of LLOV encapsidates the viral genomic RNA to form a helical NP-RNA complex, which acts as a scaffold for nucleocapsid formation and de novo viral RNA synthesis. In this study, using single-particle cryo-electron microscopy, we determined two structures of the LLOV NP-RNA helical complex, comprising a full-length and a C-terminally truncated NP. The two helical structures were identical, demonstrating that the N-terminal region determines the helical arrangement of the NP. The LLOV NP-RNA protomers displayed a structure similar to that in the Ebola and Marburg virus, but the spatial arrangements in the helix differed. Structure-based mutational analysis identified amino acids involved in the helical assembly and viral RNA synthesis. These structures advance our understanding of the filovirus nucleocapsid formation, and provide a structural basis for the development of anti-filoviral therapeutics., ヨーロッパに分布するエボラウイルス近縁ウイルスの増殖機構を解明 --広範囲の抗フィロウイルス療法の開発に期待--. 京都大学プレスリリース. 2023-04-10.
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- 2023
6. Genome-scale CRISPR‒Cas9 screen identifies novel host factors as potential therapeutic targets for SARS-CoV-2 infection
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Madoka Sakai, Yoshie Masuda, Yusuke Tarumoto, Naoyuki Aihara, Yugo Tsunoda, Michiko Iwata, Yumiko Kamiya, Ryo Komorizono, Takeshi Noda, Kosuke Yusa, Keizo Tomonaga, and Akiko Makino
- Abstract
Although many host factors important for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection have been reported, the mechanisms by which the virus interacts with host cells remain elusive. Here, we identified tripartite motif containing (TRIM) 28, TRIM33, euchromatic histone lysine methyltransferase (EHMT) 1, and EHMT2 as novel proviral factors involved in SARS-CoV-2 infection by CRISPR‒Cas9 screening. We demonstrated that TRIM28 plays a role(s) in viral particle formation and that TRIM33, EHMT1, and EHMT2 are involved in viral transcription and replication using cells with suppressed gene expression. UNC0642, a compound that specifically inhibits the methyltransferase activity of EHMT1/2, strikingly suppressed SARS-CoV-2 growth in cultured cells and reduced disease severity in a hamster infection model. This study suggests that EHMT1/2 may be a novel therapeutic target for SARS-CoV-2 infection.
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- 2023
7. Elucidation of the liver pathophysiology of COVID-19 patients using liver-on-a-chips
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Sayaka Deguchi, Kaori Kosugi, Rina Hashimoto, Ayaka Sakamoto, Masaki Yamamoto, Rafal P Krol, Peter Gee, Ryosuke Negoro, Takeshi Noda, Takuya Yamamoto, Yu-suke Torisawa, Miki Nagao, and Kazuo Takayama
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Baricitinib ,SARS-CoV-2 ,Remdesivir ,COVID-19 ,organs-on-a-chip ,liver-on-a-chip - Abstract
SARS-CoV-2 induces severe organ damage not only in the lung but also in the liver, heart, kidney, and intestine. It is known that COVID-19 severity correlates with liver dysfunction, but few studies have investigated the liver pathophysiology in COVID-19 patients. Here, we elucidated liver pathophysiology in COVID-19 patients using organs-on-a-chip technology and clinical analyses. First, we developed liver-on-a-chip (LoC) which recapitulating hepatic functions around the intrahepatic bile duct and blood vessel. We found that hepatic dysfunctions, but not hepatobiliary diseases, were strongly induced by SARS-CoV-2 infection. Next, we evaluated the therapeutic effects of COVID-19 drugs to inhibit viral replication and recover hepatic dysfunctions, and found that the combination of anti-viral and immunosuppressive drugs (Remdesivir and Baricitinib) is effective to treat hepatic dysfunctions caused by SARS-CoV-2 infection. Finally, we analyzed the sera obtained from COVID-19 patients, and revealed that COVID-19 patients, who were positive for serum viral RNA, are likely to become severe and develop hepatic dysfunctions, as compared with COVID-19 patients who were negative for serum viral RNA. We succeeded in modeling the liver pathophysiology of COVID-19 patients using LoC technology and clinical samples., 新型コロナウイルス感染症(COVID-19)研究のための肝臓チップの開発 --肝障害の病態解明と治療薬の評価--. 京都大学プレスリリース. 2023-03-08., Using organ-on-a-chip technology to elucidate the liver pathophysiology of COVID-19 patients. 京都大学プレスリリース. 2023-03-08.
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- 2023
8. Isoflurane induces Art2‐Rsp5‐dependent endocytosis of Bap2 in yeast
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Yasuhiro Araki, Fumi Kozu, Kanae Shirahama-Noda, Hitoshi Niwa, Shintaro Kira, and Takeshi Noda
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Saccharomyces cerevisiae Proteins ,Amino Acid Transport Systems ,QH301-705.5 ,Ubiquitin-Protein Ligases ,Saccharomyces cerevisiae ,Endocytosis ,General Biochemistry, Genetics and Molecular Biology ,Ubiquitin ,ubiquitin ,medicine ,endocytosis ,Amino acid transporter ,Biology (General) ,Research Articles ,biology ,Endosomal Sorting Complexes Required for Transport ,Isoflurane ,Chemistry ,arrestin ,Cell Membrane ,Ubiquitination ,Ubiquitin-Protein Ligase Complexes ,Transporter ,Ubiquitin ligase ,Cell biology ,Membrane protein ,anesthetic ,transporter ,Anesthetic ,biology.protein ,medicine.drug ,Research Article - Abstract
Although general anesthesia is indispensable during modern surgical procedures, the mechanism by which inhalation anesthetics act on the synaptic membrane at the molecular and cellular level is largely unknown. In this study, we used yeast cells to examine the effect of isoflurane, an inhalation anesthetic, on membrane proteins. Bap2, an amino acid transporter localized on the plasma membrane, was endocytosed when yeast cells were treated with isoflurane. Depletion of RSP5, an E3 ligase, prevented this endocytosis and Bap2 was ubiquitinated in response to isoflurane, indicating an ubiquitin‐dependent process. Screening all the Rsp5 binding adaptors showed that Art2 plays a central role in this process. These results suggest that isoflurane affects Bap2 via an Art2‐Rsp5‐dependent ubiquitination system., Bap2, an amino acid transporter localized on the plasma membrane in yeast, was endocytosed when yeast cells were treated with isoflurane, general anesthesia. Depletion of RSP5, an E3 ligase, and Art2 (arrestin) prevented this endocytosis and Bap2. Thus, isoflurane affects Bap2 via an Art2‐Rsp5‐dependent ubiquitination system.
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- 2021
9. Electrical Treeing Breakdown Characteristics of Epoxy/Spherical Boron Nitride with Card‐House Structure Composites
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Naohiro Hozumi, Tomohiro Kawashima, Takeshi Noda, and Yoshinobu Murakami
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chemistry.chemical_compound ,Thermal conductivity ,Materials science ,chemistry ,Boron nitride ,visual_art ,Composite number ,visual_art.visual_art_medium ,Electrical treeing ,Epoxy ,Electrical and Electronic Engineering ,Composite material - Published
- 2021
10. A CRISPR/Cas9‐based method for seamless N‐terminal protein tagging in <scp> Saccharomyces cerevisiae </scp>
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Shintaro Kira and Takeshi Noda
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Protein kinase complex ,Saccharomyces cerevisiae Proteins ,Expression vector ,biology ,Cas9 ,Saccharomyces cerevisiae ,Bioengineering ,Protein tag ,Computational biology ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Biochemistry ,Plasmid ,Genetics ,CRISPR ,CRISPR-Cas Systems ,Homologous recombination ,Plasmids ,RNA, Guide, Kinetoplastida ,Biotechnology - Abstract
Protein tagging is an effective method for characterizing a gene of interest. Tagging can be accomplished in vivo in Saccharomyces cerevisiae by chromosomal integration of a PCR-amplified cassette. However, common tagging cassettes are not suitable for in situ N-terminal tagging when we aim to preserve the gene's endogenous promoter. Existing methods require either two rounds of homologous recombination or a relatively complex cloning process to construct strains with N-terminal protein tags. Here, we describe a simple CRISPR/Cas9-based method for seamless N-terminal tagging of yeast genes that preserves their endogenous promoter. This method enables the generation of N-terminally tagged strains by introducing an expression vector containing the cas9 gene and a specific gRNA for cleaving the 5' end of the target gene's protein-coding sequence, along with donor DNA containing the tag sequence and homology arms. gRNA cloning was executed by inverse PCR instead of the conventional method. After verifying the tag, the Cas9 and gRNA expression plasmids were eliminated without using antibiotic-containing medium. By this method, we generated strains that express N-terminally tagged subunits of the TORC1 protein kinase complex and found that these strains are comparable to strains made by conventional methods. Thus, our method provides a cost-effective alternative for seamless N-terminal tagging in baker's yeast.
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- 2021
11. Antiviral Face Mask Functionalized with Solidified Hand Soap: Low-Cost Infection Prevention Clothing against Enveloped Viruses Such as SARS-CoV-2
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Alba Cano-Vicent, Ángel Serrano-Aroca, Yukiko Muramoto, Alberto Tuñón-Molina, Kazuo Takayama, Takeshi Noda, and Miguel Martí
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2019-20 coronavirus outbreak ,Coronavirus disease 2019 (COVID-19) ,SARS-CoV-2 ,business.industry ,Transmission (medicine) ,General Chemical Engineering ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,technology, industry, and agriculture ,Human keratinocyte ,General Chemistry ,Antimicrobial agents ,Clothing ,Virology ,Article ,Chemistry ,Viral envelope ,Biological waste ,Viruses ,parasitic diseases ,Genetics ,Infectious diseases ,Infection control ,Medicine ,business ,QD1-999 - Abstract
Infection prevention clothing is becoming an essential protective tool in the current pandemic, especially because now we know that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can easily infect humans in poorly ventilated indoor spaces. However, commercial infection prevention clothing is made of fabrics that are not capable of inactivating the virus. Therefore, viral infections of symptomatic and asymptomatic individuals wearing protective clothing such as masks can occur through aerosol transmission or by contact with the contaminated surfaces of the masks, which are suspected as an increasing source of highly infectious biological waste. Herein, we report an easy fabrication method of a novel antiviral non-woven fabric containing polymer filaments that were coated with solidified hand soap. This extra protective fabric is capable of inactivating enveloped viruses such as SARS-CoV-2 and phage Φ6 within 1 min of contact. In this study, this antiviral fabric was used to fabricate an antiviral face mask and did not show any cytotoxic effect in human keratinocyte HaCaT cells. Furthermore, this antiviral non-woven fabric could be used for the fabrication of other infection prevention clothing such as caps, scrubs, shirts, trousers, disposable gowns, overalls, hoods, aprons, and shoe covers. Therefore, this low-cost technology could provide a wide range of infection-protective tools to combat COVID-19 and future pandemics in developed and underdeveloped countries.
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- 2021
12. A circulating subset of iNKT cells mediates antitumor and antiviral immunity
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Guangwei Cui, Akihiro Shimba, Jianshi Jin, Taisaku Ogawa, Yukiko Muramoto, Hitoshi Miyachi, Shinya Abe, Takuma Asahi, Shizue Tani-ichi, Johannes M. Dijkstra, Yayoi Iwamoto, Kirill Kryukov, Yuanbo Zhu, Daichi Takami, Takahiro Hara, Satsuki Kitano, Yan Xu, Hajime Morita, Moyu Zhang, Lynn Zreka, Keishi Miyata, Takashi Kanaya, Shinya Okumura, Takashi Ito, Etsuro Hatano, Yoshimasa Takahashi, Hiroshi Watarai, Yuichi Oike, Tadashi Imanishi, Hiroshi Ohno, Toshiaki Ohteki, Nagahiro Minato, Masato Kubo, Georg A. Holländer, Hideki Ueno, Takeshi Noda, Katsuyuki Shiroguchi, and Koichi Ikuta
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Interleukin-15 ,Mice ,Immunology ,Humans ,Animals ,Natural Killer T-Cells ,Receptors, Natural Killer Cell ,Receptors, Chemokine ,General Medicine ,Antiviral Agents ,Lipids ,Granzymes - Abstract
Invariant natural killer T (iNKT) cells are a group of innate-like T lymphocytes that recognize lipid antigens. They are supposed to be tissue resident and important for systemic and local immune regulation. To investigate the heterogeneity of iNKT cells, we recharacterized iNKT cells in the thymus and peripheral tissues. iNKT cells in the thymus were divided into three subpopulations by the expression of the natural killer cell receptor CD244 and the chemokine receptor CXCR6 and designated as C0 (CD244⁻CXCR6⁻), C1 (CD244⁻CXCR6⁺), or C2 (CD244⁺CXCR6⁺) iNKT cells. The development and maturation of C2 iNKT cells from C0 iNKT cells strictly depended on IL-15 produced by thymic epithelial cells. C2 iNKT cells expressed high levels of IFN-γ and granzymes and exhibited more NK cell–like features, whereas C1 iNKT cells showed more T cell–like characteristics. C2 iNKT cells were influenced by the microbiome and aging and suppressed the expression of the autoimmune regulator AIRE in the thymus. In peripheral tissues, C2 iNKT cells were circulating that were distinct from conventional tissue-resident C1 iNKT cells. Functionally, C2 iNKT cells protected mice from the tumor metastasis of melanoma cells by enhancing antitumor immunity and promoted antiviral immune responses against influenza virus infection. Furthermore, we identified human CD244⁺CXCR6⁺ iNKT cells with high cytotoxic properties as a counterpart of mouse C2 iNKT cells. Thus, this study reveals a circulating subset of iNKT cells with NK cell–like properties distinct from conventional tissue-resident iNKT cells., 新規の循環型iNKT細胞を発見 --抗腫瘍・抗ウイルス感染効果の高い免疫細胞療法の開発への貢献に期待--. 京都大学プレスリリース. 2022-10-24.
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- 2022
13. Filovirus helical nucleocapsid structures
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Shangfan Hu and Takeshi Noda
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Structural Biology ,Radiology, Nuclear Medicine and imaging ,Instrumentation - Abstract
Filoviruses are filamentous enveloped viruses belonging to the family Filoviridae, in the order Mononegavirales. Some filovirus members, such as Ebola virus (EBOV) and Marburg virus (MARV), cause severe hemorrhagic fever in humans and non-human primates. The filovirus ribonucleoprotein complex, called the nucleocapsid, forms a double-layered helical structure in which a non-segmented, single-stranded negative-sense RNA genome is encapsidated by nucleoprotein (NP), viral protein 35 (VP35), VP24, VP30, and RNA-dependent RNA polymerase (L). The inner layer consists of the helical NP-RNA complex, acting as a scaffold for the binding of VP35 and VP24 that constitute the outer layer. Recent structural studies using cryo-electron microscopy have advanced our understanding of the molecular mechanism of filovirus nucleocapsid formation. Here, we review the key characteristics of the EBOV and MARV nucleocapsid structures, highlighting the similarities and differences between the two viruses. In particular, we focus on the structure of the helical NP-RNA complex, the RNA binding mechanism, and the NP-NP interactions in the helix. The structural analyses reveal a possible mechanism of nucleocapsid assembly and provide potential targets for the anti-filovirus drug design. Mini-abstract Filovirus nucleocapsid is responsible for viral genome transcription and replication. Here, we summarize the helical nucleocapsid structure of Ebola and Marburg viruses, focusing on the nucleoprotein-RNA helix of the nucleocapsid core structure, which has been recently determined by cryo-electron microscopy.
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- 2022
14. SARS-CoV-2 disrupts respiratory vascular barriers by suppressing Claudin-5 expression
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Rina Hashimoto, Junya Takahashi, Keisuke Shirakura, Risa Funatsu, Kaori Kosugi, Sayaka Deguchi, Masaki Yamamoto, Yugo Tsunoda, Maaya Morita, Kosuke Muraoka, Masato Tanaka, Tomoaki Kanbara, Shota Tanaka, Shigeyuki Tamiya, Nagisa Tokunoh, Atsushi Kawai, Masahito Ikawa, Chikako Ono, Keisuke Tachibana, Masuo Kondoh, Masanori Obana, Yoshiharu Matsuura, Akihiro Ohsumi, Takeshi Noda, Takuya Yamamoto, Yasuo Yoshioka, Yu-suke Torisawa, Hiroshi Date, Yasushi Fujio, Miki Nagao, Kazuo Takayama, and Yoshiaki Okada
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Tight Junction Proteins ,Multidisciplinary ,SARS-CoV-2 ,COVID-19 ,Endothelial Cells ,Humans ,Claudin-5 ,Fluvastatin - Abstract
In the initial process of coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects respiratory epithelial cells and then transfers to other organs the blood vessels. It is believed that SARS-CoV-2 can pass the vascular wall by altering the endothelial barrier using an unknown mechanism. In this study, we investigated the effect of SARS-CoV-2 on the endothelial barrier using an airway-on-a-chip that mimics respiratory organs and found that SARS-CoV-2 produced from infected epithelial cells disrupts the barrier by decreasing Claudin-5 (CLDN5), a tight junction protein, and disrupting vascular endothelial cadherin–mediated adherens junctions. Consistently, the gene and protein expression levels of CLDN5 in the lungs of a patient with COVID-19 were decreased. CLDN5 overexpression or Fluvastatin treatment rescued the SARS-CoV-2–induced respiratory endothelial barrier disruption. We concluded that the down-regulation of CLDN5 expression is a pivotal mechanism for SARS-CoV-2–induced endothelial barrier disruption in respiratory organs and that inducing CLDN5 expression is a therapeutic strategy against COVID-19., 臓器チップ技術を用いて新型コロナウイルスが血管へ侵入するメカニズムを解明 --Claudin-5発現抑制による呼吸器の血管内皮バリア破壊--. 京都大学プレスリリース. 2022-09-22., A study using an organ-on-a-chip reveals a mechanism of SARS-CoV-2 invasion into blood vessels --Disruption of vascular endothelial barrier in respiratory organs by decreasing Claudin-5 expression--. 京都大学プレスリリース. 2022-09-27.
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- 2022
15. Role of VP30 Phosphorylation in Ebola Virus Nucleocapsid Assembly and Transport
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Yuki Takamatsu, Tomoki Yoshikawa, Takeshi Kurosu, Shuetsu Fukushi, Noriyo Nagata, Masayuki Shimojima, Hideki Ebihara, Masayuki Saijo, and Takeshi Noda
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Structure and Assembly ,Virus Assembly ,Immunology ,Virion ,Biological Transport ,Hemorrhagic Fever, Ebola ,Ebolavirus ,Microbiology ,Viral Proteins ,Virology ,Insect Science ,Humans ,Phosphorylation ,Nucleocapsid ,Transcription Factors - Abstract
Ebola virus (EBOV) VP30 regulates viral genome transcription and replication by switching its phosphorylation status. However, the importance of VP30 phosphorylation and dephosphorylation in other viral replication processes such as nucleocapsid and virion assembly is unclear. Interestingly, VP30 is predominantly dephosphorylated by cellular phosphatases in viral inclusions, while it is phosphorylated in the released virions. Thus, uncertainties regarding how VP30 phosphorylation in nucleocapsids is achieved and whether VP30 phosphorylation provides any advantages in later steps in viral replication have arisen. In the present study, to characterize the roles of VP30 phosphorylation in nucleocapsid formation, we used electron microscopic analyses and live cell imaging systems. We identified VP30 localized to the surface of protrusions surrounding nucleoprotein (NP)-forming helical structures in the nucleocapsid, suggesting the involvement in assembly and transport of nucleocapsids. Interestingly, VP30 phosphorylation facilitated its association with nucleocapsid-like structures (NCLSs). On the contrary, VP30 phosphorylation does not influence the transport characteristics and NCLS number leaving from and coming back into viral inclusions, indicating that the phosphorylation status of VP30 is not a prerequisite for NCLS departure. Moreover, the phosphorylation status of VP30 did not cause major differences in nucleocapsid transport in authentic EBOV-infected cells. In the following budding step, the association of VP30 and its phosphorylation status did not influence the budding efficiency of virus-like particles. Taken together, it is plausible that EBOV may utilize the phosphorylation of VP30 for its selective association with nucleocapsids, without affecting nucleocapsid transport and virion budding processes. IMPORTANCE Ebola virus (EBOV) causes severe fevers with unusually high case fatality rates. The nucleocapsid provides the template for viral genome transcription and replication. Thus, understanding the regulatory mechanism behind its formation is important for the development of novel therapeutic approaches. Previously, we established a live-cell imaging system based on the ectopic expression of viral fluorescent fusion proteins, allowing the visualization and characterization of intracytoplasmic transport of nucleocapsid-like structures. EBOV VP30 is an essential transcriptional factor for viral genome synthesis, and, although its role in viral genome transcription and replication is well understood, the functional importance of VP30 phosphorylation in assembly of nucleocapsids is still unclear. Our work determines the localization of VP30 at the surface of ruffled nucleocapsids, which differs from the localization of polymerase in EBOV-infected cells. This study sheds light on the novel role of VP30 phosphorylation in nucleocapsid assembly, which is an important prerequisite for virion formation.
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- 2022
16. VEGF-Mediated Augmentation of Autophagic and Lysosomal Activity in Endothelial Cells Defends against Intracellular Streptococcus pyogenes
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Shiou-Ling Lu, Hiroko Omori, Yi Zhou, Yee-Shin Lin, Ching-Chuan Liu, Jiunn-Jong Wu, and Takeshi Noda
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Vascular Endothelial Growth Factor A ,Mice ,Streptococcus pyogenes ,Vascular Endothelial Growth Factors ,Sepsis ,Virology ,Autophagy ,Animals ,Endothelial Cells ,Humans ,Lysosomes ,Microbiology - Abstract
Group A Streptococcus (GAS), a deleterious human-pathogenic bacterium, causes life-threatening diseases such as sepsis and necrotic fasciitis. We recently reported that GAS survives and replicates within blood vessel endothelial cells because these cells are intrinsically defective in xenophagy. Because blood vessel endothelial cells are relatively germfree environments, specific stimulation may be required to sufficiently induce xenophagy. Here, we explored how vascular endothelial growth factor (VEGF) promoted xenophagy and lysosomal activity in endothelial cells. These effects were achieved by amplifying the activation of TFEB, a transcriptional factor crucial for lysosome/autophagy biogenesis, via cAMP-mediated calcium release. In a mouse model of local infection with GAS, the VEGF level was significantly elevated at the infection site. Interestingly, low serum VEGF levels were found in a mouse model of invasive bacteremia and in patients with severe GAS-induced sepsis. Moreover, the administration of VEGF improved the survival of GAS-infected mice. We propose a novel theory regarding GAS infection in endothelial cells, wherein VEGF concentrations in the systemic circulation play a critical role.
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- 2022
17. Acetylation of the influenza A virus polymerase subunit PA in the N‐terminal domain positively regulates its endonuclease activity
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Masahiro Nakano, Dai Hatakeyama, Takashi Kuzuhara, Hazuki Tsuneishi, Erina Nishikawa, Seiryo Ogata, Mizuki Takahira, Kyoko Makiyama, Takeshi Masuda, Ayaka Saitoh, Masaki Shoji, Tsugunori Komatsu, Takeshi Noda, Sumio Ohtsuki, Yoshihiro Kawaoka, Asuka Miyatake, and Ayana Ohkubo
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Protein subunit ,medicine.disease_cause ,Biochemistry ,Viral Transcription ,Viral Proteins ,Endonuclease ,chemistry.chemical_compound ,Transcription (biology) ,RNA polymerase ,Influenza, Human ,Influenza A virus ,medicine ,Humans ,p300-CBP Transcription Factors ,Amino Acid Sequence ,Molecular Biology ,Polymerase ,Histone Acetyltransferases ,biology ,Chemistry ,Acetylation ,Cell Biology ,RNA-Dependent RNA Polymerase ,Molecular biology ,Nucleoprotein ,Nucleoproteins ,biology.protein ,RNA, Viral ,Protein Processing, Post-Translational ,Protein Binding - Abstract
The post-translational acetylation of lysine residues is found in many nonhistone proteins and is involved in a wide range of biological processes. Recently, we showed that the nucleoprotein of the influenza A virus is acetylated by histone acetyltransferases (HATs), a phenomenon that affects viral transcription. Here, we report that the PA subunit of influenza A virus RNA-dependent RNA polymerase is acetylated by the HATs, P300/CREB-binding protein-associated factor (PCAF), and general control nonderepressible 5 (GCN5), resulting in accelerated endonuclease activity. Specifically, the full-length PA subunit expressed in cultured 293T cells was found to be strongly acetylated. Moreover, the partial recombinant protein of the PA N-terminal region containing the endonuclease domain was also acetylated by PCAF and GCN5 in vitro, which facilitated its endonuclease activity. Mass spectrometry analyses identified K19 as a candidate acetylation target in the PA N-terminal region. Notably, the substitution of the lysine residue at position 19 with glutamine, a mimic of the acetyl-lysine residue, enhanced its endonuclease activity in vitro; this point mutation also accelerated influenza A virus RNA-dependent RNA polymerase activity in the cell. Our findings suggest that PA acetylation is important for the regulation of the endonuclease and RNA polymerase activities of the influenza A virus.
- Published
- 2021
18. Annealing-Induced Structural Evolution of InAs Quantum Dots on InP (111)A Formed by Droplet Epitaxy
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Takaaki Mano, Kazuaki Sakoda, Akihiro Ohtake, Neul Ha, Takashi Kuroda, Yoshiki Sakuma, and Takeshi Noda
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Materials science ,Quantum dot ,business.industry ,Optoelectronics ,General Materials Science ,General Chemistry ,Condensed Matter Physics ,Epitaxy ,business ,Structural evolution ,Annealing (glass) - Published
- 2021
19. Cell response analysis in SARS-CoV-2 infected bronchial organoids
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Emi Sano, Tatsuya Suzuki, Rina Hashimoto, Yumi Itoh, Ayaka Sakamoto, Yusuke Sakai, Akatsuki Saito, Daisuke Okuzaki, Daisuke Motooka, Yukiko Muramoto, Takeshi Noda, Tomohiko Takasaki, Jun-Ichi Sakuragi, Shohei Minami, Takeshi Kobayashi, Takuya Yamamoto, Yasufumi Matsumura, Miki Nagao, Toru Okamoto, and Kazuo Takayama
- Subjects
Organoids ,Respiratory tract diseases ,SARS-CoV-2 ,viruses ,COVID-19 ,Humans ,Medicine (miscellaneous) ,Bronchi ,respiratory system ,General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology ,respiratory tract diseases - Abstract
The development of an in vitro cell model that can be used to study severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) research is expected. Here we conducted infection experiments in bronchial organoids (BO) and an BO-derived air-liquid interface model (BO-ALI) using 8 SARS-CoV-2 variants. The infection efficiency in BO-ALI was more than 1, 000 times higher than that in BO. Among the bronchial epithelial cells, we found that ciliated cells were infected with the virus, but basal cells were not. Ciliated cells died 7 days after the viral infection, but basal cells survived after the viral infection and differentiated into ciliated cells. Fibroblast growth factor 10 signaling was essential for this differentiation. These results indicate that BO and BO-ALI may be used not only to evaluate the cell response to SARS-CoV-2 and coronavirus disease 2019 (COVID-19) therapeutic agents, but also for airway regeneration studies., 気管支オルガノイドを用いた新型コロナウイルス研究とその創薬応用. 京都大学プレスリリース. 2022-05-30., COVID-19 Research Using Bronchial Organoids and Drug Discovery Applications. 京都大学プレスリリース. 2022-06-02.
- Published
- 2022
20. Automated amplification-free digital RNA detection platform for rapid and sensitive SARS-CoV-2 diagnosis
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Hajime Shinoda, Tatsuya Iida, Asami Makino, Mami Yoshimura, Junichiro Ishikawa, Jun Ando, Kazue Murai, Katsumi Sugiyama, Yukiko Muramoto, Masahiro Nakano, Kotaro Kiga, Longzhu Cui, Osamu Nureki, Hiroaki Takeuchi, Takeshi Noda, Hiroshi Nishimasu, and Rikiya Watanabe
- Subjects
COVID-19 Testing ,stomatognathic system ,Lab-on-a-chip ,Single-molecule biophysics ,SARS-CoV-2 ,COVID-19 ,Humans ,RNA, Viral ,Medicine (miscellaneous) ,Nanobiotechnology ,General Agricultural and Biological Sciences ,Pandemics ,General Biochemistry, Genetics and Molecular Biology - Abstract
In the ongoing COVID-19 pandemic, rapid and sensitive diagnosis of viral infection is a critical deterrent to the spread of SARS-CoV-2. To this end, we developed an automated amplification-free digital RNA detection platform using CRISPR-Cas13a and microchamber device (opn-SATORI), which automatically completes a detection process from sample mixing to RNA quantification in clinical specimens within ~9 min. Using the optimal Cas13a enzyme and magnetic beads technology, opn-SATORI detected SARS-CoV-2 genomic RNA with a LoD of, 新型コロナウイルスの超高感度・全自動迅速検出装置の開発 --汎用的な感染症診断装置としての社会実装に期待--. 京都大学プレスリリース. 2022-05-27.
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- 2022
21. Cyclin J–CDK complexes limit innate immune responses by reducing proinflammatory changes in macrophage metabolism
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Yee Kien Chong, Sarang Tartey, Yuki Yoshikawa, Koshi Imami, Songling Li, Masanori Yoshinaga, Ai Hirabayashi, Guohao Liu, Alexis Vandenbon, Fabian Hia, Takuya Uehata, Takashi Mino, Yutaka Suzuki, Takeshi Noda, Dominique Ferrandon, Daron M. Standley, Yasushi Ishihama, and Osamu Takeuchi
- Subjects
Mice ,Cyclins ,Macrophages ,Toll-Like Receptors ,Animals ,Humans ,Forkhead Transcription Factors ,Cell Biology ,Reactive Oxygen Species ,Molecular Biology ,Biochemistry ,Cyclin-Dependent Kinases ,Immunity, Innate - Abstract
Toll-like receptor (TLR) stimulation induces glycolysis and the production of mitochondrial reactive oxygen species (ROS), both of which are critical for inflammatory responses in macrophages. Here, we demonstrated that cyclin J, a TLR-inducible member of the cyclin family, reduced cytokine production in macrophages by coordinately controlling glycolysis and mitochondrial functions. Cyclin J interacted with cyclin-dependent kinases (CDKs), which increased the phosphorylation of a subset of CDK substrates, including the transcription factor FoxK1 and the GTPase Drp1. Cyclin J–dependent phosphorylation of FoxK1 decreased the transcription of glycolytic genes and Hif-1α activation, whereas hyperactivation of Drp1 by cyclin J–dependent phosphorylation promoted mitochondrial fragmentation and impaired the production of mitochondrial ROS. In mice, cyclin J in macrophages limited the growth of tumor xenografts and protected against LPS-induced shock but increased the susceptibility to bacterial infection. Collectively, our findings indicate that cyclin J–CDK signaling promotes antitumor immunity and the resolution of inflammation by opposing the metabolic changes that drive inflammatory responses in macrophages.
- Published
- 2022
22. Migration of Influenza Virus Nucleoprotein into the Nucleolus Is Essential for Ribonucleoprotein Complex Formation
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Sho Miyamoto, Masahiro Nakano, Takeshi Morikawa, Ai Hirabayashi, Ryoma Tamura, Yoko Fujita-Fujiharu, Nanami Hirose, Yukiko Muramoto, and Takeshi Noda
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assembly ,viruses ,Virology ,nucleolus ,Microbiology ,influenza virus ,Research Article ,RNP - Abstract
Influenza A virus double-helical ribonucleoprotein complex (RNP) performs transcription and replication of viral genomic RNA (vRNA). Although RNP formation occurs in the nuclei of virus-infected cells, the nuclear domains involved in this process remain unclear. Here, we show that the nucleolus is an essential site for functional RNP formation. Viral nucleoprotein (NP), a major RNP component, temporarily localized to the nucleoli of virus-infected cells. Mutations in a nucleolar localization signal (NoLS) on NP abolished double-helical RNP formation, resulting in a loss of viral RNA synthesis ability, whereas ectopic fusion of the NoLS enabled the NP mutant to form functional double-helical RNPs. Furthermore, nucleolar disruption of virus-infected cells inhibited NP assembly into double-helical RNPs, resulting in decreased viral RNA synthesis. Collectively, our findings demonstrate that NP migration into the nucleolus is a critical step for functional RNP formation, showing the importance of the nucleolus in the influenza virus life cycle.
- Published
- 2022
23. STEEP mediates STING ER exit and activation of signaling
- Author
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Chiranjeevi Bodda, Søren B. Jensen, Carsten Scavenius, Anne Troldborg, Christine Doucet, Cheng long Sun, Ramya Nandakumar, Anders Laustsen, Thaneas Prabakaran, Zong liang Gao, Samuel J. Windross, Harald Stenmark, Zheng Guo, Line S. Reinert, Sonia Assil, Rune Hartmann, Søren R. Paludan, Jinrong Huang, Martin K. Thomsen, Yan Zhang, Ryo Narita, Jan J. Enghild, Bao cun Zhang, Zhijian J. Chen, Raphaela Goldbach-Mansky, Takeshi Noda, David Olagnier, Rasmus O. Bak, Cong gang Zhang, Martin F. Berthelsen, Yujia Cai, Department of Biomedicine Aarhus, Department of Clinical Medicine (Aarhus University), Department of Chemistry and Interdisciplinary Nanoscience Center, Aarhus University, Department of Molecular Biology, University of Aarhus, Institute for Cancer Research [Oslo], Oslo University Hospital [Oslo], Centre de Biochimie Structurale [Montpellier] (CBS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Graduate School of Frontier Biosciences [Osaka], Osaka University [Osaka], National Institute of Allergy and Infectious Diseases [Bethesda] (NIAID-NIH), National Institutes of Health [Bethesda] (NIH), The Novo Nordisk Foundation (NNF18OC0030274) and the Lundbeck Foundation (R198-2015-171, R268-2016-3927), B.-c.Z. is funded by a postdoctoral grant from the Danish Council for Independent Research, Medical Sciences (5053-00083B), the postdoctoral salary to S.A. was funded by the European Union under the Horizon 2020 Research and Innovation Program and Marie Skłodowska-Curie Actions (MSCA) – international fellowship (PathAutoBio 796840), the PhD scholarship to S.J.W. was funded by the European Union under the Horizon 2020 Research and Innovation Program and the MSCA-Innovative Training Networks Programme MSCA-ITN (EDGE, 675278), the postdoctoral grant to A.T. was funded by the Lundbeck Foundation (R264-2017-3344)., and European Project: 786602,ENVISION
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0301 basic medicine ,cGAS-STING pathway ,[SDV]Life Sciences [q-bio] ,Regulator ,DNA VIRUSES ,Stimulation ,Endoplasmic Reticulum ,ADAPTER ,Mice ,0302 clinical medicine ,Gene expression ,Lupus Erythematosus, Systemic ,Immunology and Allergy ,SYNTHASE ,PHOSPHORYLATION ,Chemistry ,Nuclear Proteins ,3. Good health ,Cell biology ,TRANSLOCATION ,Protein Transport ,Membrane curvature ,symbols ,2ND-MESSENGER ,AUTOPHAGY ,Innate immunology ,medicine.symptom ,Signal Transduction ,DNA sensing ,intracellular trafficking ,Immunology ,Nerve Tissue Proteins ,Inflammation ,Article ,03 medical and health sciences ,symbols.namesake ,medicine ,Animals ,Humans ,Endoplasmic reticulum ,Membrane Proteins ,CYCLIC GMP-AMP ,Golgi apparatus ,eye diseases ,Sting ,030104 developmental biology ,SAVI ,Gene Expression Regulation ,INNATE IMMUNITY ,030215 immunology - Abstract
STING is essential for control of infections and for tumor immunosurveillance, but it can also drive pathological inflammation. STING resides on the endoplasmic reticulum (ER) and traffics following stimulation to the ERGIC/Golgi, where signaling occurs. Although STING ER exit is the rate-limiting step in STING signaling, the mechanism that drives this process is not understood. Here we identify STEEP as a positive regulator of STING signaling. STEEP was associated with STING and promoted trafficking from the ER. This was mediated through stimulation of phosphatidylinositol-3-phosphate (PtdIns(3)P) production and ER membrane curvature formation, thus inducing COPII-mediated ER-to-Golgi trafficking of STING. Depletion of STEEP impaired STING-driven gene expression in response to virus infection in brain tissue and in cells from patients with STING-associated diseases. Interestingly, STING gain-of-function mutants from patients interacted strongly with STEEP, leading to increased ER PtdIns(3)P levels and membrane curvature. Thus, STEEP enables STING signaling by promoting ER exit. STING ER exit is the rate-limiting step in STING signaling, but the mechanism that drives this process is not understood. Paludan and colleagues identify CxORF56, called STEEP here, as a positive regulator of STING signaling.
- Published
- 2020
24. Influenza A virus NS1 optimises virus infectivity by enhancing genome packaging in a dsRNA-binding dependent manner
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Michaela Weber, Masahiro Nakano, Dacquin M. Kasumba, Hiroki Kato, Takeshi Noda, Tim Wai Sha, and Takashi Fujita
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0301 basic medicine ,Viral protein ,viruses ,Genome, Viral ,Biology ,Viral Nonstructural Proteins ,medicine.disease_cause ,Recombinant virus ,Virus Replication ,Virus ,Cell Line ,Madin Darby Canine Kidney Cells ,lcsh:Infectious and parasitic diseases ,03 medical and health sciences ,Viral genome packaging ,Dogs ,Transcription (biology) ,Virology ,Influenza A virus ,medicine ,Animals ,Humans ,lcsh:RC109-216 ,RNA, Double-Stranded ,Non-structural protein 1 ,Innate immunity ,Innate immune system ,030102 biochemistry & molecular biology ,Research ,virus diseases ,Viral Genome Packaging ,dsRNA binding domain ,biochemical phenomena, metabolism, and nutrition ,Influenza a virus ,Genome packaging ,030104 developmental biology ,Infectious Diseases ,HEK293 Cells ,Viral replication ,RNA, Viral ,Virus-like production system - Abstract
Background The non-structural protein 1 (NS1) of influenza A virus (IAV) is a key player in inhibiting antiviral response in host cells, thereby facilitating its replication. However, other roles of NS1, which are independent of antagonising host cells’ antiviral response, are less characterised. Methods To investigate these unidentified roles, we used a recombinant virus, which lacks NS1 expression, and observed its phenotypes during the infection of antiviral defective cells (RIG-I KO cells) in the presence or absence of exogeneous NS1. Moreover, we used virus-like particle (VLP) production system to further support our findings. Results Our experiments demonstrated that IAV deficient in NS1 replicates less efficiently than wild-type IAV in RIG-I KO cells and this replication defect was complemented by ectopic expression of NS1. As suggested previously, NS1 is incorporated in the virion and participates in the regulation of viral transcription and translation. Using the VLP production system, in which minigenome transcription or viral protein production was unaffected by NS1, we demonstrated that NS1 facilitates viral genome packaging into VLP, leading to efficient minigenome transfer by VLP. Furthermore, the incorporation of NS1 and the minigenome into VLP were impaired by introducing a point mutation (R38A) in the double stranded RNA-binding domain of NS1. Conclusion These results suggest a novel function of NS1 in improving genome packaging in a dsRNA binding-dependent manner. Taken together, NS1 acts as an essential pro-viral regulator, not only by antagonizing host immunity but also by facilitating viral replication and genome packaging.
- Published
- 2020
25. Surface-Controlled Oriented Growth of FASnI3 Crystals for Efficient Lead-free Perovskite Solar Cells
- Author
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Takeshi Noda, Xin He, Xiangyue Meng, Tianhao Wu, Xiao Liu, Liyuan Han, Yanbo Wang, Xudong Yang, Julien Barbaud, and Jianbo Lin
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chemistry.chemical_classification ,Electron mobility ,Materials science ,business.industry ,Iodide ,Energy conversion efficiency ,chemistry.chemical_element ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,Surface energy ,0104 chemical sciences ,law.invention ,General Energy ,chemistry ,law ,Optoelectronics ,Charge carrier ,Crystallization ,0210 nano-technology ,Tin ,business ,Perovskite (structure) - Abstract
Summary Tin-based perovskites with narrow band gap and high carrier mobility are promising candidates for the preparation of efficient lead-free perovskite solar cells (PSCs). However, the poor crystallization of tin-based perovskite films with unfavorable defects and rough morphology is still the biggest challenge to achieve high power conversion efficiency. In this work, we reveal the surface-controlled growth of FASnI3 perovskite films, and then precisely control the crystallization process by reducing the surface energy of the solution-air surface with a tailor-made pentafluorophen-oxyethylammonium iodide (FOEI). The demonstrated strategy enabled us to achieve highly oriented and smooth FASnI3-FOEI films with lower defect density and longer lifetime of charge carriers. A certificated efficiency of 10.16% for tin-based PSCs was obtained from an accredited institute.
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- 2020
26. Highly Reproducible and Efficient FASnI3 Perovskite Solar Cells Fabricated with Volatilizable Reducing Solvent
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Xin He, Yanbo Wang, Hiroshi Segawa, Takeshi Noda, Xiao Liu, Tianhao Wu, Liyuan Han, and Xiangyue Meng
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Materials science ,Band gap ,Doping ,chemistry.chemical_element ,Halide ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Solvent ,Crystallinity ,Formamidinium ,Chemical engineering ,chemistry ,General Materials Science ,Physical and Theoretical Chemistry ,0210 nano-technology ,Tin ,Perovskite (structure) - Abstract
Lead-free tin halide perovskite solar cells (PSCs) have attracted great attention because of their low toxicity, ideal band gap, and high carrier mobilities. However, the efficiency and reproducibility of tin halide PSCs has been limited because of the facile oxidation of Sn2+ to Sn4+. Herein, liquid formic acid (LFA) was introduced as a reducing solvent in the FASnI3 (FA: formamidinium) perovskite precursor solution. Unlike solid reducing additives, the LFA solvent is volatile, so no residual LFA remained in the FASnI3 perovskite film. Use of the LFA solvent resulted in production of the FASnI3 perovskite film with high crystallinity, low Sn4+ content, reduced background doping, and low electronic trap density. As a result, an efficiency of over 10% was obtained for lead-free tin halide PSCs with improved reproducibility.
- Published
- 2020
27. Templated growth of FASnI3 crystals for efficient tin perovskite solar cells
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Xiangyue Meng, Takeshi Noda, Xin He, Xiao Liu, Hiroshi Segawa, Tianhao Wu, Yanbo Wang, Han Chen, Liyuan Han, Xudong Yang, and Jung-Yao Chen
- Subjects
chemistry.chemical_classification ,Materials science ,Renewable Energy, Sustainability and the Environment ,Annealing (metallurgy) ,Iodide ,Halide ,chemistry.chemical_element ,Pollution ,Formamidinium ,Nuclear Energy and Engineering ,chemistry ,Chemical engineering ,Trap density ,Environmental Chemistry ,Tin - Abstract
Tin perovskite solar cells (TPSCs) are rising as the most promising candidates for lead-free PSCs. However, the randomly crystallized tin halide perovskite with a high concentration of defects is still the bottleneck for obtaining high efficiency. Here, we report the pretreatment of FASnI3 (formamidinium, FA) perovskite film by spin-coating a solution of organic halide salt n-propylammonium iodide (PAI) before annealing. It is found that PAI can induce the templated growth of FASnI3 crystals by reconstruction of the intermediate phase, resulting in a highly crystallized FASnI3 film with preferential orientation along the (100) plane and suppressed trap density. Finally, a stabilized efficiency of 11.22% was certified at an accredited test center and the cell is stable, maintaining over 95% of its initial efficiency after 1000 hours of operation at the maximum power point.
- Published
- 2020
28. Highly efficient tin perovskite solar cells achieved in a wide oxygen concentration range
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Jihuai Wu, Liyuan Han, Takeshi Noda, Tianhao Wu, Xudong Yang, Yutaka Moritomo, Xiangyue Meng, Yanbo Wang, Hiroshi Segawa, Xiao Liu, and Xin He
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Materials science ,Fabrication ,Renewable Energy, Sustainability and the Environment ,Energy conversion efficiency ,chemistry.chemical_element ,02 engineering and technology ,General Chemistry ,Permeation ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,Oxygen ,Grain size ,0104 chemical sciences ,Chemical engineering ,chemistry ,General Materials Science ,Limiting oxygen concentration ,0210 nano-technology ,Tin ,Perovskite (structure) - Abstract
Tin perovskite solar cells (TPSCs) as one of the most promising candidates for lead-free PSCs have large potential to be industrialized in the future. However, TPSCs have often been fabricated at an extremely low oxygen concentration to avoid oxidation from Sn2+ to Sn4+, which is not applicable to their large scale production. In this work, we construct a carbonylate antioxidant capping layer atop the perovskite surface via a solvent engineering method to prevent oxygen permeation into the perovskite layer during the fabrication process of the device to enlarge the oxygen region from 0.1 ppm to 100 ppm. Meanwhile, a pinhole-free perovskite film with a large grain size, reduced Sn4+ defects and long carrier recombination lifetime was fabricated. As a result, high-performance TPSCs could be achieved in a wide range of oxygen concentration with a power conversion efficiency (PCE) of 9.47% at 0.1 ppm oxygen and 9.03% at 100 ppm oxygen, respectively; the device was stable, maintaining over 93% of its initial PCE after operating at the maximum power point under continuous light soaking (AM 1.5G, 100 mW cm−2) for 600 hours.
- Published
- 2020
29. Efficient and stable tin-based perovskite solar cells by introducing π-conjugated Lewis base
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Xin He, Xiangyue Meng, Tianhao Wu, Xudong Yang, Xiao Liu, Takeshi Noda, Liyuan Han, and Yanbo Wang
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Materials science ,Energy conversion efficiency ,chemistry.chemical_element ,02 engineering and technology ,General Chemistry ,Carrier lifetime ,Permeation ,Conjugated system ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,law.invention ,chemistry ,Chemical engineering ,law ,Molecule ,Lewis acids and bases ,Crystallization ,0210 nano-technology ,Tin - Abstract
Tin-based perovskite solar cells (TPSCs) as the most promising candidate for lead-free PSCs have incurred extensive researches all over the world. However, the crystallization process of tin-based perovskite is too fast during the solution-deposited process, resulting in abundant pinholes and poor homogeneity that cause serious charge recombination in perovskite layer. Here, we employed the π-conjugated Lewis base molecules with high electron density to systematically control the crystallization rate of FASnI3 perovskite by forming stable intermediate phase with the Sn-I frameworks, leading to a compact and uniform perovskite film with large increase in the carrier lifetime. Meanwhile, the introduction of the π-conjugated systems also retards the permeation of moisture into perovskite crystal, which significantly suppresses the film degradation in air. These benefits contributed to a stabilizing power conversion efficiency (PCE) of 10.1% for the TPSCs and maintained over 90% of its initial PCE after 1000-h light soaking in air. Also, a steady-state efficiency of 9.2% was certified at the accredited test center.
- Published
- 2019
30. Non-Woven Infection Prevention Fabrics Coated with Biobased Cranberry Extracts Inactivate Enveloped Viruses Such as SARS-CoV-2 and Multidrug-Resistant Bacteria
- Author
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Miguel Martí, José Luis Aparicio-Collado, Yukiko Muramoto, Ángel Serrano-Aroca, Alba Cano-Vicent, Kazuo Takayama, Takeshi Noda, Alberto Tuñón-Molina, and Roser Sabater i Serra
- Subjects
infection prevention clothing ,Antibiotics ,Staphylococcus aureus ,non-woven fabric ,Antimicrobial activity ,medicine.disease_cause ,Anti-Infective Agents ,Staphylococcus epidermidis ,Drug Resistance, Multiple, Bacterial ,Multidrug-resistant ,bacteriophage phi 6 ,Biology (General) ,bacteria ,Pathogen ,Spectroscopy ,Coronavirus ,biology ,Chemistry ,Infection prevention clothing ,Textiles ,General Medicine ,Antimicrobial ,Bacteriophage phi 6 ,Computer Science Applications ,Anti-Bacterial Agents ,Vaccinium macrocarpon ,Methicillin-Resistant Staphylococcus aureus ,medicine.drug_class ,QH301-705.5 ,multidrug-resistant ,Catalysis ,Article ,Microbiology ,Inorganic Chemistry ,Cranberry extract ,medicine ,Animals ,Humans ,Physical and Theoretical Chemistry ,Caenorhabditis elegans ,Molecular Biology ,QD1-999 ,Enveloped viruses ,antimicrobial activity ,Bacteria ,Plant Extracts ,SARS-CoV-2 ,Organic Chemistry ,enveloped viruses ,cranberry extract ,COVID-19 ,biology.organism_classification ,Multiple drug resistance ,Non-woven fabric ,INGENIERIA ELECTRICA - Abstract
[EN] The Coronavirus Disease (COVID-19) pandemic is demanding the rapid action of the authorities and scientific community in order to find new antimicrobial solutions that could inactivate the pathogen SARS-CoV-2 that causes this disease. Gram-positive bacteria contribute to severe pneumonia associated with COVID-19, and their resistance to antibiotics is exponentially increasing. In this regard, non-woven fabrics are currently used for the fabrication of infection prevention clothing such as face masks, caps, scrubs, shirts, trousers, disposable gowns, overalls, hoods, aprons and shoe covers as protective tools against viral and bacterial infections. However, these non-woven fabrics are made of materials that do not exhibit intrinsic antimicrobial activity. Thus, we have here developed non-woven fabrics with antimicrobial coatings of cranberry extracts capable of inactivating enveloped viruses such as SARS-CoV-2 and the bacteriophage phi 6 (about 99% of viral inactivation in 1 min of viral contact), and two multidrug-resistant bacteria: the methicillin-resistant Staphylococcus aureus and the methicillin-resistant Staphylococcus epidermidis. The morphology, thermal and mechanical properties of the produced filters were characterized by optical and electron microscopy, differential scanning calorimetry, thermogravimetry and dynamic mechanical thermal analysis. The non-toxicity of these advanced technologies was ensured using a Caenorhabditis elegans in vivo model. These results open up a new prevention path using natural and biodegradable compounds for the fabrication of infection prevention clothing in the current COVID-19 pandemic and microbial resistant era., This research was supported by the Fundacion Universidad Catolica de Valencia San VicenteMartir, Grant 2020-231-006UCV and by the Ministerio de Ciencia e Innovacion: project PID2020-119333RB-I00/AEI/10.13039/501100011033 (awarded to A.S.-A.) and project RTI2018-097862-B-C21 (awarded to R.S.i.S. including FEDER funding). CIBER-BBN is an initiative funded by the VI National R&D&I Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program. CIBER Actions are financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. This research was also supported by grants from the Japan Agency for Medical Research and Development (AMED) (20fk0108533h0001), and the JST Core Research for Evolutional Science and Technology (JPMJCR20HA). This work was supported by the Joint Usage/Research Center program of Institute for Frontier Life and Medical Sciences Kyoto University.
- Published
- 2021
31. CP100356 Hydrochloride, a P-Glycoprotein Inhibitor, Inhibits Lassa Virus Entry: Implication of a Candidate Pan-Mammarenavirus Entry Inhibitor
- Author
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Ai Hirabayashi, Thomas Strecker, Zihan Zhang, Masahiro Nakano, Allison Groseth, Yuki Takamatsu, Yukiko Muramoto, Junichi Kajikawa, Sho Miyamoto, Sarah Katharina Fehling, Shuzo Urata, Takeshi Noda, and Toru Takenaga
- Subjects
viruses ,Biology ,Lymphocytic choriomeningitis ,medicine.disease_cause ,Antiviral Agents ,Microbiology ,Virus ,Article ,pseudotyped vesicular stomatitis virus ,Lassa Fever ,Viral entry ,Virology ,Chlorocebus aethiops ,medicine ,Animals ,Humans ,ATP Binding Cassette Transporter, Subfamily B, Member 1 ,Lassa fever ,Arenaviridae ,Lassa virus ,arenavirus ,Vero Cells ,entry inhibitor ,Viral Fusion Protein Inhibitors ,Arenavirus ,virus diseases ,Virus Internalization ,medicine.disease ,biology.organism_classification ,Isoquinolines ,QR1-502 ,Entry inhibitor ,lymphocytic choriomeningitis virus ,Infectious Diseases ,Vesicular stomatitis virus ,Quinazolines ,Receptors, Virus ,Vesicular Stomatitis ,medicine.drug - Abstract
Lassa virus (LASV)—a member of the family Arenaviridae—causes Lassa fever in humans and is endemic in West Africa. Currently, no approved drugs are available. We screened 2480 small compounds for their potential antiviral activity using pseudotyped vesicular stomatitis virus harboring the LASV glycoprotein (VSV-LASVGP) and a related prototypic arenavirus, lymphocytic choriomeningitis virus (LCMV). Follow-up studies confirmed that CP100356 hydrochloride (CP100356), a specific P-glycoprotein (P-gp) inhibitor, suppressed VSV-LASVGP, LCMV, and LASV infection with half maximal inhibitory concentrations of 0.52, 0.54, and 0.062 μM, respectively, without significant cytotoxicity. Although CP100356 did not block receptor binding at the cell surface, it inhibited low-pH-dependent membrane fusion mediated by arenavirus glycoproteins. P-gp downregulation did not cause a significant reduction in either VSV-LASVGP or LCMV infection, suggesting that P-gp itself is unlikely to be involved in arenavirus entry. Finally, our data also indicate that CP100356 inhibits the infection by other mammarenaviruses. Thus, our findings suggest that CP100356 can be considered as an effective virus entry inhibitor for LASV and other highly pathogenic mammarenaviruses.
- Published
- 2021
32. Erratum for Cheng et al., 'Group A Streptococcus Induces LAPosomes via SLO/β1 Integrin/NOX2/ROS Pathway in Endothelial Cells That Are Ineffective in Bacterial Killing and Suppress Xenophagy'
- Author
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Cheng Lu Hsieh, Hiroko Omori, Chia Ling Chen, Shang Rung Wu, Chiou Feng Lin, Robert Anderson, Chih Feng Kuo, Yee Shin Lin, Jiunn Jong Wu, Takeshi Noda, Ya Na Wu, Yi Lin Cheng, and Shiou Ling Lu
- Subjects
Streptococcus pyogenes ,Chemistry ,Streptococcus ,Integrin beta1 ,Bacterial killing ,β1 integrin ,Endothelial Cells ,medicine.disease_cause ,Microbiology ,Group A ,QR1-502 ,Cell Line ,Bacterial Proteins ,Virology ,Macroautophagy ,NADPH Oxidase 2 ,Streptolysins ,Vacuoles ,medicine ,Xenophagy ,Humans ,Erratum ,Reactive Oxygen Species ,Microtubule-Associated Proteins - Abstract
Group A streptococcus (GAS) is an important human pathogen which can cause fatal diseases after invasion into the bloodstream. Although antibiotics and immune surveillance are the main defenses against GAS infection, GAS utilizes internalization into cells as a major immune evasion strategy. Our previous findings revealed that light chain 3 (LC3)-associated single membrane GAS-containing vacuoles in endothelial cells are compromised for bacterial clearance due to insufficient acidification after fusion with lysosomes. However, the characteristics and the activation mechanisms of these LC3-positive compartments are still largely unknown. In the present study, we demonstrated that the LC3-positive GAS is surrounded by single membrane and colocalizes with NADPH oxidase 2 (NOX2) complex but without ULK1, which are characteristics of LC3-associated phagocytosis (LAP). Inhibition of NOX2 or reactive oxygen species (ROS) significantly reduces GAS multiplication and enhances autolysosome acidification in endothelial cells through converting LAP to conventional xenophagy, which is revealed by enhancement of ULK1 recruitment, attenuation of p70s6k phosphorylation, and formation of the isolation membrane. We also clarify that the inactivation of mTORC1, which is the initiation signal of autophagy, is inhibited by NOX2- and ROS-activated phosphatidylinositol 3-kinase (PI3K)/AKT and MEK/extracellular signal-regulated kinase (ERK) pathways. In addition, streptolysin O (SLO) of GAS is identified as a crucial inducer of ROS for β1 integrin-mediated LAP induction. After downregulation of β1 integrin, GAS multiplication is reduced, accompanied with LAP inhibition and xenophagy induction. These results demonstrate that GAS infection preferentially induces ineffective LAP to evade xenophagic killing in endothelial cells through the SLO/β1 integrin/NOX2/ROS pathway.
- Published
- 2021
33. Switching of OAS1 splicing isoforms mitigates SARS-CoV-2 infection
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Ryo Kurosawa, Takeshi Noda, Masahiko Ajiro, Toru Takenaga, Masatsugu Denawa, Kei Iida, Yukiko Muramoto, and Masatoshi Hagiwara
- Subjects
Genetics ,Exon ,Innate immune system ,Interferon ,RNA splicing ,Sense (molecular biology) ,medicine ,RNA ,Single-nucleotide polymorphism ,Biology ,medicine.disease_cause ,Coronavirus ,medicine.drug - Abstract
BackgroundThe rapidly accumulating disease susceptibility information collected from coronavirus disease (COVID-19) patient genomes must be urgently utilized to develop therapeutic interventions for SARS-CoV-2 infection. Chromosome 12q24.13, which encodes the 2’-5’-oligoadenylate synthetase (OAS) family of proteins that sense viral genomic RNAs and trigger an antiviral response, is identified as one of the genomic regions that contains SNPs associated with COVID-19 severity. A high-risk SNP identified at the splice acceptor site of OAS1 exon 6 is known to change the proportions of the various splicing isoforms and the activity of the enzyme.MethodsWe employed in-silico motif search and RNA pull-down assay to define a factor responsible for the OAS1 splicing. Next, we rationally selected a candidate for slicing modulator to modulate this splicing.ResultsWe found that inhibition of CDC-like kinase with a small chemical compound induces switching of OAS1 splice isoforms in human lung cells. In this condition, increased resistance to SARS-CoV-2 infection, enhanced RNA degradation, and transcriptional activation of interferon β1, were also observed.ConclusionsThe results indicate the possibility of using chemical splicing modifiers aided by genome-based precision medicine to boost the innate immune response against SARS-CoV-2 infection.
- Published
- 2021
34. Next generation infection prevention clothing: Non-woven Fabrics Coated with Cranberry Extracts Capable of Inactivating Enveloped Viruses such as SARS-CoV-2 and Multidrug-resistant Bacteria
- Author
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Alberto Tuñón-Molina, Kazuo Takayama, Alba Cano-Vicent, Yukiko Muramoto, Takeshi Noda, Miguel Martí, and Ángel Serrano-Aroca
- Subjects
medicine.drug_class ,Antibiotics ,Biology ,medicine.disease_cause ,biology.organism_classification ,Antimicrobial ,medicine.disease ,Microbiology ,Staphylococcus epidermidis ,Staphylococcus aureus ,parasitic diseases ,medicine ,Infection control ,Pathogen ,Pneumonia (non-human) ,Coronavirus - Abstract
The Coronavirus Disease (COVID-19) pandemic is demanding rapid action of the authorities and scientific community in order to find new antimicrobial solutions that could inactivate the pathogen SARS-CoV-2 that causes this disease. Gram-positive bacteria contribute to severe pneumonia associated with COVID-19, and their resistance to antibiotics is increasing at an alarming rate. In this regard, non-woven fabrics are currently used for the fabrication of infection prevention clothing such as face masks, caps, scrubs, shirts, trousers, disposable gowns, overalls, hoods, aprons and shoe covers as protective tools against viral and bacterial infections. However, these non-woven fabrics are made of materials that do not possess antimicrobial activity. Thus, we have developed here non-woven fabrics with antimicrobial coatings of cranberry extracts capable of inactivating enveloped viruses such as SARS-CoV-2 and the phage phi 6, and two multidrug-resistant bacteria: the methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. The non-toxicity of these advanced technology was ensured using a Caenorhabditis elegans in vivo model. These results open up a new prevention path using natural and biodegradable compounds for the fabrication of infection prevention clothing in the current COVID-19 and future pandemics.
- Published
- 2021
35. Antimicrobial Face Shield: Next Generation of Facial Protective Equipment against SARS-CoV-2 and Multidrug-Resistant Bacteria
- Author
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Alberto Tuñón-Molina, Miguel Martí, Ángel Serrano-Aroca, Yukiko Muramoto, Takeshi Noda, and Kazuo Takayama
- Subjects
Face shield ,business.product_category ,face shield ,Antibiotics ,MRSA ,medicine.disease_cause ,Anti-Infective Agents ,Staphylococcus epidermidis ,Disk Diffusion Antimicrobial Tests ,polyethylene terephthalate ,Drug Resistance, Multiple, Bacterial ,Biology (General) ,Spectroscopy ,biology ,Polyethylene Terephthalates ,General Medicine ,Antimicrobial ,Computer Science Applications ,Bacteriophage phi 6 ,Chemistry ,surfaces,_coatings_films ,Multidrug resistant bacteria ,Staphylococcus aureus ,MRSE ,Benzalkonium Compounds ,Pneumonia (non-human) ,Methicillin-Resistant Staphylococcus aureus ,QH301-705.5 ,medicine.drug_class ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,multidrug-resistant bacteria ,benzalkonium chloride ,Catalysis ,Article ,Microbiology ,Inorganic Chemistry ,medicine ,Humans ,Physical and Theoretical Chemistry ,QD1-999 ,Molecular Biology ,Personal protective equipment ,Personal Protective Equipment ,business.industry ,SARS-CoV-2 ,Organic Chemistry ,COVID-19 ,phage phi 6 ,facial protective equipment ,medicine.disease ,biology.organism_classification ,business - Abstract
Transparent materials used for facial protection equipment provide protection against microbial infections caused by viruses and bacteria, including multidrug-resistant strains. However, transparent materials used for this type of application are made of materials that do not possess antimicrobial activity. They just avoid direct contact between the person and the biological agent. Therefore, healthy people can become infected through contact of the contaminated material surfaces and this equipment constitute an increasing source of infectious biological waste. Furthermore, infected people can transmit microbial infections easily because the protective equipment do not inactivate the microbial load generated while breathing, sneezing or coughing. In this regard, the goal of this work consisted of fabricating a transparent face shield with intrinsic antimicrobial activity that could provide extra-protection against infectious agents and reduce the generation of infectious waste. Thus, a single-use transparent antimicrobial face shield composed of polyethylene terephthalate and an antimicrobial coating of benzalkonium chloride has been developed for the next generation of facial protective equipment. The antimicrobial coating was analyzed by atomic force microscopy and field emission scanning electron microscopy with elemental analysis. This is the first facial transparent protective material capable of inactivating enveloped viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in less than one minute of contact, and the methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis. Bacterial infections contribute to severe pneumonia associated with the SARS-CoV-2 infection, and their resistance to antibiotics is increasing. Our extra protective broad-spectrum antimicrobial composite material could also be applied for the fabrication of other facial protective tools such as such as goggles, helmets, plastic masks and space separation screens used for counters or vehicles. This low-cost technology would be very useful to combat the current pandemic and protect health care workers from multidrug-resistant infections in developed and underdeveloped countries.
- Published
- 2021
36. Structural insight into Marburg virus nucleoprotein-RNA complex formation
- Author
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Yoko Fujita-Fujiharu, Yukihiko Sugita, Yuki Takamatsu, Kazuya Houri, Manabu Igarashi, Yukiko Muramoto, Masahiro Nakano, Yugo Tsunoda, Ichiro Taniguchi, Stephan Becker, and Takeshi Noda
- Subjects
Ebola virus ,Multidisciplinary ,Nucleoproteins ,Marburgvirus ,viruses ,Cryoelectron Microscopy ,General Physics and Astronomy ,RNA, Viral ,General Chemistry ,Ebolavirus ,General Biochemistry, Genetics and Molecular Biology ,Marburg virus - Abstract
The nucleoprotein (NP) of Marburg virus (MARV), a close relative of Ebola virus (EBOV), encapsidates the single-stranded, negative-sense viral genomic RNA (vRNA) to form the helical NP–RNA complex. The NP–RNA complex constitutes the core structure for the assembly of the nucleocapsid that is responsible for viral RNA synthesis. Although appropriate interactions among NPs and RNA are required for the formation of nucleocapsid, the structural basis of the helical assembly remains largely elusive. Here, we show the structure of the MARV NP–RNA complex determined using cryo-electron microscopy at a resolution of 3.1 Å. The structures of the asymmetric unit, a complex of an NP and six RNA nucleotides, was very similar to that of EBOV, suggesting that both viruses share common mechanisms for the nucleocapsid formation. Structure-based mutational analysis of both MARV and EBOV NPs identified key residues for helical assembly and subsequent viral RNA synthesis. Importantly, most of the residues identified were conserved in both viruses. These findings provide a structural basis for understanding the nucleocapsid formation and contribute to the development of novel antivirals against MARV and EBOV., 致死的な出血熱を引き起こすマールブルグウイルスの増殖機構を解明 --エボラ・マールブルグウイルスの創薬に期待--. 京都大学プレスリリース. 2022-03-07., Viruses of a feather: Similar structures in Marburg and Ebola viruses provide clues for antivirals. 京都大学プレスリリース. 2022-07-25.
- Published
- 2021
37. Structural insight into Marburg virus nucleoprotein-RNA complex formation
- Author
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Masahiro Nakano, Yoko Fujita-Fujiharu, Yugo Tsunoda, Yukiko Muramoto, Kazuya Houri, Yukihiko Sugita, Yuki Takamatsu, Stephan Becker, Takeshi Noda, and Manabu Igarashi
- Subjects
chemistry.chemical_classification ,Ebola virus ,viruses ,Complex formation ,RNA ,medicine.disease_cause ,Virology ,Nucleoprotein ,Marburg virus ,Mutational analysis ,chemistry ,medicine ,Nucleotide ,Viral rna - Abstract
The nucleoprotein (NP) of Marburg virus (MARV), a close relative of Ebola virus (EBOV), encapsidates the single-stranded, negative-sense viral genomic RNA (vRNA) to form the helical NP-RNA complex. The NP-RNA complex serves as a scaffold for the assembly of the nucleocapsid that is responsible for viral RNA synthesis. Although appropriate interactions among NPs and RNA are required for the formation of nucleocapsid, the structural basis of the helical assembly remains largely elusive. Here, we show the structure of the MARV NP-RNA complex determined using cryo-electron microscopy at a resolution of 3.1 Å. The structures of the asymmetric unit, a complex of an NP and six RNA nucleotides, was very similar to that of EBOV, suggesting that both viruses share common mechanisms for the nucleocapsid formation. Structure-based mutational analysis of both MARV and EBOV NPs identified key residues for the viral RNA synthesis as well as the helical assembly. Importantly, most of the residues identified were conserved in both viruses. These findings provide a structural basis for understanding the nucleocapsid formation and contribute to the development of novel antivirals against MARV and EBOV.
- Published
- 2021
38. Ultrastructure of influenza virus ribonucleoprotein complexes during viral RNA synthesis
- Author
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Takeshi Noda, Yukiko Muramoto, Sho Miyamoto, Noriyuki Kodera, Matthias Wolf, Masahiro Nakano, and Yukihiko Sugita
- Subjects
STRUCTURAL BASIS ,DYNAMICS ,Models, Molecular ,QH301-705.5 ,viruses ,Medicine (miscellaneous) ,Microscopy, Atomic Force ,Virus Replication ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Viral Proteins ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,POLYADENYLATION ,Transcription (biology) ,RNA polymerase ,Influenza, Human ,Influenza A virus ,medicine ,Humans ,CRYSTAL-STRUCTURE ,Biology (General) ,030304 developmental biology ,Ribonucleoprotein ,0303 health sciences ,Chemistry ,POLYMERASE ,Cryoelectron Microscopy ,CAP-BINDING ,RNA ,VIRION RNA ,Virus structures ,In vitro ,ENDONUCLEASE ,Cell biology ,Nucleoprotein ,INSIGHTS ,Ribonucleoproteins ,Nucleic Acid Conformation ,RNA, Viral ,MESSENGER-RNA ,Influenza virus ,General Agricultural and Biological Sciences ,030217 neurology & neurosurgery - Abstract
The single-stranded, negative-sense, viral genomic RNA (vRNA) of influenza A virus is encapsidated by viral nucleoproteins (NPs) and an RNA polymerase to form a ribonucleoprotein complex (vRNP) with a helical, rod-shaped structure. The vRNP is responsible for transcription and replication of the vRNA. However, the vRNP conformation during RNA synthesis is not well understood. Here, using high-speed atomic force microscopy and cryo-electron microscopy, we investigated the native structure of influenza A vRNPs during RNA synthesis in vitro. Two distinct types of vRNPs were observed in association with newly synthesized RNAs: an intact, helical rod-shaped vRNP connected with a folded RNA and a deformed vRNP associated with a looped RNA. Interestingly, the looped RNA was a double-stranded RNA, which likely comprises a nascent RNA and the template RNA detached from NPs of the vRNP. These results suggest that while some vRNPs keep their helical structures during RNA synthesis, for the repeated cycle of RNA synthesis, others accidentally become structurally deformed, which likely results in failure to commence or continue RNA synthesis. Thus, our findings provide the ultrastructural feature of vRNPs during RNA synthesis., Nakano et al. use high-speed AFM and cryo-EM to study influenza virus RNA synthesis in vitro. They observe the synthesis of two types of RNA: folded single stranded RNA associated to an intact viral ribonucleoprotein (vRNP) and looped, double stranded RNA associated with a partially rearranged vRNP structure.
- Published
- 2021
39. Quercetin in Tartary Buckwheat Induces Autophagy against Protein Aggregations
- Author
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Yuancai Liu, Shiou-Ling Lu, Guoqiang Tong, Lu Shiguang, Sumiko Ikari, Takeshi Noda, Feike Hao, and Qiang Yang
- Subjects
0301 basic medicine ,Antioxidant ,Physiology ,medicine.medical_treatment ,Clinical Biochemistry ,Aggrephagy ,P70-S6 Kinase 1 ,mTORC1 ,RM1-950 ,Biochemistry ,Article ,Dephosphorylation ,HeLa ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine ,aggrephagy ,Molecular Biology ,biology ,Chemistry ,Autophagy ,Cell Biology ,biology.organism_classification ,030104 developmental biology ,Therapeutics. Pharmacology ,Quercetin ,030217 neurology & neurosurgery ,tartary buckwheat - Abstract
Tartary buckwheat is used as an ingredient in flour and tea, as well as in traditional Chinese medicine for its antioxidant effects. Here, we found that an ethanol extract of tartary buckwheat (TBE) potently induced autophagy flux in HeLa cells by suppressing mTORC1 activity, as revealed by dephosphorylation of the mTORC1 substrates Ulk1, S6K, and 4EBP, as well as by the nuclear translocation of transcriptional factor EB. In addition to non-selective bulk autophagy, TBE also induced aggrephagy, which is defined as autophagy against aggregated proteins. Quercetin is a flavonol found at high levels in TBE. We showed that quercetin induced both non-selective bulk autophagy and aggrephagy. These effects were also observed in Huh-7 cells derived from hepatocytes. Thus, aggrephagy induction by TBE and quercetin may relieve alcoholic hepatitis, which is closely linked to the accumulation of protein aggregations called Mallory–Denk bodies.
- Published
- 2021
40. Author response for 'Acetylation of the influenza A virus polymerase subunit PA in the N-terminal domain positively regulates its endonuclease activity'
- Author
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Seiryo Ogata, Sumio Ohtsuki, Ayana Ohkubo, Asuka Miyatake, Yoshihiro Kawaoka, Masahiro Nakano, Takeshi Noda, Takashi Kuzuhara, Hazuki Tsuneishi, Masaki Shoji, Dai Hatakeyama, Mizuki Takahira, Kyoko Makiyama, Takeshi Masuda, Ayaka Saitoh, Tsugunori Komatsu, and Erina Nishikawa
- Subjects
Endonuclease ,biology ,Terminal (electronics) ,Chemistry ,Acetylation ,Protein subunit ,Influenza A virus ,medicine ,biology.protein ,medicine.disease_cause ,Molecular biology ,Polymerase ,Domain (software engineering) - Published
- 2021
41. Resistance of SARS-CoV-2 variants to neutralization by antibodies induced in convalescent patients with COVID-19
- Author
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Kaho Matsumoto, Yukiko Muramoto, Yoshio Koyanagi, Hiroshi Takahashi, Chihiro Motozono, Takeo Kuwata, Terumasa Ikeda, Kazuya Shimura, Takamasa Ueno, Yoji Nagasaki, Yosuke Maeda, Shuzo Matsushita, Takao Hashiguchi, Tsuneyuki Tatsuke, Takeshi Noda, Hajime Iwagoe, Shashwata Biswas, Kazuhiko Fujii, Tateki Suzuki, Mako Toyoda, Mikiko Shimizu, Hiroto Kishi, Noriko Kuramoto, Jiei Sasaki, Hasan Md Zahid, Chiho Onishi, Rumi Minami, Yu Kaku, and Yoko Kawanami
- Subjects
Male ,QH301-705.5 ,medicine.drug_class ,Antibodies, Viral ,medicine.disease_cause ,Monoclonal antibody ,General Biochemistry, Genetics and Molecular Biology ,Neutralization ,Cell Line ,Protein Domains ,Neutralization Tests ,biology.animal ,Report ,Humans ,Medicine ,Biology (General) ,Mink ,Neutralizing antibody ,COVID-19 Serotherapy ,mAb ,Mutation ,biology ,business.industry ,SARS-CoV-2 ,HEK 293 cells ,Immunization, Passive ,Antibodies, Monoclonal ,COVID-19 ,neutralizing antibody ,Antibodies, Neutralizing ,Virology ,HEK293 Cells ,variant ,Cell culture ,Spike Glycoprotein, Coronavirus ,biology.protein ,Antibody ,business - Abstract
Administration of plasma from convalescent patients or neutralizing monoclonal antibodies (mAbs) is potent therapeutic option for COVID-19 caused by SARS-CoV-2 infection. However, SARS-CoV-2 variants with mutations in the spike protein have emerged in many countries. To evaluate the efficacy of neutralizing antibodies induced in convalescent patients against emerging variants, we isolate anti-Spike mAbs from two convalescent patients with COVID-19 infected with prototypic SARS-CoV-2 by single cell sorting of IgG+ memory B cells. Anti-Spike antibody induction is robust in these patients and five mAbs have potent neutralizing activities. The efficacy of most neutralizing mAbs and convalescent plasma samples is maintained against B.1.1.7 and mink cluster 5 variants but is significantly decreased against variants B.1.351 from South Africa and P.1 from Brazil. However, mAbs with a high affinity to the receptor-binding domain remain effective against these neutralization-resistant variants. Rapid spread of these variants significantly impacts antibody-based therapies and vaccine strategies against SARS-CoV-2., Graphical Abstract, Kaku et al. demonstrates that efficacy of neutralizing mAbs and convalescent plasmas is maintained against SARS-CoV-2 variants B.1.1.7 from U.K. and mink cluster 5, but decreases against B.1.351 from South Africa and P.1 from Brazil. Rapid spread of these variants significantly impacts therapies and vaccine strategies against SARS-CoV-2.
- Published
- 2021
42. Modeling SARS-CoV-2 infection and its individual differences with ACE2-expressing human iPS cells
- Author
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Natsumi Mimura, Emi Sano, Yukiko Muramoto, Ayaka Sakamoto, Sayaka Deguchi, Takuya Yamamoto, Ai Hirabayashi, Takeshi Noda, and Kazuo Takayama
- Subjects
0301 basic medicine ,Coronavirus disease 2019 (COVID-19) ,Science ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,viruses ,02 engineering and technology ,Biology ,Article ,Virus ,03 medical and health sciences ,Virology ,Health Sciences ,Induced pluripotent stem cell ,Receptor ,chemistry.chemical_classification ,Budding ,Multidisciplinary ,fungi ,Cell Biology ,021001 nanoscience & nanotechnology ,Embryonic stem cell ,030104 developmental biology ,Enzyme ,chemistry ,0210 nano-technology ,hormones, hormone substitutes, and hormone antagonists - Abstract
Genetic differences are a primary reason for differences in the susceptibility and severity of COVID-19. As induced pluripotent stem (iPS) cells maintain the genetic information of the donor, they can be used to model individual differences in SARS-CoV-2 infection in vitro. We found that human iPS cells expressing the SARS-CoV-2 receptor angiotensin-converting enzyme 2 (ACE2) (ACE2-iPS cells) can be infected w SARS-CoV-2. In infected ACE2-iPS cells, the expression of SARS-CoV-2 nucleocapsid protein, budding of viral particles, and production of progeny virus, double membrane spherules, and double-membrane vesicles were confirmed. We performed SARS-CoV-2 infection experiments on ACE2-iPS/ embryonic stem (ES) cells from eight individuals. Male iPS/ES cells were more capable of producing the virus compared with female iPS/ES cells. These findings suggest that ACE2-iPS cells can not only reproduce individual differences in SARS-CoV-2 infection in vitro but also are a useful resource to clarify the causes of individual differences in COVID-19 due to genetic differences., Graphical abstract, Health Sciences ; Virology ; Cell Biology
- Published
- 2021
43. N4BP1 restricts HIV-1 and its inactivation by MALT1 promotes viral reactivation
- Author
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Naoko Misawa, Lennart Koepke, Dominik Hotter, Takuya Ichinose, Simone Joas, Elisabeth Reith, Daichi Yamasoba, Daniel Sauter, Yoshio Koyanagi, Daron M. Standley, Frank Kirchhoff, Takashi Mino, Sho Miyamoto, Osamu Takeuchi, Takuya Uehata, Takeshi Noda, Akio Yamashita, Kei Sato, Kotaro Akaki, and Tomoko Imamura
- Subjects
Microbiology (medical) ,0303 health sciences ,Messenger RNA ,030306 microbiology ,Chemistry ,Effector ,RNase P ,Immunology ,Chromosomal translocation ,Cell Biology ,Paracaspase ,Cleavage (embryo) ,Applied Microbiology and Biotechnology ,Microbiology ,Cell biology ,03 medical and health sciences ,MALT1 ,Immune system ,Genetics ,030304 developmental biology - Abstract
RNA-modulating factors not only regulate multiple steps of cellular RNA metabolism, but also emerge as key effectors of the immune response against invading viral pathogens including human immunodeficiency virus type-1 (HIV-1). However, the cellular RNA-binding proteins involved in the establishment and maintenance of latent HIV-1 reservoirs have not been extensively studied. Here, we screened a panel of 62 cellular RNA-binding proteins and identified NEDD4-binding protein 1 (N4BP1) as a potent interferon-inducible inhibitor of HIV-1 in primary T cells and macrophages. N4BP1 harbours a prototypical PilT N terminus-like RNase domain and inhibits HIV-1 replication by interacting with and degrading viral mRNA species. Following activation of CD4+ T cells, however, N4BP1 undergoes rapid cleavage at Arg 509 by the paracaspase named mucosa-associated lymphoid tissue lymphoma translocation 1 (MALT1). Mutational analyses and knockout studies revealed that MALT1-mediated inactivation of N4BP1 facilitates the reactivation of latent HIV-1 proviruses. Taken together, our findings demonstrate that the RNase N4BP1 is an efficient restriction factor of HIV-1 and suggest that inactivation of N4BP1 by induction of MALT1 activation might facilitate elimination of latent HIV-1 reservoirs. The interferon-inducible RNA-binding protein N4BP1 binds to and degrades HIV-1 mRNAs, thus inhibiting viral degradation and promoting viral latency. However, metacaspase-mediated degradation of N4BP1 following activation of CD4+ T cells facilitates HIV-1 latency reversal.
- Published
- 2019
44. Generation of a purely clonal defective interfering influenza virus
- Author
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Yutaro Yamagata, Takeshi Noda, Masahiro Nakano, Sho Miyamoto, Yukiko Muramoto, and Keiko Shindo
- Subjects
viruses ,Immunology ,Biology ,Virus Replication ,Antiviral Agents ,Microbiology ,Virus ,Madin Darby Canine Kidney Cells ,Viral Proteins ,03 medical and health sciences ,Dogs ,Orthomyxoviridae Infections ,Virology ,Influenza, Human ,Animals ,Humans ,Gene ,Infectious virus ,030304 developmental biology ,0303 health sciences ,030306 microbiology ,Antiviral therapy ,Defective Viruses ,RNA-Dependent RNA Polymerase ,In vitro ,Reverse genetics ,HEK293 Cells ,Influenza A virus ,Cell culture ,Helper virus ,RNA, Viral - Abstract
Defective interfering (DI) influenza viruses carry a large deletion in a gene segment that interferes with the replication of infectious virus; thus, such viruses have potential for antiviral therapy. However, because DI viruses cannot replicate autonomously without the aid of an infectious helper virus, clonal DI virus stocks that are not contaminated with helper virus have not yet been generated. To overcome this problem, we used reverse genetics to generate a clonal DI virus with a PB2 DI gene, amplified the clonal DI virus using a cell line stably expressing the PB2 protein, and confirmed its ability to interfere with infectious virus replication in vitro. Thus, our approach is suitable for obtaining purely clonal DI viruses, will contribute to the understanding of DI virus interference mechanisms and can be used to develop DI virus-based antivirals.
- Published
- 2019
45. Double-Sided Nonalloyed Ohmic Contacts to Si-doped GaAs for Plasmoelectronic Devices
- Author
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Takeshi Kasaya, Takaaki Mano, Takeshi Noda, Hideki T. Miyazaki, and Yoshiki Sakuma
- Subjects
Materials science ,business.industry ,General Chemical Engineering ,Si doped ,Photodetector ,General Chemistry ,Substrate (electronics) ,Laser ,Article ,law.invention ,lcsh:Chemistry ,lcsh:QD1-999 ,Cascade ,law ,Electrical resistivity and conductivity ,Optoelectronics ,Narrow range ,business ,Ohmic contact - Abstract
There is increasing demand for the ability to form ohmic contacts without lossy intermediate layers on both the top and bottom sides of metal-semiconductor-metal plasmoelectronic devices such as quantum cascade lasers and metasurface photodetectors. Although highly Si-doped n-GaAs surfaces can allow an ohmic contact without alloying, conditions for realizing nonalloyed ohmic contacts to other n-GaAs surfaces, originally buried inside but exposed by removing the substrate, have yet to be studied. We discovered that nonalloyed ohmic contacts to initially buried surfaces with a practically low contact resistivity down to 77 K can be realized by fulfilling certain requirements, specifically keeping the Si-doping concentration within a narrow range of 7.5 × 1018 to 1.25 × 1019 cm-3 and setting the growth temperature of the succeeding upper layers to a low value of 530 °C.
- Published
- 2019
46. Cobalt-doped nickel oxide nanoparticles as efficient hole transport materials for low-temperature processed perovskite solar cells
- Author
-
Md. Emrul Kayesh, Said Kazaoui, Ryuji Kaneko, Takeshi Noda, Jae-Joon Lee, Joe Otsuki, Kosuke Sugawa, Ashraful Islam, Towhid H. Chowdhury, and Guohua Wu
- Subjects
Materials science ,Renewable Energy, Sustainability and the Environment ,020209 energy ,Nickel oxide ,Doping ,chemistry.chemical_element ,Nanoparticle ,02 engineering and technology ,021001 nanoscience & nanotechnology ,chemistry ,Chemical engineering ,0202 electrical engineering, electronic engineering, information engineering ,Cobalt ions ,General Materials Science ,0210 nano-technology ,Cobalt ,Perovskite (structure) ,Electronic properties - Abstract
We have synthesized cobalt-doped NiOx nanoparticles containing 0.5, 1, 2, 5 and 10 mol% cobalt ions and have investigated their electronic properties, which can be processed into smooth and pinhole-free layers at low temperature (
- Published
- 2019
47. Study on Carrier Separation in Perovskite Solar Cells by Operando Profiling of Electrical Potential Distribution
- Author
-
Liyuan Han, Takeshi Noda, Molang Cai, Daisuke Fujita, and Nobuyuki Ishida
- Subjects
Profiling (computer programming) ,Materials science ,Distribution (number theory) ,Chemical physics ,Perovskite (structure) - Published
- 2019
48. Versatile coordination architectures of products generated by the in situ reaction of a doubly bis(2-pyridyl)pyrazolate bridged dinuclear copper(<scp>ii</scp>) complex with tetracyanoethylene
- Author
-
Motohiro Nakano, Satoshi Kawata, Yumi Hamatake, Yoji Horii, Shunya Ueno, Yuji Miyazaki, Takeshi Noda, Mikoto Uematsu, and Ryuta Ishikawa
- Subjects
Denticity ,Materials science ,Coordination polymer ,Infrared spectroscopy ,chemistry.chemical_element ,02 engineering and technology ,General Chemistry ,Tetracyanoethylene ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,01 natural sciences ,Copper ,Square pyramidal molecular geometry ,0104 chemical sciences ,law.invention ,Crystallography ,chemistry.chemical_compound ,chemistry ,law ,Molecule ,General Materials Science ,0210 nano-technology ,Electron paramagnetic resonance - Abstract
The reaction of a doubly bis(2-pyridyl)pyrazolate (bpypz−) bridged dinuclear copper(II) complex, [Cu2(bpypz)2]2+, with neutral tetracyanoethylene (TCNE) results in the formation of [Cu4(bpypz)4(TCVA)2]·2TCVA (1) in tetrahydrofuran (THF), syn-[Cu2(bpypz)2(DCNM)2] (2) in MeOH, and anti-[Cu2(bpypz)2(DCNE)2] (3) in EtOH, where TCVA− = 1,2,2-tricyanoethenolate, DCNM− = 2,2-dicyano-1-methoxyethenolate, and DCNE− = 2,2-dicyano-1-ethoxyethenolate. The ligands TCVA−, DCME−, and DCNE− in 1–3 were formed in situ during the reaction in different solvents, and the ligands were confirmed through infrared spectroscopy, single-crystal X-ray diffraction, and elemental analyses. The single-crystal X-ray structures of 1–3 show that each copper(II) centre is five-coordinate in an approximately square pyramidal geometry. 1 is a discrete dimer-of-dimers type tetranuclear complex with the copper(II) centres bridged by two trans-μ2-N,O-TCVA−. 2 and 3 are discrete dinuclear complexes. In 2, two apically monodentate N-coordinated DCNM− molecules are oriented in a cis manner to one another, consequently adopting a syn-orthogonal configuration. A weak interaction exists between each copper(II) centres and the O atoms of the apically coordinated DCNM− molecules of the neighbouring copper(II) dinuclear units, resulting in a pseudo one-dimensional ladder-like coordination polymer arrangement. In 3, two apically monodentate N-coordinated DCNE− molecules are oriented in a mutually trans manner across the basal plane of [Cu2(bpypz)2]2+, thereby adopting an anti-orthogonal configuration. Continuous wave X-band electron paramagnetic resonance spectral measurements on finely ground polycrystalline samples of 1–3 confirm a thermally depopulated S = 1 triplet state arising from antiferromagnetically coupled two S = 1/2 spins with orthorhombic g values. Variable-temperature dc magnetic susceptibility measurements of 1–3 indicate significant antiferromagnetic coupling (quantified by the isotropic spin Hamiltonian, H = −2JS1·S2, for the two copper(II) centres: J = −217.7, −210.3, and −188.5 cm−1 for 1, 2 and 3, respectively) for the doubly bpypz− bridged copper(II) centres.
- Published
- 2019
49. Atomic-scale characterization of highly doped Si impurities in GaAs using scanning tunneling microscopy
- Author
-
Nobuyuki Ishida, Takaaki Mano, and Takeshi Noda
- Subjects
General Physics and Astronomy ,Surfaces and Interfaces ,General Chemistry ,Condensed Matter Physics ,Surfaces, Coatings and Films - Published
- 2022
50. Coadditive Engineering with 5-Ammonium Valeric Acid Iodide for Efficient and Stable Sn Perovskite Solar Cells
- Author
-
Jae-Joon Lee, Ashraful Islam, Ryuji Kaneko, Takeshi Noda, Md. Emrul Kayesh, Kiyoto Matsuishi, and Said Kazaoui
- Subjects
chemistry.chemical_classification ,Materials science ,Valeric acid ,Renewable Energy, Sustainability and the Environment ,Hydrogen bond ,Energy conversion efficiency ,Iodide ,Energy Engineering and Power Technology ,Crystal growth ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,chemistry.chemical_compound ,Fuel Technology ,chemistry ,Octahedron ,Chemical engineering ,Chemistry (miscellaneous) ,Materials Chemistry ,SN2 reaction ,0210 nano-technology ,Perovskite (structure) - Abstract
Sn-based perovskite solar cells (PSCs) featuring high performance and long-term stability are very challenging because Sn2+ is relatively prone to oxidation. Here, we have performed coadditive engineering with 5-ammonium valeric acid iodide (5-AVAI) for FASnI3-based perovskite films. From the morphological, structural, and elemental analyses, we observed that 5-AVAI affects the crystal growth of perovskites through its hydrogen bond with I– of the SnI64– octahedral. As a result, pinhole-free homogeneous and stable Sn-based perovskite films form over a large area with lower Sn4+ content. This made us able to enhance the power conversion efficiency (PCE) for Sn-based PSCs up to 7% in a 0.25 cm2 aperture area. Most importantly, the 5-AVAI added PSCs showed a record stability and maintained their initial PCE under 1 sun continuous illumination at maximum power point tracking for 100 h.
- Published
- 2018
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