start-ver=1.4 cd-journal=joma no-vol=10 cd-vols= no-issue=1 article-no= start-page=19087 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20201105 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Cell cycle dependence of apoptosis photo-triggered using peptide-photosensitizer conjugate en-subtitle= kn-subtitle= en-abstract= kn-abstract=Investigation of the relevance between cell cycle status and the bioactivity of exogenously delivered biomacromolecules is hindered by their time-consuming cell internalization and the cytotoxicity of transfection methods. In this study, we addressed these problems by utilizing the photochemical internalization (PCI) method using a peptide/protein-photosensitizer conjugate, which enables immediate cytoplasmic internalization of the bioactive peptides/proteins in a light-dependent manner with low cytotoxicity. To identify the cell-cycle dependent apoptosis, a TatBim peptide-photosensitizer conjugate (TatBim-PS) with apoptotic activity was photo-dependently internalized into HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (Fucci2). Upon irradiation, cytoplasmic TatBim-PS internalization exceeded 95% for all cells classified in the G(1), S, and G(2)/M cell cycle phases with no significant differences between groups. TatBim-PS-mediated apoptosis was more efficiently triggered by photoirradiation in the G(1)/S transition than in the G(1) and S/G(2)/M phases, suggesting high sensitivity of the former phase to Bim-induced apoptosis. Thus, the cell cycle dependence of Bim peptide-induced apoptosis was successfully investigated using Fucci2 indicator and the PCI method. Since PCI-mediated cytoplasmic internalization of peptides is rapid and does not span multiple cell cycle phases, the Fucci-PCI method constitutes a promising tool for analyzing the cell cycle dependence of peptides/protein functions. en-copyright= kn-copyright= en-aut-name=KimHyungjin en-aut-sei=Kim en-aut-mei=Hyungjin kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=WatanabeSho en-aut-sei=Watanabe en-aut-mei=Sho kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=WatanabeKazunori en-aut-sei=Watanabe en-aut-mei=Kazunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= affil-num=1 en-affil=Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=2 en-affil=Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=3 en-affil=Department of Applied Chemistry, Kindai University kn-affil= affil-num=4 en-affil=Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= affil-num=5 en-affil=Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University kn-affil= en-keyword=Biological techniques kn-keyword=Biological techniques en-keyword=Biotechnology kn-keyword=Biotechnology END start-ver=1.4 cd-journal=joma no-vol=383 cd-vols= no-issue=2 article-no= start-page=111556 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2019 dt-pub=20191015 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Mechanical strain attenuates cytokine-induced ADAMTS9 expression via transient receptor potential vanilloid type 1 en-subtitle= kn-subtitle= en-abstract= kn-abstract= The synovial fluids of patients with osteoarthritis (OA) contain elevated levels of inflammatory cytokines, which induce the expression of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) and of the matrix metalloproteinase (MMP) in chondrocytes. Mechanical strain has varying effects on organisms depending on the strength, cycle, and duration of the stressor; however, it is unclear under inflammatory stimulation how mechanical strain act on. Here, we show that mechanical strain attenuates inflammatory cytokine-induced expression of matrix-degrading enzymes. Cyclic tensile strain (CTS), as a mechanical stressor, attenuated interleukin (IL)-1ƒΐ and tumor necrosis factor (TNF)-ƒΏ-induced mRNA expression of ADAMTS4, ADAMTS9, and MMP-13 in normal chondrocytes (NHAC-kn) and in a chondrocytic cell line (OUMS-27). This effect was abolished by treating cells with mechano-gated channel inhibitors, such as gadolinium, transient receptor potential (TRP) family inhibitor, ruthenium red, and with pharmacological and small interfering RNA-mediated TRPV1 inhibition. Furthermore, nuclear factor ƒΘB (NF-ƒΘB) translocation from the cytoplasm to the nucleus resulting from cytokine stimulation was also abolished by CTS. These findings suggest that mechanosensors such as the TRPV protein are potential therapeutic targets in treating OA. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=ShinaokaAkira en-aut-sei=Shinaoka en-aut-mei=Akira kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=Kumagishi-ShinaokaKanae en-aut-sei=Kumagishi-Shinaoka en-aut-mei=Kanae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=AsanoKeiichi en-aut-sei=Asano en-aut-mei=Keiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=HatipogluOmer Faruk en-aut-sei=Hatipoglu en-aut-mei=Omer Faruk kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=InagakiJunko en-aut-sei=Inagaki en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=TakahashiKen en-aut-sei=Takahashi en-aut-mei=Ken kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=OohashiToshitaka en-aut-sei=Oohashi en-aut-mei=Toshitaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=NishidaKeiichiro en-aut-sei=Nishida en-aut-mei=Keiichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= en-aut-name=NaruseKeiji en-aut-sei=Naruse en-aut-mei=Keiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=10 ORCID= en-aut-name=HirohataSatoshi en-aut-sei=Hirohata en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=11 ORCID= affil-num=1 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Human Morphology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=3 en-affil=Department of Human Morphology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= affil-num=6 en-affil=Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=7 en-affil=Department of Cardiovascular Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=8 en-affil=Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=9 en-affil=Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=10 en-affil=Department of Cardiovascular Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=11 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue= article-no= start-page=12501 end-page=12501 dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=201608 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs en-subtitle= kn-subtitle= en-abstract= kn-abstract=The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4?h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at ?405?nm at 125?mW?cm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19?s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KanzakiShigeto en-aut-sei=Kanzaki en-aut-mei=Shigeto kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NishimuraSae en-aut-sei=Nishimura en-aut-mei=Sae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KunihiroYoshio en-aut-sei=Kunihiro en-aut-mei=Yoshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=WatanabeKazunori en-aut-sei=Watanabe en-aut-mei=Kazunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=2 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=3 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=4 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=5 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=6 en-affil=Department of Biomedical Engineering, Okayama University kn-affil=‰ͺŽR‘εŠw‘εŠw‰@Ž©‘R‰ΘŠwŒ€‹†‰Θ END start-ver=1.4 cd-journal=joma no-vol=5 cd-vols= no-issue= article-no= start-page=18577 end-page=18577 dt-received= dt-revised= dt-accepted= dt-pub-year=2015 dt-pub=201512 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=The molecular mechanism of photochemical internalization of cell penetrating peptide-cargo-photosensitizer conjugates. en-subtitle= kn-subtitle= en-abstract= kn-abstract=In many drug delivery strategies, an inefficient transfer of macromolecules such as proteins and nucleic acids to the cytosol often occurs because of their endosomal entrapment. One of the methods to overcome this problem is photochemical internalization, which is achieved using a photosensitizer and light to facilitate the endosomal escape of the macromolecule. In this study, we examined the molecular mechanism of photochemical internalization of cell penetrating peptide-cargo (macromolecule)-photosensitizer conjugates. We measured the photophysical properties of eight dyes (photosensitizer candidates) and determined the respective endosomal escape efficiencies using these dyes. Correlation plots between these factors indicated that the photogenerated (1)O2 molecules from photosensitizers were highly related to the endosomal escape efficiencies. The contribution of (1)O2 was confirmed using (1)O2 quenchers. In addition, time-lapse fluorescence imaging showed that the photoinduced endosomal escape occurred at a few seconds to a few minutes after irradiation (much longer than (1)O2 lifetime), and that the pH increased in the endosome prior to the endosomal escape of the macromolecule. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=MikiShunya en-aut-sei=Miki en-aut-mei=Shunya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=KobayashiShouhei en-aut-sei=Kobayashi en-aut-mei=Shouhei kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HaraguchiTokuko en-aut-sei=Haraguchi en-aut-mei=Tokuko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=NakataEiji en-aut-sei=Nakata en-aut-mei=Eiji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HirakawaKazutaka en-aut-sei=Hirakawa en-aut-mei=Kazutaka kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=SumitaKensuke en-aut-sei=Sumita en-aut-mei=Kensuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=WatanabeKazunori en-aut-sei=Watanabe en-aut-mei=Kazunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= en-aut-name=OkazakiShigetoshi en-aut-sei=Okazaki en-aut-mei=Shigetoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=9 ORCID= affil-num=1 en-affil=Department of Medical Bioengineering, Okayama University kn-affil= affil-num=2 en-affil=Department of Medical Bioengineering, Okayama University kn-affil= affil-num=3 en-affil=Advanced ICT Research Institute Kobe, NICT kn-affil= affil-num=4 en-affil=Advanced ICT Research Institute Kobe, NICT kn-affil= affil-num=5 en-affil=Institute of Advanced Energy, Kyoto University kn-affil= affil-num=6 en-affil=Department of Applied Chemistry and Biochemical Engineering, Graduate School of Engineering, Shizuoka University kn-affil= affil-num=7 en-affil=Department of Medical Bioengineering, Okayama University kn-affil= affil-num=8 en-affil=Department of Medical Bioengineering, Okayama University kn-affil=‰ͺŽR‘εŠw‘εŠw‰@Ž©‘R‰ΘŠwŒ€‹†‰Θ affil-num=9 en-affil=Department of Medical Spectroscopy, Hamamatsu University School of Medicine kn-affil= END start-ver=1.4 cd-journal=joma no-vol=7 cd-vols= no-issue= article-no= start-page=12501 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2016 dt-pub=201608 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs en-subtitle= kn-subtitle= en-abstract= kn-abstract= The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4?h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at ?405?nm at 125?mW?cm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19?s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KanzakiShigeto en-aut-sei=Kanzaki en-aut-mei=Shigeto kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=NishimuraSae en-aut-sei=Nishimura en-aut-mei=Sae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=KunihiroYoshio en-aut-sei=Kunihiro en-aut-mei=Yoshio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=WatanabeKazunori en-aut-sei=Watanabe en-aut-mei=Kazunori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=2 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=3 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=4 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=5 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= affil-num=6 en-affil=Department of Biomedical Engineering, Okayama University kn-affil= en-keyword=Molecular engineering kn-keyword=Molecular engineering en-keyword=Optogenetics kn-keyword=Optogenetics END start-ver=1.4 cd-journal=joma no-vol=21 cd-vols= no-issue=1 article-no= start-page=225 end-page=227 dt-received= dt-revised= dt-accepted= dt-pub-year=2011 dt-pub=201101 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Antisense effect of pyrrolidine-based oxy-peptide nucleic acids in Escherichia coli en-subtitle= kn-subtitle= en-abstract= kn-abstract=To investigate the antisense effect of a pyrrolidine-based oxy-peptide nucleic acid (POPNA), we carried out the LacZ reporter assay using a 12-mer trans-l-POPNA conjugated with a cell-penetrating peptide (antisense reagent). The antisense effect of the conjugated POPNA (inhibition of LacZ activity) was comparable to that shown by a Nielsen-type peptide nucleic acid. Furthermore, the conjugated POPNA could switch the LacZ activity over a wide range of ambient temperatures. en-copyright= kn-copyright= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KuramiShunsuke en-aut-sei=Kurami en-aut-mei=Shunsuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology affil-num=2 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology affil-num=3 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology affil-num=4 en-affil= kn-affil=Department of Medical and Bioengineering, Graduate School of Natural Science and Technology en-keyword=Peptide nucleic acid kn-keyword=Peptide nucleic acid en-keyword=Cell-penetrating peptide kn-keyword=Cell-penetrating peptide en-keyword=Antisense effect kn-keyword=Antisense effect END start-ver=1.4 cd-journal=joma no-vol=19 cd-vols= no-issue=13 article-no= start-page=3410 end-page=3413 dt-received= dt-revised= dt-accepted= dt-pub-year=2009 dt-pub=20090701 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Carrier PNA for shRNA delivery into cells en-subtitle= kn-subtitle= en-abstract= kn-abstract=A peptide nucleic acid (PNA)-cell-penetrating peptide (CPP) conjugate (carrier PNA) was used as 'bridgebuilder' to connect a CPP with an shRNA. The carrier PNA successfully formed a hybrid with an shRNA bearing complementary dangling bases and the shRNA was introduced into cells by the carrier PNA, and RNAi was induced by the shRNA. en-copyright= kn-copyright= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=KuboTakanori en-aut-sei=Kubo en-aut-mei=Takanori kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MatsuzakiRino en-aut-sei=Matsuzaki en-aut-mei=Rino kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=EndohTamaki en-aut-sei=Endoh en-aut-mei=Tamaki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University affil-num=2 en-affil= kn-affil=Faculty of Pharmacy, Yasuda Womenfs University affil-num=3 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University affil-num=4 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University affil-num=5 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University affil-num=6 en-affil= kn-affil=Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University en-keyword=Peptide nucleic acid kn-keyword=Peptide nucleic acid en-keyword=Cell-penetrating peptide kn-keyword=Cell-penetrating peptide en-keyword=RNA interference kn-keyword=RNA interference END start-ver=1.4 cd-journal=joma no-vol=66 cd-vols= no-issue=51 article-no= start-page=9659 end-page=9666 dt-received= dt-revised= dt-accepted= dt-pub-year=2010 dt-pub=20101218 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Synthesis of pyrrolidine-based oxy-peptide nucleic acids carrying four types of nucleobases and their transport into cytoplasm en-subtitle= kn-subtitle= en-abstract= kn-abstract=We synthesized 16 pyrrolidine-based oxy-peptide nucleic acid (POPNA) monomers carrying four different nucleobases onto four different stereoisomers of pyrrolidine rings. Using these monomers, we prepared POPNA oligomers, which formed sequence-specific hybrids with DNAs. The oligomer configurations influenced the hybrid stability. The oligomers were not taken into CHO cells. However, they could enter the cell cytoplasm when mixed with the influenza virus hemagglutinin peptide-arginine heptamer conjugate. en-copyright= kn-copyright= en-aut-name=KitamatsuMizuki en-aut-sei=Kitamatsu en-aut-mei=Mizuki kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=TakahashiAkiko en-aut-sei=Takahashi en-aut-mei=Akiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=SisidoMasahiko en-aut-sei=Sisido en-aut-mei=Masahiko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= affil-num=1 en-affil= kn-affil=Department of Bioscience and Biotechnology, Okayama University affil-num=2 en-affil= kn-affil=Department of Bioscience and Biotechnology, Okayama University affil-num=3 en-affil= kn-affil=Department of Bioscience and Biotechnology, Okayama University affil-num=4 en-affil= kn-affil=Department of Bioscience and Biotechnology, Okayama University en-keyword=Peptide nucleic acid kn-keyword=Peptide nucleic acid en-keyword=Solid-phase peptide synthesis kn-keyword=Solid-phase peptide synthesis en-keyword=Cell-penetrating peptide kn-keyword=Cell-penetrating peptide en-keyword=Confocal laser scanning microscopy kn-keyword=Confocal laser scanning microscopy END start-ver=1.4 cd-journal=joma no-vol=21 cd-vols= no-issue=9 article-no= start-page=3140 end-page= dt-received= dt-revised= dt-accepted= dt-pub-year=2020 dt-pub=20200429 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Induction of CEMIP in Chondrocytes by Inflammatory Cytokines: Underlying Mechanisms and Potential Involvement in Osteoarthritis en-subtitle= kn-subtitle= en-abstract= kn-abstract=In patients with osteoarthritis (OA), there is a decrease in both the concentration and molecular size of hyaluronan (HA) in the synovial fluid and cartilage. Cell migration-inducing hyaluronidase 1 (CEMIP), also known as hyaluronan (HA)-binding protein involved in HA depolymerization (HYBID), was recently reported as an HA depolymerization-related molecule expressed in the cartilage of patients with OA. However, the underlying mechanism of CEMIP regulation is not well understood. We found that CEMIP expression was transiently increased by interleukine-1 beta (IL-1 beta) stimulation in chondrocytic cells. We also observed that ERK activation and NF-kappa B nuclear translocation were involved in the induction of CEMIP by IL-1 beta. In addition, both administration of HA and mechanical strain attenuated the CEMIP induction in IL-1 beta-stimulated chondrocytes. In conclusion, we clarified the regulatory mechanism of CEMIP in chondrocytes by inflammatory cytokines and suggested the potential involvement in osteoarthritis development. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=HatipogluOmer F. en-aut-sei=Hatipoglu en-aut-mei=Omer F. kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=AsanoKeiichi en-aut-sei=Asano en-aut-mei=Keiichi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=InagakiJunko en-aut-sei=Inagaki en-aut-mei=Junko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=NishidaKeiichiro en-aut-sei=Nishida en-aut-mei=Keiichiro kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=HirohataSatoshi en-aut-sei=Hirohata en-aut-mei=Satoshi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= affil-num=2 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= affil-num=3 en-affil=Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=4 en-affil=Department of Cell Chemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=5 en-affil=Department of Orthopaediac Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences kn-affil= affil-num=6 en-affil=Department of Medical Technology, Graduate School of Health Sciences, Okayama University kn-affil= en-keyword=cell migration-inducing hyaluronidase 1 (CEMIP) kn-keyword=cell migration-inducing hyaluronidase 1 (CEMIP) en-keyword=chondrocyte kn-keyword=chondrocyte en-keyword=hyaluronan kn-keyword=hyaluronan en-keyword=mechanical strain kn-keyword=mechanical strain en-keyword=nuclear factor kappa B (NF-kappa B) kn-keyword=nuclear factor kappa B (NF-kappa B) en-keyword=osteoarthritis kn-keyword=osteoarthritis END start-ver=1.4 cd-journal=joma no-vol=24 cd-vols= no-issue=3 article-no= start-page=241 end-page=247 dt-received= dt-revised= dt-accepted= dt-pub-year=2007 dt-pub=200703 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Alternative Leader-Exon Usage in Mouse IGF-I mRNA Variants: Class 1 and Class 2 IGF-I mRNAs en-subtitle= kn-subtitle= en-abstract= kn-abstract=The mouse IGF-I gene contains six exons, and exon 1 and exon 2 gene are considered to be leader exons. The regulatory mechanism of alternative usage of the leader exons is unclear in mice. The present study, was aimed at clarifying changes in class 1 (derived from exon 1) and class 2 (derived from exon 2) IGF-I mRNA expression in mice under various conditions. Both class 1 and class 2 IGF-I mRNAs were expressed in the mouse uterus, liver and kidney, and class 1 IGF-I mRNA was the major transcript in all organs studied. In the uterus, both class 1 and class 2 IGF-I mRNA expression changed markedly during the estrous cycle, with the highest level at proestrus, but in the liver and kidney there were no significant changes in IGF-I mRNA expression during the estrous cycle. Estrogen treatment increased both class 1 and class 2 IGF-I mRNA levels in the uterus of ovariectomized mice, but class 1 mRNA expression increased more in response to estrogen treatment than class 2 mRNA expression. These findings suggest that estrogen stimulates IGF-I gene expression in, uterine cells, and that a promoter involved in transcription of class 1 IGF-I mRNA is more responsive to estrogen. In conclusion, the present study revealed that two leader exons of mouse IGF-I gene are used in the uterus, liver and kidney. IGF-I mRNA levels of both classes changed during the estrous cycle in the uterus, but not in the liver or kidney. Estrogen increased IGF-I mRNA levels of both classes in the uterus. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OtsukiMariko en-aut-sei=Otsuki en-aut-mei=Mariko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MurakamiYousuke en-aut-sei=Murakami en-aut-mei=Yousuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=HirataKensaku en-aut-sei=Hirata en-aut-mei=Kensaku kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=TakeuchiSakae en-aut-sei=Takeuchi en-aut-mei=Sakae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=TakahashiSumio en-aut-sei=Takahashi en-aut-mei=Sumio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= affil-num=1 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=2 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=5 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=6 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University en-keyword=IGF-I kn-keyword=IGF-I en-keyword=leader exon kn-keyword=leader exon en-keyword=estrogen kn-keyword=estrogen en-keyword=uterus kn-keyword=uterus en-keyword=mouse kn-keyword=mouse END start-ver=1.4 cd-journal=joma no-vol=22 cd-vols= no-issue=9 article-no= start-page=1011 end-page=1021 dt-received= dt-revised= dt-accepted= dt-pub-year=2005 dt-pub=200509 dt-online= en-article= kn-article= en-subject= kn-subject= en-title= kn-title=Organ-Specific and Age-Dependent Expression of Insulin-like Growth Factor-I (IGF-I) mRNA Variants: IGF-IA and IB mRNAs in the Mouse en-subtitle= kn-subtitle= en-abstract= kn-abstract=Insulin-like growth factor-I (IGF-I) gene generates several IGF-I mRNA variants by alternative splicing. Two promoters are present in mouse IGF-I gene. Each promoter encodes two IGF-I mRNA variants (IGF-IA and IGF-IB mRNAs). Variants differ by the presence (IGF-IB) or absence (IGF-IA) of a 52-bp insert in the E domain-coding region. Functional differences among IGF-I mRNAs, and regulatory mechanisms for alternative splicing of IGF-I mRNA are not yet known. We analyzed the expression of mouse IGF-IA and IGF-IB mRNAs using SYBR Green real-time RT-PCR. In the liver, IGF-I mRNA expression increased from 10 days of age to 45 days. In the uterus and ovary, IGF-I mRNA expression increased from 21 days of age, and then decreased at 45 days. In the kidney, IGF-I mRNA expression decreased from 10 days of age. IGF-IA mRNA levels were higher than IGF-IB mRNA levels in all organs examined. Estradiol-17 beta (E2) treatment in ovariectomized mice increased uterine IGF-IA and IGF-IB mRNA levels from 3 hr after injection, and highest levels for both mRNAs were detected at 6 hr, and relative increase was greater for IGF-IB mRNA than for IGF-IA mRNA. These results suggest that expression of IGF-I mRNA variants is regulated in organ-specific and age-dependent manners, and estrogen is involved in the change of IGF-I mRNA variant expression. en-copyright= kn-copyright= en-aut-name=OhtsukiTakashi en-aut-sei=Ohtsuki en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=1 ORCID= en-aut-name=OtsukiMariko en-aut-sei=Otsuki en-aut-mei=Mariko kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=2 ORCID= en-aut-name=MurakamiYousuke en-aut-sei=Murakami en-aut-mei=Yousuke kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=3 ORCID= en-aut-name=MaekawaTetsuya en-aut-sei=Maekawa en-aut-mei=Tetsuya kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=4 ORCID= en-aut-name=YamamotoTakashi en-aut-sei=Yamamoto en-aut-mei=Takashi kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=5 ORCID= en-aut-name=AkasakaKoji en-aut-sei=Akasaka en-aut-mei=Koji kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=6 ORCID= en-aut-name=TakeuchiSakae en-aut-sei=Takeuchi en-aut-mei=Sakae kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=7 ORCID= en-aut-name=TakahashiSumio en-aut-sei=Takahashi en-aut-mei=Sumio kn-aut-name= kn-aut-sei= kn-aut-mei= aut-affil-num=8 ORCID= affil-num=1 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=2 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=3 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=4 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=5 en-affil= kn-affil=Department of Mathematical and Life Science, Graduate School of Science, Hiroshima University affil-num=6 en-affil= kn-affil=Misaki Marine Biological Station, University of Tokyo affil-num=7 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University affil-num=8 en-affil= kn-affil=Department of Biology, Faculty of Science, Okayama University en-keyword=insulin like growth factor-I (IGF-I) kn-keyword=insulin like growth factor-I (IGF-I) en-keyword=uterus kn-keyword=uterus en-keyword=estradiol kn-keyword=estradiol en-keyword=mouse kn-keyword=mouse END