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981.
982.
Masaya Mitsumoto Kanna Sugaya Kazuki Kazama Ryosuke Nakano Takahiro Kosugi Takeshi Murata Nobuyasu Koga 《International journal of molecular sciences》2021,22(23)
G-protein coupled receptors (GPCRs) are known for their low stability and large conformational changes upon transitions between multiple states. A widely used method for stabilizing these receptors is to make chimeric receptors by fusing soluble proteins (i.e., fusion partner proteins) into the intracellular loop 3 (ICL3) connecting the transmembrane helices 5 and 6 (TM5 and TM6). However, this fusion approach requires experimental trial and error to identify appropriate soluble proteins, residue positions, and linker lengths for making the fusion. Moreover, this approach has not provided state-targeting stabilization of GPCRs. Here, to rationally stabilize a class A GPCR, adenosine A2A receptor (A2AR) in a target state, we carried out the custom-made de novo design of α-helical fusion partner proteins, which can fix the conformation of TM5 and TM6 to that in an inactive state of A2AR through straight helical connections without any kinks or intervening loops. The chimeric A2AR fused with one of the designs (FiX1) exhibited increased thermal stability. Moreover, compared with the wild type, the binding affinity of the chimera against the agonist NECA was significantly decreased, whereas that against the inverse agonist ZM241385 was similar, indicating that the inactive state was selectively stabilized. Our strategy contributes to the rational state-targeting stabilization of GPCRs. 相似文献
983.
Kazuhiro Kiyohara Daisuke Uta Yuuya Nagaoka Yurika Kino Hideki Nonaka Midori Ninomiya-Baba Takuya Fujita 《International journal of molecular sciences》2022,23(4)
Several of the drugs currently available for the treatment of premature ejaculation (PE) (e.g., local anesthetics or antidepressants) are associated with numerous safety concerns and exhibit weak efficacy. To date, no therapeutics for PE have been approved in the United States, highlighting the need to develop novel agents with sufficient efficacy and fewer side effects. In this study, we focused on the histamine H3 receptor (H3R) as a potential target for the treatment of PE and evaluated the effects of imetit (an H3R/H4R agonist), ciproxifan (an H3R antagonist), and JNJ-7777120 (an H4R antagonist) in vivo. Our in vivo electrophysiological experiments revealed that imetit reduced mechanical stimuli-evoked neuronal firing in anesthetized rats. This effect was inhibited by ciproxifan but not by JNJ-7777120. Subsequently, we evaluated the effect of imetit using a copulatory behavior test to assess ejaculation latency (EL) in rats. Imetit prolonged EL, although this effect was inhibited by ciproxifan. These findings indicate that H3R stimulation suppresses mechanical stimuli-evoked neuronal firing in the spinal–penile neurotransmission system, thereby resulting in prolonged EL. To our knowledge, this is the first report to describe the relationship between H3R and PE. Thus, H3R agonists may represent a novel treatment option for PE. 相似文献
984.
Daisuke Yamada Tomoka Takao Masahiro Nakamura Toki Kitano Eiji Nakata Takeshi Takarada 《International journal of molecular sciences》2022,23(5)
Stem cell-based therapies and experimental methods rely on efficient induction of human pluripotent stem cells (hPSCs). During limb development, the lateral plate mesoderm (LPM) produces limb-bud mesenchymal (LBM) cells that differentiate into osteochondroprogenitor cells and form cartilage tissues in the appendicular skeleton. Previously, we generated PRRX1-tdTomato reporter hPSCs to establish the protocol for inducing the hPSC-derived PRRX1+ LBM-like cells. However, surface antigens that assess the induction efficiency of hPSC-derived PRRX1+ LBM-like cells from LPM have not been identified. Here, we used PRRX1-tdTomato reporter hPSCs and found that high pluripotent cell density suppressed the expression of PRRX1 mRNA and tdTomato after LBM-like induction. RNA sequencing and flow cytometry suggested that PRRX1-tdTomato+ LBM-like cells are defined as CD44high CD140Bhigh CD49f−. Importantly, other hPSC lines, including four human induced pluripotent stem cell lines (414C2, 1383D2, HPS1042, HPS1043) and two human embryonic stem cell lines (SEES4, SEES7), showed the same results. Thus, an appropriate cell density of hPSCs before differentiation is a prerequisite for inducing the CD44high CD140Bhigh CD49f− PRRX1+ LBM-like cells. 相似文献
985.
Erina Shiraishi Keishi Ishida Daisuke Matsumaru Akiko Ido Youhei Hiromori Hisamitsu Nagase Tsuyoshi Nakanishi 《International journal of molecular sciences》2021,22(24)
Propolis is a resinous mixture produced by bees from their secretions and plant material, so its composition varies depending on its botanical origin. Propolis has several beneficial bioactivities, but its skin sensitization properties have long been suspected. Nevertheless, the skin sensitization potency of Brazilian green propolis (BGP) has not been scientifically evaluated. Here, we used scientifically reliable tests to evaluate it. In vitro antigenicity test based on the human cell line activation test (OECD TG 442E) was performed by measuring the expression of CD54 and CD86, which are indicators of the antigenicity of test substances, on THP-1 and DC2.4 cells. BGP did not affect the expression of either marker on THP-1 cells, but upregulated the expression of CD86 on DC2.4 cells, suggesting that BGP may be a skin sensitizer. Then, we performed local lymph node assay (LLNA, OECD TG 429) as a definitive in vivo test. LLNA showed that 1.70% BGP primed skin sensitization and is a “moderate sensitizer”. Our results indicate scientific proof of the validity of arbitrary concentrations (1–2%), which have been used empirically, and provide the first scientific information on the safe use of BGP. 相似文献
986.
Yujiro Hayashi Junichiro Yamaguchi Kazuhiko Hibino Tatsunobu Sumiya Tatsuya Urushima Mitsuru Shoji Daisuke Hashizume Hiroyuki Koshino 《Advanced Synthesis \u0026amp; Catalysis》2004,346(12):1435-1439
trans‐4‐tert‐Butyldimethylsiloxy‐L ‐proline displays a greater catalytic activity than the parent proline without compromising the enantioselectivity, which widens the substrate scope in the α‐aminoxylation of carbonyl compounds, as well as O‐nitroso‐aldol/Michael, and Mannich reactions. 相似文献
987.
Daisuke Ueda 《Journal of Electroceramics》1999,3(2):105-113
GaAs microwave monolithic ICs with novel on-chip BST (Ba
X
Sr1 –X
TiO3) capacitors are demonstrated. MOD (Metal Organic Decomposition) technique was employed to make the BST thin film. The fabricated BST capacitor has the dielectric constant as high as 300, which is 50 times higher than the conventional Si3N4 one. The implemented GaAs MMIC with on-chip BST capacitor provides both high gain and low power dissipation characteristics. These MMIC can be packaged in a compact outline with reduced pin counts. BST capacitors also show the suppressed harmonics due to their low-pass filtering characteristics of the frequency roll-off around 2 GHz. The present technology will reduce the system size for a variety of the mobile communication systems. 相似文献
988.
Kaho Kanno Tomohisa Sakaue Mika Hamaguchi Kenji Namiguchi Daisuke Nanba Jun Aono Mie Kurata Junya Masumoto Shigeki Higashiyama Hironori Izutani 《International journal of molecular sciences》2021,22(19)
The characterization of aortic valve interstitial cells (VICs) cultured under optimal conditions is essential for understanding the molecular mechanisms underlying aortic valve stenosis. Here, we propose 2% hypoxia as an optimum VIC culture condition. Leaflets harvested from patients with aortic valve regurgitation were digested using collagenase and VICs were cultured under the 2% hypoxic condition. A significant increase in VIC growth was observed in 2% hypoxia (hypo-VICs), compared to normoxia (normo-VICs). RNA-sequencing revealed that downregulation of oxidative stress-marker genes (such as superoxide dismutase) and upregulation of cell cycle accelerators (such as cyclins) occurred in hypo-VICs. Accumulation of reactive oxygen species was observed in normo-VICs, indicating that low oxygen tension can avoid oxidative stress with cell-cycle arrest. Further mRNA quantifications revealed significant upregulation of several mesenchymal and hematopoietic progenitor markers, including CD34, in hypo-VICs. The stemness of hypo-VICs was confirmed using osteoblast differentiation assays, indicating that hypoxic culture is beneficial for maintaining growth and stemness, as well as for avoiding senescence via oxidative stress. The availability of hypoxic culture was also demonstrated in the molecular screening using proteomics. Therefore, hypoxic culture can be helpful for the identification of therapeutic targets and the evaluation of VIC molecular functions in vitro. 相似文献
989.
Hideyuki Yoshikawa Koichi Kondo Shigeyoshi Yoshida Daisuke Shindo Masanori Abe 《IEEJ Transactions on Electrical and Electronic Engineering》2007,2(4):445-449
Lorentz TEM observations of magnetic domain wall motion, as well as TEM observations of grain boundaries, were performed on spin‐sprayed ferrite films #1 (Ni0.17Zn0.22Fe2.61O4) and #2 (Ni0.19Zn0.20Co0.03Fe2.58O4), both 0.5 µm in thickness. They exhibit much higher natural resonance frequencies than the bulk ferrite and thus have been applied to gigahertz noise suppressors. Films #1 and #2 exhibit prominent and weak in‐plane uniaxial magnetic anisotropy, respectively, which is induced along the liquid flow direction during spin‐spraying. Both films have columnar crystallites with 100‐200 nm widths aligned perpendicular to the film plane, and the boundaries of the crystallites have no pores or impurity phases. Therefore, the crystallites are magnetically exchange‐coupled, which is responsible for the unusually high permeability and high natural resonance frequencies of the films. Under zero bias magnetic field, film #1 exhibits mosaic‐shaped magnetic domains, whereas film #2 exhibits magnetic domains elongated along the easy magnetization axis, both several hundred nanometers in width. For both films the domain structure remains unchanged when an in‐plane bias DC magnetic field,Hdc, of up to 10 Oe is applied along the hard axis. Under a stronger Hdc, the domain structure prominently changes, and the domain walls disappear when Hdc exceeds ∼100 Oe. This confirms our previous finding that the initial permeability is ascribed only to magnetization rotation, with no contribution from domain wall motion [J. Magn. Magn. Mater., 278 , 256 (2004)]. © 2007 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc. 相似文献
990.
The effects of the molecular aggregation structure on the rheological properties of thermoplastic polyurethane (TPU) were investigated. The TPU was composed of poly{(tetramethylene adipate)-co-(hexamethylene adipate)} glycol as the soft segments, 4,4′-diphenylmethane diisocyanate and 1,4-butanediol as the hard segments. The TPU sheets prepared by injection molding were annealed at various temperatures from 23 to 120 °C to vary the molecular aggregation structure. Glass transition temperature of the soft segment and melting points of the hard segment domains of the TPUs decreased and increased, respectively, with increasing annealing temperature. The results of DSC, solid-state NMR spectroscopy and dynamic viscoelastic measurements revealed that the degree of micro-phase separation of the TPUs becomes stronger with increasing annealing temperature due to the progress of formation of well-organized hard segment domains. The dynamic temperature sweep experiments for molten TPUs revealed that the temperature at critical gel point, which is defined as the temperature at which the dynamic storage modulus coincides with the loss storage modulus, in the cooling process increased with the progress of aggregation of the hard segments in the TPUs observed in the solid state. The uniaxial elongational viscosity measurements showed that TPUs exhibited an obvious strain hardening behavior with strain rate owing to residual hard segment domains at an operating temperature. It was revealed that the formation of well-organized hard segment domains had a profound effect on the rheological properties of TPUs, in particular on their elongational viscosity. 相似文献