31 results on '"Mukai SA"'
Search Results
2. Liquid/liquid dynamic phase separation induced by a focused laser
- Author
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30371735, Mukai, SA, Magome, N, Kitahata, H, Yoshikawa, K, 30371735, Mukai, SA, Magome, N, Kitahata, H, and Yoshikawa, K
- Published
- 2003
3. A solid-in-oil-in-water emulsion: An adjuvant-based immune-carrier enhances vaccine effect.
- Author
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Tahara Y, Mizuno R, Nishimura T, Mukai SA, Wakabayashi R, Kamiya N, Akiyoshi K, and Goto M
- Subjects
- Antigens, Emulsions, Humans, Vaccination, Water, Adjuvants, Immunologic, Cancer Vaccines
- Abstract
The biomaterial-based immunoengineering has become one of the most attractive research fields in the last decade. In the present study, a solid-in-oil-in-water (S/O/W) emulsion encapsulating antigen in the oil phase of an oil-in-water (O/W) emulsion was prepared as a novel vaccine carrier consisting of similar materials to the emulsion adjuvant of which the safety, immunogenicity and vaccination efficacy have been already confirmed in human. Direct observation by high-resolution confocal laser scanning microscopy and small angle X-ray scattering analysis showed that the antigens were dispersed inside of the oil phase of the S/O/W emulsion as solid-state particles. The S/O/W emulsion robustly produced antigen-specific antibodies and enhanced the antitumor effects in a therapeutic cancer vaccination compared with free antigens or the O/W emulsion in vivo. This result is in good agreement with the activation effect of antigen-specific cytotoxic T lymphocytes and antigen presentation by the S/O/W emulsion, indicating that the S/O/W emulsion consisting of already approved materials is a promising vaccine carrier to produce both humoral and cellular immunity., (Copyright © 2022 Elsevier Ltd. All rights reserved.)
- Published
- 2022
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4. Magnetically Navigated Protein Transduction In Vivo using Iron Oxide-Nanogel Chaperone Hybrid.
- Author
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Kawasaki R, Sasaki Y, Nishimura T, Katagiri K, Morita KI, Sekine Y, Sawada SI, Mukai SA, and Akiyoshi K
- Subjects
- Molecular Chaperones, Nanogels, Ferric Compounds, Magnetics
- Abstract
Systems for "protein transduction," intracellular delivery of functional proteins, are needed to address deliverability challenges of protein therapeutics. However, in vivo protein transduction remains challenging because of instability in serum, extracellular protease digestion and rapid excretion from the bloodstream. Here, a magnetically guided in vivo protein transduction using magnetic nanogel chaperone (MC) composed of iron oxide nanoparticles and a polysaccharide nanogel, a protein carrier inspired by "catch and release" mechanisms of molecular chaperones is demonstrated. The MC system enables efficient delivery of anti-cancer proteins, saporin and RNaseA, into cultured tumor lines and inhibits cell proliferation, mainly via apoptosis. Magnetic in vivo protein transduction via intravenous whole body administration is demonstrated in a fibrosarcoma model. By in vivo optical imaging, MC accumulated in tumor tissues under magnetic field three times more than without irradiation. With subcutaneous injection, saporin is delivered by MC to the cytoplasm in magnetically targeted tissues. In an oral cancer model, MC-delivered magnetically targeted saporin decreased tumor volume without significant body weight changes and no regrowth of tumor at 3 months after complete regression. Protein transduction with MC shows promise for cancer therapeutics and, potentially, for regenerative medicine and other biomedical applications., (© 2021 Wiley-VCH GmbH.)
- Published
- 2021
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5. Synergistic anti-tumor efficacy by combination therapy of a self-assembled nanogel vaccine with an immune checkpoint anti-PD-1 antibody.
- Author
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Miura R, Sawada SI, Mukai SA, Sasaki Y, and Akiyoshi K
- Abstract
Therapeutic strategies for cancer involving immune checkpoint inhibitors (ICIs) have been gaining widespread attention, but their efficacy remains limited. Thus, combination of ICI therapies with other therapeutic modalities may be required to improve their outcomes. In this study, we examined the improved efficacy of a CHP nanogel-based vaccine delivery system after combination with ICI therapy. For this, we evaluated the therapeutic efficacy of combining an anti-PD-1 antibody as an ICI with an OVA antigen-complexed CHP nanogel vaccine delivery system in a mouse E.G7-OVA tumor model. Mice were subcutaneously inoculated with E.G7-OVA tumor cells on one side of the back, and subcutaneously injected with OVA or the OVA/CHP nanogel vaccine on the other side of the back. Anti-PD-1 antibody was administered at defined intervals. Tumor volume, immune responses, and tumor-infiltrating cells were evaluated. Mice treated with OVA vaccine alone showed weak tumor suppression compared with untreated control mice. Mice receiving combined OVA/CHP nanogel vaccine and anti-PD-1 antibody therapy exhibited strong tumor growth suppression and markedly improved survival, suggesting that PD-1 signaling blockade by the anti-PD-1 antibody enhanced the anti-tumor efficacy of the OVA vaccine. Furthermore, tumor-infiltrating cells and immune responses were increased in the combined therapy group. No serious side effects were observed for any of the treatments. Taken together, the immune system activation induced by the CHP nanogel vaccine was synergistically enhanced by the anti-PD-1 antibody. The present findings suggest the potential for enhanced therapeutic efficacy by combining the CHP nanogel vaccine delivery system with ICI therapy for various cancer types., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)
- Published
- 2020
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6. Antigen Delivery to Antigen-Presenting Cells for Adaptive Immune Response by Self-Assembled Anionic Polysaccharide Nanogel Vaccines.
- Author
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Miura R, Sawada SI, Mukai SA, Sasaki Y, and Akiyoshi K
- Subjects
- Animals, Antigen-Presenting Cells drug effects, Dendritic Cells drug effects, Dendritic Cells immunology, Drug Delivery Systems, Epitopes, Female, Immunoglobulin G blood, Lymph Nodes drug effects, Lymph Nodes immunology, Mice, Mice, Inbred C57BL, Nanogels chemistry, Ovalbumin administration & dosage, Ovalbumin pharmacokinetics, Polysaccharides chemistry, RAW 264.7 Cells, Vaccines pharmacology, Adaptive Immunity drug effects, Antigen-Presenting Cells immunology, Vaccines administration & dosage, Vaccines chemistry
- Abstract
Although current vaccine technology induces sufficient antibody responses to prophylactically ward off viral infections, an anticancer vaccine that directs the patient's immune system to directly fight extant malignant cells will require inducing Th1 and cytotoxic T lymphocyte responses in addition to antibody-mediated activities. Thus, new mechanisms are necessary to deliver antigen to cells in the lymphatic system that will induce these responses. To this end, we have developed a cholesterol-bearing pullulan (CHP) self-assembly nanogel of less than 100 nm, which we have now further modified to be anionic by carboxyl group substitution. Overall, the nanogel-protected antigens during transport to the lymphatic system and converting the vehicle to an anionic charge improved interactions with antigen-presenting cells. We further show that these modified nanogels are a more efficient system for delivering antigen to antigen-presenting cells, particularly langerin-expressing cells, and that this induced significant adaptive immunity. Therefore, we think that this technology could be used to improve anticancer immunotherapies.
- Published
- 2020
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7. Supramacromolecular injectable hydrogels by crystallization-driven self-assembly of carbohydrate-conjugated poly(2-isopropyloxazoline)s for biomedical applications.
- Author
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Nishimura T, Sumi N, Mukai SA, Sasaki Y, and Akiyoshi K
- Subjects
- Crystallization, Biomedical Technology methods, Carbohydrates chemistry, Hydrogels chemistry, Oxazoles chemistry
- Abstract
An approach for the preparation of self-healing and injectable hydrogels based on the crystallization-driven self-assembly of carbohydrate-conjugated poly(2-isopropyloxazoline)s is reported. Hydrogelation does not require any organic solvents, as the polymers dissolve in water below their lower critical solution temperatures. The transplanted hydrogels cause no significant foreign-body response. Considering the simplicity of the method and the biocompatibility of the resulting injectable hydrogels, crystallization-driven hydrogelation of poly(oxazoline)-based polymers may potentially be used in a wide range of biomedical applications.
- Published
- 2019
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8. Magnetically Navigated Intracellular Delivery of Extracellular Vesicles Using Amphiphilic Nanogels.
- Author
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Mizuta R, Sasaki Y, Kawasaki R, Katagiri K, Sawada SI, Mukai SA, and Akiyoshi K
- Subjects
- Animals, Cell Differentiation, Mesenchymal Stem Cells cytology, Neurons, PC12 Cells, Rats, Surface-Active Agents, Drug Delivery Systems methods, Extracellular Vesicles metabolism, Magnetics, Nanogels chemistry
- Abstract
Various cells in vivo secrete exosomes consisting of lipid bilayers. They carry mRNAs and miRNAs capable of controlling cellular functions and can be used as drug delivery system nanocarriers. There is the current need to further improve the efficiency of exosome uptake into target cells. In this study, we prepared a hybrid of exosomes and magnetic nanoparticles, which could be guided to target cells by a magnetic field for efficient uptake. Magnetic nanogels were prepared and hybridized to fluorescently labeled exosomes isolated from PC12 cells. By applying a magnetic field to a hybrid with magnetic nanogel, exosomes were efficiently transferred into target cells as confirmed by confocal laser microscopy. Finally, we found that differentiation of adipose-derived stem cells to neuron-like cells was enhanced by magnetic induction of the exosome-magnetic nanogel hybrid, indicating maintenance of the intrinsic functions of the exosomes in the differentiation of adipose-derived stem cells.
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- 2019
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9. Nanogel Tectonics for Tissue Engineering: Protein Delivery Systems with Nanogel Chaperones.
- Author
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Hashimoto Y, Mukai SA, Sasaki Y, and Akiyoshi K
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- Animals, Antigens, Neoplasm chemistry, Antigens, Neoplasm immunology, Antigens, Neoplasm metabolism, Bone Diseases therapy, Bone Diseases veterinary, Bone Morphogenetic Protein 2 chemistry, Bone Morphogenetic Protein 2 metabolism, Bone Regeneration, Drug Carriers chemistry, Fibroblast Growth Factors chemistry, Fibroblast Growth Factors metabolism, Humans, Membrane Proteins chemistry, Membrane Proteins immunology, Membrane Proteins metabolism, Mice, Molecular Chaperones chemistry, Molecular Chaperones metabolism, Nanogels, Polyethylene Glycols therapeutic use, Polyethyleneimine therapeutic use, Polysaccharides chemistry, beta-Cyclodextrins chemistry, Polyethylene Glycols chemistry, Polyethyleneimine chemistry, Tissue Engineering
- Abstract
Amphiphilic polysaccharide self-assembled (SA) nanogels are promising protein carriers owing to their chaperone-like activity that allows them to nanoencapsulate proteins within their polymer networks. The chaperoning function is an important concept that has led to breakthroughs in the development of effective protein drug delivery systems by stabilizing formulations and controlling the quality of unstable proteins. Recently, nanogel-tectonic materials that integrate SA nanogels as building blocks have been designed as new hydrogel biomaterials. This article describes recent progress and applications of SA nanogel tectonic materials as protein delivery systems for tissue engineering., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2018
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10. Self-Assembled Polypeptide Nanogels with Enzymatically Transformable Surface as a Small Interfering RNA Delivery Platform.
- Author
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Nishimura T, Yamada A, Umezaki K, Sawada SI, Mukai SA, Sasaki Y, and Akiyoshi K
- Subjects
- Animals, Cations chemistry, Cell Line, Tumor, Lysine chemistry, Mice, Molecular Weight, Nanogels, Polysaccharides chemistry, Vascular Endothelial Growth Factor A chemistry, alpha-Amylases chemistry, Nanoparticles chemistry, Peptides chemistry, Polyethylene Glycols chemistry, Polyethyleneimine chemistry, Polymers chemistry, RNA, Small Interfering chemistry
- Abstract
Nanometer-size gel particles, or nanogels, have potential for delivering therapeutic macromolecules. A cationic surface promotes cellular internalization of nanogels, but undesired electrostatic interactions, such as with blood components, cause instability and toxicities. Poly(ethylene glycol) coating has been used to shield charges, but this decreases delivery efficiency. Technical difficulties in synthesis and controlling molecular weights make it unfeasible to, instead, coat with biodegradable polymers. Our proposed solution is cationized nanogels enzymatically functionalized with branched polysaccharide chains, forming a shell to shield charges and increase stability. Biodegradation of the polysaccharides by an endogenous enzyme would then expose the cationic charges, allowing cellular internalization and cargo delivery. We tested this concept, preparing maltopentaose functionalized cholesteryl poly(l-lysine) nanogel and using tandem enzymatic polymerization with glycogen phosphorylase and glycogen branching enzyme, to add branched amylose moieties, forming a CbAmyPL nanogel. We characterized CbAmyPL nanogels and investigated their suitability as small interfering RNA (siRNA) carriers in murine renal carcinoma (Renca) cells. The nanogels had neutral ζ potential values that became positive after degradation by α-amylase. Foster resonance energy transfer demonstrated that the nanogels formed stable complexes with siRNA, even in the presence of bovine serum albumin and after α-amylase exposure. The nanogels, with or without α-amylase, were not cytotoxic. Complexes of CbAmyPL with siRNA against vascular endothelial growth factor (VEGF), when incubated alone with Renca cells decreased VEGF mRNA levels by only 20%. With α-amylase added, however, VEGF mRNA knockdown by the siRNA/nanogels complexes was 50%. Our findings strongly supported the hypothesis that enzyme-responsive nanogels are promising as a therapeutic siRNA delivery platform.
- Published
- 2017
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11. Self-Assembled Nanogels of Cholesterol-Bearing Hydroxypropyl Cellulose: A Thermoresponsive Building Block for Nanogel Tectonic Materials.
- Author
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Tahara Y, Sakiyama M, Takeda S, Nishimura T, Mukai SA, Sawada SI, Sasaki Y, and Akiyoshi K
- Abstract
Hydroxypropyl cellulose (HPC) is a fascinating polysaccharide to use in developing a nanogel to be a thermoresponsive building unit for nanogel tectonic materials. Cholesterol-bearing HPC (Ch-HPC) self-assembled to form nanogels through hydrophobic interactions of the cholesteryl groups in water. Ch-HPC nanogels had a lower critical solution temperature in line with that of native HPC. The particle size of Ch-HPC nanogels was reversibly controlled by the temperature and salting-out effect. The thermoresponsive property was also observed in Ch-HPC nanogel-cross-linked macrogels. These results suggest that a Ch-HPC nanogel is an attractive building block for thermoresponsive nanogel tectonic materials.
- Published
- 2016
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12. Nanoscopic Structural Investigation of Physically Cross-Linked Nanogels Formed from Self-Associating Polymers.
- Author
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Sekine Y, Endo H, Iwase H, Takeda S, Mukai SA, Fukazawa H, Littrell KC, Sasaki Y, and Akiyoshi K
- Abstract
The detailed structure of a nanogel formed by self-association of cholesterol-bearing pullulans (CHPs) was determined by contrast variation small-angle neutron scattering. The decomposition of scattering intensities into partial scattering functions of each CHP nanogel component, i.e., pullulan, cholesterol, and the cross-term between the pullulan and the cholesterol, allows us to investigate the internal structure of the nanogel. The effective spherical radius of the skeleton formed by pullulan chains was found to be 8.1 ± 0.3 nm. In the CHP nanogel, there are about 19 cross-linking points where a cross-linking point is formed by aggregation of trimer cholesterol molecules, and the spatially inhomogeneous distribution of the cross-linking points in the nanogel can be represented by the mass fractal dimension of 2.6. The average radius of gyration of the partial chains can also be determined to be 1.7 ± 0.1 nm by analyzing the extracted cross-correlation between the cross-linker and the tethered polymer chain quantitatively, and the size agrees with the value assuming random distribution of the cross-linkers on the chains. As the result, the complex structure of the nanogels is coherently revealed at the nanoscopic level.
- Published
- 2016
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13. Magnetically Guided Protein Transduction by Hybrid Nanogel Chaperones with Iron Oxide Nanoparticles.
- Author
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Kawasaki R, Sasaki Y, Katagiri K, Mukai SA, Sawada S, and Akiyoshi K
- Subjects
- Animals, Cattle, Fluorescent Dyes chemistry, Glucans chemistry, HeLa Cells, Humans, Insulin chemistry, Insulin metabolism, Magnetics, Microscopy, Confocal, Molecular Chaperones chemistry, Molecular Chaperones metabolism, Nanogels, Proteins chemistry, Serum Albumin, Bovine chemistry, Serum Albumin, Bovine metabolism, Drug Carriers chemistry, Ferric Compounds chemistry, Metal Nanoparticles chemistry, Polyethylene Glycols chemistry, Polyethyleneimine chemistry, Proteins metabolism
- Abstract
Protein pharmaceuticals show great therapeutic promise, but effective intracellular delivery remains challenging. To address the need for efficient protein transduction systems, we used a magnetic nanogel chaperone (MC): a hybrid of a polysaccharide nanogel, a protein carrier with molecular chaperone-like properties, and iron oxide nanoparticles, enabling magnetically guided delivery. The MC complexed with model proteins, such as BSA and insulin, and was not cytotoxic. Cargo proteins were delivered to the target HeLa cell cytosol using a magnetic field to promote movement of the protein complex toward the cells. Delivery was confirmed by fluorescence microscopy and flow cytometry. Delivered β-galactosidase, inactive within the MC complex, became enzymatically active within cells to convert a prodrug. Thus, cargo proteins were released from MC complexes through exchange interactions with cytosolic proteins. The MC is a promising tool for realizing the therapeutic potential of proteins., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
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- 2016
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14. Advanced Artificial Extracellular Matrices Using Amphiphilic Nanogel-Cross-Linked Thin Films To Anchor Adhesion Proteins and Cytokines.
- Author
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Hashimoto Y, Mukai SA, Sawada SI, Sasaki Y, and Akiyoshi K
- Abstract
A novel type of nanogel-cross-linked (NanoClik) film composed of acryloyl-modified cholesterol-bearing pullulan nanogels with pentaerythritol tetra(mercaptoethyl)polyoxyethylene as a cross-linker is created through the Michael addition coupled with solvent evaporation. Tensile testing and atomic force microscopy show that the elastic property of the NanoClik films can be controlled by changing the cross-linker concentration. The NanoClik films strongly absorb proteins after simple immersion in solutions of functional proteins, including the hormone insulin, cytokine bone morphogenetic protein-2 (BMP-2), and vitronectin. The amphiphilic nanogels in the films induce this absorption by acting as anchoring and loading proteins. Mouse embryo fibroblast cells adhere to and proliferate on the NanoClik films anchoring vitronectin, while NanoClik films loaded with BMP-2 strongly increase the differentiation of human mesenchymal stem cells into osteoblasts. These results suggest that the NanoClik films act as a novel artificial extracellular matrix that enables the reservation of various biological proteins to the nanogels.
- Published
- 2016
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15. Engineering hybrid exosomes by membrane fusion with liposomes.
- Author
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Sato YT, Umezaki K, Sawada S, Mukai SA, Sasaki Y, Harada N, Shiku H, and Akiyoshi K
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- Animals, Blotting, Western, Exosomes chemistry, Flow Cytometry, Fluorescence Resonance Energy Transfer, Fluorescent Dyes chemistry, Freezing, HeLa Cells, Humans, Liposomes chemistry, Membrane Fusion, Mice, Microscopy, Confocal, Nanoparticles chemistry, Nanoparticles metabolism, RAW 264.7 Cells, Receptor, ErbB-2 immunology, Receptor, ErbB-2 metabolism, Exosomes metabolism, Liposomes metabolism
- Abstract
Exosomes are a valuable biomaterial for the development of novel nanocarriers as functionally advanced drug delivery systems. To control and modify the performance of exosomal nanocarriers, we developed hybrid exosomes by fusing their membranes with liposomes using the freeze-thaw method. Exosomes embedded with a specific membrane protein isolated from genetically modified cells were fused with various liposomes, confirming that membrane engineering methods can be combined with genetic modification techniques. Cellular uptake studies performed using the hybrid exosomes revealed that the interactions between the developed exosomes and cells could be modified by changing the lipid composition or the properties of the exogenous lipids. These results suggest that the membrane-engineering approach reported here offers a new strategy for developing rationally designed exosomes as hybrid nanocarriers for use in advanced drug delivery systems.
- Published
- 2016
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16. A light sensitive self-assembled nanogel as a tecton for protein patterning materials.
- Author
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Nishimura T, Takara M, Mukai SA, Sawada S, Sasaki Y, and Akiyoshi K
- Subjects
- Spectrophotometry, Ultraviolet, Gels, Light, Nanostructures, Proteins chemistry
- Abstract
A self-assembled nanogel is constructed from light-sensitive cholesteryl pullulan (Ls-CHP) by using photo-labile ortho-nitrobenzyl (o-NB) units. The nanogel-based film is obtained by evaporation of an Ls-CHP nanogel solution. Exposure of the resulting nanogel-based film to light with a mask resulted in a patterned film that can encapsulate FITC-insulin.
- Published
- 2016
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17. Exosomes as nanocarriers for systemic delivery of the Helicobacter pylori virulence factor CagA.
- Author
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Shimoda A, Ueda K, Nishiumi S, Murata-Kamiya N, Mukai SA, Sawada S, Azuma T, Hatakeyama M, and Akiyoshi K
- Subjects
- Antigens, Bacterial genetics, Bacterial Proteins genetics, Biological Transport, Biomarkers, Cell Line, Chromatography, Liquid, Helicobacter Infections complications, Helicobacter Infections microbiology, Humans, Protein Transport, Stomach Neoplasms etiology, Tandem Mass Spectrometry, Virulence Factors, Antigens, Bacterial metabolism, Bacterial Proteins metabolism, Exosomes metabolism, Helicobacter pylori physiology
- Abstract
CagA, encoded by cytotoxin-associated gene A (cagA), is a major virulence factor of Helicobacter pylori, a gastric pathogen involved in the development of upper gastrointestinal diseases. Infection with cagA-positive H. pylori may also be associated with diseases outside the stomach, although the mechanisms through which H. pylori infection promotes extragastric diseases remain unknown. Here, we report that CagA is present in serum-derived extracellular vesicles, known as exosomes, in patients infected with cagA-positive H. pylori (n = 4). We also found that gastric epithelial cells inducibly expressing CagA secrete exosomes containing CagA. Addition of purified CagA-containing exosomes to gastric epithelial cells induced an elongated cell shape, indicating that the exosomes deliver functional CagA into cells. These findings indicated that exosomes secreted from CagA-expressing gastric epithelial cells may enter into circulation, delivering CagA to distant organs and tissues. Thus, CagA-containing exosomes may be involved in the development of extragastric disorders associated with cagA-positive H. pylori infection.
- Published
- 2016
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18. Comprehensive study of liposome-assisted synthesis of membrane proteins using a reconstituted cell-free translation system.
- Author
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Niwa T, Sasaki Y, Uemura E, Nakamura S, Akiyama M, Ando M, Sawada S, Mukai SA, Ueda T, Taguchi H, and Akiyoshi K
- Subjects
- Computational Biology methods, Escherichia coli genetics, Escherichia coli metabolism, Escherichia coli Proteins genetics, Escherichia coli Proteins metabolism, Hydrophobic and Hydrophilic Interactions, Protein Folding, Solubility, Cell-Free System metabolism, Liposomes metabolism, Membrane Proteins genetics, Membrane Proteins metabolism, Protein Biosynthesis genetics
- Abstract
Membrane proteins play pivotal roles in cellular processes and are key targets for drug discovery. However, the reliable synthesis and folding of membrane proteins are significant problems that need to be addressed owing to their extremely high hydrophobic properties, which promote irreversible aggregation in hydrophilic conditions. Previous reports have suggested that protein aggregation could be prevented by including exogenous liposomes in cell-free translation processes. Systematic studies that identify which membrane proteins can be rescued from irreversible aggregation during translation by liposomes would be valuable in terms of understanding the effects of liposomes and developing applications for membrane protein engineering in the context of pharmaceutical science and nanodevice development. Therefore, we performed a comprehensive study to evaluate the effects of liposomes on 85 aggregation-prone membrane proteins from Escherichia coli by using a reconstituted, chemically defined cell-free translation system. Statistical analyses revealed that the presence of liposomes increased the solubility of >90% of the studied membrane proteins, and ultimately improved the yields of the synthesized proteins. Bioinformatics analyses revealed significant correlations between the liposome effect and the physicochemical properties of the membrane proteins.
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- 2015
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19. Nanocarrier-Integrated Microspheres: Nanogel Tectonic Engineering for Advanced Drug-Delivery Systems.
- Author
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Tahara Y, Mukai SA, Sawada S, Sasaki Y, and Akiyoshi K
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- Animals, Drug Liberation, Drug Stability, Injections, Mice, Models, Molecular, Molecular Conformation, Drug Carriers chemistry, Engineering, Hydrogels chemistry, Microspheres, Nanostructures chemistry
- Abstract
A nanocarrier-integrated bottom-up method is a promising strategy for advanced drug-release systems. Self-assembled nanogels, which are one of the most beneficial nanocarriers for drug-delivery systems, are tectonically integrated to prepare nanogel-crosslinked (NanoClik) microspheres. NanoClik microspheres consisting of nanogel-derived structures (observed by STED microscopy) release "drug-loaded nanogels" after hydrolysis, resulting in successful sustained drug delivery in vivo., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2015
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20. Glyco Star Polymers as Helical Multivalent Host and Biofunctional Nano-Platform.
- Author
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Nishimura T, Mukai SA, Sawada SI, and Akiyoshi K
- Abstract
A series of amylose-based star polymers (1, 2, 4, and 8 arms) as a new glyco biomaterial was synthesized by a click reaction and enzymatic polymerization of specific primers with phosphorylase. The molecular weights were controlled by the enzymatic reaction. Further polymerization resulted in a viscous solution and, especially, for the 8-arm primer, a hydrogel was obtained due to effective cross-linking between the multiarmed structures. The star polymers with a degree of polymerization of about 60 per arm acted as an allosteric multivalent host for hydrophobic molecules by helical formation. A cationic 8-arm star polymer catalyzed DNA strand exchange as a nucleic acid chaperone. Amylose-based star polymers are promising building blocks for producing advanced hybrid glyco biomaterials.
- Published
- 2015
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21. Nanogel tectonic porous gel loading biologics, nanocarriers, and cells for advanced scaffold.
- Author
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Hashimoto Y, Mukai SA, Sawada S, Sasaki Y, and Akiyoshi K
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- Animals, Cattle, Cholesterol chemistry, Cross-Linking Reagents chemistry, Elastic Modulus, Female, Implants, Experimental, Liposomes, Mice, Mice, Inbred BALB C, NIH 3T3 Cells, Nanogels, Polysaccharides chemistry, Porosity, Subcutaneous Tissue, Drug Carriers chemistry, Fibroblasts cytology, Polyethylene Glycols chemistry, Polyethyleneimine chemistry, Tissue Scaffolds chemistry
- Abstract
We developed a new self-assembled amphiphilic nanogel-crosslinked porous (NanoCliP) gel that can trap proteins, liposomes, and cells. The NanoCliP gel was prepared by Michael addition of a self-assembled nanogel of acryloyl group-modified cholesterol-bearing pullulan to pentaerythritol tetra (mercaptoethyl) polyoxyethylene, followed by freezing-induced phase separation. Dynamic rheological analysis revealed that the storage modulus (G') of the NanoCliP gel was approximately 10 times greater than that of a nonporous nanogel-crosslinked gel. Two-photon excitation deep imaging revealed that the NanoCliP gel comprises interconnected pores of several hundred micrometers in diameter. The NanoCliP gel trapped proteins and liposomes via hydrophobic interactions because its amphiphilic nanogels exhibit chaperone-like activity. Mouse embryonic fibroblasts penetrated the interconnected pores and adhered to the porous surface of fibronectin-complexed NanoCliP gel. In vivo, the NanoCliP gel enhanced cell infiltration, tissue ingrowth, and neovascularization without requiring exogenous growth factors, suggesting that the NanoCliP gel is a promising scaffold for tissue engineering., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2015
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22. Amylose-Based Cationic Star Polymers for siRNA Delivery.
- Author
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Nishimura T, Umezaki K, Mukai SA, Sawada S, and Akiyoshi K
- Subjects
- Animals, Cations chemistry, Cell Line, Tumor, Cell Survival drug effects, Mice, RNA, Small Interfering genetics, RNA, Small Interfering pharmacology, Amylose chemistry, Drug Carriers chemistry, Polymers chemistry, RNA, Small Interfering chemistry, RNA, Small Interfering pharmacokinetics
- Abstract
A new siRNA delivery system using a cationic glyco-star polymer is described. Spermine-modified 8-arm amylose star polymer (with a degree of polymerization of approximately 60 per arm) was synthesized by chemoenzymatic methods. The cationic star polymer effectively bound to siRNA and formed spherical complexes with an average hydrodynamic diameter of 230 nm. The cationic 8-arm star polymer complexes showed superior cellular uptake characteristics and higher gene silencing effects than a cationic 1-arm polymer. These results suggest that amylose-based star polymers are a promising nanoplatform for glycobiomaterials.
- Published
- 2015
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23. Cycloamylose-nanogel drug delivery system-mediated intratumor silencing of the vascular endothelial growth factor regulates neovascularization in tumor microenvironment.
- Author
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Fujii H, Shin-Ya M, Takeda S, Hashimoto Y, Mukai SA, Sawada S, Adachi T, Akiyoshi K, Miki T, and Mazda O
- Subjects
- Animals, Carcinoma, Renal Cell blood supply, Carcinoma, Renal Cell pathology, Cell Line, Tumor, Drug Delivery Systems, Gene Expression Regulation, Neoplastic, Genetic Therapy, Humans, Kidney Neoplasms blood supply, Kidney Neoplasms pathology, Mice, Nanogels, Neoplasms, Experimental, Neovascularization, Pathologic pathology, Organ Specificity, Polyethylene Glycols chemistry, Polyethyleneimine chemistry, Tumor Microenvironment, Carcinoma, Renal Cell therapy, Cyclodextrins administration & dosage, Kidney Neoplasms therapy, Neovascularization, Pathologic therapy, RNA, Small Interfering administration & dosage, Vascular Endothelial Growth Factor A antagonists & inhibitors
- Abstract
RNAi enables potent and specific gene silencing, potentially offering useful means for treatment of cancers. However, safe and efficient drug delivery systems (DDS) that are appropriate for intra-tumor delivery of siRNA or shRNA have rarely been established, hindering clinical application of RNAi technology to cancer therapy. We have devised hydrogel polymer nanoparticles, or nanogel, and shown its validity as a novel DDS for various molecules. Here we examined the potential of self-assembled nanogel of cholesterol-bearing cycloamylose with spermine group (CH-CA-Spe) to deliver vascular endothelial growth factor (VEGF)-specific short interfering RNA (siVEGF) into tumor cells. The siVEGF/nanogel complex was engulfed by renal cell carcinoma (RCC) cells through the endocytotic pathway, resulting in efficient knockdown of VEGF. Intra-tumor injections of the complex significantly suppressed neovascularization and growth of RCC in mice. The treatment also inhibited induction of myeloid-derived suppressor cells, while it decreased interleukin-17A production. Therefore, the CH-CA-Spe nanogel may be a feasible DDS for intra-tumor delivery of therapeutic siRNA. The results also suggest that local suppression of VEGF may have a positive impact on systemic immune responses against malignancies., (© 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.)
- Published
- 2014
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24. Nanogel-based immunologically stealth vaccine targets macrophages in the medulla of lymph node and induces potent antitumor immunity.
- Author
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Muraoka D, Harada N, Hayashi T, Tahara Y, Momose F, Sawada S, Mukai SA, Akiyoshi K, and Shiku H
- Subjects
- Amino Acid Sequence, Animals, Antigens, Neoplasm chemistry, Antigens, Neoplasm immunology, CD8-Positive T-Lymphocytes immunology, Cancer Vaccines chemistry, Cell Line, Tumor, Cell Proliferation, Female, Glucans chemistry, Mice, Molecular Sequence Data, T-Lymphocytes immunology, Toll-Like Receptors agonists, Cancer Vaccines immunology, Drug Carriers chemistry, Hydrogels chemistry, Lymph Nodes immunology, Macrophages immunology, Nanoparticles chemistry
- Abstract
Because existing therapeutic cancer vaccines provide only a limited clinical benefit, a different vaccination strategy is necessary to improve vaccine efficacy. We developed a nanoparticulate cancer vaccine by encapsulating a synthetic long peptide antigen within an immunologically inert nanoparticulate hydrogel (nanogel) of cholesteryl pullulan (CHP). After subcutaneous injection to mice, the nanogel-based vaccine was efficiently transported to the draining lymph node, and was preferentially engulfed by medullary macrophages but was not sensed by other macrophages and dendritic cells (so-called "immunologically stealth mode"). Although the function of medullary macrophages in T cell immunity has been unexplored so far, these macrophages effectively cross-primed the vaccine-specific CD8(+) T cells in the presence of a Toll-like receptor (TLR) agonist as an adjuvant. The nanogel-based vaccine significantly inhibited in vivo tumor growth in the prophylactic and therapeutic settings, compared to another vaccine formulation using a conventional delivery system, incomplete Freund's adjuvant. We also revealed that lymph node macrophages were highly responsive to TLR stimulation, which may underlie the potency of the macrophage-oriented, nanogel-based vaccine. These results indicate that targeting medullary macrophages using the immunologically stealth nanoparticulate delivery system is an effective vaccine strategy.
- Published
- 2014
- Full Text
- View/download PDF
25. Anomalous long-range repulsion between silica surfaces induced by density inhomogeneities in supercritical ethanol.
- Author
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Mukai SA, Koyama T, Tsujii K, and Deguchi S
- Abstract
Anomalous long-range repulsion, extending over several micrometres, emerged between silica surfaces around the ridge of density fluctuations in supercritical ethanol at temperatures and pressures near the gas/liquid critical point (T(c) = 241 °C, P(c) = 6.14 MPa). Analysis shows that augmentation of ethanol density around silica surfaces in the presence of density fluctuations facilitates dissociation of silanol groups, leading to long-range electrostatic repulsion in the nonpolar medium.
- Published
- 2014
- Full Text
- View/download PDF
26. Non-engineered nanoparticles of C₆₀.
- Author
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Deguchi S, Mukai SA, Sakaguchi H, and Nonomura Y
- Subjects
- Microscopy, Electron, Scanning, Hydrocarbons chemistry, Nanoparticles
- Abstract
We discovered that rubbing bulk solids of C₆₀ between fingertips generates nanoparticles including the ones smaller than 20 nm. Considering the difficulties usually associated with nanoparticle production by pulverisation, formation of nanoparticles by such a mundane method is unprecedented and noteworthy. We also found that nanoparticles of C₆₀ could be generated from bulk solids incidentally without deliberate engineering of any sort. Our findings imply that there exist highly unusual human exposure routes to nanoparticles of C₆₀, and elucidating formation mechanisms of nanoparticles is crucial in assessing their environmental impacts.
- Published
- 2013
- Full Text
- View/download PDF
27. Microbial growth at hyperaccelerations up to 403,627 x g.
- Author
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Deguchi S, Shimoshige H, Tsudome M, Mukai SA, Corkery RW, Ito S, and Horikoshi K
- Subjects
- Acceleration, Bacteria cytology, Escherichia coli growth & development, Exobiology, Fungi cytology, Gram-Negative Bacteria growth & development, Gram-Positive Bacteria growth & development, Hydrostatic Pressure, Paracoccus denitrificans growth & development, Saccharomyces cerevisiae growth & development, Stress, Mechanical, Bacteria growth & development, Extraterrestrial Environment, Fungi growth & development, Hypergravity
- Abstract
It is well known that prokaryotic life can withstand extremes of temperature, pH, pressure, and radiation. Little is known about the proliferation of prokaryotic life under conditions of hyperacceleration attributable to extreme gravity, however. We found that living organisms can be surprisingly proliferative during hyperacceleration. In tests reported here, a variety of microorganisms, including Gram-negative Escherichia coli, Paracoccus denitrificans, and Shewanella amazonensis; Gram-positive Lactobacillus delbrueckii; and eukaryotic Saccharomyces cerevisiae, were cultured while being subjected to hyperaccelerative conditions. We observed and quantified robust cellular growth in these cultures across a wide range of hyperacceleration values. Most notably, the organisms P. denitrificans and E. coli were able to proliferate even at 403,627 × g. Analysis shows that the small size of prokaryotic cells is essential for their proliferation under conditions of hyperacceleration. Our results indicate that microorganisms cannot only survive during hyperacceleration but can display such robust proliferative behavior that the habitability of extraterrestrial environments must not be limited by gravity.
- Published
- 2011
- Full Text
- View/download PDF
28. Anomalously stable dispersions of graphite in water/acetone mixtures.
- Author
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Nonomura Y, Morita Y, Deguchi S, and Mukai SA
- Subjects
- Particle Size, Surface Properties, Acetone chemistry, Graphite chemistry, Water chemistry
- Abstract
Highly stable dispersions of graphite are obtained in water/acetone mixtures, while graphite precipitates promptly in water and in acetone. We found that acetone facilitates disintegration of the graphite particles and their aggregates during a sonication process. Fine graphite particles are stabilised by the electrostatic repulsion due to negative charges on the graphite particle surfaces., (Copyright © 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
29. Comment on "Effect of pressure on colloidal behavior in hydrothermal water".
- Author
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Deguchi S and Mukai SA
- Subjects
- Chemistry, Physical methods, Diamond chemistry, Kinetics, Models, Theoretical, Pressure, Temperature, Colloids chemistry, Nanoparticles chemistry, Water chemistry
- Published
- 2009
- Full Text
- View/download PDF
30. Supercritical ethanol--a fascinating dispersion medium for silica nanoparticles.
- Author
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Ghosh SK, Deguchi S, Mukai SA, and Tsujii K
- Subjects
- Chromatography, Supercritical Fluid, Models, Chemical, Particle Size, Pressure, Solubility, Temperature, Ethanol chemistry, Nanoparticles chemistry, Silicon Dioxide chemistry
- Abstract
For the first time, the dispersion stability of silica nanoparticles has been investigated in high-temperature and high-pressure ethanol by measuring the hydrodynamic diffusion coefficient of the particles by means of dynamic light scattering. The silica nanoparticles remain stable in ethanol within a wide temperature range of 24-304 degrees C at 12.3 MPa, and they start to aggregate at T >or= 305 degrees C. Numerical analysis reveals that the net interparticle repulsive potential barrier decreases dramatically with increasing temperature due to the changes in the properties of the medium. We observed that particles remain highly stable in the nonpolar supercritical ethanol in the temperature regime 241-304 degrees C, where the DLVO potential barrier is only 5-2 k(B)T. The dispersion stability of silica nanoparticles at this low potential barrier in high-temperature and high-pressure ethanol, especially in the supercritical ethanol, is fascinating. The silica-ethanol system might be a unique and special example in the colloidal dispersions. Results suggest that silica nanoparticles may be used as a model colloid to investigate the colloidal transport phenomena in the supercritical ethanol.
- Published
- 2007
- Full Text
- View/download PDF
31. Stabilization of C60 nanoparticles by protein adsorption and its implications for toxicity studies.
- Author
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Deguchi S, Yamazaki T, Mukai SA, Usami R, and Horikoshi K
- Subjects
- Adsorption drug effects, Animals, Cattle, Dose-Response Relationship, Drug, Drug Stability, Flocculation, Fullerenes pharmacology, Humans, Particle Size, Phosphates chemistry, Serum Albumin chemistry, Sodium Chloride chemistry, Time Factors, Toxicity Tests methods, Toxicity Tests trends, Ultrafiltration methods, Fullerenes chemistry, Nanoparticles chemistry, Proteins chemistry
- Abstract
Dispersion stability of nanoparticles of C60 under a model condition simulating a physiological environment was studied by dynamic light scattering. Although the C60 nanoparticles at a concentration of 9.3 x 10(-6) M (6.7 microg/mL) coagulated and precipitated out rapidly in phosphate buffered saline, coagulation was suppressed completely when HSA was present at concentrations above 1 mg/mL. DLS results show that the HSA molecules adsorb onto the surfaces of the C60 nanoparticles, thereby forming a protective layer, and prevent salt-induced coagulation. DLS results also indicate that the HSA molecules take an expanded conformation on the surface. Our findings suggest that C60 nanoparticles can be stabilized in the physiological environment even if they are not deliberately stabilized by using stabilizers and are of significant implications for the on-going efforts to evaluate the cytotoxicity of C60 nanoparticles in which no such effect has been considered.
- Published
- 2007
- Full Text
- View/download PDF
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