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＠TakekiHamasaki, TaichiKashiwagi, TakaakiKomatsu, ShigeruKabayama, NoboruNakamichi,KiichiroTeruya, SanetakaShirahata
, A new colorimetric method for determining antioxidant levels using 3,5-dibromo-4- nitrosobenzene sulfonate (DBNBS)., MethodsX, 10.1016/j.mex.2022.101797, 9, 2022.07, We describe here a novel assay that determines the total a+ntioxidative activities of known antioxidants and antioxidants in beverages. The method employs the substrate 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) that yields the colored product 3,5,3’,5’-tetrabromoazobenzene sulfate sodium salt (azo-TBBS). The amounts of azo-TBBS are measured using HPLC and then used to calculate total antioxidative capacity (TAC) values. We first show that the TAC values measured using the new DBNBS system were significantly higher compared with the control. The assay was validated through further analysis of 56 compounds, including previously characterized antioxidants. The data are consistent with published values. Here we describe in detail the application of the DBNBS method to the measurement of the TAC values of eight beverages, including wines and fruit juices. The DBNBS assay employs a readily applicable protocol that sensitively determines the levels of antioxidants in foodstuffs.. |
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Taichi Kasiwagi, Hanxu Yan, Takeki Hamasaki, Kinjo Tomoya, Noboru Nakamichi, Kiichiro Teruya, Shigeru Kabayama, Sanetaka Shirahata, Electrochemically Reduced Water Protects Neural Cells from Oxidative Damage, OXIDATIVE MEDICINE AND CELLULAR LONGEVITY, 10.1155/2014/869121, 2014.10, Aging-related neurodegenerative disorders are closely associated with mitochondrial dysfunction and oxidative stresses and their incidence tends to increase with aging. Brain is the most vulnerable to reactive species generated by a higher rate of oxygen consumption and glucose utilization compared to other organs. Electrochemically reduced water (ERW) was demonstrated to scavenge reactive oxygen species (ROS) in several cell types. In the present study, the protective effect of ERW against hydrogen peroxide (H2O2) and nitric oxide (NO) was investigated in several rodent neuronal cell lines and primary cells. ERW was found to significantly suppress H2O2 (50-200 mu M) induced PC12 and SFME cell deaths. ERW scavenged intracellular ROS and exhibited a protective effect against neuronal network damage caused by 200 mu M H2O2 in N1E-115 cells. ERW significantly suppressed NO-induced cytotoxicity in PC12 cells despite the fact that it did not have the ability to scavenge intracellular NO. ERW significantly suppressed both glutamate induced Ca2+ influx and the resulting cytotoxicity in primary cells. These results collectively demonstrated for the first time that ERW protects several types of neuronal cells by scavenging ROS because of the presence of hydrogen and platinum nanoparticles dissolved in ERW.. |
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Takeki Hamasaki, Noboru Nakamichi, Kiichiro Teruya, Sanetaka Shirahata, Removal Efficiency of Radioactive Cesium and Iodine Ions by a Flow-Type Apparatus Designed for Electrochemically Reduced Water Production., PLoS ONE, 10.1371/journal.pone.0102218, 9, 7, 2014.06, The Fukushima Daiichi Nuclear Power Plant accident on March 11, 2011 attracted people's attention, with anxiety over possible radiation hazards. Immediate and long-term concerns are around protection from external and internal exposure by the liberated radionuclides. In particular, residents living in the affected regions are most concerned about ingesting contaminated foodstuffs, including drinking water. Efficient removal of radionuclides from rainwater and drinking water has been reported using several pot-type filtration devices. A currently used flow-type test apparatus is expected to simultaneously provide radionuclide elimination prior to ingestion and protection from internal exposure by accidental ingestion of radionuclides through the use of a micro-carbon carboxymethyl cartridge unit and an electrochemically reduced water production unit, respectively. However, the removability of radionuclides from contaminated tap water has not been tested to date. Thus, the current research was undertaken to assess the capability of the apparatus to remove radionuclides from artificially contaminated tap water. The results presented here demonstrate that the apparatus can reduce radioactivity levels to below the detection limit in applied tap water containing either 300 Bq/kg of 137Cs or 150 Bq/kg of 125I. The apparatus had a removal efficiency of over 90% for all concentration ranges of radio-cesium and -iodine tested. The results showing efficient radionuclide removability, together with previous studies on molecular hydrogen and platinum nanoparticles as reactive oxygen species scavengers, strongly suggest that the test apparatus has the potential to offer maximum safety against radionuclide-contaminated foodstuffs, including drinking water. Removal Efficiency of Radioactive Cesium and Iodine Ions by a Flow-Type Apparatus Designed for Electrochemically Reduced Water Production. Available from: https://www.researchgate.net/publication/264010807_Removal_Efficiency_of_Radioactive_Cesium_and_Iodine_Ions_by_a_Flow-Type_Apparatus_Designed_for_Electrochemically_Reduced_Water_Production [accessed May 2, 2017].. |
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Sanetaka Shirahata, Takeki Hamasaki, Tomoya Kinjyo, Noboru Nakamichi, Shigeru Kabayama, Kiichiro Teruya, The Mechanism of Redox Regulation in Animal Cells by Hydrogen and Pt Nanoparticles as Antioxidants, IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL, 10.1371/journal.pone.0171192, 12, 2, 2014.03. |
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Kosuke Tomimatsu, Matsumoto Shin-ei, Hayato Tanaka, Makiko Yamashita, Hidekazu Nakanishi, Kiichiro Teruya, Saiko Kazuno, Kinjo Tomoya, Takeki Hamasaki, Ken-ichi Kusumoto, Shigeru Kabayama, Yoshinori Katakura, Sanetaka Shirahata, A rapid screening and production method using a novel mammalian cell display to isolate human monoclonal antibodies., Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2013.10.007, 441, 1, 59-64, 2013.10, Antibody display methods are increasingly being used to produce human monoclonal antibodies for disease therapy. Rapid screening and isolation of specific human antibody genes are valuable for producing human monoclonal antibodies showing high specificity and affinity. In this report, we describe a novel mammalian cell display method in which whole human IgG is displayed on the cell surface of CHO cells. Cells expressing antigen-specific human monoclonal IgGs with high affinity on the cell surface after normal folding and posttranscriptional modification were screened using a cell sorter. The membrane-type IgG-expressing CHO cells were then converted to IgG-secreting cells by transfection with a plasmid coding Cre recombinase. This mammalian cell display method was applied to in vitro affinity maturation of monoclonal C9 IgG specific to the human high-affinity IgE receptor (Fc epsilon RI alpha). The CDR3 of the C9 heavy chain variable region gene was randomly mutated and inserted into pcDNA5FRT/IgG. A C9 IgG (CDRH3r)-expressing CHO cell display library consisting of 1.1 x 10(6) independent clones was constructed. IgG-displaying cells showing high reactivity to Fc epsilon RI alpha antigen were screened by the cell sorter, resulting in the establishment of a CHO cell line producing with higher reactivity than the parent C9 IgG. (C) 2013 Elsevier Inc. All rights reserved.. |
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Yuping Li, Takeki Hamasaki, Kiichiro Teruya, Noboru Nakamichi, Xbigniew Gadek, Taichi Kasiwagi, Hanxu Yan, Kinjo Tomoya, Takaaki Komatsu, Yoshitoki Ishii, Sanetaka Shirahata, Suppressive effects of natural reduced waters on alloxan-induced apoptosis and type 1 diabetes mellitus, CYTOTECHNOLOGY, 10.1007/s10616-011-9414-, 64, 3, 2012.03, Insulin-producing cells express limited activities of anti-oxidative enzymes. Therefore, reactive oxygen species (ROS) produced in these cells play a crucial role in cytotoxic effects. Furthermore, diabetes mellitus (DM) development is closely linked to higher ROS levels in insulin-producing cells. Hita Tenryosui Water(A (R)) (Hita T. W., Hita, Japan) and Nordenau water (Nord. W., Nordenau, Germany), referred to as natural reduced waters (NRWs), scavenge ROS in cultured cells, and therefore, might be a possibility as an alternative to conventional pharmacological agents against DM. Therefore, this study aimed to investigate the role of NRWs in alloxan (ALX)-induced beta-cell apoptosis as well as in ALX-induced diabetic mice. NRWs equally suppressed DNA fragmentation levels. Hita T. W. and Nord. W. ameliorated ALX-induced sub-G(1) phase production from approximately 40% of control levels to 8.5 and 11.8%, respectively. NRWs restored serum insulin levels (p |
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Kinjo Tomoya, Jun Ye, Hanxu Yan, Takeki Hamasaki, Hidekazu Nakanishi, Kazuko Toh, Noboru Nakamichi, Shigeru Kabayama, Kiichiro Teruya, Sanetaka Shirahata, Suppressive effects of electrochemically reduced water on matrix metalloproteinase-2 activities and in vitro invasion of human fibrosarcoma HT1080 cells, CYTOTECHNOLOGY, 10.1007/s10616-012-9469-7, 64, 3, 357-371, 2012.03, It has been demonstrated that hydrogen peroxide (H2O2) is directly associated with elevated matrix metalloproteinase-2 (MMP-2) expression in several cell lines. Electrochemically reduced water (ERW), produced near the cathode during electrolysis, and scavenges intracellular H2O2 in human fibrosarcoma HT1080 cells. RT-PCR and zymography analyses revealed that when HT1080 cells were treated with ERW, the gene expression of MMP-2 and membrane type 1 MMP and activation of MMP-2 was repressed, resulting in decreased invasion of the cells into matrigel. ERW also inhibited H2O2-induced MMP-2 upregulation. To investigate signal transduction involved in MMP-2 downregulation, mitogen-activated protein kinase (MAPK)-specific inhibitors, SB203580 (p38 MAPK inhibitor), PD98059 (MAPK/extracellular regulated kinase kinase 1 inhibitor) and c-Jun NH2-terminal kinase inhibitor II, were used to block the MAPK signal cascade. MMP-2 gene expression was only inhibited by SB203580 treatment, suggesting a pivotal role of p38 MAPK in regulation of MMP-2 gene expression. Western blot analysis showed that ERW downregulated the phosphorylation of p38 both in H2O2-treated and untreated HT1080 cells. These results indicate that the inhibitory effect of ERW on tumor invasion is due to, at least in part, its antioxidative effect.. |
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Sanetaka Shirahata, Takeki Hamasaki, Kiichiro Teruya, Advanced research on the health benefit of reduced water, TRENDS IN FOOD SCIENCE & TECHNOLOGY, 10.1016/j.tifs.2011.10.009, 23, 2, 124, 2012.02, In Japan, research on functional water, especially on reduced water, is developing at a rapid pace. Reduced water such as electrochemically reduced water and natural reduced water can scavenge reactive oxygen species in cultured cells. Reduced waters are expected to have preventive and positive effects on oxidative stress-related diseases such as diabetes, cancer, arteriosclerosis, neurodegenerative diseases, and side effects of hemodialysis. It has been suggested that the active agents in reduced water are hydrogen (atoms and molecules), mineral nanoparticles, and mineral nanoparticle hydrides.. |
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Hanxu Yan, Kinjo Tomoya, Huaize Tian, Takeki Hamasaki, Kiichiro Teruya, Shigeru Kabayama, Sanetaka Shirahata, Mechanism of the Lifespan Extension of Caenorhabditis elegans by Electrolyzed Reduced Water-Participation of Pt Nanoparticles, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 10.1271/bbb.110072, 75, 7, 1295, 2011.06, Electrolyzed reduced water (ERW) contains a large amount of molecular hydrogen and a small amount of Pt nanoparticles (Pt NPs). We have found that ERW significantly extended the lifespan of Caenorhabditis elegans in a novel culture medium designated Water Medium. In this study, we found that synthetic Pt NPs at ppb levels significantly extended the nematode lifespan and scavenged reactive oxygen species (ROS) in the nematode induced by paraquat treatment. In contrast, a high concentration of dissolved molecular hydrogen had no significant effect on the lifespan of the nematode. These findings suggest that the Pt NPs in ERW, rather than the molecular hydrogen, extend the longevity of the nematode, at least partly by scavenging ROS.. |
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K. Yamamoto, Y. Tsuji, Y. Aso, T. Hamasaki, Y. Katakura, S. Shirahata, Effect of diazinon exposure on antioxidant reactions in the silkmoth, Bombyx mori, JOURNAL OF APPLIED ENTOMOLOGY, 10.1111/j.1439-0418.2010.01549.x, 135, 4, 320-325, 2011.05, We investigated the effects of exposure to diazinon, an organophosphate insecticide, on the expression of antioxidant and detoxification factors in the silkmoth. Exposure to diazinon resulted in induction of mRNA encoding manganese containing superoxide dismutase (SOD) and omega-class glutathione S-transferases (GST), whereas no changes were observed in catalase, other class of GST and acetylcholinesterase. Liquid chromatography showed that the amount of reactive oxygen species was increased, whereas the amount of glutathione was decreased in the silkmoth fat body after exposure to diazinon. These results suggest that SOD and omega-class GSTs can contribute to organophosphate resistance in Lepidopterans.. |
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Ye J, Li Y, Hamasaki T, Nakamichi N, Komatsu T, Kashiwagi T, Teruya K, Nishikawa R, Kawahara T, Osada K, Toh K, Abe M, Tian H, Kabayama S, Otsubo K, Morisawa S, Katakura Y, Shirahata S. , Inhibitory effect of electrolyzed reduced water on tumor angiogenesis., Biol. Pharm. bull. 31, 19-26. (2008)., 2008.07. |
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Hamasaki T, Aramaki S, Imada T, Teruya K, Kabayama S, Katakura Y, Otubo K, Morisawa S, Shirahata S. , Induction of caspase-3-dependent apoptosis by electrolyzed reduced water/platinum nanoparticles in cancer cells., Cell Technology for Cell Products 87-89. (2007)., 2007.07. |
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Ye J, Li Y, Hamasaki T, Nakamichi N, Kawahara T, Osada K, Teruya K, Kato Y, Toh K, Abe M, Katakura Y, Noguchi K, Shirahata S. , Catalyser-21(TM), a mineral water derived from leaf soil, inhibits tumor cell invasion and angiogenesis., Cytotechnology 55, 61-70. (2007)., 2007.07. |
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Hamasaki T, Kashiwagi T, Aramaki S, Imada T, Komatsu T, Li Y, Teruya K, Katakura Y, Kabayama S, Otsubo K, Morisawa S, Shirahata S., Suppression of cell growth by platinum nanocolloids as scavengers against reactive oxygen species. , Animal Cell Technology Meets Genomics 249-251. (2005), 249-251, 2005.07. |
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Kato Y, Ohta M, Kamei K, Wakita H, Kawahara T, Yamano Y, Hamasaki T, Teruya K, Osada K, Nishikawa R, Noguchi K, Shirahata S. , Structure analysis of complex carbohydrate exhibiting IL-12 inducing activity derived from mineral water in natural leaf soil. , Bunsekikagaku 54, 391-397. (2005)., 10.2116/bunsekikagaku.54.391, 54, 5, 391-397, 2005.07. |
