Ōsaka, Japan
Ōsaka, Japan

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Huang K.-C.,National Taiwan University Hospital | Hsu S.-P.,National Taiwan University Hospital | Hsu S.-P.,Far Eastern Memorial Hospital | Yang C.-C.,Far Eastern Memorial Hospital | And 6 more authors.
Nephrology Dialysis Transplantation | Year: 2010

Background. T-cell damage by increased oxidative stress in end-stage renal disease (ESRD) patients undergoing chronic haemodialysis (HD) led to the increased T-cell apoptosis and the alteration of surface markers and Th1/Th2 ratio in CD4+ T lymphocytes. Antioxidant electrolysed-reduced water (ERW) was used as the dialysate in ESRD patients undergoing chronic HD to test for improved oxidative stress-related T-cell apoptosis, alterations of surface markers and intracellular cytokine profile.Methods. We evaluated apoptosis formation by annexin V, CD25-related surface markers, and cytokine ratio of Th1/Th2 in CD4+ T lymphocytes and Tc1/Tc2 in CD8+ T lymphocytes of 42 ESRD patients haemodialysed with ERW for 1 year.Results. In comparison to 12 healthy individuals, the ESRD patients had more T-cell apoptosis and less CD3+, CD4+ and CD8+ T cells and CD25/CD69/CD94/CD3+ phenotypes at baseline. Lower intracellular IL-2 and IFN-γ levels in the Th1/CD4+ and Tc1/CD8+ cells and higher intracellular IL-4, IL-6 and IL-10 levels in the Th2/CD4 + and Tc2/CD8+ cells were also noted in the ESRD patients. After a 1-year ERW treatment, the patients had a decrease in T-cell apoptosis and increases in CD3+, CD4+ and CD8+ cell numbers and CD25/CD69/CD94/CD3+ phenotypes in the T cells. The intracellular IL-2 and IFN-γ levels in the Th1/Tc1 cells significantly (P < 0.05) increased and the intracellular IL-4, IL-6 and IL-10 levels in the Th2/Tc2 cells decreased. Furthermore, the Th1/Th2 and Tc1/Tc2 cytokine ratios were improved toward a normal status.Conclusion. One-year ERW treatment effectively ameliorated T-cell apoptosis, altered CD25-related surface markers and intracellular cytokine profile in the HD patients. © The Author [2009]. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.


Kinjo T.,Kyushu University | Ye J.,Kyushu University | Ye J.,Xiamen University | Yan H.,Kyushu University | And 7 more authors.
Cytotechnology | Year: 2012

It has been demonstrated that hydrogen peroxide (H 2O 2) 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 H 2O 2 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 H 2O 2-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 NH 2-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 H 2O 2-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. © Springer Science+Business Media B.V. 2012.


Yan H.,Kyushu University | Tian H.,Kyushu University | Kinjo T.,Kyushu University | Hamasaki T.,Kyushu University | And 5 more authors.
Bioscience, Biotechnology and Biochemistry | Year: 2010

Electrolyzed reduced water (ERW) has attracted much attention because of its therapeutic effects. In the present study, a new culture medium, which we designated Water medium, was developed to elucidate the effects of ERW on the lifespan of Caenorhabditis elegans. Wild-type C. elegans had a significantly shorter lifespan in Water medium than in conventional S medium. However, worms cultured in ERW-Water medium exhibited a significantly extended lifespan (from 11% to 41%) compared with worms cultured in ultrapure water-Water medium. There was no difference between the lifespans of worms cultured in ERW-S medium and ultrapure water-S medium. Nematodes cultured in ultrapure water-Water medium showed significantly higher levels of reactive oxygen species than those cultured in ultrapure water-S medium. Moreover, ERW-Water medium significantly reduced the ROS accumulation induced in the worms by paraquat, suggesting that ERW-Water medium extends the longevity of nematodes at least partly by scavenging ROS.


Spulber S.,Karolinska Institutet | Edoff K.,Karolinska Institutet | Hong L.,Karolinska Institutet | Morisawa S.,Nihon Trim Co Ltd. | And 2 more authors.
PLoS ONE | Year: 2012

Molecular hydrogen has been shown to have neuroprotective effects in mouse models of acute neurodegeneration. The effect was suggested to be mediated by its free-radical scavenger properties. However, it has been shown recently that molecular hydrogen alters gene expression and protein phosphorylation. The aim of this study was to test whether chronic ad libitum consumption of molecular hydrogen-enriched electrochemically reduced water (H-ERW) improves the outcome of lipopolysaccharide (LPS)-induced neuroinflammation. Seven days after the initiation of H-ERW treatment, C57Bl/6 mice received a single injection of LPS (0.33 mg/kg i.p.) or an equivalent volume of vehicle. The LPS-induced sickness behaviour was assessed 2 h after the injection, and recovery was assessed by monitoring the spontaneous locomotor activity in the homecage for 72 h after the administration of LPS. The mice were killed in the acute or recovery phase, and the expression of pro- and antiinflammatory cytokines in the hippocampus was assessed by real-time PCR. We found that molecular hydrogen reduces the LPS-induced sickness behaviour and promotes recovery. These effects are associated with a shift towards anti-inflammatory gene expression profile at baseline (downregulation of TNF- α and upregulation of IL-10). In addition, molecular hydrogen increases the amplitude, but shortens the duration and promotes the extinction of neuroinflammation. Consistently, molecular hydrogen modulates the activation and gene expression in a similar fashion in immortalized murine microglia (BV-2 cell line), suggesting that the effects observed in vivo may involve the modulation of microglial activation. Taken together, our data point to the regulation of cytokine expression being an additional critical mechanism underlying the beneficial effects of molecular hydrogen. © 2012 Spulber et al.


Yan H.,Kyushu University | Kinjo T.,Kyushu University | Tian H.,Kyushu University | Hamasaki T.,Kyushu University | And 3 more authors.
Bioscience, Biotechnology and Biochemistry | Year: 2011

Electrolyzed reduced water (ERW) contains a large amount of molecular hydrogen and asmall 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.


Nakanishi H.,Kyushu University | Hamasaki T.,Kyushu University | Kinjo T.,Kyushu University | Yan H.,Kyushu University | And 4 more authors.
Nano Biomedicine and Engineering | Year: 2013

Prolonged exposure to excessive reactive oxygen species (ROS) increases risk factorsfor many diseases. Therefore, elimination of ROS as well as prevention of its production becomes critically important. In the present study, we evaluated the levels of cytotoxicity and ROS scavenging activity induced by synthetic platinum nanoparticles (PtNPs). Average size of synthesized PtNPs was 2.2 nm. Synthetic PtNPs were found to scavenge both induced and endogenous H2O2 significantly in L6 rat skeletal muscle cells at a very low concentration (10-2 mg/l). To investigate the mechanism of action, the hierarchical oxidative stress model was used as an experimental model. To evaluate this possibility, we assessedglutathione concentration and gene levels of several antioxidant enzymes in PtNPs-treated(10-3-10 mg/l) L6 cells. Reduced glutathione (GSH) was increased in the range of 10-3-1 mg/l, but not in the 10 mg/l PtNP-treated cells. The GSH/GSSG ratio increased significantly at 1 mg/l and decreased in the 10 mg/l PtNPtreated cells. Most of the gene transcripts for oxidative stress inducible heme oxygenase-1 (HO-1), glutathione reductase (GR), copper-zinc superoxide dismutase (CuZn-SOD), manganese superoxide dismutase (Mn-SOD), glutathione peroxidase (GPx), and catalase were increased significantly by PtNPs at 10-1-10 mg/l.Such upregulatory effects induced by synthetic PtNPs at high concentrations (1-10 mg/l) in L6 cells can be explained by the hierarchical oxidative stress model. However, the cellular responses induced by low levels (10-3-10-2 mg/l) of PtNPs could not be fully explained by this model. © 2013 H. Nakanishi et al.


Hamauzu Y.,Kochi University | Ishikawa K.,Kochi University | Morisawa S.,Nihon Trim Co.
Environmental Control in Biology | Year: 2014

This study was intended to verify the effects of H2-enriched electrolyzed water in hydroponic nutrient solutions, as well as its mechanism. In the experiment, a "Reduced Vegetable Water Purifier" H 2-enriched electrolyzed water producing system, which was supplied by Nihon Trim Co., Ltd., was used. According to the measurements in which ORP electrodes were used as an indicator of oxidation-reduction power, the H 2-enriched electrolyzed water showed superior reduction power, reaching levels as low as -113 mV. The germination and early growth tests performed on the Komatsuna plants revealed that the roots grew significantly longer in the H2-enriched electrolyzed water than in the control. Komatsuna plants were cultivated by the NFT methods in a nutrient solution prepared using H2-enriched electrolyzed water. This study demonstrates that the nutrient solution in which H2-enriched electrolyzed water was used promoted the growth of leaves and roots of Komatsuna plants.


There is provided an apparatus for producing water for preparation of a dialysis solution, including an electrolytic water generation device that performs electrolysis using a solid polymer membrane, and a reverse osmosis membrane treatment device, the electrolytic water generation device being capable of reducing drained water as much as possible, and performing intense electrolysis to increase the amount of dissolved hydrogen without raising pH.


Patent
Nihon Trim Co. | Date: 2015-05-13

There is provided an apparatus (1) for producing water for preparation of a dialysis solution, including an electrolytic water generation device (2) that performs electrolysis using a solid polymer membrane (5), and a reverse osmosis membrane treatment device (3), the electrolytic water generation device being capable of reducing drained water as much as possible, and performing intense electrolysis to increase the amount of dissolved hydrogen without raising pH.


PubMed | Nihon Trim Co. and Kyushu University
Type: | Journal: Oxidative medicine and cellular longevity | Year: 2014

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 (50-200M) induced PC12 and SFME cell deaths. ERW scavenged intracellular ROS and exhibited a protective effect against neuronal network damage caused by 200M 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 Ca(2+) 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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