Abstract: Anti-diabetes effect of water containing hydrogen molecule and Pt nanoparticles

No abstract available

Suppressive Effect of Electrolyzed-Reduced Water on the Growth of Cancer Cells and Microorganisms

We investigated the suppressive effect of electrolyzed reduced water (ERW) on the growth of not only various human cancer cells but also microorganisms such as gram-negative Escherichia coli. ERW suppressed the growth of cancer cells, especially in soft-agar culture. The suppressive effect of ERW on the growth of cancer cells depended upon cell types and malignancy of cancer cells and the production methods of ERW. We assumed that scavenging intracellular reactive oxygen species (ROS) by ERW resulted in impairing the tumor phenotypes such as rapid proliferation and anchorage-independent growth without affecting serious damage to normal cells. We also found that ERW exhibited weak microbicidal effect, especially in low cell densities of microorganisms. It may contribute to prevent the rot of food or improve the intestinal microflora to prevent abnormal fermentation.

Abstract: The neuroprotective effects of electrolyzed reduced water and its model water containing molecular hydrogen and Pt nanoparticles

No abstract available

Protective mechanism of reduced water against alloxan-induced pancreatic beta-cell damage: Scavenging effect against reactive oxygen species

Reactive oxygen species (ROS) cause irreversible damage to biological macromolecules, resulting in many diseases. Reduced water (RW) such as hydrogen-rich electrolyzed reduced water and natural reduced waters like Hita Tenryosui water in Japan and Nordenau water in Germany that are known to improve various diseases, could protect a hamster pancreatic beta cell line, HIT-T15 from alloxan-induced cell damage. Alloxan, a diabetogenic compound, is used to induce type 1 diabetes mellitus in animals. Its diabetogenic effect is exerted via the production of ROS. Alloxan-treated HIT-T15 cells exhibited lowered viability, increased intracellular ROS levels, elevated cytosolic free Ca(2+) concentration, DNA fragmentation, decreased intracellular ATP levels and lowering of glucose-stimulated release of insulin. RW completely prevented the generation of alloxan-induced ROS, increase of cytosolic Ca(2+) concentration, decrease of intracellular ATP level, and lowering of glucose-stimulated insulin release, and strongly blocked DNA fragmentation, partially suppressing the lowering of viability of alloxan-treated cells. Intracellular ATP levels and glucose-stimulated insulin secretion were increased by RW to 2-3.5 times and 2-4 times, respectively, suggesting that RW enhances the glucose-sensitivity and glucose response of beta-cells. The protective activity of RW was stable at 4 degrees C for over a month, but was lost by autoclaving. These results suggest that RW protects pancreatic beta-cells from alloxan-induced cell damage by preventing alloxan-derived ROS generation. RW may be useful in preventing alloxan-induced type 1-diabetes mellitus.

Electrolyzed Reduced Water Induces Differentiation in K-562 Human Leukemia Cells

Electrolyzed reduced water (ERW) is known to scavenge reactive oxygen species (ROS) which is related to malignant tumor phenotypes. We attempted to clarify the effect of ERW on tumor phenotypes of K562 human leukemia cells. Treatment of K562 cells with ERW resulted in growth arrest, morphological changes, and expression of CD41, a cell marker of megakaryocyte differentiation. However, the morphology of N-acetylcysteine (NAC)-treated cells was rather similar to that of non-treated control K562 cells. These results suggested that ERW, but not NAC can differentiate K562 cells into megakaryocytes. The induction of megakaryocytes from K562 cells by ERW was preceded by a rapid rise in the activity of MEK (MAP kinase /extra-cellular regulated kinases) that leads to sustained activation of ERK (extra-cellular regulated kinases; MAPK). However, In NAC-treated K562 cells, ERK activation was only transient. The different persistency of ERK activation induced by ERW and NAC might affect the cell fate.

Electrolyzed Reduced Water Supplemented with Platinum Nanoparticles Suppresses Promotion of Two-stage Cell Transformation

In the two-stage cell transformation theory, cancer cells first receive initiation, which is mainly caused by DNA damage, and then promotion, which enhances transformation. Murine Balb/c 3T3 cells are widely used for transformation experiments because they lose contact inhibition ability when transformed. Electrolyzed reduced water (ERW), which is produced near a cathode during electrolysis of water, is an alkaline drinking water that is beneficial to health. ERW contains a high concentration of dissolved hydrogen and scavenge reactive oxygen species (ROS), along with a small amount of platinum (Pt) nanoparticles (Pt nps) derived from Pt-coated titanium electrodes. Pt nps stably disperse in aqueous solution for a long time, and convert hydrogen molecules to active hydrogen (atomic hydrogen) that can scavenge ROS. Therefore, ERW supplemented with synthesized Pt nps is a model strong reduced water. This is the first report that ERW supplemented with synthesized Pt nps strongly prevents transformation of Balb/c 3T3 cells. ERW was prepared by electrolysis of 0.002 M NaOH solution using a batch-type electrolysis device. Balb/c 3T3 cells were treated with 3-methyl cholanthrene (MCA) as an initiation substance, followed by treatment with phorbol-12-myristate-13-acetate (PMA) as a promotion substance. MCA/PMA-induced formation of a transformation focus was strongly suppressed by ERW supplemented with Pt nps but not by ERW or Pt nps individually. ERW supplemented with Pt nps suppressed transformation at the promoter stage, not at initiation, suggesting that ERW supplemented with Pt nps suppressed the PMA-induced augmentation of intracellular ROS. ERW supplemented with Pt nps is a potential new antioxidant against carcinogenesis.

Suppression of two-stage cell transformation by electrolyzed reduced water containing platinum nanoparticles

According to the two-stage cell transformation theory, cancer cells first receive initiation, which is mainly caused by DNA damage and then promotion, which enhance transformation. Since murine Balb/c 3T3 cells lose contact inhibition by cell transformation, the cells have been widely used for transformation experiments. Electrolyzed reduced water (ERW) is a health beneficial alkaline drinking water containing high concentration of dissolved hydrogen, and can scavenge intracellular reactive oxygen species (ROS). ERW contains a small amount of platinum (Pt) nano- particles as atomic hydrogen donor and ROS-scavenger. Therefore, ERW supplemented with Pt nanoparticles (ERW/Pt) can be considered as a model of strong ERW. Here, we report that ERW/Pt can prevent transformation of Balb/c 3T3 cells. ERW was prepared by electrolysis of 0.002 M NaOH. Balb/c 3T3 cells were treated with 3-methyl cholantrene (MCA) as an initiation compound, followed by the treatment with phorbol-12-myristate-13-acetate (PMA) as a promotion compound. The cell transformation induced by MCA/PMA was strongly suppressed by ERW/Pt treatment, especially at the stage of promotion. Analysis of intracellular ROS level showed that ERW/Pt could decrease excess intracellular ROS induced by PMA. These results suggested that ERW/Pt suppressed cell transformation at promotion stage by its ROS scavenging effect.

Inhibitory effect of electrolyzed reduced water on tumor angiogenesis

Vascular endothelial growth factor (VEGF) is a key mediator of tumor angiogenesis. Tumor cells are exposed to higher oxidative stress compared to normal cells. Numerous reports have demonstrated that the intracellular redox (oxidation/reduction) state is closely associated with the pattern of VEGF expression. Electrolyzed reduced water (ERW) produced near the cathode during the electrolysis of water scavenged intracellular H(2)O(2) and decreased the release of H(2)O(2) from a human lung adenocarcinoma cell line, A549, and down-regulated both VEGF transcription and protein secretion in a time-dependent manner. To investigate the signal transduction pathway involved in regulating VEGF expression, mitogen-activated kinase (MAPK) specific inhibitors, SB203580 (p38 MAPK inhibitor), PD98059 (ERK1/2 inhibitor) and JNKi (c-Jun N-terminal protein kinase inhibitor) were applied. The results showed that only PD98059 blocks VEGF expression, suggesting an important role for ERK1/2 in regulating VEGF expression in A549 cells. As well, ERW inhibited the activation of extracellular signal-regulated kinase (ERK) in a time-dependent manner. Co-culture experiments to analyze in vitro tubule formation assay revealed that A549 cell-derived conditioned medium significantly stimulated the formation of vascular tubules in all analyzed parameters; tubule total area, tubule junction, number of tubules, and total tubule length. ERW counteracted the effect of A549 cell-conditioned medium and decreased total tube length (p<0.01). The present study demonstrated that ERW down-regulated VEGF gene transcription and protein secretion through inactivation of ERK.

Extension of the lifespan of Caenorhabditis elegans by the use of electrolyzed reduced water

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.

Anti-Diabetes Effects of Hita Tenryou-Sui Water®, a Natural Reduced Water

It has been reported that a natural reduced water derived from a deep well in Japan (trademark, Hita Tenryousui water®; Nakanoshima, Hita city, Oita, Japan) scavenged intracellular reactive oxygen species (ROS) and protected a hamster pancreas β cell line HIT-T15 from oxidative damage by alloxan, a type 1 diabetes inducer. Here we demonstrated that the water also suppressed the fasted blood glucose levels of the alloxan-induced type 1-diabetes mice. Protein tyrosine phosphatase (PTP) is a redox-regulatable signal enzyme and activation of PTP in Type 2-diabetes patients is noted to be responsible to inactivation of insulin receptor. Hita Tenryousui water® suppressed the PTP activity, leading the activation of insulin receptor. This water stimulated glucose uptake into myotubes and showed a tendency to improve the impaired sugar tolerance of Type 2-diabetes model mice. An open clinical test of Hita Tenryousui water® against 65 patients with hyperglycemia and 50 patients with hyperlipemia for 2 months resulted in significant improvement of impaired blood sugar, plasma triglycerol and total cholesterol levels (P<0.05). These facts suggest that daily intake of natural reduced water will be beneficial to prevent and improve diabetes mellitus.