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.
The purpose of this study was to evaluate the cleaning effect of root canal walls using strong acid electrolytic water (SAEW) as a root canal irrigant, and to investigate the influence of SAEW on the root canal dentin by micro-hardness test. Forty-three single-rooted, single-canaled teeth were instrumented using standard step-back technique with K-files. Irrigation was performed using distilled water, 5.25% NaOCl and 3% H(2)O(2), SAEW, or 15% EDTA solution in five groups. Samples were prepared to be examined under a scanning electron microscope (SEM) and micro Vickers hardness (H(V)) test machine. Our results showed that the root cleaning effects of the combined use of SAEW and NaOCl solution as root canal irrigants were equivalent to those in the group with NaOCl and 15% EDTA. When SAEW was used for 1 min under ultrasonic vibration, no decreases in the hardness of dentin inside the root canal were detected.
The generation of reactive oxygen species is thought to cause extensive oxidative damage to various biomolecules such as DNA, RNA, and protein. In this study, the preventive, suppressive, and protective effects of in vitro supplementation with electrolyzed-reduced water on H2O2-induced DNA damage in human lymphocytes were examined using a comet assay. Pretreatment, cotreatment, and posttreatment with electrolyzed-reduced water enhanced human lymphocyte resistance to the DNA strand breaks induced by H2O2 in vitro. Moreover, electrolyzed-reduced water was much more effective than diethylpyrocarbonate-treated water in preventing total RNA degradation at 4 and 25 degrees C. In addition, electrolyzed-reduced water completely prevented the oxidative cleavage of horseradish peroxidase, as determined using sodium dodecyl sulfate-polyacrylamide gels. Enhancement of the antioxidant activity of ascorbic acid dissolved in electrolyzed-reduced water was about threefold that of ascorbic acid dissolved in nonelectrolyzed deionized water, as measured by a xanthine-xanthine oxidase superoxide scavenging assay system, suggesting an inhibitory effect of electrolyzedreduced water on the oxidation of ascorbic acid.
This study investigated the antibacterial effect of electrolyzed water on oral bacteria both in vitro and in vivo. Tap water was electrolyzed in a water vessel using platinum cell technology. The electrolyzed tap water (called Puri-water) was put in contact with five major periodontopathogens or toothbrushes contaminated with these bacteria for 30 sec. In addition, Puri-water was used as a mouthwash for 30 sec in 16 subjects and the antibacterial effect on salivary bacteria was evaluated. Puri-water significantly reduced the growth of all periodontopathogens in culture and on toothbrushes, and that of aerobic and anaerobic bacteria in saliva, when compared to the effect of tap water. It also significantly reduced mutans streptococci growing on mitis salivarius-bacitracin agar. Our results demonstrate that the electrolyzed tap water is effective as a mouthwash and for toothbrush disinfection.
Mineral-induced alkaline-reduced water (MRW) is generated by the chemical reaction of water with alkaline earth metals and characterized by high pH and low oxidation-reduction potential. As ROS are believed to have a role in the pathogenesis of obesity, we attempted to determine the effect of MRW on obesity in Sprague-Dawley (SD) rats fed on a high-fat diet. The body weight of the MRW group was significantly lower than that of the control group in most periods of the examination (P<0.05). Serum level of triglycerides (P<0.05) and fat deposition in the livers of the MRW group were found to have been significantly reduced. This suggests that MRW down-regulates lipid metabolism, thereby suppressing obesity. Possible mechanisms of MRW related to reactive oxygen species were also discussed. Our results suggest that MRW is effective in the alleviation of obesity in SD rats fed on high-fat diet.
Oxidative stress is produced under diabetic conditions and is likely involved in progression of pancreatic beta-cell dysfunction found in diabetes. Both an increase in reactive oxygen free radical species (ROS) and a decrease in the antioxidant defense mechanism lead to the increase in oxidative stress in diabetes. Electrolyzed reduced water (ERW) with ROS scavenging ability may have a potential effect on diabetic animals, a model for high oxidative stress. Therefore, the present study examined the possible anti-diabetic effect of ERW in two different diabetic animal models. The genetically diabetic mouse strain C57BL/6J-db/db (db/db) and streptozotocin (STZ)-induced diabetic mouse were used as insulin deficient type 1 and insulin resistant type 2 animal model, respectively. ERW, provided as a drinking water, significantly reduced the blood glucose concentration and improved glucose tolerance in both animal models. However, ERW fail to affect blood insulin levels in STZ-diabetic mice whereas blood insulin level was markedly increased in genetically diabetic db/db mice. This improved blood glucose control could result from enhanced insulin sensitivity, as well as increased insulin release. The present data suggest that ERW may function as an orally effective anti-diabetic agent and merit further studies on its precise mechanism.
Chronic hemodialysis (HD) patients increase erythrocyte susceptibility to hemolysis and impair cell survival. We explored whether electrolyte-reduced water (ERW) could palliate HD-evoked erythrocyte impairment and anemia. Forty-three patients undergoing chronic HD were enrolled and received ERW administration for 6 month. We evaluated oxidative stress in blood and plasma, erythrocyte methemoglobin (metHb)/ferricyanide reductase activity, plasma metHb, and proinflammatory cytokines in the chronic HD patients without treatment (n=15) or with vitamin C (VC)- (n=15), vitamin E (VE)-coated dialyzer (n=15), or ERW treatment (n=15) during an HD course. The patients showed marked increases (15-fold) in blood reactive oxygen species, mostly H(2)O(2), after HD without any treatment. HD resulted in decreased plasma VC, total antioxidant status, and erythrocyte metHb/ferricyanide reductase activity and increased erythrocyte levels of phosphatidylcholine hydroperoxide (PCOOH) and plasma metHb. Antioxidants treatment significantly palliated single HD course-induced oxidative stress, plasma and RBC PCOOH, and plasma metHb levels, and preserved erythrocyte metHb /ferricyanide reductase activity in an order VC>ERW>VE-coated dialyzer. However, ERW had no side effects of oxalate accumulation easily induced by VC. Six-month ERW treatment increased hematocrit and attenuated proinflammatory cytokines profile in the HD patients. In conclusion, ERW treatment administration is effective in palliating HD-evoked oxidative stress, as indicated by lipid peroxidation, hemolysis, and overexpression of proinflammatory cytokines in HD patients.
Alkalin-reduced water (ARW) is known to exert several anti-cancer effects, as well as to scavenge reactive oxygen species (ROS) and reduce blood-glucose levels. This study was performed in order to determine the effects of ARW on the control of spontaneous diabetes in Otsuka Long-Evans Tokushima Fatty (OLETF) rats. We assigned 16 male OLETF rats (4 wk) to two groups: an experimental group, which was given ARW, and a control group, which received laboratory tap water. From week 6 to 32, the body weight, lipid composition, and glucose levels in the blood of the rats were measured. The glucose levels of both groups tended to increase. However, the ARW group’s glucose levels were significantly lower than those of the control group after 12 weeks (p<0.05). The total cholesterol and triglyceride levels in the ARW group were found to be significantly lower than those of the control group during the experimental period. These results suggest that ARW spurred the growth of OLETF rats during the growth stage, and that long-term ingestion of ARW resulted in a reduction in the levels of glucose, triglycerides, and total cholesterol in the blood.
The study examined changes in the relevant tests parameters of 320 diabetes patients (176 female, 144 male) drinking natural reduced water from the ‘Nordenau Spring’ in Nordenau, Germany as well as the correlation of these changes with the fluctuation of the reactive oxygen species concentration in their blood. The average age of the test persons was 71.8 years old and the daily consumption of reduced water was as much as two liters. The average duration of stay in Nordenau lasted 6 days. The diagnostic parameters such as blood sugar, HbA1c, cholesterol, HDL, LDL, triglyceride and potassium were tested twice – at the beginning and at the end of the participants stay in Nordenau. Seventy-six of the test persons repeated the same procedure two or more times in the following months and years. Additionally a random sample of reactive oxygen species concentration in the blood of 50 patients had been taken in order to find out its possible causal connections to the diabetes relevant tests parameters. Blood sugar and HbA1c had been considered as the substantial tests parameters in order to break down the whole group into responder and non-responder. Two hundred and thirty six tested persons that is to say (73.8%) had been assigned to the responder group and 84 (26.2%) to non- responder. Similar ratio of 37 responders (74%) and 13 non-responders (26%) respectively had been obtained in the random sample group with an additional free oxygen radicals test (FORT). The survey pointed out the causal connections between the changes of blood sugar and HbA1c values and the fluctuation of reactive oxygen species concentration in the blood of the tested persons.
The in vivo effects of drinking a water product, which had been confirmed to have antioxidant activi- ties in vitro, were preliminarily studied by monitoring the blood concentrations of oxidative stress marker substances in the two group subjects who ingested the same quantity of the water product and a tap water solution during the same time. The results indicated that hydrogen gas and reductive vanadium ions as the components responsible for the antioxidant activities in vitro cannot enhance the scavenging ability for re- active oxygen species in vivo after being drunk and absorbed into the human body, although it was sug- gested that an ingestion of a greater quantity of water than usual gives a slight reduction, overall, in the oxi- dative stress.