Electrolyzed Reduced Water
Research Articles

The information provided here and throughout this site is intended for informational and educational purposes and is not meant to cure or treat any disease or illness. Consult your doctor for specialized medical advice. Please note that the following are scientific papers and as such the language used is not friendly. For more down to earth and simple to understand information on the benefits of alkaline water please visit our health books page. These books document much of the available research but put them in language most people can easily understand. Our Alkaline Water Video page is also an excellent source for learning about alkaline water. The comments from health professionals who have personal experience with alkaline water and administering it to their patients should also be considered in drawing an opinion about benefits of drinking alkaline water.

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Links to Research on Alkaline Water

Links to Research on Alkaline Water

Proc Nutr Soc. 2010 Feb;69(1):166-73. Epub 2009 Dec 3.
Postgraduate Symposium: Positive influence of nutritional alkalinity on bone health.
Wynn E, Krieg MA, Lanham-New SA, Burckhardt P. University Hospital (CHUV), 1011 Lausanne, Switzerland.

There is growing evidence that consumption of a Western diet is a risk factor for osteoporosis through excess acid supply, while fruits and vegetables balance the excess acidity, mostly by providing K-rich bicarbonate-rich foods. Western diets consumed by adults generate approximately 50-100 mEq acid/d; therefore, healthy adults consuming such a diet are at risk of chronic low-grade metabolic acidosis, which worsens with age as a result of declining kidney function. Bone buffers the excess acid by delivering cations and it is considered that with time an overstimulation of this process will lead to the dissolution of the bone mineral content and hence to reduced bone mass. Intakes of K, Mg and fruit and vegetables have been associated with a higher alkaline status and a subsequent beneficial effect on bone health. In healthy male volunteers an acid-forming diet increases urinary Ca excretion by 74% and urinary C-terminal telopeptide of type I collagen (C-telopeptide) excretion by 19% when compared with an alkali (base-forming) diet. Cross-sectional studies have shown that there is a correlation between the nutritional acid load and bone health measured by bone ultrasound or dual-energy X-ray absorptiometry. Few studies have been undertaken in very elderly women (>75 years), whose osteoporosis risk is very pertinent. The EVAluation of Nutrients Intakes and Bone Ultra Sound Study has developed and validated (n 51) an FFQ for use in a very elderly Swiss population (mean age 80.4 (sd 2.99) years), which has shown intakes of key nutrients (energy, fat, carbohydrate, Ca, Mg, vitamin C, D and E) to be low in 401 subjects. A subsequent study to assess net endogenous acid production (NEAP) and bone ultrasound results in 256 women aged > or = 75 years has shown that lower NEAP (P=0.023) and higher K intake (P=0.033) are correlated with higher bone ultrasound results. High acid load may be an important additional risk factor that may be particularly relevant in very elderly patients with an already-high fracture risk. The latter study adds to knowledge by confirming a positive link between dietary alkalinity and bone health indices in the very elderly. In a further study to complement these findings it has also been shown in a group of thirty young women that in Ca sufficiency an acid Ca-rich water has no effect on bone resorption, while an alkaline bicarbonate-rich water leads to a decrease in both serum parathyroid hormone and serum C-telopeptide. Further investigations need to be undertaken to study whether these positive effects on bone loss are maintained over long-term treatment. Mineral-water consumption could be an easy and inexpensive way of helping to prevent osteoporosis and could be of major interest for long-term prevention of bone loss. PMID: 19954569
Source: http://www.ncbi.nlm.nih.gov/pubmed/19954569

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Journal of the Internation Society of Sports Nutrition, 2010 September 13;7(1):29.
Acid-base balance and hydration status following consumption of mineral-based alkaline bottled water. Heil DP.

ABSTRACT:
BACKGROUND: The present study sought to determine whether The consumption of a mineral-rich alkalizing bottled water could improve both acid-base balance and hydration status in young healthy adults under free-living conditions. The mineral-rich alkalizing bottled water contains a naturally high mineral content along with Alka-PlexLiquidTM, a dissolved supplement that increases the mineral content and gives the water an alkalizing pH of 10.0.

METHODS: Thirty-eight subjects were matched by gender and self-reported physical activity (SRPA, hrs/week) and then split into Control (12 women, 7 men; Mean+/-SD: 23+/-2 yrs; 7.2+/-3.6 hrs/week SRPA) and Experimental (13 women, 6 men; 22+/-2 yrs; 6.4+/-4.0 hrs/week SRPA) groups. The Control group consumed non-mineralized placebo bottled water over a 4-week period while the Experimental group consumed the placebo water during the 1st and 4th weeks and the mineral-rich alkalizing bottled water during the middle 2-week treatment period. Fingertip blood and 24-hour urine samples were collected three times each week for subsequent measures of blood and urine osmolality and pH, as well as total urine volume. Dependent variables were analyzed using multivariate repeated measures ANOVA with post-hoc focused on evaluating changes over time within Control and Experimental groups (alpha = 0.05). RESULTS: There were no significant changes in any of the dependent variables for the Control group. The Experimental group, however, showed significant increases in both the blood and urine pH (6.23 to 7.07 and 7.52 to 7.69, respectively), a decreased blood and increased urine osmolality, and a decreased urine output (2.51 to 2.05 L/day), all during the second week of the treatment period (P<0.05). Further, these changes reversed for the Experimental group once subjects switched to the placebo water during the 4th week.

CONCLUSIONS: Consumption of mineral-rich alkalizing bottled water was associated with improved acid-base balance (i.e., an alkalization of the blood and urine) and hydration status when consumed under free-living conditions. In contrast, subjects who consumed the placebo bottled water showed no changes over the same period of time. These results indicate that the habitual consumption of mineral-rich alkalizing bottled water may be a valuable nutritional vector for influencing both acid-base balance and hydration status in healthy adults. PMID: 20836884
Source: http://www.ncbi.nlm.nih.gov/pubmed/20836884


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Biomed Res. 2009 Oct;30(5):263-9.
Electrolyzed-reduced water inhibits acute ethanol-induced hangovers in Sprague-Dawley rats.
Park SK, Qi XF, Song SB, Kim DH, Teng YC, Yoon YS, Kim KY, Li JH, Jin D, Lee KJ. Department of Environmental Medical Biology, Wonju College of Medicine, Yonsei University, Wonju, Gangwon, Republic of Korea.

Ethanol consumption disturbs the balance between the pro- and anti-oxidant systems of the organism, leading to oxidative stress. Electrolyzed-reduced water (ERW) is widely used by people in East Asia for drinking purposes because of its therapeutic properties including scavenging effect of reactive oxygen species. This study was performed to investigate the effect of ERW on acute ethanol-induced hangovers in Sprague-Dawley rats. Alcohol concentration in serum of ERW-treated rats showed significant difference at 1 h, 3 h and 5 h respectively as compared with the rats treated with distilled water. Both alcohol dehydrogenase type 1 and acetaldehyde dehydrogenase related with oxidation of alcohol were significantly increased in liver tissue while the level of aspartate aminotransferase and alanine aminotransferase in serum was markedly decreased 24 h after pre-oral administration of ERW. Moreover, oral administration of ERW significantly activated non-ezymatic (glutathione) and enzymatic (glutathione peroxidase, glutathione-S-transferase, Cu/Zn-superoxide dismutase and catalase) antioxidants in liver tissues compared with the control group. These results suggest that drinking ERW has an effect of alcohol detoxification by antioxidant mechanism and has potentiality for relief of ethanol-induced hangover symptoms. PMID: 19887722
Source: http://www.ncbi.nlm.nih.gov/pubmed/19887722

Microwaterman comment: This is not an excuse to get drunk, but it does give insight into why one of the first videos I saw on microwater from Japan showed the use of alkaline ice used with whisky. 

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Alkaline Ionized Water Is Effective Against Constipation and Diarrhea.
Presented at the 25th General Assembly of Japan Medical Congress, the most prominent in Japan.

It was verified as "effective" by double blind clinical tests conducted for the first time in the world.
Alkaline ionized water has been widely used in households since it was approved by the Pharmaceutical Affiars Law in 1965. In the wake of increasing public interest in drinking water and health in recent years medical and scientific verification of its effect has long been awaited. As a result of our repeated clinical test for the last 6 months, it was verified that alkaline ionized water is effective against various abdominal symptoms through the epoch-making double blind clinical tests conducted for the first time in the world that endorse the validity of the tests. Dr. Hokudou presented it at the 25th General Assembly of Japan Medical Congress (4/2/99) representing the Committee of Alkaline Ionized Water Electrolyzer. This conference is the most prominent in Japan that has been held every 4 years since 1903. This year, the total number of attendees was 26,000 including 22,300 doctors. The following symptoms were examined in the clinical tests; chronicle diarrhea, constipation, abnormal intestinal fermentation, hyperchylia.

Got Rid of Abdominal Disorders.
Results of preliminary open clinical tests
25 volunteers who suffered from abdominal discomfort took 1000 liter/day alkaline ionized water for 2 weeks. 88% of them felt somehow improved. No one got worse.

Improves Constipation
Alkaline ionized water and clean water was given to 163 volunteers who suffered from abdominal discomfort in the amount of 500+ml/day for 4 weeks. Overall rate of improvement was significantly higher for those who were given alkaline ionized water.

As a result of comparative double blind clinical tests, alkaline ionized water was proved to be effective against constipation, too, and most importantly, its safety was verified. It is considered to be a quite opposite direction from using general medicine where side effects are not avoidable to some extent. The effectiveness of alkaline ionized water is very high at 80.5% which is higher, although not significantly, than that of clean water.

Read the full report click here

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Food Chem Toxicol. 2009 Aug;47(8):2031-6. Epub 2009 May 27.
Hepatoprotective effect of electrolyzed reduced water against carbon tetrachloride-induced liver damage in mice.
Tsai CF, Hsu YW, Chen WK, Chang WH, Yen CC, Ho YC, Lu FJ. Institute of Medicine, College of Medicine, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Rd., Taichung 402, Taiwan.

The study investigated the protective effect of electrolyzed reduced water (ERW) against carbon tetrachloride (CCl(4))-induced liver damage. Male ICR mice were randomly divided into control, CCl(4), CCl(4)+silymarin, and CCl(4)+ERW groups. CCl(4)-induced liver lesions include leukocytes infiltration, hepatocyte necrosis, ballooning degeneration, mitosis, calcification, fibrosis and an increase of serum alanine aminotransferase (ALT), and aminotransferase (AST) activity. In addition, CCl(4) also significantly decreased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). By contrast, ERW or silymarin supplement significantly ameliorated the CCl(4)-induced liver lesions, lowered the serum levels of hepatic enzyme markers (ALT and AST) and increased the activities of SOD, catalase, and GSH-Px in liver. Therefore, the results of this study show that ERW can be proposed to protect the liver against CCl(4)-induced oxidative damage in mice, and the hepatoprotective effect might be correlated with its antioxidant and free radical scavenging effect.
PMID: 19477216

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Biological & Pharmaceutical Bulletin. 2009 Mar;32(3):456-62.
The immunological effects of electrolyzed reduced water on the Echinostoma hortense infection in C57BL/6 mice.
Lee KJ, Jin D, Chang BS, Teng YC, Kim DH. Department of Environmental Medical Biology, Wonju College of Medicine, Yonsei University, Gang Won, South Korea.

Electrolyzed reduced water (ERW) is widely used for drinking by people in Asia. The purpose of this study was to examine the immunological effect of ERW on the immunity of animals by supplying ERW to C57BL/6 mice infected with Echinostoma hortense metacercariae. In the non-infected groups, interleukin (IL)-4 (p < 0.001), IL-5, IL-10, IL-1beta, tumor necrosis factor (TNF)-alpha and immunoglobulin (Ig) A expression of the group fed ERW (ERW group) increased in small intestine compared with the normal control group. In the case of infected groups, the group fed ERW (ERW+E. hortense group) showed the result that IL-4, IL-5, IL-10 and Ig A expression increased, but IL-1beta and TNF-alpha (p < 0.001) decreased, and the number of goblet cells (p < 0.001) and helix pomatia agglutinin (HPA) positive cells increased compared with the group without feeding ERW. However, adult worm recovery rate was markedly increased (p < 0.05). On the other hand, the expression of all the cytokines except IL-10 in spleen was mildly increased but not significant statistically, and there was no significant difference in the numerical changes of white blood cell (WBC). These results indicate that feeding ERW may have influence on the local immune response (Th-1 type cytokines such as IL-1beta, TNF-alpha) in the small intestine but not on the systemic immune response.
PMID: 19252295

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Biological & Pharmaceutical Bulletin. 2009 Feb;73(2):280-7. Epub 2009 Feb 7.
Enhanced induction of mitochondrial damage and apoptosis in human leukemia HL-60 cells due to electrolyzed-reduced water and glutathione.
Tsai CF, Hsu YW, Chen WK, Ho YC, Lu FJ. Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.

Electrolzyed-reduced water (ERW) is a higher pH and lower oxidation-reduction potential water. In the present study, we examined the enhanced effect of ERW in the apoptosis of leukemia cells (HL-60) induced by glutathione (GSH). An enhanced inhibitory effect on the viability of the HL-60 cells was observed after treatment with a combination of ERW with various concentrations of GSH, whereas no cytotoxic effect in normal peripheral blood mononuclear cells was observed. The results of apoptotic related protein indicated that the induction of HL-60 cell death was caused by the induction of apoptosis through upregulation of Bax and downregulation of Bcl-2. The results of further investigation showed a diminution of intracellular GSH levels in ERW, and combination with GSH groups. These results suggest that ERW is an antioxidant, and that ERW, in combination with GSH, has an enhanced apoptosis-inducing effect on HL-60 cells, which might be mediated through the mitochondria-dependent pathway. PMID: 19202298
Source: http://www.ncbi.nlm.nih.gov/pubmed/19202298. Complete study in PDF

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Cytotechnology. 2002 Nov;40(1-3):139-49.
Protective mechanism of reduced water against alloxan-induced pancreatic beta-cell damage: Scavenging effect against reactive oxygen species.
Li Y, Nishimura T, Teruya K, Maki T, Komatsu T, Hamasaki T, Kashiwagi T, Kabayama S, Shim SY, Katakura Y, Osada K, Kawahara T, Otsubo K, Morisawa S, Ishii Y, Gadek Z, Shirahata S. Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, Japan.

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. PMID: 19003114

Microwaterman comment: Our Alkaline Water Videos page is another great source of information on Electrolyzed Reduced Water. 

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Cytotechnology. 2005 Jan;47(1-3):97-105.
Electrolyzed Reduced Water Supplemented with Platinum Nanoparticles Suppresses Promotion of Two-stage Cell Transformation.
Nishikawa R, Teruya K, Katakura Y, Osada K, Hamasaki T, Kashiwagi T, Komatsu T, Li Y, Ye J, Ichikawa A, Otsubo K, Morisawa S, Xu Q, Shirahata S. Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, 812-8581, Higashi-ku, Fukuoka, Japan.

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. PMID: 19003049


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Biological & Pharmaceutical Bulletin 2007 Feb;30(2):234-6
Preservative effect of electrolyzed reduced water on pancreatic beta-cell mass in diabetic db/db mice.
Kim MJ, Jung KH, Uhm YK, Leem KH, Kim HK.
Department of Obesity Management, Graduate School of Obesity Science, Dongduk Women's University, Seoul, South Korea. mijakim@dongduck.ac.jp

Oxidative stress is produced under diabetic conditions and involved in progression of pancreatic beta-cell dysfunction. 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 genetically diabetic mouse strain C57BL/6J-db/db (db/db). ERW with ROS scavenging ability reduced the blood glucose concentration, increased blood insulin level, improved glucose tolerance and preserved beta-cell mass in db/db mice. The present data suggest that ERW may protects beta-cell damage and would be useful for antidiabetic agent. PMID: 17268057

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Life Science. 2006 Nov 10;79(24):2288-92. Epub 2006 Aug 2.
Anti-diabetic effects of electrolyzed reduced water in streptozotocin-induced and genetic diabetic mice.
Kim MJ, Kim HK. Department of Obesity management, Graduate School of Obesity Science, Dongduk Women's University, 23-1 Wolkgukdong, Seoul, 136-714, South Korea. mijakim@dongduk.ac.kr

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.
PMID: 16945392

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Kidney Int. 2006 Jul;70(2):391-8. Epub 2006 Jun 7.
Electrolyzed-reduced water reduced hemodialysis-induced erythrocyte impairment in end-stage renal disease patients.
Huang KC, Yang CC, Hsu SP, Lee KT, Liu HW, Morisawa S, Otsubo K, Chien CT. Department of Family Medicine, National Taiwan University College of Medicine and National Taiwan University Hospital, Taipei, Taiwan.

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. PMID: 16760903

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Kidney Int. 2003 Aug;64(2):704-14.
Reduced hemodialysis-induced oxidative stress in end-stage renal disease patients by electrolyzed reduced water.
Huang KC, Yang CC, Lee KT, Chien CT. Department of Family Medicine, National Taiwan University College of Medicine and National Taiwan University Hospital, Taipei, Taiwan.

BACKGROUND: Increased oxidative stress in end-stage renal disease (ESRD) patients may oxidize macromolecules and consequently lead to cardiovascular events during chronic hemodialysis. Electrolyzed reduced water (ERW) with reactive oxygen species (ROS) scavenging ability may have a potential effect on reduction of hemodialysis-induced oxidative stress in ESRD patients. METHODS: We developed a chemiluminescence emission spectrum and high-performance liquid chromatography analysis to assess the effect of ERW replacement on plasma ROS (H2O2 and HOCl) scavenging activity and oxidized lipid or protein production in ESRD patients undergoing hemodialysis. Oxidized markers, dityrosine, methylguanidine, and phosphatidylcholine hydroperoxide, and inflammatory markers, interleukin 6 (IL-6), and C-reactive protein (CRP) were determined.

RESULTS: Although hemodialysis efficiently removes dityrosine and creatinine, hemodialysis increased oxidative stress, including phosphatidylcholine hydroperoxide, and methylguanidine. Hemodialysis reduced the plasma ROS scavenging activity, as shown by the augmented reference H2O2 and HOCl counts (Rh2o2 and Rhocl, respectively) and decreased antioxidative activity (expressed as total antioxidant status in this study). ERW administration diminished hemodialysis-enhanced Rh2o2 and Rhocl, minimized oxidized and inflammatory markers (CRP and IL-6), and partly restored total antioxidant status during 1-month treatment.

CONCLUSION: This study demonstrates that hemodialysis with ERW administration may efficiently increase the H2O2- and HOCl-dependent antioxidant defense and reduce H2O2- and HOCl-induced oxidative stress. PMID: 12846769

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Inhibitory effect of electrolyzed reduced water on tumor angiogenesis
Biological & Pharmaceutical Bulletin. 2008 Jan;31(1):19-26

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. Graduate School of Systems Life Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8581, Japan

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, ORP) 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. PMID: 18175936  

Related Research:
Levels of Soluble Vascular Endothelial Growth Factor (VEGF) Receptor 1 in Astrocytic Tumors and Its Relation to Malignancy, Vascularity, and VEGF-A

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Anti Cancer Effect of Alkaline Reduced Water
By the Department of Parasitology, Institute of Basic Medical Sciences, Department of Microbiology, Department of Biochemistry, Wonju College of Medicine , Yonsei University Department of Biomedical Laboratory Science and Institute of Health Science, College of Health Science, Yonsei University .

Certain minerals can produce alkaline reduced water with high pH and low oxidation-reduction potential (ORP) when dissolved in water. Alkaline reduced water showed significant anticancer effect. When B16 melanoma cells were inoculated subcutaneous and intra-peritoneally, C56BL/6 mice fed with alkaline reduced water showed tumor growth delay and the survival span was significantly lengthened. Alkaline reduced water also showed the inhibition of metastasis by reducing the numbers of B16 melanoma colonies when injected through tail vein. The amount of reactive oxygen species (ROS) was very reduced when fed with alkaline reduced water except for spleen, which is a major organ for immunity. Even for normal mice, alkaline reduced water intake invoked systemic cytokines, such as, Th1 (IFN- g, IL-12) and Th2 (IL-4, IL-5), suggesting strong immuno-modulation effect. Both ROS scavenging effect and immuno-modulation effect might be responsible for anticancer effect of alkaline reduced water. Continue reading the full study

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Electrolyzed-reduced water scavenges active oxygen species and protects DNA from oxidative damage.
Biochem Biophys Res Commun. 1997 May 8;234(1):269-74.

Shirahata S, Kabayama S, Nakano M, Miura T, Kusumoto K, Gotoh M, Hayashi H, Otsubo K, Morisawa S, Katakura Y.

Institute of Cellular Regulation Technology, Graduate School of Genetic Resources Technology, Kyushu University, Fukuoka, Japan. sirahata@grt.kyushu-u.ac.jp

Active oxygen species or free radicals are considered to cause extensive oxidative damage to biological macromolecules, which brings about a variety of diseases as well as aging. The ideal scavenger for active oxygen should be 'active hydrogen'. 'Active hydrogen' can be produced in reduced water near the cathode during electrolysis of water. Reduced water exhibits high pH, low dissolved oxygen (DO), extremely high dissolved molecular hydrogen (DH), and extremely negative redox potential (RP) values. Strongly electrolyzed-reduced water, as well as ascorbic acid, (+)-catechin and tannic acid, completely scavenged O.-2 produced by the hypoxanthine-xanthine oxidase (HX-XOD) system in sodium phosphate buffer (pH 7.0). The superoxide dismutase (SOD)-like activity of reduced water is stable at 4 degrees C for over a month and was not lost even after neutralization, repeated freezing and melting, deflation with sonication, vigorous mixing, boiling, repeated filtration, or closed autoclaving, but was lost by opened autoclaving or by closed autoclaving in the presence of tungsten trioxide which efficiently adsorbs active atomic hydrogen. Water bubbled with hydrogen gas exhibited low DO, extremely high DH and extremely low RP values, as does reduced water, but it has no SOD-like activity. These results suggest that the SOD-like activity of reduced water is not due to the dissolved molecular hydrogen but due to the dissolved atomic hydrogen (active hydrogen). Although SOD accumulated H2O2 when added to the HX-XOD system, reduced water decreased the amount of H2O2 produced by XOD. Reduced water, as well as catalase and ascorbic acid, could directly scavenge H2O2. Reduced water suppresses single-strand breakage of DNA b active oxygen species produced by the Cu(II)-catalyzed oxidation of ascorbic acid in a dose-dependent manner, suggesting that reduced water can scavenge not only O2.- and H2O2, but also 1O2 and .OH.  PMID: 9169001

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Electrolyzed-reduced water protects against oxidative damage to DNA, RNA, and protein.
Appl Biochem Biotechnol. 2006 Nov;135(2):133-44.

Lee MY, Kim YK, Ryoo KK, Lee YB, Park EJ. Department of Genetic Engineering, Soonchunhyang University, Asan, Chungnam 336-600, Korea.

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. Pre-treatment, co-treatment, and post-treatment 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 electrolyzed reduced water on the oxidation of ascorbic acid.  PMID: 17159237

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Biophys Chem. 2004 Jan 1;107(1):71-82.

The mechanism of the enhanced antioxidant effects against superoxide anion radicals of reduced water produced by electrolysis.
Hanaoka K, Sun D, Lawrence R, Kamitani Y, Fernandes G.

Bio-REDOX Laboratory Inc. 1187-4, Oaza-Ueda, Ueda-shi, Nagano-ken 386-0001, Japan. hanak@rapid.ocn.ne.jp

We reported that reduced water produced by electrolysis enhanced the antioxidant effects of proton donors such as ascorbic acid (AsA) in a previous paper. We also demonstrated that reduced water produced by electrolysis of 2 mM NaCl solutions did not show antioxidant effects by itself. We reasoned that the enhancement of antioxidant effects may be due to the increase of the ionic product of water as solvent. The ionic product of water (pKw) was estimated by measurements of pH and by a neutralization titration method. As an indicator of oxidative damage, Reactive Oxygen Species- (ROS) mediated DNA strand breaks were measured by the conversion of supercoiled phiX-174 RF I double-strand DNA to open and linear forms. Reduced water had a tendency to suppress single-strand breakage of DNA induced by reactive oxygen species produced by H2O2/Cu (II) and HQ/Cu (II) systems. The enhancement of superoxide anion radical dismutation activity can be explained by changes in the ionic product of water in the reduced water. PMID: 14871602 [PubMed]

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Use of Ionized water in hypochlorhydria or achlorhydria
Prof. Kuninaka Hironage, Head of Kuninaka Hospital
"Too many fats in the diets, which lead to the deposition of cholesterol on the blood vessels, which in turn constrict the blood flow, cause most illnesses such as high blood pressure. In accordance with the theory of Professor Gato of Kyushu University on Vitamin K (because vitamin K enables the blood calcium to increase ), or The consumption of more antioxidant water, the effectiveness of the increase in the calcium in high blood pressure is most significant. The consumption of alkaline antioxidant water for a period of 2 to 3 months, I have observed the blood pressure slowly drop, due to the water's solvent ability, which dissolves the cholesterol in the blood vessels."

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Use of Ionized water for gynecological conditions
Prof. Watanabe Ifao, Watanabe Hospital
"Ionized alklaine antioxidant water improves body constituents and ensures effective healing to many illnesses. The uses of antioxidant water in gynecological patients have proved to be very effective. The main reason for its effectiveness is that this water can neutralize toxins.

When given antioxidant water to pre-eclamptic toxemia cases, the results are most significant. During my long years of servicing the pre-eclamptic toxemia cases, I found that the women with pre-eclamptic toxemia who consumed antioxidant water tend to deliver healthier babies with stronger muscles. A survey report carried out on babies in this group showed intelligence above average."


Toxin Neutralization
Prof. Kuwata Keijiroo, Doctor of Medicine

"In my opinion, the wonder of antioxidant water is the ability neutralizes toxins, but it is not a medicine. The difference is that the medicine can only apply to each and individual case, whereas the antioxidant water can be consumed generally and its neutralizing power is something which is very much unexpected. Now, in brief, let me introduce to you a heart disease case and how it was cured.

The patient was a 35 years old male suffering from vascular heart disease. For 5 years, his sickness deteriorated. He was in the Setagays Government Hospital for treatment.

During those 5 years, he had been in and out of the hospital 5 to 6 times. He had undergone high tech examinations such as angiogram by injecting VINYL via the vein into the heart. He consulted and sought treatment from many good doctors where later he underwent a major surgical operation. Upon his discharge from the hospital, he quit his job to convalesce. However, each time when his illness relapsed, the attack seemed to be even more severe.

Last year, in August, his relatives were in despair and expected he would not live much longer. It so happened at that time that the victim's relative came across antioxidant water processor. His illness responded well and he is now on the road to recovery."

(In the United States, cardiovascular diseases account for more than one-half of the approximate 2 million deaths occurring each year…. It is estimated that optimal conditioning of drinking water could reduce this cardiovascular disease mortality rate by as much as 15 percent in the United States) - Report of the Safe Drinking Water Committee of the National Academy of Sciences, 1977

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Eczema
Prof. Tamura Tatsuji, Keifuku Rehabilitation Center
"Eczema is used to describe several varieties of skin conditions, which have a number of common features. The exact cause or causes of eczema are not fully understood. I many cases, eczema can be attributed by external irritants.

Let me introduce a patient who recovered from skin disease after consuming the antioxidant water. This patient suffered 10 years of eczema and could not be cured effectively even under specialist treatment. This patient, who is 70 years of age, is the president of a vehicle spare parts company. After the war, his lower limbs suffered acute eczema, which later became chronic. He was repeatedly treated in a specialist skin hospital.

The left limb responded well to treatment, but not so on the right limb. He suffered severe itchiness, which, when scratched led to bleeding. During the last 10 years, he was seen and treated by many doctors. When I first examined him, his lower limb around the joints was covered with vesicles. Weeping occurred owing to serum exuding from the vesicles.

I advised him to try consuming antioxidant water. He bought a unit and consumed the antioxidant water religiously and used the acidic water to bathe the affected areas. After 2 weeks of treatment the vesicles dried up. The eczema was completely cleared without any relapse after 1½ month."

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Allergies
Prof. Kuninaka Hironaga, Head of Kuninaka Hospital
"Mr. Yamada, the head of Police Research Institute, suffered from severe allergy. He was treated repeatedly by skin specialist, but with no success. Then he started consuming antioxidant water. The allergy responded very well and was soon completely cured. No relapse had occurred, although he had taken all kinds of food. He was most grateful and excited about this treatment.

As for myself, I had also suffered severe allergy. Ever since I began to consume antioxidant water, the allergy has recovered. Since then, I started a research on the effectiveness of antioxidant water.

I discovered that most allergies are due to acidification of body condition and is also related to consuming too much meat and sugar. In every allergy case, the patient's antioxidant minerals are excessively low which in turn lower the body resistance significantly. The body becomes overly sensitive and develops allergy easily. To stabilize the sensitivity, calcium solution in injected into the vein. Therefore, it is clear that the antioxidant water has ionic calcium, which can help alleviate allergy.

The ionic calcium not only enhances the heart, urination, and neutralization of toxins but controls acidity. It also enhances the digestive system and liver function. This will promote natural healing power and hence increase its resistance to allergy. In some special cases of illness, which do not respond to drugs, it is found, it is found to respond well to antioxidant water."

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Digestive Problems
Prof. Kogure Keizou, Kogure Clinic of Juntendo Hospital

"The stomach is readily upset both by diseases affecting the stomach and by other general illnesses. In addition, any nervous tension or anxiety frequently causes gastric upset, vague symptoms when This information is under some strain.

The important role of antioxidant water in our stomach is to neutralize the secretion and strengthen it s functions. Usually, after consuming the antioxidant water for 1 to 3 minutes, the gastric juice increase to 1½ times. For those suffering from hypochlorhydria or achlorhydria ( low in gastric juice ) the presence of antioxidant water will stimulate the stomach cells to secrete more gastric juice. This in turn enhances digestion and absorption of minerals.

However, on the other hand, those with hyperchlorhydria ( high in gastric juice ), the antioxidant water neutralizes the excessive gastric juice. Hence, it does not create any adverse reaction.

According to the medical lecturer from Maeba University, the pH of the gastric secretion will still remain normal when antioxidant water is consumed. This proves that the ability of the antioxidant water is able to neutralize as well as to stimulate the secretion."

Microwaterman comment: I find Professor Keizou's observations to be right on the money and agrees with our own experience.

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Diabetes

Prof. Kuwata Keijiroo, Doctor of Medicine
"When I was serving in the Fire Insurance Association, I used to examine many diabetic patients. Besides treating them with drugs, I provided them with antioxidant water. After drinking antioxidant water for one month, 15 diabetic patients were selected and sent to Tokyo University for further test and observations.

Initially, the more serious patients were a bit apprehensive about the treatment. When the antioxidant water was consumed for some time, the sugar in the blood and urine ranged from a ratio of 300 mg/l to 2 mg / dc. There was a time where the patient had undergone 5 to 6 blood tests a day and detected to be within normal range. Results also showed that even 1 ½ hour after meals, the blood sugar and urine ratio was 100 mg/dc: 0 mg/dc . The sugar in the urine has completely disappeared."

NOTE:
More Americans than ever before are suffering from diabetes, with the number of new cases averaging almost 800,000 each year. The disease has steadily increased in the United States since 1980, and in 1998, 16 million Americans were diagnosed with diabetes (10.3 million diagnosed; 5.4 million undiagnosed). Diabetes is the seventh leading cause of death in the United States, and more than 193,000 died from the disease and its related complication in 1996. The greatest increase - 76 percent - occurred in people age 30 to 30.

From: U. S. Department of Health and Human Services, October 13, 2000 Fact Sheet

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Clinical Improvements Obtained From The Intake Of Reduced Water

Extracts from " Presentation At The Eight Annual International Symposium On man And His Environment in Health And Disease" on February 24th 1990, at The Grand Kempinski Hotel, Dallas, Texas, USA by Dr. H. Hayashi, M.D. and Dr. M Kawamura, M.D., on : -

THE CONCEPT OF PREHEPATIC MEDICINES

Since the introduction of alkaline ionic water in our clinic in 1985, we have had the following interesting clinical experiences in the use of this type of water. By the use of alkaline ionic water for drinking and the preparation of meals for our in-patients, we have noticed :

Declines in blood sugar levels in diabetic patients.
Improvements in peripheral circulation in diabetic gangrene.
Declines in uric acid levels in patients with gout.
Improvements in liver function exams in hepatic disorders.
Improvements in gastroduodenal ulcer and prevention of their recurrences.
Improvements in hypertension and hypotension.
Improvements in allergic disorders such as asthma, urticaria, rhinites and atopic dermatitis.
Improvements in persistent diarrhea which occurred after gastrectomy.
Quicker improvements in post operative bower paralysis.
Improvements in serum bilirubin levels in new born babies.

Being confirming clinical improvements, we have always observed changes of stools of the patients, with the colour of their feaces changing from black-brown colour to a brigher yellow-brown one, and the odour of their feaces becoming almost negligible.

The number of patients complaining of constipation also decreased markedly. The change of stool findings strongly suggests that alkaline ionic water intake can decrease the production of putrefield or pathogenic metabolites.

Devices to produce reduced water were introduced into our clinic in May 1985. Based on the clinical experiences obtained in the past 15 years, it can be said that introduction of electrolyzed-reduced water for drinking and cooking purpose for in-patients should be the very prerequisite in our daily medical practices. Any dietary recipe cannot be a scientific one if property of water is not taken by the patients is not taken into consideration.

The Ministry of Health and Welfare in Japan announced in 1965 that the intake of reduced water is effective for restoration of intestinal flora metabolism.

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Clinical evaluation of alkaline ionized water for abdominal complaints: Placebo controlled double blind tests

by Hirokazu Tashiro, Tetsuji Hokudo, Hiromi Ono, Yoshihide Fujiyama, Tadao Baba (National Ohkura Hospital, Dept. of Gastroenterology; Institute of Clinical Research, Shiga University of Medical Science, Second Dept. of Internal Medicine)

Effect of alkaline ionized water on abdominal complaints was evaluated by placebo controlled double blind tests. Overall scores of improvement using alkaline ionized water marked higher than those of placebo controlled group, and its effect proved to be significantly higher especially in slight symptoms of chronic diarrhoea and abdominal complaints in cases of general malaise. Alkaline ionized water group did not get interrupted in the course of the test, nor did it show serious side effects nor abnormal test data. It was confirmed that alkaline ionized water is safer and more effective than placebos.

Summary

Effect of alkaline ionized water on abdominal complaints was clinically examined by double blind tests using clean water as placebo. Overall improvement rate was higher for alkaline ionized water group than placebo group and the former proved to be significantly more effective than the other especially in cases of slight symptoms. Examining improvement rate for each case of chronic diarrhoea, constipation and abdominal complaints, alkaline ionized water group turned out to be more effective than placebo group for chronic diarrhoea, and abdominal complaints. The test was stopped in one case of chronic diarrhoea, among placebo group due to exacerbation, whereas alkaline ionized water group did not stop testing without serious side effects or abnormal test data in all cases. It was confirmed that alkaline ionized water is more effective than clean water against chronic diarrhoea, abdominal complaints and overall improvement rate (relief of abdominal complaints) and safer than clean water.

Introduction

Since the approval of alkaline ionized water electrolyzers by Pharmaceutical Affairs Law in 1966 for its antacid effect and efficacy against gastrointestinal disorders including hyperchylia, indigestion, abnormal gastrointestinal fermentation and chronic diarrhoea, they have been extensively used among patients. However, medical and scientific evaluation of their validity is not established. In our study, we examined clinical effect of alkaline ionized water on gastrointestinal disorders across many symptoms in various facilities. Particularly, we studied safety and usefulness of alkaline ionized water by doubleblind tests using clean water as a control group.

Test subjects and methods

163 patients (34 men, 129 women, age 21 to 72, average 38.6 years old) of indigestion, abnormal gastrointestinal fermentation (with abnormal gas emission and rugitus) and abdominal complaints caused by irregular dejection (chronic diarrhoea, or constipation) were tested as subjects with good informed consent. Placebo controlled double blind tests were conducted using alkaline ionized water and clean water at multiple facilities. An alkaline ionized water electrolyzer sold commercially was installed with a pump driven calcium dispenser in each of the subject homes. Tested alkaline ionized water had pH at 9.5 and calcium concentration at 30ppm. Each subject in placebo group used a water purifier that has the same appearance as the electrolyzer and produces clean water.

The tested equipment was randomly assigned by a controller who scaled off the key code which was stored safely until the tests were completed and the seal was opened again.

Water samples were given to each patient in the amount of 200ml in the morning with the total of 50OmI or more per day for a month. Before and after the tests, blood, urine and stool were tested and a log was kept on the subjective symptoms, bowel movements and accessory symptoms. After the tests, the results were analyzed based on the log and the test data.

Water Ionizer Test Results

1. Symptoms

Among 163 tested subjects, alkaline ionized water group included 84 and placebo group 79. Background factors such as gender, age and basal disorders did not contribute to significant difference in the results.

2. Overall improvement rate

As to overall improvement rate of abdominal complaints, alkaline ionized water group had 2 cases of outstanding improvement (2.5%), 26 cases of fair improvement (32.1%), 36 cases of slight improvement (44.4%), 13 cases of no change (16%) and 4 cases of exacerbation (4.9%), whereas placebo group exhibited 4 (5.2%), 19 (24.7%), 27 (35.1%), 25 (32.5%) and 2 cases (2.6%) for the same category. Comparison between alkaline ionized water and placebo groups did not reveal any significant difference at the level of 5% significance according to the Wilcoxon test, although alkaline ionized water group turned out to be significantly more effective than placebo group at the level of p value of 0.22.

Examining overall improvement rates by a 7, 2 test (with no adjustment for continuity) between the effective and noneffective groups, alkaline ionized water group had 64 (79%) of effective cases and 17 cases (21%) of non effective cases, whereas placebo group had 50 (64.9%) and 27 (35.1%) cases respectively. The result indicated that alkaline ionized water group was significantly more effective than placebo group at the level of p value of 0.0.48.

Looking only at 83 slight cases of abdominal complaints, overall improvement rate for alkaline ionized water groupp

(45 cases) was composed of 11 cases (242%) of fair improvement, 22 cases (48.9%) of slight improvement, 17 cases (44.7%) of no change and 3 cases (6.7%) of exacerbation, whereas placebo group (38 cases) had 3 (7.8%), 17 (44.7%), 17 (44.7%) and 1 (2.6%) cases for the same category. Alkaline ionized water group was significantly more effective than placebo group according to the comparison between the groups (p value = 0.033).

3. Improvement rate by basal symptom

4. Safety

Conclusion

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Physiological effects of alkaline ionized water: Effects on metabolites produced by intestinal fermentation

by Takashi Hayakawa, Chicko Tushiya, Hisanori Onoda, Hisayo Ohkouchi, Harul-~to Tsuge (Gifu University, Faculty of Engineering, Dept. of Food Science)

We have found that long-term ingestion of alkaline ionized water (alkaline ionized water) reduces cecal fermentation in rats that were given highly fermentable commercial diet (MF: Oriental Yeast Co., Ltd.). In this experiment, rats were fed MF and test water (tap water, alkaline ionized water with pH at 9 and 10) for about 3 months. Feces were collected on the 57th day, and the rats were dissected on the 88th day. The amount of ammonium in fresh feces and cecal contents as well as fecal free-glucose tended to drop down for the alkaline ionized water group. In most cases, the amount of free-amino acids in cecal contents did not differ significantly except for cysteine (decreased in alkaline ionized water with pH at 10) and isoleucine (increased in alkaline ionized water with pH at 10).

Purpose of tests

Testing methods

Test results and analyses

Bibliography

*2. "Official Pharmaceutical Guidelines of Japan, Vol. IT' by Japan Public Documents Association, Hirokawa PublIshin Co., 1996

*3. "Science and Technology of Functional Water" (part) by Takashi Hayakawa, Haruffito Tsuge, edited by Water Scienll cc Institute, 1999, pp.109-116

*4. 'Tasics and Effective Use of Alkaline Ionized Water" by Takashi Hayakawa, Haruhito Tsuge, edited by Tetsuji Hc kudou, 25th General Assembly of Japan Medical Congress 'Tunctional Water in Medical Treatment", Administratio~ Offices, 1999, pp. 10- 11

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Effects of alkaline ionized water on formation & maintenance of osseous tissues

Effects of calcium alkaline ionized water on formation and maintenance of osseous tissues in rats were examined. In the absence of calcium in the diet, no apparent calcification was observed with only osteoid formation being prominent. Striking differences were found among groups that were given diets with 30% and 60% calcium. Rats raised by calcium ionized water showed the least osteogenetic disturbance. Tibiae and humeri are more susceptible to calcium deficiency than femora. Theses results may indicate that calcium in drinking water effectively supplements osteogenesis in case of dietary calcium deficiency. The mechanism involved in osteoid formation such as absorption rate of calcium from the intestine and effects of calcium alkaline ionized drinking water on maintaining bone structure in the process of aging or under the condition of calcium deficiency is investigated.

Osteoporosis that has lately drawn public attention is defined as "conditions of bone brittleness caused by reduction in the amount of bone frames and deterioration of osseous microstructure." Abnormal calcium metabolism has been considered to be one of the factors to contribute to this problem, which in turn is caused by insufficient calcium take in, reduction in enteral absorption rate of calcium and increase in the amount of calcium in urinal discharge. Under normal conditions, bones absorb old bones by regular metabolism through osteoid formation to maintain their strength and function as supporting structure. It is getting clear that remodeling of bones at the tissue level goes through the process of activation, resorption, reversal, matrix synthesis and mineralization. Another important function of bones is storing minerals especially by coordinating with intestines and kidneys to control calcium concentration in the blood. When something happens to this osteo metabolism, it results in abnormal morphological changes. Our analyses have been focusing mostly on the changes in the amount of bones to examine effects of calcium alkaline ionized water on the reaction system of osteo metabolism and its efficiency. Ibis time, however, we studied it further from the standpoint of histology. In other words, we conducted comparative studies on morphological and kinetic changes of osteogenesis by testing alkaline ionized water, tap water and solution of lactate on rats.

Three week old male Wistar rats were divided into 12 groups by conditions of feed and drinking water. Feeds were prepared with 0%, 30%, 60% and 100% of normal amount of calcium and were given freely. Three types of drinking water, tap water (city water, about 6ppm of Ca), calcium lactate solution (Ca=40ppm) and alkaline ionized water (Ca =40ppm, pH=9, produced by an electrolyzer NDX 4 LMC by Omco OMC Co., Ltd.) were also given keely. Rats' weight, amount of drinking water and feed as well as the content of Ca in drinking water were assayed every day. On the 19th and 25th days of testing, tetracycline hydrochloride was added to the feed for 48 hours so as to bring its concentration to 30mg/kg. On the 30th day, blood samples were taken under Nembutal anesthesia, and tibiae, humeri and femora were taken out to make non decalcified samples. Their conditions of osteoid formation and rotation were observed using Villanueva bone stain and Villanueva goldner stain.

Three groups that were given different types of drinking water and the same amount of Ca in the feed were compared to find out no significant difference in the rate of weight gain and intakes of feed and drinking water. Alkaline ionized water group had significantly greater amount of tibiae and humeri with higher concentration of calcium in the bones.

The group of 0% calcium in the feed saw drastic increase in the amount of osteoid. There was not much difference by types of drinking water. Almost no tetracycline was taken into tibiae and humeri, although a small amount was identified in ferora. As a result, osteogenesis went as far as osteoid formation, but it was likely that decalcification has not happened yet, or most of newly formed bones were absorbed.

As to the groups of 30% and 60% calcium in the feed, increase in the area of tetracycline take in was more identifiable with higher clarity in descending order of alkaline ionized water, calcium lactate solution and tap water groups. Especially in case of tap water group, irregularity among the areas of tetracycline take in was distinctive. The group of 100% calcium in the feed saw some improvements in osteogenesis in descending order of alkaline ionized water, calcium lactate solution and tap water. In any case, bone formation seemed to be in good condition at near normal level.

Alkaline ionized water was regarded to be effective for improvements of osteogenesis under the conditions of insufficient calcium in the feed. Also, the extent. of dysosteogenesis differed by the region. That is, tibiae and humeri tend to have more significant dysosteogenesis than femora.

In addition, there is a possibility that osteo metabolism varies depending on enteral absorption rate of calcium, adjustment of discharge from kidneys and functional adjustment of accessory thyroid in the presence of alkaline ionized water. We are now studying its impact on calcium concentration in the blood. We are also examining whether it is possible to deter bone deterioration by testing on fast aging mouse models.

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ACIDIC WATER

Comparison of electrolyzed oxidizing water with various antimicrobial interventions to reduce Salmonella species on poultry.
Poult Sci. 2002 Oct;81(10):1598-605.
Fabrizio KA, Sharma RR, Demirci A, Cutter CN. Department of Food Science, The Pennsylvania State University, University Park 16802, USA.

Foodborne pathogens in cell suspensions or attached to surfaces can be reduced by electrolyzed oxidizing (EO) water; however, the use of EO water against pathogens associated with poultry has not been explored. In this study, acidic EO water [EO-A; pH 2.6, chlorine (CL) 20 to 50 ppm, and oxidation-reduction potential (ORP) of 1,150 mV], basic EO water (EO-B; pH 11.6, ORP of -795 mV), CL, ozonated water (OZ), acetic acid (AA), or trisodium phosphate (TSP) was applied to broiler carcasses inoculated with Salmonella Typhimurium (ST) and submerged (4 C, 45 min), spray-washed (85 psi, 25 C, 15 s), or subjected to multiple interventions (EO-B spray, immersed in EO-A; AA or TSP spray, immersed in CL). Remaining bacterial populations were determined and compared at Day 0 and 7 of aerobic, refrigerated storage. At Day 0, submersion in TSP and AA reduced ST 1.41 log10, whereas EO-A water reduced ST approximately 0.86 log10. After 7 d of storage, EO-A water, OZ, TSP, and AA reduced ST, with detection only after selective enrichment. Spray-washing treatments with any of the compounds did not reduce ST at Day 0. After 7 d of storage, TSP, AA, and EO-A water reduced ST 2.17, 2.31, and 1.06 log10, respectively. ST was reduced 2.11 log10 immediately following the multiple interventions, 3.81 log10 after 7 d of storage. Although effective against ST, TSP and AA are costly and adversely affect the environment. This study demonstrates that EO water can reduce ST on poultry surfaces following extended refrigerated storage. PMID: 12412930


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Treatment of Escherichia coli (O157:H7) inoculated alfalfa seeds and sprouts with electrolyzed oxidizing water.
Int J Food Microbiol. 2003 Sep 15;86(3):231-7.

Department of Agricultural and Biological Engineering, Pennsylvania State University, University Park, PA 16802, USA.

Electrolyzed oxidizing water is a relatively new concept that has been utilized in agriculture, livestock management, medical sterilization, and food sanitation. Electrolyzed oxidizing (EO) water generated by passing sodium chloride solution through an EO water generator was used to treat alfalfa seeds and sprouts inoculated with a five-strain cocktail of nalidixic acid resistant Escherichia coli O157:H7. EO water had a pH of 2.6, an oxidation-reduction potential of 1150 mV and about 50 ppm free chlorine. The percentage reduction in bacterial load was determined for reaction times of 2, 4, 8, 16, 32, and 64 min. Mechanical agitation was done while treating the seeds at different time intervals to increase the effectiveness of the treatment. Since E. coli O157:H7 was released due to soaking during treatment, the initial counts on seeds and sprouts were determined by soaking the contaminated seeds/sprouts in 0.1% peptone water for a period equivalent to treatment time. The samples were then pummeled in 0.1% peptone water and spread plated on tryptic soy agar with 5 microg/ml of nalidixic acid (TSAN). Results showed that there were reductions between 38.2% and 97.1% (0.22-1.56 log(10) CFU/g) in the bacterial load of treated seeds. The reductions for sprouts were between 91.1% and 99.8% (1.05-2.72 log(10) CFU/g). An increase in treatment time increased the percentage reduction of E. coli O157:H7. However, germination of the treated seeds reduced from 92% to 49% as amperage to make EO water and soaking time increased. EO water did not cause any visible damage to the sprouts. PMID: 12915034

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Inactivation of Escherichia coli (O157:H7) and Listeria monocytogenes on plastic kitchen cutting boards by electrolyzed oxidizing water.
Venkitanarayanan KS, Ezeike GO, Hung YC, Doyle MP. Department of Animal Science, University of Connecticut, Storrs 06269, USA.

One milliliter of culture containing a five-strain mixture of Escherichia coli O157:H7 (approximately 10(10) CFU) was inoculated on a 100-cm2 area marked on unscarred cutting boards. Following inoculation, the boards were air-dried under a laminar flow hood for 1 h, immersed in 2 liters of electrolyzed oxidizing water or sterile deionized water at 23 degrees C or 35 degrees C for 10 or 20 min; 45 degrees C for 5 or 10 min; or 55 degrees C for 5 min. After each temperature-time combination, the surviving population of the pathogen on cutting boards and in soaking water was determined. Soaking of inoculated cutting boards in electrolyzed oxidizing water reduced E. coli O157:H7 populations by > or = 5.0 log CFU/100 cm2 on cutting boards. However, immersion of cutting boards in deionized water decreased the pathogen count only by 1.0 to 1.5 log CFU/100 cm2. Treatment of cutting boards inoculated with Listeria monocytogenes in electrolyzed oxidizing water at selected temperature-time combinations (23 degrees C for 20 min, 35 degrees C for 10 min, and 45 degrees C for 10 min) substantially reduced the populations of L. monocytogenes in comparison to the counts recovered from the boards immersed in deionized water. E. coli O157:H7 and L. monocytogenes were not detected in electrolyzed oxidizing water after soaking treatment, whereas the pathogens survived in the deionized water used for soaking the cutting boards. This study revealed that immersion of kitchen cutting boards in electrolyzed oxidizing water could be used as an effective method for inactivating foodborne pathogens on smooth, plastic cutting boards. PMID: 10456736


The bactericidal effects of electrolyzed oxidizing water on bacterial strains involved in hospital infections.
Artif Organs. 2004 Jun;28(6):590-2.

Vorobjeva NV, Vorobjeva LI, Khodjaev EY. Department of Physiology of Microorganisms, Biology Faculty, Moscow State University, Lenin Hills 1/12, Moscow 119992, Russia. nvvorobjeva@mail.ru

The study is designed to investigate bactericidal actions of electrolyzed oxidizing water on hospital infections. Ten of the most common opportunistic pathogens are used for this study. Cultures are inoculated in 4.5 mL of electrolyzed oxidizing (EO) water or 4.5 mL of sterile deionized water (control), and incubated for 0, 0.5, and 5 min at room temperature. At the exposure time of 30 s the EO water completely inactivates all of the bacterial strains, with the exception of vegetative cells and spores of bacilli which need 5 min to be killed. The results indicate that electrolyzed oxidizing water may be a useful disinfectant for hospital infections, but its clinical application has still to be evaluated. PMID: 15153153

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Effect of electrolyzed oxidizing water and hydrocolloid occlusive dressings on excised burn-wounds in rats.
Xin H, Zheng YJ, Hajime N, Han ZG. Department of Thoracic Surgery, China-Japan Union Hospital, Jilin University, Jilin 130031, China. xinhua7254@yahoo.com.cn

OBJECTIVE: To study the efficacy of electrolyzed oxidizing water (EOW) and hydrocolloid occlusive dressings in the acceleration of epithelialization in excised burn-wounds in rats. METHODS: Each of the anesthetized Sprague-Dawley rats (n=28) was subjected to a third-degree burn that covered approximately 10% of the total body surface area. Rats were assigned into four groups: Group I (no irrigation), Group II (irrigation with physiologic saline), Group III (irrigation with EOW) and Group IV (hydrocolloid occlusive dressing after EOW irrigation). Wounds were observed macroscopically until complete epithelialization was present, then the epithelialized wounds were examined microscopically.

RESULTS: Healing of the burn wounds was the fastest in Group IV treated with hydrocolloid occlusive dressing together with EOW. Although extensive regenerative epidermis was seen in each Group, the proliferations of lymphocytes and macrophages associated with dense collagen deposition were more extensive in Group II, III and IV than in Group I. These findings were particularly evident in Group III and IV. CONCLUSIONS: Wound Healing may be accelerated by applying a hydrocolloid occlusive dressing on burn surfaces after they are cleaned with EOW. PMID: 12857518


Effect of electrolyzed water on wound healing.
Artif Organs. 2000 Dec;24(12):984-7.

Yahagi N, Kono M, Kitahara M, Ohmura A, Sumita O, Hashimoto T, Hori K, Ning-Juan C, Woodson P, Kubota S, Murakami A, Takamoto S. Department of Anesthesiology, Teikyo University Mizonokuchi Hospital, Tokyo, Japan. naokiyah@aol.com

Electrolyzed water accelerated the healing of full-thickness cutaneous wounds in rats, but only anode chamber water (acid pH or neutralized) was effective. Hypochlorous acid (HOCl), also produced by electrolysis, was ineffective, suggesting that these types of electrolyzed water enhance wound healing by a mechanism unrelated to the well-known antibacterial action of HOCl. One possibility is that reactive oxygen species, shown to be electron spin resonance spectra present in anode chamber water, might trigger early wound healing through fibroblast migration and proliferation. PMID: 11121980

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Decomposition of ethylene, a flower-senescence hormone, with electrolyzed anode water.
Biosci Biotechnol Biochem.
2003 Apr;67(4):790-6.

Harada K, Yasui K. Department of Research and Development, Hokkaido Electric Power Co., Inc., 2-1 Tsuishikari, Ebetsu, Hokkaido 067-0033, Japan. kharada@h1.hotcn.ne.jp

Electrolyzed anode water (EAW) markedly extended the vase life of cut carnation flowers. Therefore, a flower-senescence hormone involving ethylene decomposition by EAW with potassium chloride as an electrolyte was investigated. Ethylene was added externally to EAW, and the reaction between ethylen and the available chlorine in EAW was examined. EAW had a low pH value (2.5), a high concentration of dissolved oxygen, and extremely high redox potential (19.2 mg/l and 1323 mV, respectively) when available chlorine was at a concentration of about 620 microns. The addition of ethylene to EAW led to ethylene decomposition, and an equimolar amount of ethylene chlorohydrine with available chlorine was produced. The ethylene chlorohydrine production was greatly affected by the pH value (pH 2.5, 5.0 and 10.0 were tested), and was faster in an acidic solution. Ethylene chlorohydrine was not produced after ethylene had been added to EAW at pH 2.6 when available chlorine was absent, but was produced after potassium hypochlorite had been added to such EAW. The effect of the pH value of EAW on the vase life of cut carnations was compatible with the decomposition rate of ethylene in EAW of the same pH value. These results suggest that the effect of EAW on the vase life of cut carnations was due to the decomposition of ethylene to ethylene chlorohydrine by chlorine from chlorine compounds. PMID: 12784619

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