Deuterium Depleted Water Abstracts 1


Anti-aging effects of deuterium depletion on Mn-induced toxicity in a C. elegans model.
            (Avila et al., 2012) Download
Work with sub-natural levels of deuterium (D) in animals has demonstrated an anti-cancer effect of low D-concentration in water. Our objective was to investigate whether deuterium-depleted water (DDW) can overturn reverse manganese (Mn)-induced reduction in life span, using the Caenorhabditis elegans (C. elegans) as a model system. DDW per se had no effect on worm's life span 48 h after treatment; however, it reversed the Mn-induced decrease in C. elegans life span. Mn reduced DAF-16 levels, a transcription factor strongly associated with life-span regulation. Low D-concentration (90 ppm) restored the Mn-induced changes in DAF-16 to levels indistinguishable from controls, suggesting DDW can regulate the DAF-16 pathway. We further show that insulin-like receptor DAF-2 levels were unaltered by Mn exposure, tAKT levels increased, whilst superoxide dismutase (SOD-3) levels were decreased by Mn. DDW (90 ppm) restored the levels of tAKT and superoxide dismutase (SOD) to control values without changing DAF-2 levels. Treatment of Mn exposed worms with DDW (90 ppm) restored life-span, DAF-16 and SOD-3 levels to control levels, strongly suggesting that low D concentrations can protect against Mn toxic effects.

Technologies For Obtaining Deuterium Depleted Water
            (Barishev et al., 2013) Download
The principal manufacturers of deuterium depleted water currently apply the method of distillation in rectifying columns. The disadvantage of this method is the low separation coefficient. Multiple stages are needed in order to significantly reduce the deuterium content which makes the method expensive. We have designed the electrolytical method with a recuperation unit which allows reducing by 4-6 power consumption required for producing light water comparing to rectification methods applied nowadays.


Submolecular regulation of cell transformation by deuterium depleting water exchange reactions in the tricarboxylic acid substrate cycle.
            (Boros et al., 2016) Download
The naturally occurring isotope of hydrogen ((1)H), deuterium ((2)H), could have an important biological role. Deuterium depleted water delays tumor progression in mice, dogs, cats and humans. Hydratase enzymes of the tricarboxylic acid (TCA) cycle control cell growth and deplete deuterium from redox cofactors, fatty acids and DNA, which undergo hydride ion and hydrogen atom transfer reactions. A model is proposed that emphasizes the terminal complex of mitochondrial electron transport chain reducing molecular oxygen to deuterium depleted water (DDW); this affects gluconeogenesis as well as fatty acid oxidation. In the former, the DDW is thought to diminish the deuteration of sugar-phosphates in the DNA backbone, helping to preserve stability of hydrogen bond networks, possibly protecting against aneuploidy and resisting strand breaks, occurring upon exposure to radiation and certain anticancer chemotherapeutics. DDW is proposed here to link cancer prevention and treatment using natural ketogenic diets, low deuterium drinking water, as well as DDW production as the mitochondrial downstream mechanism of targeted anti-cancer drugs such as Avastin and Glivec. The role of (2)H in biology is a potential missing link to the elusive cancer puzzle seemingly correlated with cancer epidemiology in western populations as a result of excessive (2)H loading from processed carbohydrate intake in place of natural fat consumption.

Deuterium-depleted water inhibits human lung carcinoma cell growth by apoptosis.
            (Cong et al., 2010) Download
To investigate the in vivo and in vitro inhibitory effects of deuterium-depleted water (DDW) on human lung cancer and the possible mechanisms underlying these effects, we cultured and treated human lung carcinoma cell line A549 and human embryonic lung fibroblasts HLF-1 with various concentrations of DDW from 2 to 72 h. Cellular growth inhibition rates were determined using the 3-(4, 5-dimethyldiazol-2-yl)-2, 5-diphenyltetrazolium-bromide) (MTT) proliferation assay. A549 cells were treated with 50±5 ppm DDW, and the morphology and structure of cells were observed by scanning electron microscopy (SEM). We observed alterations in the cellular skeleton by transmission electron microscopy (TEM) and changes in cell cycle by flow cytometry. Our data showed that DDW significantly inhibited the proliferation of A549 cells at a specific time point, and cells demonstrated the characteristic morphological changes of apoptosis under SEM and TEM. The length of the S phase increased significantly in cells treated with 50 ppm DDW, whereas the G0 to G1 phase and G2 to M phase were decreased. We observed DDW-induced cellular apoptosis using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and DNA fragment analyses. In addition, we established a tumor transplantion model by injecting H460 tumor cells into subcutaneous tissue of BALB/c mice treated with DDW for 60 days. We determined the tumor inhibition rate of treated and control groups and found that the tumor weight was significantly decreased and the tumor inhibition rate was approximately 30% in the DDW group. We conclude that DDW is a promising new anticancer agent with potential for future clinical application.

Content of deuterium in biological fluids and organs: Influence of deuterium depleted water on D/H gradient and the process of adaptation.
            (Dzhimak et al., 2015) Download
It is found that consumption of deuterium depleted water reduces not only the content of deuterium in biological fluids but also more than 2 times reduces the D/H gradient value along the line: mixed saliva > blood plasma. The experimental data showed that a physiological solution prepared on deuterium depleted water during induced apoptosis activates the DNA repair system, significantly reducing the number of single-stranded DNA breaks, which, in general, indicates an increase in the efficiency of defensive systems of the cell.

Revealing water's secrets: deuterium depleted water.
            (Goncharuk et al., 2013) Download
BACKGROUND:  The anomalous properties of water have been of great interest for generations of scientists. However the impact of small amount of deuterium content which is always present in water has never been explored before. For the first time the fundamental properties of deuterium depleted (light) water at 4°C and 20°C are here presented. RESULTS:  The obtained results show the important role of the deuterium in the properties of bulk water. At 4°C the lowest value of the kinematic viscosity (1.46 mm2/s) has been found for 96.5 ppm D/H ratio. The significant deviation in surface tension values has been observed in deuterium depleted water samples at the both temperature regimes. The experimental data provides direct evidence that density, surface tension and viscosity anomalies of water are caused by the presence of variable concentration of deuterium which leads to the formation of water clusters of different size and quantity. CONCLUSIONS:  The investigated properties of light water reveal the origin of the water anomalies. The new theoretical model of cluster formation with account of isotope effect is proposed.


Deuterium depleted water effects on survival of lung cancer patients and expression of Kras, Bcl2, and Myc genes in mouse lung.
            (Gyöngyi et al., 2013) Download
Although advances in cancer therapies continue to develop, the shortness of the survival of lung cancer patients is still disappointing. Therefore, finding new adjuvant strategies is within the focus of cancer cure. Based on observations that deuterium depletion inhibits the growth of cancer cell lines and suppresses certain proto-oncogenes, we have conducted a clinical study in 129 patients with small cell and nonsmall cell lung cancers who consumed deuterium-depleted drinking water (DDW) as a nontoxic agent in addition to conventional chemotherapy and radiotherapy. Median survival time (MST) was 25.9 mo in males and 74.1 mo in female patients; the difference between genders was statistically significant (p < 0.05). Median survival of subjects with brain metastasis was 27.1 mo. Cumulative 5-yr survival probabilities were 19%, 52%, and 33% in males, females, and all patients with brain metastasis, respectively. Gene expression analysis in mouse lung indicated that DDW attenuates 7,12-dimethylbenz(a)anthracene (DMBA)-induced expression of Bcl2, Kras, and Myc in females. In conclusion, DDW counteracts the DMBA-induced overexpression of Bcl2, Kras and Myc genes in mouse lung, and it may extend survival of lung cancer patients as a nontoxic anticancer dietary supplement, especially for women with tumors overexpressing cancer-related genes, because MST of DDW-consuming group was 2-4 times longer than it is generally observed in lung cancer patients.

A retrospective evaluation of the effects of deuterium depleted water consumption on 4 patients with brain metastases from lung cancer.
            (Krempels et al., 2008) Download
HYPOTHESES:  Because of the number of sufferers and high mortality rate, the standard care and new therapeutic options in the treatment of brain metastasis from lung cancer are the subject of intense research. A new concept based on the different chemical and physical behavior of protium and deuterium affecting cell signaling and tumor growth has been introduced in the treatment of cancer patients. The aim of this study was to investigate the impact of deuterium depleted water (DDW) consumption in addition to conventional forms of therapy on the survival of lung cancer patients with brain metastasis. STUDY DESIGN:  A series of 4 case histories was retrospectively evaluated. The patients were diagnosed with brain metastasis deriving from a primary lung tumor and started consuming DDW at the time of or after the diagnosis of the brain metastasis, which was inoperable or the surgical intervention did not result in complete regression. The primary objective was survival. METHODS:  The daily water intake of the patients was replaced with DDW, which complemented the conventional forms of treatment. Patients were consuming DDW for at least 3 months. The treatment was continued with DDW of 10 to 15 to 20 ppm lower deuterium (D) content every 1 to 2 months and thus a gradual decrease was maintained in the D-concentration in the patient's body. RESULTS:  DDW consumption integrated into conventional treatments resulted in a survival time of 26.6, 54.6, 21.9, and 33.4 months in the 4 patients, respectively. The brain metastasis of 2 patients showed complete response (CR), whereas partial response (PR) was detected in 1 patient, and the tumor growth was halted (no change or NC) in 1 case. The primary tumor of 2 patients indicated CR, and the lung tumor in 2 patients showed PR. CONCLUSIONS:  DDW was administered as an oral anticancer agent in addition to conventional therapy, and noticeably prolonged the survival time of all 4 lung cancer patients with brain metastasis. We suggest that DDW treatment, when integrated into other forms of cancer treatment, might provide a new therapeutic option.

Deuterium-depleted water has stimulating effects on long-term memory in rats.
            (Mladin et al., 2014) Download
Deuterium-depleted water (DDW) is a water which has a 6-7-fold less concentration of the naturally occurring deuterium (20-25ppm vs. 150ppm). While administrated for a longer period, it may reduce the concentration of deuterium throughout the body, thus activating cellular mechanisms which are depending on protons (channels, pumps, enzyme proteins). The aim of the present work was to study, for the first time in our knowledge, the possible influence of deuterium-depleted water (DDW) chronic administration in normal Wistar rats, as compared to a control group which received distilled water, on spatial working memory and the locomotor activity (as studied through Y-maze) or both short-term and long-term spatial memory (assed in radial 8 arms-maze task). Our results presented here showed no significant modifications in terms of spatial working memory (assessed through spontaneous alternation percentage) and locomotor activity (expressed through the number of arm entries) in Y-maze, as a result of DDW ingestion. Also, no significant differences between the DDW and control group were found in terms of the number of working memory errors in the eight-arm radial maze, as a parameter of short-term memory. Still, we observed a significant decrease for the number of reference memory errors in the DDW rats. In this way, we could speculate that the administration of DDW may generate an improvement of the reference memory, as an index of long-term memory. Thus, we can reach the conclusion that the change between the deuterium/hydrogen balance may have important consequences for the mechanisms that govern long-term memory, as showed here especially in the behavioral parameters from the eight-arm radial maze task.


Biological effects of deuteronation: ATP synthase as an example.
            (Olgun, 2007) Download
BACKGROUND:  In nature, deuterium/hydrogen ratio is approximately 1/6600, therefore one of approximately 3300 water (H2O) molecules is deuterated (HOD + D2O). In body fluids the ratio of deuterons to protons is approximately 1/15000 because of the lower ionization constant of heavy water. The probability of deuteronation rather than protonation of Asp 61 on the subunit c of F0 part of ATP synthase is also approximately 1/15000. The contribution of deuteronation to the pKa of Asp 61 is 0.35. THEORY AND DISCUSSION:  In mitochondria, the release of a deuteron into the matrix side half-channel of F0 is likely to be slower than that of a proton. As another example, deuteronation may slow down electron transfer in the electron transport chain (ETC) by interfering with proton coupled electron transport reactions (PCET), and increase free radical production through the leakage of temporarily accumulated electrons at the downstream complexes. CONCLUSION:  Deuteronation, as exemplified by ATP synthase and the ETC, may interfere with the conformations and functions of many macromolecules and contribute to some pathologies like heavy water toxicity and aging.

Deuteronation and aging.
            (Olgun et al., 2007) Download
Deuterium has one proton and one neutron in its atomic nucleus, but hydrogen has only proton. The natural abundance of deuterium is 1 per approximately 6600 hydrogen atoms. Therefore deuterated water (both HOD + D(2)O [heavy water]) abundance is 1 per approximately 3300 water molecules. One dissociation product of deuterated and heavy water is deuteron (proton + neutron, D(+), H(2)OD(+)/D(3)O(+)). Because heavy water has a lower ionization constant than water, the D(+)/H(+) ratio is approximately 1/15,000 in biological fluids. O-D bond length is shorter than O-H, and D-O-D angle is lesser than H-O-H. Once a deuteron exchanges with proton on the water-exposed surface of a macromolecule, it can lead to a conformational change and the reverse exchange will be less likely. Deuteron bonds are stronger than proton bonds. Therefore an increase of deuteronated macromolecules can be expected in due course of time. In order to test this hypothesis, we conducted a pilot study and measured the D/H ratio in the tails of three Sprague-Dawley rats at different ages (4 weeks, 5 weeks, and >1-year old) by elemental analysis coupled with isotope ratio mass spectrometry (EA-IRMS) technique. To prevent the effect of daily water consumption, the homogenized tails were lyophilized before analysis. The results, as mean of several measurements, of 4 weeks, 5 weeks, and >1-year-old rats were per thousand-94 +/- 9.56, per thousand-101.71 +/- 6.89, per thousand-83.68 +/- 3.46 delta((2)H) relative to VSMOW, respectively. Although there is a slight increase in >1-year-old rat, the difference among the animals was not significant. We propose that, before reaching to a final conclusion about the accumulation of deuterium with aging, the measurements should be done not in whole tissue samples but in purified macromolecules from a larger set of animals.

Effect of deuterium-depleted water on selected cardiometabolic parameters in fructose-treated rats.
            (Rehakova et al., 2016) Download
Deuterium-depleted water (DDW) has a lower concentration of deuterium than occurs naturally (less than 145 ppm). While effects of DDW on cancer started to be intensively studied, the effects on cardiovascular system are completely unknown. Thus, we aimed to analyze the effects of DDW (55+/-5 ppm) administration to 12-week-old normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) treated with 15 % fructose for 6 weeks. Blood pressure (BP) and selected biochemical parameters were measured together with determination of nitric oxide synthase (NOS) activity and iNOS and eNOS protein expressions in the left ventricle (LV) and aorta. Neither DDW nor fructose had any significant effect on BP in both strains. DDW treatment decreased total cholesterol and triglyceride levels in WKY, but it was not able to prevent increase in the same parameters elevated due to fructose treatment in SHR. Both fructose and DDW increased insulin level in WKY. Fructose did not affect NOS activity either in WKY or SHR. DDW increased NOS activity in LV of both WKY and SHR, while it decreased NOS activity and iNOS expression in the aorta of SHR with or without fructose treatment. In conclusion, DDW treatment significantly modified biochemical parameters in WKY together with NOS activity elevation in the heart. On the other hand, it did not affect biochemical parameters in SHR, but decreased NOS activity elevated due to iNOS upregulation in the aorta.

In vitro assessment of antineoplastic effects of deuterium depleted water.
            (Soleyman-Jahi et al., 2014) Download
BACKGROUND:  In vitro, in vivo and clinical studies have demonstrated anti-cancer effects of deuterium depleted water (DDW). The nature of this agents action, cytotoxic or cytostatic, remains to be elucidated. We here aimed to address the point by examining effects on different cell lines. MATERIALS AND METHODS:  3-(4, 5-dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) -based cytotoxicity analysis was conducted for human breast, stomach, colon, prostate cancer and glioblastoma multiforme cell lines as well as human dermal fibroblasts. The cell lines were treated with decreasing deuterium concentrations of DDW alone, paclitaxel alone and both. One way analysis of variance (ANOVA) was used for statistical analysis. RESULTS:  Treatment with different deuterium concentrations of DDW alone did not impose any significant inhibitory effects on growth of cell lines. Paclitaxel significantly decreased the survival fractions of all cell lines. DDW augmented paclitaxel inhibitory effects on breast, prostate, stomach cancer and glioblastoma cell lines, with influence being more pronounced in breast and prostate cases. CONCLUSIONS:  DDW per se does not appear to have inhibitory effects on the assessed tumor cell lines as well as normal fibroblasts. As an adjuvant, however, DDW augmented inhibitory effects of paclitaxel and thus it could be considered as an adjuvant to conventional anticancer agents in future trials.

Naturally occurring deuterium is essential for the normal growth rate of cells.
            (Somlyai et al., 1993) Download
The role of naturally occurring D in living organisms has been examined by using deuterium-depleted water (30-40 ppm D) instead of water containing the natural abundance of D (150 ppm). The deuterium-depleted water significantly decreased the growth rate of the L929 fibroblast cell line, and also inhibited the tumor growth in xenotransplanted mice. Eighty days after transplantation in 10 (59%) out of 17 tumorous mice the tumor, after having grown, regressed and then disappeared. We suggest that the naturally occurring D has a central role in signal transduction involved in cell cycle regulation.

Deuterium content of water increases depression susceptibility: the potential role of a serotonin-related mechanism.
            (Strekalova et al., 2015) Download
Environmental factors can significantly affect disease prevalence, including neuropsychiatric disorders such as depression. The ratio of deuterium to protium in water shows substantial geographical variation, which could affect disease susceptibility. Thus the link between deuterium content of water and depression was investigated, both epidemiologically, and in a mouse model of chronic mild stress. We performed a correlation analysis between deuterium content of tap water and rates of depression in regions of the USA. Next, we used a 10-day chronic stress paradigm to test whether 2-week deuterium-depleted water treatment (91 ppm) affects depressive-like behavior and hippocampal SERT. The effect of deuterium-depletion on sleep electrophysiology was also evaluated in naïve mice. There was a geographic correlation between a content of deuterium and the prevalence of depression across the USA. In the chronic stress model, depressive-like features were reduced in mice fed with deuterium-depleted water, and SERT expression was decreased in mice treated with deuterium-treated water compared with regular water. Five days of predator stress also suppressed proliferation in the dentate gyrus; this effect was attenuated in mice fed with deuterium-depleted water. Finally, in naïve mice, deuterium-depleted water treatment increased EEG indices of wakefulness, and decreased duration of REM sleep, phenomena that have been shown to result from the administration of selective serotonin reuptake inhibitors (SSRI). Our data suggest that the deuterium content of water may influence the incidence of affective disorder-related pathophysiology and major depression, which might be mediated by the serotoninergic mechanisms.

Deuterium-depleted water (DDW) inhibits the proliferation and migration of nasopharyngeal carcinoma cells in vitro.
            (Wang et al., 2013) Download
Recent studies have demonstrated that natural water that has 65% of the deuterium concentration depleted, can exhibit anti-tumor properties. However, the anti-tumor effects of DDW on various nasopharyngeal carcinoma (NPC) cells have not previously been reported. In the present study, NPC cell lines and normal preosteoblast MC3T3-E1 cells were grown in RPMI1640 media containing different deuterium concentrations (50-150 ppm). The effects of DDW on the proliferation and migration of NPC and MC3T3-E1 cells were investigated using the MTT, plate colony formation, and Transwell assays, as well as Boyden chamber arrays, flow cytometry (FCM), western blot and immunofluorescence. We found that DDW was an effective inhibitor of NPC cell proliferation, plated colony formation, migration and invasion. In contrast, the growth of normal preosteoblast MC3T3-E1 cells was promoted when they were cultured in the presence of DDW. Cell cycle analysis revealed that DDW caused cell cycle arrest in the G1/S transition, reduced the number of cells in the S phase and significantly increased the population of cells in the G1 phase in NPC cells. Western blot analysis revealed that treatment with DDW significantly increased the expression of NADPH:quinone oxidoreductase-1 (NQO1), while immunofluorescence assay analysis revealed that treatment with DDW decreased the expression of PCNA and matrix metalloproteinase 9 (MMP9) in NPC cells. These results demonstrated that DDW is a novel, non-toxic adjuvant therapeutic agent that suppresses NPC cell proliferation, migration, and invasion by inducing the expression of NQO1 and causing cell cycle arrest, as well as decreasing PCNA and MMP9 expression.

Effects of low-level deuterium enrichment on bacterial growth.
            (Xie and Zubarev, 2014) Download
Using very precise (±0.05%) measurements of the growth parameters for bacteria E. coli grown on minimal media, we aimed to determine the lowest deuterium concentration at which the adverse effects that are prominent at higher enrichments start to become noticeable. Such a threshold was found at 0.5% D, a surprisingly high value, while the ultralow deuterium concentrations (≤0.25% D) showed signs of the opposite trend. Bacterial adaptation for 400 generations in isotopically different environment confirmed preference for ultralow (≤0.25% D) enrichment. This effect appears to be similar to those described in sporadic but multiple earlier reports. Possible explanations include hormesis and isotopic resonance phenomena, with the latter explanation being favored.



Avila, DS, et al. (2012), ‘Anti-aging effects of deuterium depletion on Mn-induced toxicity in a C. elegans model.’, Toxicol Lett, 211 (3), 319-24. PubMed: 22561170
Barishev, MG, et al. (2013), ‘Technologies For Obtaining Deuterium Depleted Water’, Membranes, 3 (1), 523-26. PubMed:
Boros, LG, et al. (2016), ‘Submolecular regulation of cell transformation by deuterium depleting water exchange reactions in the tricarboxylic acid substrate cycle.’, Med Hypotheses, 87 69-74. PubMed: 26826644
Cong, FS, et al. (2010), ‘Deuterium-depleted water inhibits human lung carcinoma cell growth by apoptosis.’, Exp Ther Med, 1 (2), 277-83. PubMed: 22993540
Dzhimak, SS, AA Basov, and MG Baryshev (2015), ‘Content of deuterium in biological fluids and organs: Influence of deuterium depleted water on D/H gradient and the process of adaptation.’, Dokl Biochem Biophys, 465 370-73. PubMed: 26728727
Goncharuk, VV, et al. (2013), ‘Revealing water’s secrets: deuterium depleted water.’, Chem Cent J, 7 (1), 103. PubMed: 23773696
Gyöngyi, Z, et al. (2013), ‘Deuterium depleted water effects on survival of lung cancer patients and expression of Kras, Bcl2, and Myc genes in mouse lung.’, Nutr Cancer, 65 (2), 240-46. PubMed: 23441611
Krempels, K, I Somlyai, and G Somlyai (2008), ‘A retrospective evaluation of the effects of deuterium depleted water consumption on 4 patients with brain metastases from lung cancer.’, Integr Cancer Ther, 7 (3), 172-81. PubMed: 18815148
Mladin, C, et al. (2014), ‘Deuterium-depleted water has stimulating effects on long-term memory in rats.’, Neurosci Lett, 583 154-58. PubMed: 25263786
Olgun, A, et al. (2007), ‘Deuteronation and aging.’, Ann N Y Acad Sci, 1100 400-3. PubMed: 17460204
Olgun, A (2007), ‘Biological effects of deuteronation: ATP synthase as an example.’, Theor Biol Med Model, 4 9. PubMed: 17316427
Rehakova, R, et al. (2016), ‘Effect of deuterium-depleted water on selected cardiometabolic parameters in fructose-treated rats.’, Physiol Res, 65 (Supplementum 3), S401-7. PubMed: 27775425
Soleyman-Jahi, S, et al. (2014), ‘In vitro assessment of antineoplastic effects of deuterium depleted water.’, Asian Pac J Cancer Prev, 15 (5), 2179-83. PubMed: 24716953
Somlyai, G, et al. (1993), ‘Naturally occurring deuterium is essential for the normal growth rate of cells.’, FEBS Lett, 317 (1-2), 1-4. PubMed: 8428617
Strekalova, T, et al. (2015), ‘Deuterium content of water increases depression susceptibility: the potential role of a serotonin-related mechanism.’, Behav Brain Res, 277 237-44. PubMed: 25092571
Wang, H, et al. (2013), ‘Deuterium-depleted water (DDW) inhibits the proliferation and migration of nasopharyngeal carcinoma cells in vitro.’, Biomed Pharmacother, 67 (6), 489-96. PubMed: 23773852
Xie, X and RA Zubarev (2014), ‘Effects of low-level deuterium enrichment on bacterial growth.’, PLoS One, 9 (7), e102071. PubMed: 25033078