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The role of naturally occurring D (deuterium, heavy isotope of hydrogen) in living organisms has been examined by using deuterium-depleted water (DDW) containing 30-100 ppm D instead of water containing D at its natural abundance of D (150 ppm).

It has been discovered in the early 1990s that decreasing the deuterium (heavy isotope of hydrogen) concentration of the living organism below normal physiological level delays or halts the proliferation of tumorous cell lines. Deuterium-depleted water (DDW) significantly decreased the growth rate of L929 fibroblast, HT-29 colon, A4, MDA and MCF-7 breast, PC-3 prostate, M19 melanoma cell lines in vitro.

In subsequent in vitro and in vivo toxicology studies DDW proved to be completely safe and non-toxic. DDW caused tumor regression in vivo in xenotransplanted mice (MDA and MCF-7, human breast; PC-3), in dogs and cats with spontaneous tumors and induced apoptosis in vitro and in vivo.

In order to reveal the molecular background of the inhibitory effect of DDW COX-2 gene expression was investigated in healthy myometrial and HT-29 colon tumor cell lines. It was found that deuterium depletion inhibited COX-2 expression and the inhibitory effect correlated with the D concentration.

Deuterium depletion also inhibited the expression of the oncogenes c-myc and H-ras and p53 suppressor gene in the different organs of carcinogen exposed mice.

The application of DDW also proved to influence the expression of genes encoding different kinases (detected using nanocapillary quantitative real-time PCR analysis technique) and to modify the activity of amilorid- sensitive Na+/H+ antiport system.

We suppose that the naturally occurring D is the key element of a hitherto unknown sub-molecular regulatory system (SMRS).
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