How is Aqua-Rx™ Produced?
Cavitation such as found in Sonoluminescence.

Researchers at the Oak Ridge National Laboratory (ORNL), Rensselaer Polytechnic Institute and the Russian Academy of Sciences have reported results that suggest the possibility of nuclear reactions during the explosive collapse of bubbles in liquid, a process known as cavitation. The bubbles, which grow in the presence of sound waves, collapse to produce locally high pressure and temperatures. When a sound wave propagates through a liquid, the molecules in the liquid are subjected to positive and negative pressures. During the negative phase of the wave, tiny bubbles in the liquid can grow dramatically (up to a factor of 1000 in volume), since the pressure is below the vapor pressure. When the positive pressure phase of the sound wave passes, the bubble collapses, and the energy accumulated in the bubble during growth is released. This process is known as "acoustic cavitation".

Temperatures in the collapsing bubbles can reach 10,000,000 Kelvin, sufficient to influence chemical reactions. These pressures and temperatures can be sufficiently high to result in light emissions, called sonoluminescence, from the collapsing bubbles. This suggests the possibility of producing densities and temperatures necessary for nuclear reactions. In particular, a long-sought goal of sonoluminescence research has been the possibility of achieving nuclear fusion reaction conditions.

 

Aqua-Rx™ Molecularly Redefined Water –
U.S. Patent 6,521,248 and 7,198,254

A water molecule is made up of a single oxygen atom and two hydrogen atoms, but single molecules of water do not exist in liquid water. Liquid water is made up of H2O molecules that are held together in clusters that range from five molecules, the theoretical smallest number occurring as liquid water, to several hundred molecules. Some H2O molecules and clusters are bonded with molecules of other substances, and are referred to as “bound water.”

The Company’s proprietary physics process is used to reorganize large molecular clusters into a smaller, higher energy state spatial organization of molecules. The molecularly redefined water is stable and exists at a meta-stable energy state. This is validated by the water’s statistically significant higher boiling point and viscosity.

Cell Hydrated with Aqua-Rx™

The study of the molecular structure of liquid water is a relatively new field, but one that has become the focus of intensive scientific study by a number of educational institutions, including Cambridge University and the University of South Banks. More information about structured water can be found at the Cluster Database website maintained by Cambridge University (www.cam.ac.uk, using the search term: “cluster”, select the “Cambridge Cluster Database”) and at the University of South Banks’ website (www.sbu.ac.uk/water).

Human cells use water as a core component of their most critical functions, such as exchanging oxygen for toxins (including free-radicals), creating energy and repairing cells. Cells exchange water through active protein channels known as aquaporin. The body must break down large clusters and bound water before the water can be used since aquaporin channels only allow single molecules to pass through the cell membrane. Aquaporin dilate to only 3 Angstroms in diameter, just large enough to let single H2O molecules into the cell.

Research and Studies

Validation of Aqua-Rx™’s Unique Structure
by General Physics Institute, Moscow, Russia

In September 2002, Dr. A.F. Bunkin, Dr. A.A. Nyrmatov, and Dr. S.M. Pershin of the General Physics Institute in Moscow tested Aqua-Rx™ water comparing it to other types of water utilizing modified Raman spectroscopy laser technology. This laser technology was developed in 2001 by the General Physics Institute and an article about this technology was published in the Physics of Vibration scientific journal, Vol. 9 No.3 pp 141-144, 2001, published by Allerton Press of New York. The article is entitled, “Modification of Spontaneous Raman Spectrum of Liquid Water Exposed To Strong Ultrasonic Field”.

Test results demonstrated Aqua-Rx™ water is a new spatial organization of water molecules. The difference between Aqua-Rx™ and other waters is that at a given temperature, Aqua-Rx™ water clusters are, on a constant basis, equal to the size of normal water clusters at 12 degrees Centigrade higher temperature. Additional tests in 2003 show that the water clusters in Aqua-Rx™ average 30% smaller than standard water clusters. The scientists at the General Physics Institute have never seen this unique physical property before in water. Drs. Bunkin, Nyrmatov and Pershin published their results in “Study of cluster molecular structures in various types of liquid waters by using spontaneous Raman spectroscopy “, Physics of Vibrations, Vol. 10 No. 2, 2002.

Boiling point and viscosity study
by Professor Andrew Kickson at major California university in San Diego

The aim of these measurements was to measure the boiling temperature of Aqua-Rx™ water at atmospheric pressure and to compare it with that measured for laboratory deionized water in a similar experiment. The Aqua-Rx™ water used was obtained from Bio-Hydration Research Lab, Inc. in 500 mL PETE bottles (produced 040604); the laboratory deionized water was obtained from a Barnstead E-pure 4 cartridge deionization system which produces Type I reagent-grade water.

The measurements were carried out in a 5 L glass beaker containing approximately 3 L of deionized water. In this beaker was placed a separate glass container (35 cm height and 8 cm diameter) that contained the sample whose boiling point was being measured. The sample was stirred using a magnetic stirrer while the whole system was heated to boiling temperature. The temperature of the boiling sample was measured using a calibrated platinum resistance thermometer (Hart 1504) immersed to a depth of about 2 cm in the liquid. The ambient atmospheric pressure was monitored using a digital quartz barometer (Paroscientific Model 740-16B).

The temperature was recorded 15 min after boiling started and again every minute for 10 minutes. Three series of measurements were made on both the Type I reagent grade water and on the Aqua-Rx™ water.

The average measured value of boiling temperature for all measurements of reagent-grade water is 100.082 ± 0.008 °C and for the Aqua-Rx™ water is 100.125 ± 0.008 °C. The boiling point of the Aqua-Rx™ water is thus 0.043 °C higher than that of the reagent-grade water, and this difference is statistically significant.

The Company supplied samples of water collected from the feed water purified by reverse osmosis and deionization and water collected immediately after the patented physics process, without the subsequent oxygenation step. Measurements were made at room temperature of the kinematic viscosity of the water samples.

A Cannon-Ubbelohde suspended meniscus viscometer (size 0) was used to measure kinematic viscosity of the samples provided at atmospheric pressure and at room temperature.

The viscometer was cleaned before use with a 50% aqua regia solution (a mixture of three parts of HCl acid and one part of HNO3 acid) to remove any traces of organic deposits. It was then rinsed with copious amounts of deionized water and dried at 110 °C. Replicate measurements were made on 5 individual samples of both water samples. The measured efflux times and the computed kinematic and dynamic viscosities were calculated.
The results show a relative difference observed at 20 °C of about 0.3 %. This is statistically significant.

 

Mutagenic Study of Human DNA

The analysis of the frequency of chromosomal aberrations in a culture of human lymphocytes is one of the main tests applied in the study of mutagenic activity of environmental factors and is approved by the (WHO) World Health Organization. The determination of sister chromatid exchange (SCE) frequency is also one of the standard tests used in the evaluation of mutagenicity. This method possesses specificity and high sensitivity in the evaluation of mutagenic properties of chemical compounds. Tests were accomplished at The Institute of Medical Genetics in Moscow by Prof. Chebotarev to determine if Aqua-Rx™ water might have an effect on the frequency of chromosomal aberrations. Conclusions: SCE frequency is 19% statistically lower (P<0.001) in Aqua-Rx™ water. Frequency of chromosomal aberrations is 64% lower (P<0.05) in Aqua-Rx™ water. Results of the study were presented at the “Medical Genetic Evaluation of the Food Products” Conference in Moscow on 27-29 May 2003.

 

Increased Intracellular pH

September 2002, Doctors Ludmila Litinskaya Ph.D. and Tatyana Ogloblina Ph.D. at the Moscow University in Russia tested the effects of Aqua-Rx™ water on intracellular pH. Pig embryo kidney cells were cultured in FDA 199 medium with 10% bovine serum containing Fluorescein diacetate fluorescent dye. One solution was made with standard double distilled lab water and one was made using Aqua-Rx™ water. pH measurements were taken using the microfluormetric method based on using FDA dye. An increase of 0.43 pH was noted on the cells grown in the Aqua-Rx™ solution, while the standard double distilled water solution showed an overall decrease in intracellular pH.

 

Cell Survivability & Intracellular pH Study

In September 2002, Dr. V. Turovetsky at the Biophysics department of Moscow University accomplished a cell survivability and intracellular pH study on mice peritoneal macrophage cells to determine the effects on cell membranes under short and long term exposure to 199 solution made with double distilled lab water verses FDA 199 solution made with Aqua-Rx™ water and containing FDA dye. Conclusion: Peritoneal macrophage cells prepared using Aqua-Rx™ water demonstrated an increase in cell survivability of 266% and an increase in intracellular pH delta of 0.16 in 15 minutes, 0.32 in 190 minutes and 0.43 in 230 minutes, while the cells incubated in the medium prepared in the double distilled solution had an increase of 0.01 in 15 minutes, 0.03 in 190 minutes and 0.06 in 230 minutes. There was a substantial increase in intracellular pH for Aqua-Rx™ over double distilled water. This increase in intracellular pH is most likely responsible for the dramatic increase in cell survivability.

 

Solubility of Calcium Oxalate Monohydrate

Calcium Oxalate Monohydrate (COM) is the major component of 85% of human kidney stones. A test was accomplished at Moscow University by Prof. Rashkovich to determine if Aqua-Rx™ water might have an effect on dissolving kidney stones compared to distilled water. It has long been known that water does not dissolve calcium oxalate type kidney stones. A 30-micron long COM crystal was tested by flowing distilled lab water over the crystal at 125/liters/hr flow rate. The distilled lab water crystal showed no loss of material. A 30 micron COM crystal exposed to the same flow rate using Aqua-Rx™ water demonstrated a loss of 25 to 35 % of the calcium oxalate crystal in 121 minutes of exposure. Conclusion: Aqua-Rx™ water dissolves COM crystals, the major component of human kidney stones, and explains the consumer reports of passing kidney stones shortly after starting to drink Aqua-Rx™ water. Prof. Rashkovich has completed follow-up studies and presented his findings in a lecture to N.A.S.A. in March 2003 in Huntsville, Alabama. This study was published in Elsevier Science’s Journal of Crystal Growth 261 (2004) pp 539-548.

 

Rat Brain Tissue Study

Experiments were conducted at The Institute of Higher Nervous Activity and Neuro-physiology at the Academy of Sciences of Russia in Moscow on brain tissue sections of Wistar rats, 1 month of age. Rat brain was isolated and placed into cold artificial cerebrospinal (AC) fluid prepared with double distilled water and Aqua-Rx™. The dynamics of induced response amplitude were recorded for each solution. After one hour, the amplitude decreased to 31.7% of the initial value for the double distilled solution. For the Aqua-Rx™ solution, the amplitude was measured at 102% after 1 hour and 94.8% after 2 hours. The study was stopped after 2 hours on the Aqua-Rx™ solution, as the test unit was out of solution. How long the rat brain tissue would have continued to be viable will be the subject of future studies.

 

Culturing Viability of Primary T Cell Long Term Using DMEM Culture Medium

This study compared the long-term viability of human Primary T cells prepared in cell culture media using soluble DMEM powder culture medium from Life Technologies, using Aqua-Rx™ and Standard Lab water to compare long-term cell viability over a 24-day time period. Results: After long-term culturing ex-vivo (24 days), cells cultured in the Aqua-Rx™ solution demonstrated 87.9% viability compared to 57.1% for primary T cells in the Standard Lab Water solution.

Application: Aqua-Rx™ improves viability of primary T cells during long-term culturing ex-vivo. Currently, primary T cells exposed to professional antigen presenting cells (APC), specifically, dendritic cells are under investigation for adoptive immunotherapy of cancer (Rosenberg et al. 2002). Ex vivo culturing is a crucial step in protocols designed to obtain preparative quantities of antigen-primed T cells applicable for re-infusion therapy. Improving conditions of culturing indicates that Aqua-Rx™ could substantially increase the possibility of success of re-infusion therapies.

 

Gas Solubility Study

Aqua-Rx™ exhibits an increased carbon dioxide (CO2) transport capacity. The test involved titration of 100 ml of Aqua-Rx™ and 100 ml of Milli-Q water. Both were mixed with 20 mM of NaHCO3 to a pH of approximately 8.57 to 8.60. Then the solutions were titrated using a 0.1 HCL calibrated solution in 20 uL units allowing 24 hours for the pH to reach a stable equilibrium. The pH results were recorded until a pH of 8.91 was registered on the Milli-Q water at which time the Aqua-Rx™ water read a pH of 9.28. This .37 higher pH reading demonstrates an increased carbon dioxide exchange capacity of the Aqua-Rx™ water as compared to the lab standard Milli-Q water.

Aqua-Rx™’s ability to absorb and exchange more CO2 gas than normal water has major relevance for biological systems. The ability of Aqua-Rx™ to increase CO2 exchange may increase the storage time of donor organs. Typically, the excess CO2 in the cells leads to an acidic shift in the pH that can cause problems for cell function and decrease organ preservation time.

 

Aqua-Rx™ Water Comparison to Reverse Osmosis Deionized Water (RODI)

The purpose of this research was to evaluate the effectiveness of Aqua-Rx™ Water for formulating cosmetics verses regular reverse osmosis deionized water (RODI). Identical oil in water emulsions were prepared, one with RODI Water and another with Aqua-Rx™ Water. All mixing and batching procedures were identical for both preparations. The final products were compared in three separate studies as follows for emulsion stability, ability to increase skin moisturization and penetration enhancement of an ancillary ingredient.

 

Product Stability Testing

A simple oil/water emulsion was made with Aqua-Rx™ Water verses RODI Water with a 50% decrease in levels of emulsifying agents. Emulsion stability was graded by microscopic examination and short term (4weeks) accelerated aging at room temperature (RT), 40C and 50C. Initial microscopic evaluation of the samples showed uniform dispersion of oil droplets and visually appeared as a very thin white stable emulsion with a viscosity of 600 cPs. Over the time period tested, the sample made with RODI Water fell apart after 5 days at 50C, 14 days at 40C and maintained continuity after 28 days at room temperature. Conversely, the Aqua-Rx™ Water showed slight instability after 10 days at 50C and was 100% stable at 40C and room temperature after 28 days. Based on the data obtained, it appears that the Aqua-Rx™ Water allows for better product stability when used in a conventional cosmetic lotion.

 

Moisturization Testing

Two oil/water emulsions, one made with Aqua-Rx™ Water and the other RODI Water, were applied to the forearms of 5 subjects. Moisture content of the skin was measured via electrical conductance with a Novameter at baseline, 15 minute, 30 minutes, 1 hour, 2 hour, and 4 hours post application. During the course of testing the sample made with Aqua-Rx™ Water consistently demonstrated higher conductance readings at the time points measured indicating increase hydration over the sample made with RODI Water.

 

Penetration

A basic emulsion, containing 10% lidocaine, was made with Aqua-Rx™ Water verses RODI Water. Lidocaine numbs the skin in a penetration/time dependant manner that can be monitored via a pin prick to the test site. Therefore, enhanced penetration can be measured by decreased time for onset of numbness. A 5cm x 5 cm test site was applied with the samples on the forearms of 5 subjects and allowed to dry for 15 minutes. Subjects were asked to evaluate numbing, via rating pain sensation associated with a pin prick, on a 0 to 10 scale (10 being no numbing and 0 being 100% numbing) at baseline, 15 minute, 30 minutes, 1 hour, 2 hour, and 4 hours post application. Aqua-Rx™ Water demonstrated a higher degree of numbing within 15 minutes of applications when compared to the 10% Lidocaine Cream made with the RODI Water. Based on data obtained Aqua-Rx™ Water can enhance skin penetration of an ancillary ingredient.

Nanosizing

As the old saying goes, oil and water don’t mix. In chemistry handbooks, pure olive oil is listed as 0.00% soluble in water. However, using API’s patented processing device, the Company has processed Aqua-Rx™ water and oil into solution with no other ingredients. A nanosized solution of .42% by weight of olive oil in Aqua-Rx™ has average particles that are less than 150 nanometers. The solution is stable with no creaming or settling out over a year period at 50 C. The Company has also nanosized other substances in Aqua-Rx™ including cranberry seed oil, zinc oxide, and titanium dioxide.