MRET Water Activator

Water is the foundation of all life on earth. Every part of the body is dependent on water. The functions of our glands and organs will eventually deteriorate if they are not nourished with good clean water. Water is an excellent solvent, and assimilation of vital nutrients and proteins occurs through absorption of water. Metabolism, digestion, blood flow and cellular reproduction depend on proper water supply.

If your body isn't properly hydrated, cells will draw water from your bloodstream, which causes your heart to work harder. Meanwhile, your kidneys can't properly purify your blood, and when that happens, some of the kidneys' workload is passed along to the liver and other organs, which can cause them to be severely stressed. Additionally, you can develop a number of minor health conditions such as constipation, dry and itchy skin, acne, nosebleeds, urinary tract infection, coughs, sneezing, sinus pressure, and headaches. Thus, clean water is essential to health. The water that we drink must be metabolically restructured before it is bio-available to our body. MRET Water Activator does it for us.

Molecular Resonance Effect Technology (MRET) is patented by Igor Smirnov, Ph.D., US Patent No.6022479 issued in 2002. During the process of activation, the subtle low frequency electromagnetic field is imprinted into the water. It closely resembles the natural geomagnetic field found near the healing water springs. MRET Activated Water has modified molecular structuring, physical and electrodynamic characteristics, and is compatible with the cell water dynamic multilayer molecular structuring described by Dr. Gilbert N. Ling. Such similarity contributes to easy bio-availability and fast assimilation of MRET water in biological systems. Due to its “super liquidity”, MRET water easily penetrates through small pores and capillaries in the body, improves hydration, and stimulates the proper function of cells in biological systems. MRET Water acts as a communication medium among the cells. It has many special physiological properties that make it ideal to help the body to restore its quota of properly structured bio-available water.

MRET Water supports the process of cellular regeneration by means of the hydration of cells with MRET Activated Water. It results in the proper function of cells and cellular structures, which leads to normalization of metabolism and general revitalization of the body. MRET Water prevents cellular dehydration, and slows down the process of aging closely related to the dehydration of cells. It improves the nutrient transport and assimilation, as well as the detoxification of the body. The research conducted on MRET Water confirmed its antibacterial effect, as well as its ability to substantially increase the main factors of natural cellular resistance to infections and inflammations such as phagocytosis and immune responses, to inhibit the growth of mutated cells, and to enhance the vitality of healthy cells.

MRET Activator is a household appliance, not a medical device. Any pure drinking water or other liquid food substances like milk, oil, juice or wine can be activated. You also can use MRET water for cooking and keep cooked food in the refrigerator, and to spray your skin and hair. The research on MRET water confirmed that it had the best properties beneficial for health right after the activation period, and kept a substantial part of the structure after the storage at room temperature for at least 24 hours, and refrigeration for 45 days.

MRET WATER ACTIVATOR

Today, everybody is aware of the importance of water for health. It is well known that the body of an average matured person consists of 70% of water (actually from 45% for old people up to 85% for babies), and the brain contains 90% liquid. The disputes regarding the optimal amount of water for daily consumption have ceased, and now most of the proponents refer to 1.5-2 liters a day. The question regarding what kind of water is good for health is still under discussion. There are a lot of devices on the market which are claimed as producing the water beneficial for health. How to make a right choice?

The beneficial, rejuvenating and healing properties of water from several water springs have been known for centuries. In the last decades, serious scientific studies were conducted on biochemical, physical and microbiological properties of water. The short overview presented below will help to give answers to the questions that worry a lot of people:

• What properties of water make the water beneficial for health?
• Why does the water from certain natural water springs have beneficial, healing and rejuvenating properties?
• Is it possible to have a source of such water at home?

Speaking about healing water, people usually talk about such properties as the content of minerals, Redox potential (reduction-oxidation reaction), balanced pH index, and in more recent years about the structure of water. The first three parameters are well known, and rather easy to measure in appropriate laboratories. The application of the term “structured water” and the description of properties of such water require serious scientific investigation.

By definition, structured water is the water in which part of molecules are stuck together forming repeating geometrical structures or clusters, and other free water molecules move around these structures. The more molecules of water form structures, and the longer they do not break up, the more effective is the process of water structuring. For example, ice water remains structured only within several minutes of defrosting. The water inside and around the cells in biological systems has specific dynamic, multilayer structuring, as was established in the last decades by world renowned microbiologists Dr. Gilbert Ling and Dr. Drost-Hansen. Only such structured water can effectively influence all life supporting processes, both inside and outside the cells. For example, it has a unique capacity to create a protective water "environment" around the big biological molecules and around the cells, thus, for instance, preventing the red blood cells from sticking together. Only the water with the structure and properties similar to the structured water in our organism is beneficial for the body, and can produce healing effects.

During the process of the structuring of regular drinking water, our organism spends a lot of energy. In the modern world, we are subjected to numerous stresses, and as a result, our organism suffers from a lack of energy. Therefore, the body cannot always produce the necessary amount of structured water. In turn, a lack of structured water leads to the distortion of the work of all systems in the body, and results in an even greater shortage of energy.

How MRET Activator works

The MRET water activator changes the bond angle of H2O from 104.5° to 114.5° by utilizing a patented technology. The bond angle of 114.5° is found in intracellular water molecules. If we can make this change to the water that we drink even before we consume it, hydration becomes more efficient.

MRET WATER ACTIVATING TECHNOLOGY

can help to break this vicious cycle.

MRET water activator structures the water before you drink it. Thus, you get a unique source of water with these beneficial, healing and rejuvenating properties at home. During the 30 minutes, MRET activator will produce for you and your family 2 liters of structured water with beneficial properties, which were extensively studied and confirmed by numerous biological and physical experiments at Certified Research Institutions and Universities. MRET WATER ACTIVATOR is a household appliance, not a medical device.

MOLECULAR RESONANCE EFFECT TECHNOLOGY (MRET)

In February of 2000, Igor Smirnov, Ph. D. was granted the U.S. Patent N 6022479 for the invention of Molecular Resonance Effect Technology and the development of a device for the activation of water and liquid substances based on this technology.

The process of MRET activation changes the structure of water and other liquids creating the long-range dynamic multilayer water molecular structures similar to the structures of cell water in biological systems. During the process of activation the water is subjected to the subtle low-frequency electromagnetic field comparable with geomagnetic field of the Earth in the areas around healing water springs. This spontaneous electromagnetic field is generated by unique MRET polymer compound with fractal structure which has properties compatible with the properties of the cellular fractal structures in living tissues. Therefore, such field can enter into a resonance with the electromagnetic signals of the cells in living organisms. The field generated by MRET polymer affects the water in the same way as the signals generated by the fractal structures of living tissues affect the cell water. As a result they structure MRET water similar to the cell water structuring. It is biologically safe process because MRET water is produced with the help of non-contact process from drinking water and remains chemically pure and non toxic. It can keep its structural basis and significant part of beneficial properties for a long time (not less than 24 hours at a room temperature and not less than 45 days refrigerated at a temperature of +4 degrees of Celsius).

Microbiological and physical properties of MRET activated water were studied in leading Research Institutions and Universities, presented at numerous International Scientific Conferences and published in scientific books and magazines.*

The structural changes in the water after MRET activation were confirmed by Nuclear Magnetic Resonance and High Voltage Photography testing. The researches of anomalous viscosity and electrodynamic characteristics of MRET water confirmed the long-range dynamic multilayer structuring of MRET water similar to cell water. Bioelectrical Impedance analysis showed the biological compatibility of MRET water with cell water as well as significantly enhanced hydration capacities of water after the process of MRET activation and its easy absorption by the tissues of the body.

The incredible health benefits of MRET water were demonstrated in the process of several biological investigations such as Oncology Research on 500 mice, Staphylococcal Infection Research on 400 mice, numerous experiments on Antibacterial Sterilizing Effects of MRET water and its Stimulating Effect on Metabolic (biological) Activity of Complex Microbiological Associations similar to microbiological flora in the intestine, Inhibition of Growth of Mutated Cells, Enhanced Morphology of Red Blood Cells and significant Rebounding Effect on White Blood Cells.**

As a summary it is possible to say that based on the research results the consumption of MRET water leads to the general revitalization of the body: it enhances the natural immune response including the main factors of natural cellular resistance to infections and inflammations such as phagocytosys (engulfing of alien cells and resistance of the body to the penetration and reproduction of bacteria, viruses and other pathogenic microorganisms).

Unique properties and benefits of MRET activated water based on research results in animal models and in vitro

• MRET water has a unique long-range dynamic structuring similar to cell water. It is compatible with cell water and as a result it has the enhanced hydration capacities and it is easy absorbed by the tissues of the body. MRET water penetrates into the cells and eliminates from the cells about three times faster than regular water.
• Improves nutrient transport and assimilation due to the fact of faster hydration.
• Accelerates and improves the processes of extermination of waste products out of the body and the detoxification of the body due to the enhanced cellular hydration.
• The intensified hydration with the structured MRET water leads to the improvement of metabolism and of the process of regeneration of healthy cells. It also helps to slow down the process of ageing because this process is closely related to the cells dehydration. Therefore, the application of MRET water in cosmetic purposes improves the skin, strengthens and makes shine the hair.
• MRET water has antibacterial effect. It inhibits the growth of bacteria and mutated cells and supports the life activity of normal healthy cells.
• Aids in promoting healthy intestinal flora.
• Enhances the natural immune response, including the factors of natural resistance of the body against the penetration and reproduction of bacteria, viruses and other pathogenic microorganisms, such as phagocytosis, thus strengthening the resistance of the body to infections and inflammations.
• MRET water enhances the morphology of red blood cells and improves the process of rebounding of white blood cells.

The SYSTEM of MRET WATER ACTIVATION is successfully applied in the USA, Canada, in a number of the European countries, Singapore, Malaysia, Thailand, China, India, Australia and Southern Africa.

MRET WATER ACTIVATOR can be used in a mode "WORK" for the total of 7,200 hours or for 14 400 half-hour operation cycles. It makes 2 years in case of 10 hours average daily usage (20 cycles or 40 liters), 4 years – in case of 5 hours average daily usage (10 cycles or 20 liters), most common 5 years – in case of 4 hours average daily usage (8 cycles or 16 liters), etc. After the specified term the activator has to be replaced with the new one.

You can drink MRET water following the principle "Listen to your body." At the initial stage during the period of extraction of the toxins or in case of health problems it is recommended to begin the consumption of water starting with 0.5 glasses a day, gradually increasing the dosage up to 3 glasses a day. After the adaptation of the body to MRET water it is possible to continue to drink it as much as you like and also to prepare tea, coffee, soups and other liquid products on MRET water. Your pets will enjoy drinking MRET water. You can use it to water plants and flowers.

MAINTENANCE and INSTRUCTIONS

MRET WATER ACTIVATOR can be used in a mode "WORK" for the total of 7,200 hours or for 14,400 half-hour operation cycles. It makes 2 years in case of 10 hours average daily usage (20 cycles or 40 liters), 4 years – in case of 5 hours average daily usage (10 cycles or 20 liters), most common 5 years – in case of 4 hours average daily usage (8 cycles or 16 liters), etc. After the specified term the activator has to be replaced with the new one.

Pour the drinking water (milk, juice or other liquid products) in the pitcher. Install the activator on the top of the pitcher thus that the bottom part of the activator does not touch the liquid (2-5 centimeters above the liquid). Plug the adaptor in power lines. The blinking red light shows that MRET activator works. The process of optimal activation takes 30 minutes. Then switch off the adapter from power lines. The water (milk, juice, etc.) is ready for consumption. It is also possible to place the activator on any glasses or jars with up to 2 liters volume.

* MRET water studies at Research Institutions and Universities:

• AltheaDx technology, San Diego, USA
• Engene Biotech, Inc., Rancho Santa Fe, USA
• Nu Mega Resonance Lab, San Diego, USA
• USACAI Environmental Lab, Carlsbad, USA
• Meixa Tech Laboratory, Cardiff, USA
• Geotechnical & Environmental Sciences Lab, San Diego, USA
• Elixir-Health Ltd., Singapore
• Kyiv National Shevchenko University, Ukraine
• Kyiv Institute of Experimental Pathology, Oncology and Radiobiology of Ukrainian Academy of Science
• Kyiv Institute of Microbiology and Virology of Ukrainian Academy of Science
• Kyiv Institute of Plants Genetics of Ukrainian Academy of Sciences
• Moscow State University, Russia

BIOLOGICAL TESTS RESULTS

MRET ACTIVATED WATER AND ITS SUCCESFUL APPLICATION FOR PREVENTIVE TREATMENT AND ENHANCED TUMOR RESISTANCE IN ONCOLOGY

The goal of this investigation was to study on mice the effect of MRET activated water as a potential agent for the prevention and treatment of two kinds of oncology diseases (laboratory models of Ehrlich's ascites tumor and Sarcoma ascites form). Significant positive effect of MRET activated water regarding the tumor resistance on animals was observed in the experiments conducted on 500 mice (22 groups with 20 mice in each group and 10 groups with 5 mice in each group). The best results were observed in the groups of mice on MRET water activated for 30 minutes (optimal regime of activation). The substantial anti-tumor efficacy was confirmed by very high level of reduction of Total Number of Viable Tumor Cells by 76% in “preventive treatment” and by 55% in “therapeutic treatment” regime. The life span of mice which received optimal activated water increased by 61% in “preventive treatment” and by 43% in “therapeutic treatment” regime. The significant anti-tumor effect of MRET Activated Water on mice was close to the action of the chemotherapy agents and allowed to avoid the side effects that typically follow chemotherapy treatment in oncology.

Fig 1: The effect of preventive (1–5) and therapeutic (6–10) application of MRET activated water on average total number of viable cells in an ascetic tumor, obtained from mice inoculated intraperitoneally with tumor cells of Ehrlich carcinoma.

Fig 2: The change of the percentage increase of life span of tumor-bearing mice with ascitic Ehrlich carcinoma which received different types of MRET activated water in “preventive treatment” and “therapeutic treatment” regimes. The digits under the charts correspond with the duration of water activation in minutes.

Fig 3: The effect of MRET activated water on cytotoxic activity of lymphocytes containing NK-cells. Activated water was applied for mice without tumors in two regimes (for 21 and 14 days), called “preventive” (1–5) and “therapeutic” (6–10).

Pic: The appearance of mice from “control” (A) and “preventive treatment” groups (optimal activation time 30 minutes) (B) on the 18th day after ascitic Ehrlich carcinoma cell inoculation.

The results of investigation of the application of water activated by non-ionizing Molecular Resonance Effect Technology (MRET) process for prevention treatment and enhancement of tumor resistance of animals to two types of oncology diseases in vivo on 500 mice are presented. The research conducted on physical parameters of water confirmed that MRET activation process contributed to substantial modification of the basic physical-molecular properties of distilled water (substantial reduction of viscosity as a function of applied tangent pressure, as well as substantial decrease of electrical conductivity and dielectric permittivity as functions of the frequencies of applied electromagnetic field). The significant positive effect of MRET activated water on tumor resistance of animals was observed in the process of this investigation in all groups of mice on different fractions of activated water. The best results were observed in the groups of mice on MRET water activated for 30 minutes (optimal regime of activation). The results were better in “preventive treatment” regime compare to “therapeutic treatment” regime. Additionally, this investigation confirmed that the long-term preservation of activated water at low temperature (around 0°C) for 45 days decreased its anti-tumor efficacy but left it on the significantly high level compare to other fractions. The test results point at the dual mechanism of MRET water effect on tumors: the prevention and inhibition of tumor growth together with the reduction of quantity of viable tumor cells. The significant anti-tumor effect of MRET activated distilled water on mice was close to the action of the chemotherapy agents and allowed to avoid the side effects that typically follow chemotherapy treatment of oncology.

In the process of investigation of cytotoxic activity of NK-cells the significant increase of lymphocyte cytotoxicity levels was observed when donor mice were treated with MRET water activated for 30 minutes. The results also showed that the extension of the application of MRET water from 14 days to 21 days significantly increased the value of cytotoxicity index. It is possible to admit that the extension of time of application of MRET water will lead to higher levels of enhancement in NK-cells activity. Thus, the application of MRET activated water can be quite promising approach for non-drug stimulation of NK-cells immunization vaccines.

The detailed description of this investigation is presented in “BENEFITS MRET WATER”.

THE EFFECT OF MRET ACTIVATED MEDIUM ON CELL MORPHOLOGY

The in vitro investigations on normal PMBC cells (peripheral blood mononuclear cell) and on HeLa cancer cells (cell line ATCC # CCL-2 cervical adenocarcinoma) and were conducted under the supervision of Patrick Pezzoli, Ph.D. at AltheaDx Technologies, San Diego, USA. The experiments analyzed: cells lysed at 0 hour, cells cultured for 24 hours in untreated medium and cells cultured for 24 hours in medium treated with MRET activator for 30 minutes. DNA samples from each batch were processed and the resultant data was analyzed using Affymetrix Genotyping Console 3.0 to obtain genotype calls and copy number calls. Cell counts and % viability were obtained using the Trypan Blue exclusion technique.

The experimental data revealed no difference between the zero hour (control), MRET treated and untreated samples in term of genotypes and copy number calls. Thus, MRET activation of water based medium did not induce any changes in cells on genetic level.

The studies showed that in MRET activated medium the viability of normal cells (PBMC) was higher (Table 1), and the viability of cancer cells (HeLa) was lower (Table 2) compared to their viability in untreated medium.

Table 1: PBMC cell counts and % viability

Table 2: HeLa cell counts and % viability

For normal cells (PBMC) the changes in cell counts were similar for untreated and MRET treated medium (Fig 1). Thus, MRET treatment did not affect the growth of normal cells.

For cancer cells (HeLa) the experimental data revealed significant inhibition of cancer cells growth in MRET treated medium. The growth of viable cancer cells was inhibited by 54% in MRET treated medium compared to untreated medium (Fig 2).

Fig 1: Viable PBMC cell counts after 24 hours of incubation	Fig 2: Viable HeLa cell counts after 24 hours of incubation

The results of AltheaDx research on HeLa cancer cells in vitro support the results obtained earlier in the investigation regarding the effects of MRET water on tumor resistance in animal model. The study on 500 mice was conducted under the supervision of Professor V. Vysotskii, S. Olishevsky, Ph.D. and Y. Yanish, Ph.D. at Kyiv Institute of Experimental Pathology, Oncology and Radiobiology of Ukrainian Academy of Science. It showed substantial inhibition of growth of viable tumor cells following the consumption of MRET water. In the course of this investigation the groups of mice in “preventive regime” ingested MRET water for 2 weeks prior to the inoculation of Ehrlich carcinoma cancer cells and for 3 weeks after the inoculation. The groups of mice in “therapeutic regime” ingested MRET water only during 3 weeks after the inoculation of Ehrlich carcinoma cancer cells. Following the consumption of MRET water activated for 30 minutes (the optimal time of activation) the growth of viable tumor cells was inhibited by 76% in “preventive regime” and by 55% in “therapeutic regime.”

As a conclusion it is possible to say that the studies conducted at AltheaDx Technology confirmed that MRET activated water did not affect the cells on genetic level; it affected the morphology of normal cells in positive way increasing their viability and promoted significant inhibition of growth of cancer cells.

MRET Rebounding Effect on White Blood Cell Counts

A clinical observation was conducted on a patient undergoing chemotherapy treatment for metastasized naso-pharyngeal cancer. On a regular basis, WBC counts usually decrease to very low level. The rebounding of WBC from this low range takes about 3-5 weeks. The ingestion of MRET water prevented the decrease of WBC counts to their lowest level and helped to regain the pre-chemotherapy level within unusually short period of time of 3 days.

Thus, the clinical observation revealed that on the 10th day after the beginning of chemotherapy (in 3 days after WBC counts dropped to 10% of pre-chemotherapy level) WBC counts rebounded to 93% of their pre-chemotherapy level. In compliance with Student’s Distribution criteria the rebounding of WBC was higher than 70% of pre-chemotherapy level with p=0.05.

ANTIBACTERIAL EFFECT OF MRET WATER

Testing conducted at C.A.I. Environmental Laboratory, Carlsbad, USA revealed the significant reduction of the amount of total coliforms following the process of MRET activation. In the rainwater activated for 30 minutes the amount of total coliforms decreased by 86% after the process of water activation. This test confirms that MRET water acts as antibacterial agent, providing sterilization effect.

The effect of MRET activated water on Escherichia coli K-12 and on Complex Microbiological Associations

The research was conducted at Kiev Institute of Microbiology and Virology of Ukrainian Academy of Science.

I. MRET activated water inhibitory effect on growth of conditionally pathogenic bacteria Escherichia coli K-12 in aerobic environment

The investigation revealed the significant effect of MRET-activated nutrient medium in aerobic environment on the process of growth and reproduction of E.coli microorganisms, their division, the size of colonies and the modification of forms of culture cells. It was observed that at low initial concentration of cells of investigated culture Escherichia coli K-12 MRET nutrient medium activated during 30 minutes and 60 minutes inhibited the growth of culture 27 and 303 times respectively during the 25 hours of experiment.

Initial view of Petri dishes with different fractions of nutrient medium at the beginning of experiment is presented on Fig 1.

Fig 1: Petri dishes at the beginning of experiment. Identical very small amount of Escherichia coli K-12 cells was introduced on a surface of non-activated nutrient medium of control dishes and on a surface of dishes with nutrient medium MRET-activated for 30 and 60 minutes (tact=0.5h and tact=1.0h) in aerobic environment. There are no colonies of microorganisms in Petri dishes in the beginning of experiment.

Petri dishes with grown colonies and statistical parameters of the colonies after 23 hours of experiments are presented on Fig 2: (a) non-activated nutrient medium (control); (b) nutrient medium activated for 30 minutes; (c) nutrient medium activated for 60 minutes. Petri dishes after 29 hours of experiment are shown on Fig 3. The significant inhibition of growth of E.coli in activated samples was revealed and it confirmed the strong bacteriostatic effect of MRET-activated medium.

(a) Control: Number of colonies NC = 1.7x108 CFU/ml. Average diameter of grown colonies d = 1.1 mm.	(b) MRET-activated, tact = 30 minutes: Number of colonies N0.5 = 6.4x106 CFU/ml. Average diameter of grown colonies d = 1.8 mm.
(c) MRET-activated, tact = 60 minutes: Number of colonies N1.0 = 5.2x105 CFU/ml. Average diameter of grown colonies is d = 1.5 mm.	Fig 2: Petri dishes after 23 hours of experiment.

Fig 3: Selected Petri dishes with grown colonies of E.coli K-12 after 29 hours of experiment.

This experiment shows that MRET-activation process has very strong bacteriostatic effect on conditionally pathogenic E.coli microorganisms and that the inhibition of E.coli growth is more effective when activation time is increased. It was observed that at low initial concentration of cells of E.coli in nutrient medium MRET-activation during 30 minutes and 60 minutes period of time inhibited the culture growth N_C/N_0.5 = 27 and N_C/N_1.0 = 303 times respectively after 25 hours of experiment (Fig 4). Consequently, the level of bacteriostatic activity was 96% in 30 minutes activated nutrient medium and 99.7% in 60 minutes activated medium. Thus, the direct correlation between bacteriostatic activity of MRET-activated nutrient medium and the time of activation was confirmed.

Inhibition of E.coli bacteria growth in aerobic environment

Fig 4: The inhibition of E.coli growth in aerobic environment: 303 times (t_act=1.0 hour) and 27 times (t_act=0.5hour) after 25 hours of experiment (blue color - control, red - 30 minutes and green - 60 minutes MRET-activated medium).

This experiment also revealed the strong effect of MRET-activated water on the process of division of E.coli microorganisms, the modification of forms of culture cells and the size of colonies. It was observed that one of the reasons of abnormally low growth of E.coli bacteria was related to the modification of the process of cell division in MRET-activated nutrient medium. In the process of growth and reproduction a large number of cells did not separate from each other and the linear line-ups consisting of 2-3 sequentially paired cells were formed. The culture cells grown in non-activated and MRET activated medium are shown on Fig 5 and Fig 6 respectively.

Fig 5: Cells of Escherichia coli K-12 grown in non-activated medium.	Fig 6: Cells of Escherichia coli K-12 grown in MRET-activated medium (tact =1 hour).

II. The effect of MRET water on metabolic activity of E.coli bacteria in aerobic and anaerobic environment

Reductant activity is an integral characteristic of metabolic activity of microorganisms and it is measured with the help of Sodium Resasurine color indicator in the percentage degree of discoloration (purple = 0%, red = 50%, transparent = 100%). The reductant activity of E.coli bacteria reduced up to 3 times in 30 minutes MRET-activated water and up to 1.6 times in 60 minutes activated water during the first 6 hours of experiment in aerobic environment (Fig 7 and 8).

K – Control; 0.5 – 0.5 hour MRET-activated water; 1.0 – 1 hour MRET-activated water
RC = 0.2, R0.5 = 0.1, R1.0 = 0.4 K0.5 = 0.5, K1.0 = 2.0 DC = D0.5 = D1.0	RC = 25, R0.5 = 9, R1.0 = 19 K0.5 = 0.36, K1.0 = 0.76 DC = D0.5 = D1.0

Fig 7: Comparative test on metabolic (reductant) activity of E.coli (control samples, 30 and 60 minutes MRET- activated water) in aerobic environment: R - reductant activity (in Control, 0.5 hour and 1.0 hour MRET-activated water), K - relative reductant activity, D - optical density.

Relative Reductant Activity of E.coli bacteria in aerobic environment

Fig 8: Relative Reductant Activity of E.coli in MRET water activated for 0.5 hour (K_0.5=R_0.5/R_C - red color) and for 1.0 hour (K_1.0=R_1.0/R_C - green color) compare to control non-activated samples (K_C=1 - blue color) in aerobic environment.

This experiment showed that there was no direct correlation between the inhibition of metabolic (reductant) activity and the inhibition of growth of E.coli (bacteriostatic activity) in MRET activated water. The bacteriostatic effect is substantially higher in 60 minutes activated water and the inhibition of reductant activity during the first 3 hours is higher in 30 minutes activated water. Thus, this experiment revealed that the optimum time of activation for the maximum inhibition of metabolic activity of E.coli bacteria in aerobic environment is 30 minutes. The same optimum time of activation was found in the process of another investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in vivo on 500 mice for two types of cancer conducted at Kiev Institute of Experimental Pathology, Oncology and Radiobiology of Ukrainian Academy of Science.

Taking in consideration that a small population of pathogenic bacteria, such as E.coli, is usually presented in complex microbial associations in the intestine of the body, the test on metabolic activity of E.coli bacteria in anaerobic environment was conducted. Anaerobic environment simulates the environmental conditions similar to the conditions in the intestine of humans and animals. The investigation showed that the reductant activity of E.coli bacteria in anaerobic environment practically did not change (Fig 9 and 10).

K – Control; 0.5 – 0.5 hour MRET-activated water; 1.0 – 1 hour MRET-activated water; K_STER – reference to sterility

Fig 9: Comparative test on metabolic (reductant) activity of E.coli (control samples, 0.5 hour and 1.0 hour MRET- activated water, K_STER - reference to sterility) in anaerobic environment: R - reductant activity (in Control, 0.5 hour and 1.0 hour MRET-activated water), K - relative reductant activity, D - optical density, V - gas volume in the bottles.

Reductant Activity of E.coli bacteria in anaerobic environment

Fig 10: Reductant Activity of E.coli in 30 and 60 minutes MRET-activated water and in Control non-activated samples (R_0.5 - red color, R_1.0 - green color, R_C - blue color) in anaerobic environment.

This experiment revealed that the process of MRET activation did not have any significant effect on reductant activity of E.coli bacteria in anaerobic environment.

III. The stimulating effect of MRET water on metabolic activity of Complex Microbial Associations in anaerobic environment

In order to simulate the environmental conditions similar to the conditions in the intestine of humans and animals the test on metabolic activity of microbial associations was conducted in anaerobic environment. It was found that MRET-activated water substantially increased reductant activity of complex microbial associations during the first several hours of experiment (Fig 11).

1.0 – 1 hour MRET-activated water; 0.5 – 0.5 hour MRET-activated water; K – Control

Fig 11: Comparative test on metabolic (reductant) activity of microbial associations (1.0 hour and 0.5 hour MRET- activated water and Control samples) in anaerobic environment: R - reductant activity (in Control, 0.5 hour and 1.0 hour MRET-activated water), K - relative reductant activity, V - gas volume in the bottles.

This experiment revealed that the optimum time of activation for the maximum increase of metabolic activity of microbial associations in anaerobic environment was 30 minutes. The same optimum time of activation was found in the process of inhibition of metabolic activity of E.coli in aerobic environment and in another investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in vivo.

CONCLUSIONS

This investigation revealed that at low initial concentration of cells of conditionally pathogenic microbiological culture Escherichia coli K-12 in water based nutrient medium activated for 30 minutes and 60 minutes the growth of culture was inhibited 27 and 303 times respectively after the 25 hours of experiment in aerobic environment. This experiment also revealed the strong effect of MRET activated water on the process of division of E.coli microorganisms, the modification of forms of culture cells and the size of colonies. It was observed that one of the reasons of abnormally low growth of E.coli population was related to the modification of the process of cell division in MRET-activated nutrient medium. These results allow admitting that the process of MRET activation and the sterilization effect of MRET water can be applied in food industry and for water purification.

The second stage of investigation revealed that the metabolic (reductant) activity of E.coli bacteria reduced up to 3 times in 30 minutes activated water and up to 1.6 times in 60 minutes activated water during the first 6 hours of experiment in aerobic environment. Another experiment showed that the process of MRET-activation did not affect the reductant activity of E.coli bacteria in anaerobic environment and, consequently, should not affect a small population of conditionally pathogenic bacteria, such as E.coli, usually presented in microbial associations in the intestine of the body.

In order to simulate the environmental conditions similar to the conditions in the intestine of humans and animals the test on metabolic activity of complex microbial associations was conducted in anaerobic environment. It was discovered that MRET activated process substantially increased reductant activity of complex microbial associations during the first hours of experiment. The same 30 minutes optimum time of activation was observed in the process of inhibition of metabolic activity of conditionally pathogenic E.coli bacteria in aerobic environment and for the maximum increase of metabolic activity of complex microbial associations in anaerobic environment (presented in the intestine). The previous investigation regarding the application of MRET activated water for preventive treatment and enhancement of tumor resistance in oncology in vivo on 500 mice also showed the best results on 30 minutes MRET-activated water. Thus, this investigation shows that the ingestion of MRET water is beneficial for the process of digestion and can enhance metabolism of the body.

The Effect of MRET Activated Water on Staphylococcal Infection in vivo in Animal Model (on the cells of immune system) and in vitro on the Culture of Staphylococcus aureus Wood-46

I. The Effect of MRET Water on Staphylococcal Infection in vivo in Animal Model

The research was conducted at Kiyv National Shevchenko University, Ukraine on 400 mice. The mice-male of line BALB in the age of 11 – 13 weeks and of the weight 18 – 21 grams were used. In the process of investigation all mice were divided in three groups. Prior to the inoculation of Staphylococcus aureus Wood-46 culture one group of mice consumed MRET water during 4 weeks (Group 1), another group consumed MRET water during 2 weeks (Group 2), the control group consumed non-activated ordinary distilled water. During the following 2 weeks of experiment the first two groups continued to consume MRET water and the control group consumed ordinary distilled water. The first line of experiments was conducted on 225 mice in order to analyze the persistence of pathogen in homogenate of kidneys of mice comparing 5 groups of mice (two Group 1 and two Group 2 on 15 minutes and 30 minutes MRET activated water and Control). After preliminary experiments the optimal 30 minutes MRET activated water (distilled) was chosen for the main line of investigation.

RESULTS:

1.  Significant protective properties of MRET water.

The significant protective properties of MRET water were confirmed by substantial decrease of Staphylococcus CFU (colony forming units) in homogenate of kidneys of mice on MRET water compare to control group of mice following the intra-peritoneal staphylococcal infection after the first 24 hours. The analysis of data in the beginning of experiments leads to the conclusion that significant decrease of pathogen’s colonies in homogenate of kidneys of mice on MRET water begins only after 24 hours following the inoculation of Staphylococcus culture. The results on 30 minutes activated water were much better than on 15 minutes activated water and all further experiments were conducted on 30 minute activated water.

2. The consumption of MRET water eliminated the death rate from 30% (control group) to 0% (MRET group) during the first 9 days of experiment.

There was no case of animal death in all investigated groups within the first 24 hours after intra-peritoneal inoculation of Staphylococcus culture, which is a pretty standard result. During the next 8 days 30% of animals died in control group which also is a standard result. But there were no death cases in both groups of mice that ingested MRET activated water and it is a remarkable result. Nevertheless, the main consequences of Staphylococcus infection do not manifest in death of animals as in case of oncology diseases. Staphylococcus virus affects the live systems and organs of the body. These pathogenic microorganisms cause inflammations, suppurations, abscesses, furuncles, quinsy, cepsical conditions, etc. That’s why a detailed investigation of the process of stimulation by MRET water of phagocytes and of lymphoid organs of immune system of mice infected with Staphylococcus aureus culture was conducted and is presented in this report.

3. The development of the local acute inflammation is essentially suppressed in case of ingestion of MRET activated water.

The local inflammation was induced with the help of the inoculation of Staphylococcus aureus culture into the hinder left paw. The ordinary inflammatory reaction was observed in the group of mice on non-activated water: the intensive reddening of the hinder left paw (Fig 1). Both groups of mice on MRET water did not develop any reddening the hinder left paw inoculated with Staphylococcus aureus culture (Fig 2). The results of this experiment confirm the fact of the substantial inhibition of inflammatory infection in case of the regular consumption of MRET water.

Fig 1: The view of paws of a mouse on non-activated water (reddenning of the injection paw) in 24 hours after the injection of Staphylococcus culture.	Fig 2: The view of paws of a mouse on MRET activated water (no reddenning of the injection paw) in 24 hours after the injection of Staphylococcus culture.

4. The consumption of MRET water stimulates the activity of phagocytic system and the level of natural resistance of animals to pathogenic microorganisms.

For the following series of experiments the inoculation of Staphylococcus aureus Wood-46 was conducted intra-peritoneal in dose LD30 in order to spread infection all over the body.

The phagocytic system is one of the main factors of natural non-specific cellular resistance to infections and inflammations. It is the first line of protection of an organism against penetration and reproduction of pathogenic microorganisms. The protective role of phagocytic cells is based on their capacity to identify, engulf and utilize the alien agents penetrated into internal environment of a macro-organism. Phagocytosis is the main mechanism of natural resistance especially at the first stage of contagious process; it is a regular part of formation of the specific immune response.

The most common methodology applied in the studies of the functional activity of phagocytes is the examination of their phagocytic (engulfing of alien cells) and oxygen-dependent bactericidal activity. Phagocytic activity of neutrophils and macrophages is estimated based on Index of Phagocytosis (percentage of the phagocytes which engulfed test-bacteria) and on Phagocytic Number (average number of test-bacteria engulfed by one phagocyte). The cultures of Staphylococcus aureus and Latex are usually used as test-bacteria. The oxygen-dependent bactericidal activity of phagocytes is studied with the help of NBT-test: an oxygen-dependent reduction of Nitro Blue Tetrazolium into an insoluble Diformazan of Nitro Blue Tetrazolium derivative by phagocytes. With the help of NBT-test it is possible to distinguish the activated phagocytes from the non-activated ones.

MRET water stimulated the phagocytic capacities of neutrophils of a peripheral blood and peritoneal macrophages increasing their phagocytic activity, particularly Phagocytic Indexs (Fig 3A) and Phagocytic Number (Fig 3B). It also stimulated their oxygen-dependent bactericidal activity, particularly the increase of quantity of NBT-positive phagocytes (Fig 4).

Fig 3A: Phagocytic Index of neutrophils and macrophages in two weeks of experiment (object of phagocytosis - Staphylococcus aureus): 1 – Control group; 2 – Mice on MRET water (preventive for 4 weeks); 3 – Mice on MRET water (preventive for 2 weeks).

Fig 3B: Phagocytic Number of neutrophils and macrophages in two weeks of experiment (object of phagocytosis – Staphylococcus aureus): 1 – Control group; 2 – Mice on MRET water (preventive for 4 weeks); 3 – Mice on MRET water (preventive for 2 weeks).

Fig 4: Oxygen-depended bactericidal activity (NBT-test) of neutrophils and macrophages in two weeks of experiment:
1 – Control group; 2 – Mice on MRET water (preventive for 4 weeks); 3 – Mice on MRET water (preventive for 2 weeks).

These experiments confirmed the increase of effective potentials of phagocytes which constitute one of the main factors of natural protection of an organism and initiate the immune response.

The analysis of data in the beginning of experiments leads to the conclusion that significant intensification of phagocytic and bactericidal activity of macrophages and neutraphils of mice on MRET water begins only after 24 hours following the intra-peritoneal inoculation of Staphylococcus culture. At the end of two weeks of experiment the mean values of studied parameters in both groups of mice on MRET water substantially increased compare to the control group. The differences in mean values of the parameters of functional activity of phagocytes of groups of mice consuming MRET water compare to the control group of mice on non-activated water were statistically significant with p<0.05 (for Phagocytic Index and NBT-test). These results confirm the significant intensification of phagocytic and bactericidal activity and of immune system response following the consumption of MRET water.

The differences in mean values of studied parameters for the groups of mice on MRET water compare to each other were statistically insignificant, which confirms the similarity of the level of beneficial effect of MRET water in both groups. This fact also confirms that the regular consumption of MRET water provides health benefits in rather short time (2 weeks in case of the animal mice model).

5.   The consumption of MRET water substantially enhances the immune activity of lymphoid organs.

By the end of another series of experiments in both groups of mice on MRET water was observed substantial statistically significant (p<0.05) increase of the weight and the cellularity (quantity of cells) of a spleen and lymph nodes as well as the insignificant increase of weight and cellularity of thymus (Fig 5 and Fig 6).

Fig 5: The weight of lymphoid organs in two weeks of experiment: 1 – Control group; 2 – Mice on MRET water (preventive for 4 weeks); 3 – Mice on MRET water (preventive for 2 weeks).

Fig 6: The cellularity of lymphoid organs in two weeks of experiment: 1 – Control group; 2 – Mice on MRET water (preventive for 4 weeks); 3 – Mice on MRET water (preventive for 2 weeks).

These results confirm the fact of significant intensification of immune system response in animals on MRET water subject to Staphylococcus infection. The difference in studied parameters between the groups of mice on MRET water (4 weeks and 2 weeks of preventive consumption of MRET water) was insignificant which confirms quite fast beneficial effect of MRET water on the immune activity of lymphoid organs.

In the beginning of experiment the cellularity and the weight of lymphoid organs in MRET groups did not show the distinct tendency to modifications. It is reasonable to admit that the consumption of MRET water affects the weight and the cellularity of lymphoid organs only during the infection period.

CONCLUSIONS:

The consumption of MRET activated water significantly enhances the factors of natural resistance of the body which constitute the first line of protection of an organism against the penetration and reproduction of pathogenic microorganisms.

The analysis of data in the beginning of experiment leads to the conclusion that significant changes in all studied parameters of mice on MRET water (decrease of pathogen colonies in homogenate of kidneys, increase of the weight and the cellularity of lymphoid organs, intensification of the phagocytic and bactericidal activity of macrophages and neutraphils) begins only after 24 hours following the inoculation of Staphylococcus culture. Another words the consumption of MRET water increases the potentials of immune capacities of the body to counteract the infections without any changes in the vital parameters of immune organs and functions prior to the penetration of infectious pathogens in the body.

At the end of two weeks of experiment the mean values of studied parameters in both groups of mice on MRET water (preventive for 4 and 2 weeks respectively) significantly increased compare to the control group. The differences in mean values of the studied parameters of the groups of mice consuming MRET water compare to the control group of mice on non-activated water were statistically significant with p<0.05 (for most of the parameters). These results confirm the significant intensification of phagocytic activity and of immune system response following the consumption of MRET water.

The differences in mean values of studied parameters for the groups of mice on MRET water compare to each other were statistically insignificant, which confirms the similarity of the level of the beneficial effect of MRET water in both groups. This fact also confirms that the regular consumption of MRET water provides health benefits in rather short period of time (2 weeks in case of the animal mice model).

II. The Effect of MRET Activation on the Process of Growth of Staphylococcal Culture in Nutrient Medium

The following experiments were designed to study the effect of MRET activation on the process of growth and development of Staphylococcus aureus Wood-46 culture in vitro in nutrient medium (MPA – meat-peptone agar). The bacterial culture was grown overnight to stationary phase and then platted on MPA in the form of suspension at different inoculation densities. The MPA with culture was MRET activated during the different periods of time (activation for 15 minutes, 30 minutes, 45 minutes, and 60 minutes respectively) following the requirements of sterility. Petri dishes with activated and non-activated medium (MPA with culture) were covered with glass caps (aerobic environment) and placed in the thermostat for cultivation at a temperature of 37oC during 18-24 hours.

After the cultivation the morphological and tinctorial properties of cultures were observed and the numbers of colonies grown on MPA were counted. The bacteriostatic activity of MRET activated nutrient medium (MPA) was measured as an Index of Bacteriostatic Activity (IBA). An Index of Bacteriostatic Activity is defined as a coefficient of the inhibition of growth and reproduction of pathogens in bacteriostatic medium, particularly in MRET activated nutrient medium. It is calculated as reduction of the number of colonies (CFU – Colony Forming Units) in MRET activated medium related to the control samples not exposed to the activation:

IBA = (N_control – N_act)/N_control
where N – number of bacteria colonies (CFU) in Control (non-activated) and MRET activated nutrient medium, respectively.

In order to verify the sterility of experiments Petri dishes with nutrient medium (MPA) without staphylococcal culture were exposed to the process of activation and then were kept in the thermostat. No colonies of culture were observed that confirms the sterility of environment.

RESULTS:

Following the investigation the direct correlation between times of activation (tact), initial concentrations of staphylococcal culture (N₀) and a number of colonies grown on MRET activated medium were observed. The results are presented below in the form of a series of photos of Petri dishes with the colonies grown on MPA surfaces and the following diagrams based on the data of these experiments (Fig 7 – 12).

In the process of investigation the effect of MRET activation on the growth of staphylococcal culture at rather small initial concentration of pathogens was analyzed. The data corresponding to higher initial concentrations N0 > 10³ bacteria/ml were not analyzed due to the difficulties related to calculation of very high values of a number of colonies, despite the fact of the high bacteriostatic activity of MRET activated nutrient medium in case of high initial concentrations.

The highly significant bacteriostatic effect of 92 – 93% was observed after MRET activation for 30 minutes and more of cultures with initial concentration N₀ = 10³ bacteria/ml (Fig 7 and 8) and of 70 – 90% with initial concentration of N₀ = 10² bacteria/ml (Fig 9 and 10). In case of cultures with low initial concentration N₀ = 10 bacteria/ml the bacteriostatic activity in 15 minutes activated nutrient medium exceeded 93% and in 30 minutes activated nutrient medium was observed 100% inhibition of staphylococcal colonies (Fig 11 and 12).

N0 = 103 bacteria/ml, Control	N0 = 103 bacteria/ml, tact = 15 min
N0 = 103 bacteria/ml, tact = 30 min	N0 = 103 bacteria/ml, tact = 45 min
N0 = 103 bacteria/ml, tact = 60 min

Fig 7: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureusWood-46 with initial concentration of culture N₀ = 10³ bacteria/ml.

Fig 8: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureus Wood-46 with initial concentration of culture N₀ = 10³ bacteria/ml. IBA – Index of Bacteriostatic Activity (reduction of the number of colonies related to the control samples not exposed to activation).

N0 = 102 bacteria/ml, Control	N0 = 102 bacteria/ml, tact = 15 min
N0 = 102 bacteria/ml, tact = 30 min	N0 = 102 bacteria/ml, tact = 45 min
N0 = 102 bacteria/ml, tact = 60 min

Fig 9: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureus Wood-46 with initial concentration of culture N₀ = 10² bacteria/ml.

Fig 10: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureus Wood-46 with initial concentration of culture N₀ = 102 bacteria/ml. IBA – Index of Bacteriostatic Activity (reduction of the number of colonies related to the control samples not exposed to activation).

N0 = 10 bacteria/ml, Control	N0 = 10 bacteria/ml, tact = 15 min

Fig 11: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureus Wood-46 with initial concentration of culture N₀ = 10 bacteria/ml.

Fig 12: The effect of time duration of MRET activation on the inhibition of growth of culture of Staphylococcus aureus Wood-46 with initial concentration of culture N₀ = 10 bacteria/ml. IBA – Index of Bacteriostatic Activity (reduction of the number of colonies related to the control samples not exposed to activation).

CONCLUSIONS:

1. MRET activation of the water based nutrient medium with suspended staphylococcal culture leads to the origination of high bacteriostatic activity of nutrient medium which depends on the time duration of activation and initial concentration of culture cells.

2. The bacteriostatic activity increases following the increase of time of activation (the times of activation up to 60 minutes were studied).

3. The efficacy of bacteriostatic activity increases following the decrease of initial concentration of the suspension of staphylococcal culture. The process of MRET activation is most effective on culture suspensions with the concentration not more than 10³ bacteria/ml.

4. The results of investigation provide the evidence regarding the high efficacy of MRET activation on the inhibition of growth of colonies and reproduction of staphylococcal microorganisms in vitro. These results allow admitting that the process of MRET activation and the sterilization effect of MRET water can be applied in food industry and for water purification.

Live Blood Cell Analysis

In this particular case, the blood sample of the subject before Activated Water ingestion shows free radical attack on cells. Note the “spiky” blood cells that take almost 90% of specimen. 30 minutes later, after Activated Water ingestion, the blood specimen shows non free radical environment.

In this particular case, the blood sample of the subject before Activated Water ingestion shows so called Rouleau formation when blood cells are stacked, forming a worm-like pattern. 30 minutes later, after Activated Water ingestion, most of the worm-like patterns broke up, forming individual patterns of nicely round spheres.

MRET water effect on seed germination and growth of plants

The examinations of influence of MRET activated water on plants was conducted during 3 months period at Kiev Institute of Plants Genetics of Ukrainian Academy of Sciences under supervision of Prof. V. Vysotskii, Kiev National University. MRET activated water accelerated the process of seed germination of several plants (particularly of cabbage, pumpkin, string bean, garden radish and peas) and enhanced their growth cycle.

These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat or prevent disease.