Applications of H2B: Scientific Overview
We have conducted a number of pilot studies and experiments to explore the potential of H2B in a wide range of applications. Here we provide a brief overview of some of those efforts.
We also append a selection of relevant published studies on molecular hydrogen.
WHY H2B AFFECTS WEIGHT CONTROL
Remarkably, H2B facilitates the creation of no – or low calorie, delicious foods and drinks that are an effective substitute for high calorie foods and drinks. This breakthrough should be of benefit to weight management and anti-aging programs:
· being anti-inflammatory through H2 delivery,
· feeling of fullness generated by H2B matrix expansion,
· fullness due to an H2 headspace effect,
· having a desirable texture imparted by H2B,
· having extended stability of H2 in foods and drinks,
· H2B is compatible with most foods and drinks.
Here, is a partial list of great tasting H2B drinks and foods that can be consumed – having little or no calories:
· Chocolate Smoothie
· Blueberry Protein Breakfast Smoothie
· Lemon Flavored Vegetarian Protein Powdered Drink
· Transforming Marketed Drinks and Powders into Delicious H2 Deserts
· Enriched Tomato Soup
· Medicinal teas synergistic with H2.
ORAL DRUG - H2 DELIVERY
Due to the proven effectiveness of H2B to both generate and sustain H2 in formulations, as well as serving as a sustained drug delivery system, numerous H2 – proven drug products are now available for therapeutics - by the oral route. Two examples are provided below:
Example 1: Treatment of Gingivitis with a Good Tasting H2 – Chlorhexidine Formula
It is well known that gingivitis (i.e., gum inflammation) leads to periodontal disease. It is the leading cause of tooth loss in both humans and animals.
Chlorhexidine has been used in the oral cavity to treat gingivitis. This treatment has been only partially effective. There is a need for both better sustained delivery systems of antimicrobials and for more effective, longer lasting anti-inflammatory therapies. A recent publication attests to the antibacterial activity of H2 water against oral bacteria (1).
Using H2B, a formula containing 0.12% chlorhexidine gluconate and 22.1% H2 was prepared. A small Phase 1 therapeutic trial was carried out. The product was rubbed into the gums. Overnight, the soreness in the gums was partially relieved. With repeated use, the symptoms of gingivitis were totally relieved.
Lee, S.H., Bach, D.H. (2013) Antibacterial Activity of Hydrogen – Rich Water Against Oral Bacteria. Int. J. Oral Biol. 38 (2), 81-85.
Example 2: Sustained Release of H2 and Bismuth Subsalicylate in the G-I Tract
There are diseases and conditions of the upper G-I tract where sustained delivery of both effective drugs and H2 would have an advantage over current treatments. Conditions such as gastritis, ulcers, gastroesophageal reflux disease (GERD), Helicobacter pylori infection, bacterial overgrowth, yeast overgrowth - could be better treated. Some of the drugs that could be delivered from H2 – rich H2B formulations are: misoprostol, H2 receptor antagonists, antibiotics, antifungals, anti-inflammatories, antacids and bismuth subsalicylate.
Using H2B, a formulation releasing 1.4 ppm H2 and containing 1.1 g./100 mL of bismuth subsalicylate was prepared. It was consumed by a volunteer suffering from GERD. His heartburn was rapidly relieved – with no side effects. The formulation was judged to taste very good.
Bismuth subsalicylate, in combination with 250 mg of metronidazole and 500 mg of tetracycline has been used to eradicate Helicobacter pylori. It is feasible to incorporate both these drugs - along with 1.1 g. bismuth subsalicylate into an H2 generating and sustaining H2B formulation. The H2B formulation will spontaneously and rapidly form in the stomach
– floating and having a long retention time. Five - to 10 - hour delivery of H2 and the drugs is achievable.
DERMATOLOGY AND SKIN ANTI-AGING APPLICATIONS
Skin conditions such as wound healing, acne, dermatitis, psoriasis, irritated skin, sunburn, infections, etc., are associated with inflammation. Drugs and treatments for skin diseases can cause inflammation. Combining topical delivery of H2 with standard therapies for skin conditions should reduce inflammation. H2B facilitates sustained topical delivery of H2 and drugs. Two Examples are as follows:
Example 1: Anti-acne H2 Formulation
An anti-acne, keratolytic preparation consisting of H2B and 2% salicylic acid, in distilled water was prepared. After expansion, the formulation was found to contain 25.4% of H2. Sites were delineated on the skin of the forearm. The formulation was spread uniformly over the test site. The concentration of H2 being delivered to the skin surface was determined. Salicylic acid on the skin surface and in the stratum corneum was observed with a Wood’s Light (UVA). Tape stripping the skin on the site allows for determining the layers of stratum corneum penetrated by salicylic acid. It was found that the H2B formulation delivered salicylic acid deeply - through 9-10 layers of stratum corneum.
Immediately after application, it was determined that 432 ppb H2 was in contact with the skin surface. Within 10 minutes this value dropped to 250 ppb. At 20 minutes it was 167 ppb. At 30 minutes, it had dropped to zero. A substantial portion of the H2 in contact with the skin surface was absorbed into the skin. The stratum corneum, the top barrier of the skin, is hydrophobic. H2 is also hydrophobic. Hydrophobic entities associate with each other, in an aqueous environment.
Example 2: H2 Moisturizing Lotion for Dry, Aging skin
As skin ages from environmental damage, it is subject to sub-chronic inflammation affecting loss of hydroscopic macromolecules – thus, reducing its moisture holding capacity. A preparation that will reduce sub chronic inflammation - and moisturize, should slow down skin aging. By sustained topical delivery of H2, H2B makes this possible. A H2-generating moisturizing lotion was prepared: 1.6% H2B, 3% lactic acid, 20% glycerin, 78.4% distilled water. The lotion released 815 ppb H2. The site was treated topically with 0.15 g./square cm. of the lotion. Immediately after treatment, the H2 on the skin surface was 804 ppb. H2 fluctuated and did not show a progressive loss up to 60 minutes. H2 measurements showed the presence of significant H2 on the skin surface - up to 210 minutes.
This persistence of H2 on the skin surface was surprising. The lotion rapidly loses most of its water as it dries on the skin surface.
Such persistence may be explained by:
1) H2B deposited on the skin surface, continues to react with water held on the skin surface by the hydroscopic effects of glycerin and lactic acid;
2) The stratum corneum has a reservoir effect for hydrophobic molecules. H2 is hydrophobic. It may contribute to the H2 detected on the skin surface due to back diffusion of H2 from the stratum corneum.
AQUACULTURE APPLICATION OF H2B
H2 promotes the health and growth of both animal and plant life. Aquatic life in the form of plants, invertebrates (e.g., shrimp, crabs, etc.) and fish should also benefit from the anti-oxidant and enhanced bioavailability effects of H2. H2 reduces reactive oxygen species (ROS). ROS affect the consumption of energy that could otherwise be used for growth and fighting disease.
Example 1: Sustained Delivery of H2 to an Aquarium or Fish Farm
An experiment was designed to determine if an H2B formulation, that generates H2, can deliver and sustain H2 in a model aquatic farm. Phosphate buffered saline (PBS) in distilled water(DW) was prepared, pH 7.6. Also, a formulation of 4.1% H2B and 0.5 % ascorbic acid in DW – was prepared. The H2 released was determined to be 1.5 ppm.
The 125 mL formulation was placed in the 1,000 mL PBS solution. Being less dense than PBS, it floated on top of the 1,000 mL container. The PBS was constantly circulated.
At baseline, there was 0 ppb H2 in the PBS solution, pH of 7.6. After 15 minutes, the H2 in the PBS remained at 0 ppb, pH of 7.6. By 4.7 hours, there was 562 ppb H2 in the PBS, pH of 7.7. At 13.8 hours, H2 in the PBS had risen to 691 ppb, pH of 7.7. At 28.3 hours, the H2 in the PBS had risen to 1245 ppb, pH 7.6. Thereafter, H2 in the PBS was sustained at a level of 1202 ppb, pH 7.3 until 109 hours. At 123 hours there was 1165 ppb H2, pH 6.8. The pH drop probably indicates bacterial growth in the simulated tank. This experiment shows the feasibility of delivering H2 to an aquarium or aquatic farm. It also points to routine steps that must be taken to insure the maintenance of water quality
ENHANCED INSECT FARMING WITH H2
As the world’s population increases, it becomes increasingly difficult to provide food, particularly in the form of complete protein. Insects are an excellent source of complete protein and some vitamins. They eat almost anything. They are ‘natural’ cleaners of the environment. Insect food can also serve as food for lower animals. Although insects breed and grow rapidly, a biotechnology that would increase protein yield would be of economic significance. Since H2 has been shown to increase reproduction and growth in many life forms (1, 2), there is no reason to doubt its growth promoting effects for insect populations. Insect cells are also powered by mitochondria.
An insect farm was modeled to determine if wildtype insects would be attracted to an H2B formulation generating H2 . The formula was as follows: 4.1% of H2B and 5% of fructose were mixed with 5% acetic acid. The H2 content of the formula was found to be 800 ppb, pH of 3.35. The container was placed outside of the laboratory. Over the course of 24-hours, bees, flies, ants and other un-identified insects appeared to be attracted to the container. The purpose of placing insect food in the H2-rich H2B, is to attract insects to the formulation, thereby exposing the insects to a higher concentration of H2. As the insects ingest both the insect food and the H2B, they will come into close contact with H2 - which will be both ingested and absorbed into their body parts – in contact with the formulation.
Huang, Lei (2016) Molecular hydrogen: a therapeutic antioxidant and beyond. Medical Gas Research 6(4): 219-222.
Zeng, J. et. Al. (2014) Progress in the study of biological effects of hydrogen on higher plants and its promising application in agriculture. Medical Gas Research 4 15.
H2 MICROBUBBLES IN H2B SOLUTIONS IN RELATION TO MEDICAL ULTRASONICS
Due to the discovery of the remarkable sustainability of H2 in aqueous H2B, a study was carried out to determine if stable microbubbles are present in an aqueous H2B formulation. The formulation was filtered through a 400 - mesh filter. All microbubbles passing through the 400 Mesh filter were 37 microns - or smaller. 20 uL of filtered solution was injected onto the sintered glass space adjacent to the glass coverslip covering the grid on a hemocytometer. The grid was observed under the 40/0.065 magnification on an OMAX® Binocular Compound Microscope Model: M83EZ. At the 4.5 - hour observation, a wide range of sizes of microbubbles were observed. The smaller sizes ranged between 2-10 microns. At 8.5-hours, the filtered formulation was again observed. A wide range of microbubbles of assorted sizes was again observed. The microbubbles have a sustainability of at least 8.5 -hours.
There is convincing evidence that the persistence of H2 in non-viscous solutions containing H2 is due to the stabilization of microbubbles by H2B.
The size of the H2 microbubbles in H2B - generating systems can be selected by molecular sieving, centrifugation or size exclusion chromatography.
The ability to generate sustainable H2 microbubbles, in aqueous solution, and their delivery to the systemic circulation - has important health benefits regarding enhancement of ultrasonic medicine:
· Sharper imaging of organs and the circulatory system.
· Drug delivery to un-accessible areas, including crossing the blood – brain barrier.
· Enhancement of non-invasive treatment of prostate and breast cancer.
· Enhancement of histotripsy (tissue fractionation using high frequency ultrasound).
· Sonothrombolysis (destruction of blood clots using ultrasound).
· Enhanced delivery of DNA, RNA and proteins into cells of targeted tissues.
· Enhancement of CRISPR - Cas9 biotechnology.
· Vascular permeability enhancement.
· Enhanced lithotripsy - the disintegration of kidney and bladder stones.
· The benefits listed above, plus the anti-inflammatory effects due to H2.
[Index of studies to follow]