CH718405A2 - Process for the production of atomic H-water with the addition of citrulline as a rehydrating, antioxidant drink in aluminum bottles and stand-up pouches using electrolysis with boron-doped full diamond and titanium electrodes. - Google Patents

Abstract

Method and equipment for the production and for the oral uptake and intracellular invasion of active negatively charged hydrogen (hydride) in body cells as an efficient antioxidant by means of water electrolysis with boron-doped diamond and titanium carbon fibers contact - electrodes for the production of active hydrogen water through the generation of excess electrons in the electrolyzed water and for microclusterization of the water for improved and faster absorption and invasion of active negatively charged atomic hydrogen as an antioxidant in body cells with additional injection addition of gaseous medicinal hydrogen and citrulline and, if required, natural fruit extract flavors.

Description

TECHNICAL AREA

Process for the production of atomic hydride H-water with the addition of citrulline, as a rehydrating, antioxidant drink in aluminum bottles and stand-up bags, by means of electrolysis with boron-doped full diamond and titanium contact electrodes.

STATE OF THE ART

[0002] Previously, hydrogen dissolved in water as an antioxidant could not be introduced directly into body cells.

With the new technology of producing hydride ions H- using boron-doped diamond and titanium - contact electrodes - electrolysis of water, which is also microclusterized by electrolysis and then additionally injected with medical hydrogen gas, it is possible assets Atomic negatively charged hydrogen as an antioxidant, to be taken orally as a drink and channeled directly into a body cell and cell organelles using the microclustered water (1 to 3 molecules per cluster). In order to break down molecular hydrogen into atoms, energy of about 4.5 eV per molecule, or more precisely 436.22 kJ/mol, must be expended; when they combine to form hydrogen molecules (H2), this energy is released again:

Active atomic hydrogen (hydride) with 1 proton and 2 electrons in the shell is, as an electron donor, the chemically strongest known antioxidant that can eliminate harmful oxidative radicals, so-called ROS (Reactive Oxidizing Species) from cells and the human body. without subsequently becoming an oxidative radical itself, as is the case with antioxidant vitamins such as vitamins C and E etc. The hydride ion H atom is the revolution of hydrogen. Water enriched with negatively charged hydrogen H has an enormous antioxidant effect, which can contribute to a healthier lifestyle and can build up the physical immune system to the maximum in order to also treat many diseases preventively and curatively. Hydrogen-enriched hydride water with the addition of citrulline provides greater health and well-being. In our everyday life we come into contact with numerous chemical compounds and eat foods that have a high oxidative effect. Water enriched with negatively charged hydrogen (hydride ion) has a high antioxidant effect that can contribute to a healthier lifestyle.

1 glass of Hydridion water, according to the description, contains the same antioxidant potential as 100 glasses of orange juice.

Active atomic hydrogen is a negatively charged hydrogen atom with a proton and 2 electrons instead of just one and is therefore negatively charged.

Active atomic hydrogen thus consists of a proton and 2 electrons. The extra electron is provided to the hydrogen by the electrolysis of water using diamond and titanium carbon fiber contact electrodes. Thanks to the electrical overpotential of 2 to 3 V with boron-doped diamond electrodes, more electrons can be supplied to the water than the water can absorb, which leads to an excess of electrons. The water is thus electrically charged and the excess electrons are absorbed by the atomic hydrogen, which is created by the new special electrolysis technology and the additionally injected medical hydrogen gas. H+ thus becomes H- with a proton and 2 electrons, which leads to the formation of negatively charged active atomic hydrogen, also called nascent hydrogen, which can donate a reducing electron as the best antioxidant. Redox reaction 2H2O + 4H+ o O2+ 4H+ + 2H2 AE = 1.229 V 4H+ plus 4e = 4H- 4H+ plus 4e = 4H-

[0008] Thanks to the microclusterized water, these negatively charged H ions are transported into the cells through the positively charged aquaporine locks embedded in the cell membranes and can even penetrate into the organelles of a cell as an antioxidant.

It should also be briefly mentioned that active, atomic hydrogen also has numerous other amazing and positive properties beyond its effect as a powerful antioxidant.

For example, negatively charged active hydrogen has a balancing effect on the acid-base ratio, which prevents viruses, bacteria and fungi from multiplying and spreading in the body. Hydridion water therefore has a preventive and curative effect against the nCorona virus Covid 19, which is proven by a number of control placebo studies.

Active negative hydrogen improves the nutrient supply and detoxification of the body and also supplies oxygen in the mitochondria for the ADP-ATP cycle and thus increases our energy potential (the amount of ATP has been increased fivefold in tests)

[0012] It ensures that the intracellular and extracellular irrigation of the cells increases significantly. The hydration state of a cell is directly related to the aging process. dr Alexis Carrell, Nobel Laureate in Medicine, said: “The cell doesn't age, it just dries up.

[0013] Active atomic negative hydrogen provides the necessary negatively charged hydrogen ions to ensure effective communication at the cellular level.

CITRULINE

[0014] Citrulline is a non-protinogenic amino acid produced by plants and animals. Citrulline is named after the watermelon (Citrullus vulgaris) in which it is concentrated. It is also found in other cucurbits. Citrulline is a homologue of L-homocitrulline derived from L-lysine by F - carbamolization.

physiology

Citrulline is increasingly excreted in the urine by humans when they are suffering from so-called nitrosative stress. Nitrosative stress is an endogenous exposure in the human body to the nitric oxide radical. In the urea cycle, arginine then reacts more with oxygen, producing NO and citrulline. Therefore, citrulline is used as a marker for nitrosative stress in medical diagnostics.

citrullination

The enzymatic conversion of arginine into citrulline is referred to as citrullination. Peptidylargenin deiminases (PAD) catalyze this post-translational modification that occurs on certain proteins and peptides. These citrullinated proteins or peptides can be regarded as foreign by the immune system and can be attacked by antibodies, for example. For example, citrullinated peptides play an important role in the pathogenesis of rheumatoid arthritis.

PRESENTATION OF THE INVENTION

INTRODUCTION

The object of the invention is to describe a new, innovative, inexpensive, efficient, biological and toxic residue-free method for efficiently supplying cells with active, atomic and negatively charged hydrogen as an electron donor and antioxidant. The basis of the new, perfected invention is the combined use of microclusterized, electrolyzed water produced with full diamond and titanium contact electrode electrolysis with an excess of electrons, which is also mixed with medicinal hydrogen as a gas and citrulline (2 gr. per 500 ml) is enriched.

HYDROGEN

Biological Significance

[0018] Hydrogen, in the form of a wide variety of compounds, is essential for all known living beings. First and foremost is water, which serves as a medium for all cellular processes and for all transport of substances. Along with carbon, oxygen, nitrogen (and more rarely other elements) it is part of those molecules from organic chemistry without which any known form of life is simply impossible.

Active atomic hydrogen also plays active roles in the organism, such as in some coenzymes such as nicotinamide adenine dinucleotide (NAD/NADH), which serve as reduction equivalents (or "proton transporters") in the body and participate in redox reactions. In the mitochondria, the powerhouses of the cell, the transfer of hydrogen cations (protons) between different molecules of the so-called respiratory chain serves to provide a potential, a proton gradient, for the generation of high-energy compounds such as adenosine triphosphate (ATP). During photosynthesis in plants and bacteria, the hydrogen from the water is needed to convert the fixed carbon dioxide into carbohydrates.

Based on the mass, hydrogen is the third most important element in the human body: in a person with a body weight of 70 kg, around 7 kg (=10% by weight) can be attributed to the hydrogen contained. Only carbon (approx. 20% by weight) and oxygen (approx. 63% by weight) make up an even larger proportion by weight. In terms of the number of atoms, the very light hydrogen is by far the most common atom in the body of every living being. (The 7 kg in humans correspond to 3.5 · 10<3>mole of hydrogen each with 2 · 6 · 10<23>atoms, that is around 4.2 · 10<27>hydrogen atoms).

Medical importance

In biological systems, atomic, negatively charged hydrogen reacts with reactive oxygen species and thus acts as an antioxidant. In animal experiments, the enrichment of drinking water with molecular hydrogen after kidney transplantation leads to better survival of the transplant, a reduced incidence of chronic damage to the transplant, a reduction in the concentration of reactive oxygen species and an inhibition of signaling pathways that increase inflammatory activity ( pro-inflammatory signaling pathways).

[0022] Active hydrogen delays the aging process of body cells.

ELECTROLYSIS OF WATER

The electrolysis of water consists of two partial reactions that take place at the two electrodes. The electrodes are immersed in water, producing oxygen and hydrogen gas.

Positively charged hydronium ions (H3O+) migrate in the electrical field to the negatively charged electrode (cathode), where they each absorb an electron. This creates hydrogen atoms, which combine with another H atom (hydridion) formed by reduction to form a hydrogen molecule. Water molecules remain. 2H3O+ + 2e- o H2+ 2H2O The separated, gaseous hydrogen rises at the cathode.

The negatively charged hydroxide ions migrate to the positively charged electrode (anode).

[0026] Each hydroxide ion gives up an electron to the plus pole, so that oxygen atoms are formed, which unite to form oxygen molecules.

The remaining H+ ions are immediately neutralized by hydroxide ions to form water molecules. 4OH- o O2+ 2H2O + 4e-

Here, too, the separated oxygen rises as a colorless gas at the anode. The overall reaction equation for the electrolysis of water is: 4H3O+ + 4OH- or 2H2+ O2+ 6H2O

The hydronium and hydroxide ions on the left come from the autoprotolysis of water: 8H2O o 4H3O+ + 4O

The electrolysis equation can therefore also be written as follows: 8H2O o 2H2+ O2+ 6H2O or after shortening the water: 2H2O o 2H2+ O2 redox reaction 2H2O + 4H+ o O2+ 4H+ + 2H2 AE = 1.229 V 4H+ plus 4e = 4H- 4H+ plus 4e = 4H-

Thanks to the excess electrons in the water, which occur in the full diamond and titanium and carbon fiber contact electrodes electrolysis because of the electrical overpotential when using boron-doped diamond electrodes in stack formation, the atomic hydrogen can have an additional electron and becomes active hydrogen, i.e. a reducing hydrogen atom that can lose 1 electron.

MICROCLUSTERIZATION

In addition, during electrolysis with diamond electrodes, the water clusters are cut from about 18 water molecules in a chain to about 3 molecules electrochemically by supplying energy. This fact leads to a 3- to 4-fold increased absorption of the water provided with active hydrogen via the aquapores of the cell membranes in body cells, where the active hydrogen can act as an antioxidant without becoming a reactive oxidant itself, but only to water.

ELECTROLYTIC MICROCLUSTERIZED WATER ADULT

Electrolytically microclusterized, chemically active water has a negative ORP (Oxidation Reduction Potential) of about 300 to 400 mV, which means that the water solution can give off electrons.

SOLUTION TO THE TASK

The solution to the problem is defined by the features of the independent patent claims.

The new optimized invention shows both a new, innovative, inexpensive, efficient, biological and toxic residue-free method for the efficient supply of cells with active, atomic, reducing hydrogen as an antioxidant as well as synergetic citrulline, and the device for Production of active hydrogen and microclustered water as a means of transport into the body cells.

The basis of the optimized invention is the combined use of full diamond and titanium contact electrodes produced, microclusterized electrolyzed reverse osmotic water with an excess of electrons, which, by absorbing electrons from the atomic water, produce active negatively charged hydrogen as an electron donor and at the same time the electrolyzed water with the excess of electrons converts the injected gaseous medical hydrogen to negatively charged atomic hydrogen and also interacts with the NO nitrogen oxide originating from citrulline.

According to the invention, the new method shows the technical execution, and the application, regarding the use and the combination of boron-doped diamond electrodes electrolysis technology with an electrical overpotential of 2 to 3 volts for the production of excess electrons in the Water, which can be absorbed by atomic hydrogen, as well as the microclusterization of the water, which can be introduced as a transport medium for the active hydrogen via the aquaporins in the cell membranes into the body cells and cell organelles. Active, negatively charged, atomic hydrogen is very unstable and can be made more durable thanks to the excess electrons in microclustered electrolytic water.

The invention forms an integrated system in which the technical components for the production of the microclustered water and the production of an electrode excess in the water for the formation of active negatively charged hydrogen are combined.

In intensive experiments, the degree of microclusterization (3 molecules per cluster), the optimal concentrations of active negatively charged hydrogen in the water and the dosage of additionally injected hydrogen gas as well as the absorption efficiency and absorption speed of active negative charged hydrogen in body cells and determined the specific parameters for dosing as an antioxidant and for citrulline.

To the inventor's knowledge, the optimized process technology described here for the residue-free, clean, efficient and cost-effective production and invasion of active negative hydrogen water with the synergistic interaction of citrulline in human, animal or plant cells is not yet known, which Patentability of this innovative invention is credibly proven.

CARRYING OUT THE INVENTION

The invention will be discussed using the example of a production plant for active hydrogen water with the method described and the integrated new and optimized electrolysis technology used therein, with the addition devices for hydrogen gas. The following components are necessary to carry out the process, see: 1. For example, 1 electrolysis generator with reverse osmotic water pre-filter, with 1 pressure generation pump and with an hourly capacity of approx. 1000 liters with preferably 4 atm pressure generation and refrigeration unit for water cooling parts corrosion-free, with electric Drive 220 V/50/60 Hz, with built-in electrolytic cells with boron-doped diamond and titanium - carbon fiber contact electrodes with electronics for control and transformer for minimum 50 V and 12 Amp DC for electrical supply of the electrodes and for dipole reversal for the automatic cleaning of the electrodes. 2. Electrolysis generator with preferably several multi-chamber electrolysis cells, preferably with boron-doped diamond and titanium - carbon fiber contact electrodes with an electrical overvoltage of 2 to 3 volts, with an upstream pressure reducing valve of 2 to 3 bar. 3. 1 water tank with a capacity of approx. 1000-2000 liters for the treatment of chilled water with level control. 4. Cooling unit with compressor for water cooling. 5. An electrically controlled gas valve for injecting gaseous hydrogen from a pressure bottle with an adjustable manometer and dosing unit for injecting medical hydrogen.

As a first step, the hydrogen-water production plant is connected to the 230 V power supply.

The hydrogen-water production plant is then connected to a water source (tap) with a hose and pressure control valve (2 to 3 bar max.). Then a hydrogen gas bottle battery (consists of several 300 liter bottles) is connected and the injection pressure is set to 4 bar.

For increased concentration of active hydrogen according to claim 1 and 2, the electrolyzed water after the electrolysis preferably 30 mg (30 ppm) of hydrogen gas per liter are added, which are automatically metered in by the hydrogen injection device. In addition, the water is charged with 4 grams of CITRULLIN powder per liter as an additional antioxidant and flavoring.

Subsequent to the production, the active hydrogen water is packaged sterile inline on a stand-up pouch line or a lightweight aluminum bottle filling system.

The newly developed gas-tight bags and lightweight aluminum bottles guarantee a shelf life of at least 1 year.

The recommended drink daily amount per person is 1 liter per 30 kg body weight.

Abstract / Studies

[0048] COVID-19 is caused by a viral infection and can be a fatal disease for immunocompromised people. HYDIDION CITRULLINE WATER is a functional health drink infused with molecular hydrogen and contains the nitric oxide-producing amino acid citrulline. Numerous preclinical and clinical studies have shown that the gaseous signaling molecules molecular hydrogen and nitric oxide have antioxidant, anti-inflammatory and immunomodulatory benefits. Although nitric oxide can kill pathogens directly, it is critical for immune function, can inhibit viral replication and can also cause fatal cell injury under stressful conditions. In contrast, molecular hydrogen has a very high safety profile and helps regulate nitric oxide production and metabolism, mitigating its harmful effects. Both molecules are used separately in clinical trials for COVID-19 patients. This article provides a brief review of the scientific literature on these two molecules, then presents preliminary data on HYDRIDION CITRULLINE WATER, including increased blood flow and wound healing. Based on the pre-clinical, clinical and preliminary data, HYDRIDION-CITRULLINE WATER is recommended for clinical tests/applications in addition to its frequent therapeutic medical use. H2 and NO • work together to suppress COVID-19-induced oxidative stress and inflammation, which helps prevent multiple organ failure and death. H2 and NO • work together to suppress COVID-19-induced oxidative stress and inflammation, which helps prevent multiple organ failure and death.

introduction

Coronavirus disease-19 (COVID-19) is a highly infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has led to the current 2019-20 coronavirus pandemic [1 ]. Although the virus can damage many organs, the lungs are the most affected. In addition to flu-like symptoms such as fever, fatigue, and shortness of breath, COVID-19 can also lead to pneumonia, multiple organ failure, and even death [1]. These consequences are consequences of pathogen-induced cellular aberrations that arise from and lead to oxidative and inflammatory stress responses. Even if the immune system is able to overcome the infection, the associated damage to multiple organs can lead to additional burden and increased mortality. Accordingly, a favorable regulation of the immune system and the associated inflammatory and oxidative reaction is optimal. There is currently no vaccine or other clearly effective medical intervention to treat COVID-19 [1]. Among many interventions and drugs tested, at least four clinical trials are investigating the use of molecular hydrogen in patients with COVID-19 and reporting positive preliminary results (RefA). HYDRIDION CITRULLINE WATER is a novel zero calorie functional beverage containing nitric oxide stimulating citrulline infused with molecular hydrogen. The concentration of H2 is above 2 ppm, determined by gas chromatography (H2 Analytics, Las Vegas, USA; SRI 8610C; California, USA). All ingredients have a high safety profile and are GRAS (Generally Recognized as Safe) marked by the USFDA. Although HYDRIDION CITRULLINE WATER is primarily considered a "healthy drink" and is regularly used by doctors to treat numerous medical diseases. HYDRIDION CITRULLINE WATER serves as an effective way to improve human physiology including the cardiovascular and immune systems. Accordingly, it can play an important role in the global fight against communicable and non-communicable diseases including COVID-19. We briefly review some of the molecular hydrogen and nitric oxide/citrulline literature related to this global COVID-19 pandemic. Finally, we provide some preliminary data for improved blood flow after consuming HYDRIDION CITRULLINE WATER, as well as several favorable case reports

HYDRIDION - hydrogen and immune function

Molecular hydrogen (H2) is an odorless, easily diffusible, flammable gas and is naturally produced by intestinal bacteria [2]. A 1975 publication in Science by researchers from Texas A&M and Baylor University reported that hyperbaric hydrogen is an effective treatment for squamous cell carcinoma in albino mice [3]. Later, a 2007 publication by Nature Medicine showed that H2 inhalation significantly suppressed brain damage in a rat stroke model by suppressing oxidative stress and inflammation [4]. Subsequent scientific publications (~1500) and human clinical studies (~80) from universities and institutions around the world have further confirmed the initial finding that H2 has therapeutic effects [5]. Molecular hydrogen is mainly administered to humans either by inhalation or dissolved in water to create hydrogen-rich water (HRW) [5]. In Japan, molecular hydrogen has been approved as an advanced drug for the treatment of post-cardiac arrest syndrome, for which a multi-center clinical trial with 360 patients is being conducted [6]. H2 has antioxidant, anti-inflammatory, anti-allergic and anti-apoptotic protective effects. Although the exact molecular mechanisms are not yet known, H2 provides this protective effect by modulating signal transduction, affecting gene expression, and modulating protein phosphorylation cascades [5]. These beneficial effects of molecular hydrogen can clearly qualify it as an optimal therapy against COVID-19 and similar diseases. In fact, H2 has already been suggested to be an effective treatment for multi-organ dysfunction induced by influenza and other viral infectious diseases based on the current animal and human publications. For example, in zebrafish exposed to the bacterium Aeromonas hydrophila, HRW downregulated pro-inflammatory genes (IL-1β, IL-6 and NF-κB) and upregulated the anti-inflammatory cytokine IL-10. In addition, HRW increased the survival of the zebrafish and effectively blocked the replication of the bacterium [8]. Similarly, lipopolysaccharide (LPS) is induced in a rat model. In systemic inflammation, HRW (Hydrogen Rich Water) significantly reduced pro-inflammatory plasma mediators (i.e. TNF-α, IL-6 prostaglandin E2) and increased the anti-inflammatory cytokine IL-10. Symptomatically, H2 prevented LPS-induced hypotension, fever [9] and neuroinflammation and promoted recovery from LPS-induced disease [10].

[0051] Hydridion H- therapy effectively suppressed radiation-induced damage to the immune system, such as preventing the decline in splenocyte counts and CD4 + and CD8 + T lymphocyte counts. In addition, hydrogen prevented the radiation-induced imbalance between Th1, T2, Th17 and Treg cells and other cytokines (TNF-α, transforming growth factor-β1, GM-CSF, IL-1β, IL-4, IL-5), IL-10, IL-17, IL-23) [11]. Similarly, in a human study of 55 stage IV colorectal cancer patients, molecular hydrogen inhalation restored depleted CD8+ T cells [12]. These cytotoxic T lymphocytes target cells infected with cancer and viruses [13]. Another randomized human clinical trial in patients with chronic hepatitis B (n=60) found that taking HRW for 6 weeks reduced oxidative stress and tended to suppress viral DNA levels and improve liver function [14 ]. Oral HRW has also been shown to reverse the immediate-type allergic reaction in mice and prevent mast cell degranulation by suppressing NADPH oxidase activity [15]. Researchers subsequently reported that HRW reduced LPS/interferon-gamma-induced NO • production in macrophages, which form a molecular basis for the functional interaction between these two gaseous signaling molecules [16]. The mechanisms responsible for these anti-inflammatory and antioxidant effects appear to involve, at least in part, hydrogen's ability to significantly repress the induction of the NLRP3 inflammasome [17] and to activate the Nrf2 antioxidant pathway [5] and the cytoprotective heme oxygenase. 1 [18]. In addition to the high cellular bioavailability of the H2 molecule, its immunomodulating activity coupled with its ability to restore homeostasis to dysregulation of cytokines (i.e. prevention/mitigation of cytokine storm) provides a significant therapeutic advantage over other medical interventions [5] .

Nitric Oxide and Immune Function

Like molecular hydrogen, nitric oxide (NO •) is a gaseous signaling molecule. Nitric oxide has long been recognized as one of the most versatile and important regulators of the immune system. It is involved in the pathogenesis and control of autoimmune processes and infectious diseases [19]. In addition, it has been shown to effectively inhibit the replication cycle of severe acute respiratory syndrome coronavirus in vitro [20]. Congruently, nitric oxide is clinically evaluated in COVID-19 patients (RefB). Nitric oxide is mainly produced by nitric oxide synthase (NOS), of which there are three isoforms, namely inducible NOS (iNOS), neuronal NOS (nNOS) and endothelial NOS (eNOS) [21]. The amino acid arginine is metabolized by this enzyme to form NO • and citrulline. NO • levels gradually decrease with age; up to 75% in 70-80 year olds compared to healthy 20 year olds [22]. Excessive inflammatory conditions lead to multiple organ dysfunction due to blood flow maldistribution and microcirculation disorders [21]. This is largely due to NO • deficiency at needed sites with excess production at unneeded sites. The availability of arginine is often significantly impaired in inflammatory diseases such as septic patients [21]. Arginine deficiency can occur due to its increased catabolism and limited availability of citrulline, which is used for arginine de novo synthesis. Because arginine is rapidly metabolized even under normal conditions, research shows that citrulline supplementation is more effective at increasing plasma arginine and NO • levels than arginine supplementation [23]. Low levels of citrulline are associated with acute respiratory distress syndrome and can also result in NOS decoupling, causing further oxidative and cellular damage [24]Combination of hydride ion hydrogen and nitric oxide. Nitric oxide, as a reactive free radical, promotes either cell survival or cell death, depending on concentration, location, and time of production. Importantly, molecular hydrogen regulates nitric oxide (NO•) levels by both reducing excess production (e.g., suppressing iNOS activity) and increasing production (e.g., increasing eNOS activity) [5 ]. Therefore, NO • regulation is crucial to potentiate their beneficial therapeutic effects and mitigate their harmful effects. NO • reacts almost instantaneously with superoxide (O2 • -) to form pernicious peroxynitrite (ONOO-), an extremely oxidative and cytotoxic molecule [25]. Reaction with O2 • - directly decreases the availability of circulating NO •, further aggravating the diseased condition [25]. The radicals O2 • - and NO • play important roles in the immune system, including killing pathogens, inhibiting their replication and regulating the inflammatory response. However, under stressful conditions such as COVID-19 and its progression to pneumonia, multi-organ failure and other sequelae, these molecules lose their critical regulation and contribute to the etiology and pathology of these diseases that form both ONOO- and extremely toxic hydroxyl radical (•OH) [25 ]. Molecular hydrogen has been shown to favorably regulate O2 • and NO • production by influencing NADPH oxidase and NOS enzymes (iNOS, eNOS and nNOS) [5]. In addition, H2 can effectively reduce the toxic hydroxyl and peroxynitrite oxidants [4]. In addition, the combination therapy of NO • and H2 showed a synergistic effect in an acute lung injury model, as can occur in COVID-19. The treatment significantly attenuated the recruitment, inflammation and apoptosis of lung neutrophils induced by LPS and polymicrobial sepsis [26]. In addition, H2 prevented NO • -induced damage, as evidenced by the elimination of nitrotyrosine levels with NO • therapy alone [26]. This lends credence to the concept that oral ingestion of HydroShot, a hydrogen-infused functional drink containing NO • -producing citrulline, can provide significant benefits to the immune system.

Preliminary data on HYDRIDION CITRULLINE WATER

Increased Nitric Oxide Production: Due to the nitric oxide stimulating ability of HYDRIDION CITRULLINE WATER, its intake accordingly increases endogenous nitric oxide production, as shown in Figure 2 after 30 minutes, followed by a slight gradual upward trend after 1, 3, 5 and 8 Hours after the baseline test (Figure 2a). Since the test strips are not quantitative and have upper and lower limits, the absolute changes, peak and duration are unknown. However, it is clear that after taking HYDRIDION-CITRULLIN - WATER, the nitric oxide levels were increased to optimal levels for at least eight hours.

[0054] Increased nitric oxide production after ingestion of the beverage mix. It decreased nitric oxide levels; Optimal nitric oxide levels at 0.5, 1, 3, 5 and 8 hours after ingestion. Increased nitric oxide production after ingestion of the drink mix corresponds to the mean percentage increase in FeNO • by 12 subjects after ingestion of the product at 45 minutes compared to baseline Similarly, elevated nitric oxide levels are also observed when using the FDA-approved NIOX VERO machine (Aerocrine AB, Solna, Sweden). NIOX VERO is a medical device for measuring fractionated nitric oxide (FeNO •), which is widely used to diagnose and treat asthma in children and adults [27]. The baseline FeNO • of 12 subjects (six men and six women; age 45.3 ± 21.1) was obtained, and then they ingested H2Bev H2-infused NO • -stimulating product, and their FeNO • levels became 45 Measured again minutes later. The product resulted in an average increase in FeNO • levels of 202.49%. Interestingly, NO • levels are slightly reduced when HRW is taken alone. This may be due to the ability of hydrogen to affect the regulation of NO • production and reduce its over-expression. Asthmatics have abnormally elevated NO • levels due to an overactive immune response. Molecular hydrogen has been shown to reduce airway inflammation in allergic asthmatic mice [28]. Anecdotally, asthmatic consumers have reported beneficial relief from molecular hydrogen uptake, but cite greater therapeutic effects from ingestion of the NO • -stimulating product. For example, a 12-year-old woman reported an average baseline peak flow meter reading of 220 L/min. However, after 15 minutes of product intake, the flow meter increased by 15%. Finally, after three months of daily use, the baseline flow meter increased to 350 mL/min (160% increase). Circulation: As mentioned earlier, nitric oxide also promotes blood flow by inducing vasodilation. Optimal blood perfusion is critical to normal organ function, wound healing, and exercise performance. Blood flow supplies cells with oxygen, nutrients, hormones, signaling metabolites, etc. and is necessary to remove harmful metabolic waste. Similar to nitric oxide levels, blood flow also decreases with age [29], which can lead to neurodegeneration, neuropathy [30] and immune system impairment [31]. Consistent with the previous data on increased nitric oxide production, Figure 3 shows the average preliminary data (triplicate measurements) of blood flow after ingestion of HYDROGEN_CITRULLINE WATER. Blood flow increased significantly from baseline after 15 minutes and remained elevated above baseline for 15 hours compared to controls. The peak increase (231%) occurred approximately five hours after dosing.

Figure 3. Changes in blood flow over 20 hours after ingestion of HydroShot compared to its baseline and control. HydroShot increased blood flow by 231% at five hours, followed by a gradual decline towards baseline. Control blood flow values did not change significantly from his baseline baseline The increased blood flow was also reflected in thermal imaging of the hand, as shown in Figure 4. The baseline temperature of the hand averaged 96.8 degrees and rose to 99.2 degrees F after 30 minutes. Figure 4. Thermal image of hand temperature after ingestion of HydroShot. Base temperature was 96.8 F(A); 30 minutes later the temperature rose to 99.2 F (B). The increased blood flow is likely due to a combination of molecular hydrogen and the other ingredients (e.g. citrulline) in the HydroShot. The increase in blood flow and reflective thermal imaging was only slightly noticeable and for a shorter duration when either hydrogen alone or citrulline alone was ingested (data not shown). This could indicate that there is a synergistic effect between these two clinically relevant molecules. Wound Healing: Consistent with increased blood flow, there have been several reports of increased healing after taking HydroShot. 5 shows a 72-year-old man with skin cancer who underwent Mohs surgery followed by 28 radiation treatments. He drank 8 ounces of HYDROGEN CITRULLINE WATER twice a day while undergoing Mohs surgery, radiation and recovery. As a result, the skin healed faster. Radiation-induced skin discoloration was predicted to heal in approximately two to three months, but instead in just 10 days.

Effects of daily intake of HYDROGEN CITRULLINE WATER during Mohs surgery, radiation and recovery. HYDROGEN CITRULLINE WATER accelerated healing in 10 days compared to the predicted two to three monthsA man who required a skin graft after surgery demonstrated the following results. Over 6 months the first two skin grafts failed and the third also began to fail. It was recommended that an amputation be performed the following week. However, the patient began taking HYDROGEN CITRULLINE WATER daily and there was noticeable improvement the following day. Shortly thereafter, the skin graft took place and was successful. The tunnel hole healed completely after 7 weeks of daily intake of HYDROGEN CITRULLINE WATER.

Discussion and Summary

The biomedical effects of negatively charged atomic hydrogen and nitric oxide are well documented in the scientific literature. In fact, these molecules have been shown in preclinical and clinical studies to exert a wide range of therapeutic effects. Consistent with our data, previous research has shown that high concentration HRW can improve endothelial function and significantly increase flow-mediated brachial artery dilatation [32]. Similarly, the enhanced healing effects of HydroShot are corroborated by previous research where administering HRW via tube feeding to heavily hospitalized elderly patients with pressure ulcers resulted in a reduction in wound size and faster recovery [33]. However, when compared to the preliminary data of HYDROGEN CITRULLINE WATER, these results suggest that the addition of citrulline significantly increases the therapeutic effects (i.e. blood flow and wound healing) of the beverage. At the cellular level, COVID-19 and related diseases are characterized by an impaired inflammatory response of the immune system, excessive oxidative stress and impaired cellular function. Nitric Oxide plays a dual role in actively enhancing the immune system, destroying pathogens, inhibiting their replication, as well as promoting oxidative cell damage and promoting excessive inflammation. However, since nitric oxide plays a key role in healthy cell function, inhibiting or activating the NOS enzymes through pharmacological intervention similarly has both beneficial and detrimental consequences [25]. In contrast, HYDRIDION - Hydrogen is a novel and safe medical gas that achieves therapeutic effects by regulating redox status and inflammatory response [5]. In addition to the clinically relevant antioxidant and anti-inflammatory effects of H2, it can also regulate nitric oxide production and mitigate its adverse effects [5]. Inhalation of H2 gas is currently used in at least four clinical studies [RefA]. dr Zhong Nanshan, the epidemiologist who discovered the SARS coronavirus in 2003, recently recommended molecular hydrogen inhalation for COVID-19 patients based on the preliminary clinical data [RefC]. Hydrogen treatment resulted in a significant improvement in dyspnea in most patients with COVID-19 in the pilot study. Accordingly, hydrogen inhalation is included as a recommendation of the Protocol of the National Health Commission of the People's Republic of China for COVID-19 [RefD]. Although their research looks at molecular hydrogen inhalation, ingestion of HRW is also effective, and at least in some cases more effective [34]. Furthermore, the addition of citrulline to increase nitric oxide levels, both of which may be deficient in late stages of COVID-19, confirms the observed therapeutic effects in the preliminary data presented. Due to the urgency of the COVID-19 pandemic and the simplicity, safety and clinical viability of HYDRIDION CITROLLINE WATER, it is recommended that HYDRIDION CITROLLINE WATER be used as a healthy beverage and clinically evaluated for its potential use in the treatment of COVID -19 patients.

author contributions

[0058] TWL helped analyze the data and organized and prepared the manuscript. MLM provided medical advice, technical insights and helped with data testing. KHR developed the hydrogen products, participated in data collection and provided insight into the manuscript. All authors have read and approved the manuscript. 1. Hui DS (2020) The ongoing epidemic threat to global health from novel coronaviruses in 2019-nCoV - The recent 2019 novel coronavirus outbreak in Wuhan, China. Int J Infect Dis 91:264- 266. 2. Suzuki Y (2009) Are the effects of alpha-glucosidase inhibitors on cardiovascular events associated with elevated levels of hydrogen gas in the gastrointestinal tract? FEBS Letters 583:2157- 2159. 3. Dole M, Wilson FR, Fife WP (1975) Hyperbaric hydrogen therapy: a potential treatment for cancer. Science 190:152-1 154. 4. Ohsawa I (2007) Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13:688-8 694. 5. 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Claims (10)

1. Process and equipment for the production and oral uptake and intracellular invasion of active negatively charged hydrogen in body cells as an efficient antioxidant by means of water electrolysis with boron-doped diamond and titanium contact electrodes for the production of active hydrogen water (hydride water) by generating excess electrons in the electrolyzed water and microclustering the water for improved and faster absorption and invasion of active negatively charged atomic hydrogen as an antioxidant in body cells with additional injection addition of gaseous medicinal hydrogen. And citrulline powder. 2. Device for the production of active hydrogen by means of the electrolysis of water with boron-doped diamond electrodes with the addition of citrulline powder and/or fruit extract flavors and gaseous hydrogen gas as required, according to claim 1, characterized in that that a device for carrying out the method has the following individual components: 3. A hydrogen production plant with reverse osmotic water pre-filter, with 1 pressure generation pump approx. 1000 liters per hour with preferably 4 atm pressure generation and refrigeration unit for water cooling. Corrosion-free parts, with electric drive 220 V/50/60 Hz, with electrolytic cells installed after the pump with boron-doped diamond and titanium and carbon fiber contact electrodes with electronics for control and transformer for 50 V and 7.8 Amp DC for electrical supply of the electrodes and for dipole reversal for the automatic cleaning of the electrodes. 4. Electrolysis generator with preferably several multi-chamber electrolysis cells, preferably with boron-doped diamond and titanium - contact electrodes with an electrical overvoltage of 2 to 3 volts, with an upstream pressure reducing valve of 2 to 3 bar. 5. 1 water tank with a capacity of approx. 1000 - 2000 liters for the treatment of chilled water with level control. 6. Cooling unit with compressor for water cooling. 7. An electrically controlled gas valve for injecting gaseous hydrogen from a pressure bottle and dosing unit for adding magnesium. 8. Electronically controlled switch buttons for normal electrolyzed water, chilled electrolyzed water and chilled electrolyzed water with addition of hydrogen, gas and magnesium. 9. For increased production of active hydrogen according to claim 1 and 2, the electrolyzed water after the electrolysis is preferably 30 mg Hydrogen gas and if required also CO2 from a battery of gas cylinders with adjustable pressure gauge which is injected at preferably 30 mg per liter, which is automatically metered in-line by the hydrogen and CO2 injector nozzles 10. Sterile in-line filling in a gas-tight stand-up pouch of any size using a fully automatic bag filling machine or using a bottle filler in lightweight aluminum bottles. The gas-tight stand-up pouch consists of 4 layers of laminate foil, with one laminate layer being aluminum foil.