Antibacterial and Antiviral Effects of Water Catholyte, Oxidal with Methylene Blue and Colloidal Silver

Coronavirus COVID-19 is an RNA virus with size 120 nm. It began spreading at the end of 2019 and at the beginning of 2020 in China, and it leads to a severe infectious disease 2019-nCoV. It resembles severe acute respiratory syndrome-related coronavirus (SARSr-CoV), and the new virus is labelled as SARS-CoV-2. Its genome is single-chained with positive strand. It contains a nucleotide with spiral symmetry. The coronavirus SARSr-CoV gets replicated via copying of the genetic material using the enzyme RNA-dependent RNA polymerase.The author considers a complex method of influence over SARS-CoV that includes intake of alkaline water Catholyte with negative oxidation reduction potential (ORP), food supplement Oxidal with Methylene blue and Colloidal Silver.


Introduction
Phylogenetic and comparative analysis of SARS-CoV E shows that TMD consists of the two nonpolar, neutral amino acids, valine and leucine. They lead to a strong hydrophobicity to the E protein. In SARSr-CoV there is an interaction between E and S proteins.The interaction is via disulfide bonds. The C-proteins are positively charged. Viral infections occur mainly by C-terminus of target proteins. The water Catholyte is obtained during electrolysis in the cathode section of the electrolyzer. The following reaction is valid for the interaction of Catholyte and the virus: 2Н2О + 2е -→ H2 + 2ОН -In this reaction, is possible the free electrons are released that neutralize the positively charged C-proteins, which are responsible for the viral infections. The doses are high and the achieved effects are significant. Methylene blue has inhibiting action on viral replication (Oz et al., 2011). Its chemical formula is as follows: C16H18ClN13S. There have been a number of successful studies of Methylene blue (MB) with West Nile virus, AIDS -related Kaposi's sarcoma and RNA viruses. There are also positive results with progeria and they show the potential of Methylene blue for interaction with the cell membrane with effects on ATP and mitochondria. Oxidal is not need photoactivation. The colloidal silver Ag + has got inhibiting (diminishing) effect over respiratory enzymes of the micro-organisms by building into the reaction center of the enzymes. Thus it prevents the further alteration of the enzymes (Dondysh, 1964). Colloidal silver makes physical changes in the bacterial membrane, like the membrane damage, which can lead to cellular contents leakage and bacterial death. interact with, and potentially disturb the functioning of biomolecules such as proteins and enzymes. The coronavirus replicase was recently predicted to employ a variety of RNA processing enzymes. The colloidal silver Ag + inhibits such copying, and in this way are neutralized the effects of SARSr-CoV-2. The research from 2005 г. has shown that SARS-CoV has observed in water and is possible to exist 14 days (Wang, X. et al., 2005). (Gundy, P., Gerba, Ch., Pepper, J., 2008). In 2003 there was connection between water and wastewater and SARS-CoV in building in Hong Kong where 300 people (Leung et al. 2003) lived were. For that is important application of water Catholyte with antiviral effects.

Electrolyzer for water Catholyte
The electrolyzer has two parts, which are separated with semipermiable membrane. The anolyte is obtained at the anode and is acidic fraction. The catholyte is obtained at the cathode and is the alkaline fraction, When the current passes through the water there are redox reactions on the surface of the electrodes. The catholyte is with negative oxidation reduction potential (ORP) in mV. The anolyte is with positive oxidation reduction potential (ORP) in mV. The functional scheme and the reactions are shown in Figure 1.

Colloidal silver
In order to obtain colloidal silver with method of electrolysis  the following ingredients are required: 1. Silver electrode with a purity of 99.99%; 2. Tetra-n-butylammonium bromide (TBAB)-ammonium salt with bromide; 3. Acetonitrile -coloured liquid solution with chemical formula CH3CN The obtaining of colloidal silver can occur via electrolysis . The synthesis method uses tetra-n-butylammonium bromide in acetonitrile as a liquid medium for electrolysis. he method of Mosin-Ignatov obtains colloid silver with sizes of 2-7 nm in a solution of tetra-n-butylammonium bromide in acetonitrile. The anode is silver, and the cathode from graphite. During the process of electrolysis a partial dissolving of the silver anode occurs that leads to saturation of the solution with Ag + . At given parameters of the electric current and tension over the electrode, the concentration of Ag + solution is determined by the working time of the electricity source, and the quantity of the aqueous solution.  anolyte relative to the non-electrochemically activated water. Water Catholyte has structuring of hydrogen bonds among water molecules with anti inflammatory and anti tumor effects .

Antiviral effects of water Catholyte
Water Catholyte stimulates the function of mitochondria and synthesis of ATP . The presence of nascent hydrogen in Catholyte bestow an antiviral effect upon the water (Vassileva, Mehandjiev et al., 2019). There evidence for beneficial effects on Graffi tumor on hamsters with increasing of blood and immune parameters.

Antiviral effects of Oxidal
When entering mitochondria, methylene blue (MB) acts like an additional electron source. It is firstly reduced to MBH2 by the NADH-dehydrogenase of complex I, and then re-oxidized back to MB by cytochrome c. Cycling between those two forms facilitates electron transportation for ATP synthesis, the major function of the mitochondria. More importantly, it prevents electron leakage for oxidants formation, the toxic side products in mitochondria. MB also improves mitochondrial function by inducing PGC1α, a central mediator of mitochondrial biogenesis , and a few key electron transport chain (ETC) components including cytochrome oxidase (COX II and COX IV . Overall, thise data supports that MB promotes mitochondrial function and Journal of Medicine, Physiology and Biophysics www.iiste.org ISSN 2422-8427 An International Peer-reviewed Journal Vol.65, 2020 29 reduces ROS production. Currently available results for SARS-CoV, demonstrate that the virus affects the mitochondria in order to suppress the immunity of human body (Nelemans, 2019) (Schoeman, Fielding) Oxidal stimulates the mitochondrial function and ATP. The researched strains of S. aureus and E. Coli reveal higher sensitivity to Oxidal in comparison with the control antibiotic Thiamphenicol, but the values are not significantly different .

Antiviral effect of colloidal silver
Colloidal Silver in European Union is food additive. The mechanism of action is shown at Fig. 4 (Dondysh, 1964). The coronavirus gets replicated via copying of the genetic material using the enzyme RNA-dependent RNA polymerase. In such a way it prevents the further alteration of the enzyme. The results are "in vitro", on the enzyme in a solution. The colloidal silver is possible Ag + inhibits such copying, and in this way the effects of COVID-2019 are neutralized. Figure 4. Interaction of Ag + ions of microbial cell: a)-the membrane protein peptidoglycan and its interaction with Ag + ions; b)-the process of penetration into the microbial cell of Ag + ions. Dondysh, 1964) Figure 5 shows the electronic photography of colloidal silver (9) (Darroudi et al., 2011) Figure 5. Electron micrographs of colloidal silver with a size of 5 nm (Darroudi et al., 2011)

Antiviral Doses of water Catholyte, Oxidal and Colloidal Silver
When water Catholyte is used, it is recommended to drink it 1 hour before or after a meal. It is necessary for pH to be over 9.5 and ORP in the range of (-700 --400) mV, otherwise the increase of pH might decrease ORP and thus negate the effects of Catholyte.
Oxidal can be applied in rather high doses, but research with people show the optimal doses of "methylene blue" per person per day to be around 15 mg-16 mg. When colloidal silver is used, the doses need to be precisely defined. Daily dosage: 5 µg micro g per 1 кg body weight, ex.5 µg x70 kg body weight=350 µg = 0.00035 g.

Conclusions and discussion
For COVID-19 the influence is with antibodies against viral proteins binding to cell receptors (via blood, intravenously). There are applyed the folloing drugs again coronavirus SARS-CoV-2 -Remdesivir and Kaletra. The potential anti-HCoV repurposable drugs are -melatonin, mercaptopurine, and sirolimus.
The author suggests the use of three products -water Catholyte, Oxidal and Colloidal Silver for prevention and as an additional method of therapy for infectious disease 2019-nCoV caused by the virus (SARSr-CoV). The effects target the cell membrane, ATP and mitochondria.
The precise dosage leads to the following effects: 1. The negative electrons reduce the positive charge of C-protein, and thus reducing the number of penetrated viruses inside the cell. 2. The virus SARS-CoV affects the mitochondria in order to suppress the human immunity. Oxidal stimulates the function of mitochondria and ATP. 3. The colloidal silver Ag + possesses inhibiting effects over the respiratory enzymes of micro-organisms. Ag + incorporates into the reaction center of the enzyme. In such a way it prevents the further alteration of the enzyme. It is "in vitro", on the enzyme in a solution. 4. As an additional antiviral remedy, we also recommended intake of high doses of Vitamin C.
One of the researchers Wolfgang Luepke suggests hyper immunization of calves with SARS-CoV-2. In doing so antibodies serum can be obtained for therapeutic intervention in people. It can be in the form of breathing aerosol, thus accomplishing passive immunity. In the cells, antibodies are formed, and from passive the immunity becomes active.