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ANTI-INFLAMMATORY PROPERTIES OF MANUKA HONEY

Figure 1: Manuka honey colloidal nanoparticles isolated from Manuka honey and analysed by Nanosight instrument. 

Manuka honey is a remarkable product. By isolating out the colloidal nanoparticle and placing it into a spray bottle for topical application I have taken bee alchemy and made it available for human health and well-being applications.

 

I have spent 9 years of my life trying to understand the benefits of these particles and I have identified a number of interesting properties over the years predominantly from first hand use of the product. It's greatest benefit has to do with brain biochemistry of the subconscious mind. I have written a book which highlights these benefits and the scientific data to support it. I have also developed a topical regenerative spray that I use personally to help me expand my mind and connect to the oneness of everything. It is brain food after all. The subconscious minds biochemistry works as a quantum super computer at the speed of light. I will explain how this works in my model.

 

The colloidal system in Manuka honey has been shown to undergo photo-fenton based radical generation to produce superoxide and the hydroxyl radical. It results in cleavage of proteins and cross-linking of MGO to the peptide in the presence of a phenolic group from Manuka honey. This is triggered by photon capture and the photo electric effect where a mineral atom such as iron is reduced and then undergoes oxidation to produce high energy hydroxyl radicals.

Whilst I was analyzing the isolated colloidal particles using MALDI TOF MS I discovered that I did not need to use matrix ions as the phenolics present from the Manuka plant absorbed the energy from the MALDI laser resulting in the ionization of the colloidal particles. This resulted in the triggering of photo Fenton chemistry. The generation of hydroxyl radicals, 1200 electron volts of energy and a half-life of 1 billionth of a second resulted in the generation of a raft of new chemical compounds that had a wide range of biological properties. 

Methyl glyoxal + phenyllactate + Fe2+ + H2O2. 10x magnification, 357 nm excitation (UVA). Light driven chemical reactions. Light from light. The quantum nature for the basis of our reality is light (electromagnetism). UVA end of the spectrum creates a non linear energy within the body (UV catastrophe), where a small amount generates 1,000,000 free electrons in water. This creates the energy to generate cellular structures. The UVA is absorbed by aromatic rings of neurotransmitters. The coordination of iron with the pi electrons of the phenyllactic acid creates the ability of UVA light to reduce Fe3+ to Fe2+. The reduced Fe2+ reacts with hydrogen peroxide to create a hydroxyl radical. The OH* is the basis for the biological recycling system that can take matter and produce CO2 and H2O. This detoxification system is capable of inner alchemy. The 1200 ev and 1 billionth of a second half-life of the hydroxyl radical is the key to inner vitality.

colloidal particles in Manuka honey

The bee colloidal particles have phenolics containing aromatic rings with coordinated minerals that are bound to the major royal jelly proteins and apisimin. This hydrophobic protein complex is around 100 nm in scale and is recognized by the inflammatory cells and taken up by phagocytosis.

phagocytosis inhibition of sheep spleen cells

In the phagosome under acidic pH the NADPH oxidase produces hydrogen peroxide inducing the photo-Fenton reaction generating hydroxyl radicals breaking bonds and releasing high energy light.  

oxidative burst

The SEM EDS analysis of the colloidal nanoparticles isolated from Manuka honey was performed to determine the composition of minerals in the honey.

colloidal system in honey

 EDS analysis SEM honey

 

The high Fe content confirms the opportunity for induction of photo-fenton chemistry. The exposure of diluted manuka honey to UV light triggers radical chemistry leading to an increased concentration of methyl syringate.

Analysis of colloidal particle phagocytosis inhibition.

The presence of phenolics in Manuka honey has been confirmed. 

phenolics

The coordination of minerals to the pi electrons in aromatic rings indicates that Ca2+ and Fe3+ that are important in brain health biochemistry are displaced by Li+ due to its strength of interaction. 

electronegativity

The iron drives the photo-reduction / photo-oxidation cycle which facilitates a rapid transformation of structure to allow the quantum subconscious mind mechanisms to create and deconstruct biomaterials in such a rapid fashion to allow processing of an enormous amount of information in real time at the speed of light.

The above video shows how UVA light creates structure in the colloidal systems generated with methyl glyoxal + phenyllactate + Fe2+ + H2O2. 10x magnification, 357 nm excitation (UVA). The radicals generated break down matter through radicals which quantumly entangles the molecules together making them behave in a coordinated fashion. Shifting the light entering the system back to full spectrum white light releases the energy back out of the structures as photons of trapped light. In fact it behaves like a quantum super computer driven by radical chemistry and photo-fenton driven hydroxyl radical formation. The environmental energies impact the reactions performed by the paramagnetic radicals. This is the underlying regeneration system and important for adaption within biology. The role of radical chemistry and its sensitivity to the magnetic fields in the environment means radicals are the drivers of quantum evolution.

The use of MALDI TOF MS analysis on the isolated colloidal particles produced new biomaterials where apisimin was modified with MGO and phenolic triggered through photo-Fenton chemistry.

   MALDI TOF MS analysis of the colloidal particles

MSMS analysis revealed apisimin modification.

 apisimin

Proposed structures and position of cleavage of apisimin corresponding to glutamine. Apisimin N-terminal sequence KTSISVKGE is the site of cleavage because the carboxylate anion coordinates with Fe3+ as part of the complex with the phenolic and the protein.

 The cleavage appear to occur in close proximity to Ser and Glu resides within apisimin as this appears to be the location for mineral coordination.

 

MALDI TOF MS analysis of Manuka honey

Different phenolics appear to be directly linked onto the N-terminus of apisimin. This is triggered by the MALDI TOF laser photon capture and photo-Fenton production of the hydroxyl radicals. MGO reaction with lysine residues produces radicals. The phenoic adducts are observed.

The MSMS data was analyzed and demonstrated to contain the N-terminal fragment of Apisimin (sequence KTSISVKGE). The amino terminal lysine residue at position 1 was not observed which indicated that the linkage between the lysine and the phenolic was most likely to occur at this point. The link between the N-terminal lysine of apisimin and the phenolic was likely to be through a MGO linkage.

 

Some of the masses observed included 1302 Da and 1274 Da. MS/MS analysis of the 1274 Da series material was analysed using MALDI TOF MS/MS. Interestingly only a 1078.2051 Da and a 38.998 Da material was observed. 38 Da could correspond to C=O carbon monoxide. This may be able to turn off oxidative phosphorylation. As iron was present in the sample and the laser produces excess energy we expect the 38 Da material originated from the Fenton reaction. Structure of 1078.2051 M+1 ion (Mass of M = 1077.22936). The mass difference between 1274 and 1077 is 197 Da. Loss of oxygen lead to a mass of 213 Da for the M+1 ion which corresponds to trimethoxybenzoic acid (Mass = 212.2 Da). The UV / Vis spectral profile appears to correspond to methyl syringate.

C=O from MGO laser induced breakdown product MS 38 Da. The 1078 Da appeared to correspond to the peptide KTSISVKGES (Apisimin) with MGO attached, which a fragment of the parent ion was corresponding to 1273 Da. Parent ion 1273 Da corresponding to apisimin N-terminus, MGO linked (C3H4 - 40 Da) trimethoxybenzoic acid (MS 212.2 Da) MGO + Trimethoxybenzoic acid (253.2 Da) or methyl syringate (MS 212.2 Da). There are various forms of phenolic in Manuka honey and a number of peaks are observed which are thought to correspond to the various forms of phenolics linked to the end of apisimin through a MGO linkage. The position of the linkage is unknown but this is a representation of what it may look like. It may be that this occurs through the carboxylate group instead of the hydroxyls.

MSMS analysis the 1302 Da parent ion was performed. The MSMS data was analyzed and demonstrated to contain the N-terminal fragment of Apisimin (sequence KTSISVKGE). The amino terminal lysine residue at position 1 was not observed which indicated that the linkage between the lysine and the phenolic was most likely to occur at this point. The link between the N-temrinal lysine of apisimin and the phenolic was likely to be through MGO linkage.

Previously we had isolated a protein of the following molecular weight 5791.7695 Da which was 254 Da larger than apisimin 5537.061 Da. 5791-5537 = 254 M+1. Modification of 253 Da. We spent considerable time trying to understand what modification was likely to cause this change. Based on the spectra properties we believe that the structure is outlined below. The combination of the phenolic linked through MGO to the N-temrinus of Apisimin fragment or full length is suggested as the modified form of apisimin responsible for a range of interesting biological properties. Further analysis revealed other remarkable details amount this biomaterial which points clearly to a completely different hypothesis for the formation of DHA, MGO and energy production within Manuka honey which separates it from other honeys in its unique ability to treat a wide range of human diseases. It also points to a completely new mechanism for energy generation which is driven by primordial evolutionary understanding of the importance of reactive oxygen species in the generation of energy and the damage that highly purified plant based phenolics could be potentially causing due to interference with normal metabolic processes at night that are driven largely due to iron based Fenton reaction radical metabolic energy generation. This is a complete about face on current thinking as to the benefits of anti-oxidants. The information that is outlined below is shown in support of this hypothesis.

Parent ion 1273 Da corresponding to Apisimin N-terminus, MGO linked (C3H4 - 40 Da) trimethoxybenzoic acid (MS 212.2 Da) MGO + Trimethoxybenzoic acid (253.2 Da). The intact apisimin with this modification has also previously been identified. Previously a protease in honey has been identified by others however the actual protease has not been identified. One possible reason for this is due to the fact that there is no protease present in the honey and the peptides that are generated over time are due to reactive species generation. The presence of high concentrations of iron which may coordinate to the phenolic compounds present in honey and its potential presence in the pollen in Manuka honey may result in the formation of reactive oxygen species and the generation Fenton reaction to produce OH radical. This radical is able to cleave all compounds due to its high reactivity and it is short lived 10^-9 seconds. It may react with the phenolics that the iron is coordinated to and lead to its breakdown into polymeric glyoxal and then DHA and MGO.

polymeric glyoxal

Polymeric glyoxal

So it is proposed that the radicals cleave peptide bonds and the position of cleave is located near mineral coordination site due to the carboxylate of glutamine. Cleavage of apisimin by V8 protease produces a N-terminal peptide that is identical to that produced by the fenton chemistry confirming the site of cleavage.