Redox Diagnostics

One of the most popular trends in healthcare these days is vitamin infusion therapy, where a mix of vitamins, minerals and antioxidants (such as glutathione), is administered intravenously to patients.

The claims are very bold, ranging from detoxification of the body to regulation of sleep, mood and appetite and an overall superior hydration. They start from the premise that the mix of vitamins and antioxidants provide an antioxidant protection that is responsible for the plethora of benefits mentioned in the advertising.

But how can a healthcare professional claim that a mix of antioxidants delivered in bulk via intravenous delivery is good for the patient without an objective assessment of the redox status of the patient? How does the healthcare professional know what is the right dose for any patient? Or how much is too much? Is the actual ‘one size fits all’ approach the right approach?

We contend it is not and here is why:

- redox homeostasis is the most delicate and important control mechanism in a living system, given its critical role in maintaining the functional integrity of all its other components.
- a redox system is made of two major and complementary components: the pro-oxidant component and the reductive (or antioxidant) component. Redox homeostasis in the system is achieved when the two components are in balance.
- an imbalance created due to either an increase in the pro-oxidant components or a deficiency in reductive components leads to oxidative stress, a condition known to be at the root of any acute and chronic diseases.
- on the other hand, a decrease of the pro-oxidant components and/or an increase of the reductive components leads to a redox imbalance called reductive stress, an equally detrimental condition, less mentioned and explored in scientific literature.
- administration of reductive (antioxidants) substrates such as vitamin C and glutathione without measurement of patients’ redox status comports risks, since both substrates become pro-oxidants after consuming their reductive action and by reducing transition metals in an oxidized state such as Fe3+ and Cu 2+, thus initiating an oxidative chain of reactions. This very relevant scientific paper highlights the premise that IV administration of vitamin C leads to oxidative inactivation of proteins.
https://www.sciencedirect.com/science/article/abs/pii/S0002916523319828

There is a simple and effective solution to finding patients’ Redox Status in minutes, at Point-of-Care:

- FRAS5 – dedicated spectrophotometer with integrated centrifuge for in-minutes determination of redox status in plasma and saliva samples
- d-ROMs fast test – determines the pro-oxidant status in plasma samples
- PAT test – determines the antioxidant status in plasma samples
- SAT test – determines the antioxidant status in saliva samples
- Oxidative Stress Index (OSI) – an integrated index calculated from the values of d-ROMs fast test and PAT test, that allows for a simpler interpretation and communication of patients’ redox status

The device and tests are validated by over 1,800 scientific papers published worldwide.

More info here: https://redoxdiagnostics.com/fras5-2/

HYDROGEN PEROXIDE - A CRITICALLY IMPORTANT MOLECULE

As noted previously, hydrogen peroxide is one of the Reactive Oxygen Species (ROS), along with hydroxyl radical (HO·), peroxyl radical (HOO·) and superoxide anion (O-2), with a moderate oxidation power.
Despite its simplicity and relative instability, hydrogen peroxide is a versatile molecule that plays pivotal roles in multiple biological processes; therefore, its synthesis and degradation are both tightly regulated processes.

I. Hydrogen peroxide synthesis
There are multiple pathways of hydrogen peroxide synthesis, such as:
a. Dismutation of superoxide anion (O-2) by superoxide dismutase (SOD), per following reaction:

2H+ + 2O-2 -> O2 + H2O2

b. Degradation of monoamine neurotransmitters: hydrogen peroxide is a catalytic reaction product from the mitochondrial outer membrane enzymes monoamine oxidases (MAO) A and B. For example, oxidative deamination of dopamine (DA) by monoamine oxidase (MAO) produces hydrogen peroxide (H2O2)
and the reactive aldehyde DOPAL (3,4 dihydroxyphenilacetaldehyde).

c. Byproduct of fatty acids hydrolysis in the peroxisomes

Peroxisomes are oxidative organelles, whose main metabolic function is the breakdown of very long chain fatty acids through beta-oxidation. In presence of oxygen (O2), the long chain fatty acids are converted to medium chain fatty acids, that are then transported to the mitochondria for further breakdown to water and carbon dioxide.
Oxidation of long fatty acids leads to formation of hydrogen peroxide per following reaction:

R-H2 + O2 -> R + H2O2

d. During oxidative protein folding in the endoplasmic reticulum.
Oxidative protein folding in the endoplasmic reticulum (ER) is a significant source of hydrogen peroxide (H2O2). This process
involves protein disulfide isomerase (PDI) working in concert with ER oxidoreductin 1 (Ero1) to catalyze the formation of disulfide bonds. Molecular oxygen is the ultimate electron acceptor in this process and yields one H2O2molecule for every disulfide bond formed. Excessive amounts of hydrogen peroxide are transported out of the ER through the transmembrane channels Aquaporins-11 into the cytosol for degradation by glutathione (GSH).

II. Hydrogen peroxide degradation
There are several pathways of hydrogen peroxide degradation, such as:
a. In peroxisomes, hydrogen peroxide is used to oxidize other substrates in a reaction mediated by the enzyme catalase:

R-H2 + H2O2 -> R + 2H2O

If hydrogen peroxide accumulates in excess, catalase will degrade it to oxygen and water, per following reaction:

2H2O2 -> O2 + H2O

To be noted that catalase is an enzyme present only in the peroxisomes and not in the cytosol and the mitochondria.

b. In cytosol and mitochondria, excessive amounts of hydrogen peroxide are degraded by reduced glutathione (GSH) to water and oxidized glutathione (GS-SG), in a reaction catalyzed by glutathione peroxidase (GPx):
H2O2 + 2GSH -> 2H2O + GS-SG

c. In the erythrocytes and mitochondria, hydrogen peroxide is also degraded by peroxiredoxin (Prx), with formation of sulfenic acid and water:
PrxSH + H2O2 -> PrxSOH + H2O

d. In the presence of photons, hydrogen peroxide undergoes a spontaneous disproportionation to water and oxygen, per following reaction:
2H2O2 -> O2 + H2O

III. Hydrogen peroxide biological functions
Hydrogen peroxide is involved in multiple critical biological processes, such as:
a. Immune response – hydrogen peroxide is synthesized by the macrophages in response to invading pathogenic bacteria and is a critical part in the bacterial degradation through oxidative degradation of its organic substrates.
It also acts as a substrate for hypochlorous acid (HOCl) formation in the neutrophils, as part of the innate immune response, in a reaction mediated by myeloperoxidase (MPO).

b. Energy metabolism
As noted in a previous post, it is our understanding that generation of hydroxyl radical (HO·) from hydrogen peroxide by Fenton chemistry is part of a novel pathway of energy production that involves a HOMO/LUMO reaction between hydroxyl radical (HO·) and reduced glutathione (GSH, as follows:

H2O2 + Fe2+  HO· + HO- + Fe3+
2 HO· + 2 GSH  2 H20 + GS-SG + 2 photons

c. Signaling
Oxidation of the thiol group (-SH) of the methionine residue (Met-SH) to methionine sulfoxide (Met-S=O) is a signal that triggers protein phosphorylation, while reduction of the methionine sulfoxide is a signal for dephosphorylation. This process has major implications in proteins functionality.

d. Modulation of redox sensors
It is well established that reversible oxidative post-translational modifications of the cysteine residues by hydrogen peroxide represent an important mechanism that regulates protein structure and function. For example, formation of disulfide bonds between ATP synthase redox sensors from alpha subunit Cys294 and gamma subunit Cys103 is correlated with inhibition of ATP synthesis and ATP hydrolysis. Reversal of disulfide bond at Cys294 results in the recovery of ATP synthase activity.

It is apparent that in small amounts and under tight control of synthesis and degradation, hydrogen peroxide plays key roles in a multitude of physiological processes. However, accumulation of excessive amounts of hydrogen peroxide it is very damaging and is associated with the development of many chronic and degenerative conditions. We will address this topic in the next post.
Given the dual nature of hydrogen peroxide as a benefactor in small, physiological amounts and a detrimental molecule in excessive amounts, monitoring of the level of peroxides in blood samples should be a top priority in any healthcare protocol. This can be easily done in-office or in laboratory by using d-ROMs test and the dedicated instrument FRAS5. More information here https://innovaticslabs.com/d-roms-fast-test/

d-ROMs Fast Test | Innovatics Laboratories The d-ROMs fast test, developed initially by the renowned scientist Mauro Carratelli and upgraded by H&D srl, is a photometric test that allows to assess the pro-oxidant status in a biological sample, by measuring the concentration of hydroperoxides (ROOH).