Redox Diagnostics, Inc.

Today is an important moment for our company as we change the name and internet domain name under which we operate to Redox Diagnostics, Inc. and www.redoxdiagnostics.com, so that it better reflects the nature of our activities. It reaffirms our commitment to bring to the market the most advanced solutions for the assessment of redox status in biological systems. After years of research in the field of redox biology and related disciplines, we feel compelled to share the knowledge with like-minded individuals who understand and are willing to leverage on the major shift currently underway in the healthcare space. Our commercial endeavors will be matched by our educational efforts, aiming to contribute to the advancement of diagnostic and therapeutic solutions rooted in the right understanding of physiological and pathophysiological processes.

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 Cnt'd in 'Comments'

PEROXIDES FORMATION AND THEIR SIGNIFICANCE   We mentioned in a previous post that hydroxyl radical (HO·) formation is pivotal for energy production by reacting with glutathione (GSH) in a HOMO/LUMO reaction. What happens if there is an imbalance between hydroxyl radical formation and reduced glutathione availability? In absence of enough glutathione supply, hydroxyl radical (HO·), a very reactive molecule, will seek to stabilize by oxidizing organic substrates with lower reductive power, such as lipids, proteins and nucleic acids and initiates a chain reaction that leads to peroxides formation in the early stage and to other oxidation byproducts in the more advanced oxidation stages. We will discuss here the process of lipid peroxidation. The lipid peroxidation chain reaction happens in three steps: Initiation, Propagation, Termination. 1.   Initiation. In the initiation step the hydroxyl radical (HO·) extracts the hydride ion (H-) required for stabilization from a lipid (LH), leading to formation of a water molecule (H20) and a carbon-centered lipid radical, per following reaction: HO· + LH -> H20 + L·   2.   Propagation The newly formed lipid radical (L·) reacts with an oxygen molecule and generates a lipid peroxyl radical, per following reaction:   L· + O2 -> LOO· The peroxyl radical (LOO·) is seeking to stabilize as well by reacting with another lipid molecule (LH), generating a more stable molecule of lipid hydroperoxide (LOOH) and another lipid radical (L·), per following reaction: LOO· + LH -> LOOH + L· The newly formed lipid radical (L·) further undergoes the reaction with oxygen (O2)  generating a peroxyl radical (LOO·) and later another lipid radical (L·); the reaction propagates until a reductive substrate terminates it.   3.   Termination In presence of reductive substrates, such as glutathione (GSH), Vitamin C, vitamin E, the oxidation chain reaction is interrupted by stabilizing the reactive species through multiple mechanisms: a.    Lipid peroxyl radicals (L-OO·) are reduced to more stable lipid hydroperoxides (L-OOH) by vitamin C and vitamin E. b.    Lipid hydroperoxides are reduced by glutathione (GSH) to more stable and less toxic lipid alcohols or hydroxy fatty acids, in a reaction mediated by glutathione peroxidase 4 (GPX4): L-OOH + 2GSH -> L-OH + GS-SG + H2O c.    Lipid radicals (L·) are reduced to stable lipid molecules (LH) by glutathione (GSH) in a reaction mediated by glutathione peroxidase: 2L· + 2GSH -> 2LH + GS-SG   In absence of reductive substrates, the lipid peroxides (LOOH) undergo further oxidation into more advanced oxidation byproducts, such as malondialdehyde (MDA) and 4-hydroxynonenal (HNE).   Continued in 'Comments' section

Hydroxyl radical (HO·) and Glutathione (GSH) – the actual powerhouse of the mitochondria   The current understanding regarding generation and roles of Reactive Oxygen Species (ROS) is multifaceted, having roles attributed as signaling molecules and key players in the immune response and the inflammatory process. They are also largely considered as unwanted side effects of the energetic metabolism. By contrary, our research indicates that in case of their roles in the energetic metabolism, the generation of ROS is quintessential to energy production and not an unwanted side effect. It challenges the actual understanding that energy production takes place at Complex V, where ATP synthesis takes place. In our hypothesis, there is a coordinated chain of reactions involving generation of Reactive Oxygen Species (ROS) that leads to energy production, as follows: 1.   Superoxide anion (O2-) is generated per following reaction, mediated by NADPH oxidase:  2O2 + NADPH -> 2O2- + NADP+ + H+  2.    Superoxide anion reacts with two protons (H+) and generates hydrogen peroxide (H2O2) in a reaction mediated by superoxide dismutase (SOD): 2O2- + 2 H+ -> O2 + H2O2 3.   In presence of ferrous iron (Fe2+), hydrogen peroxide undergoes the Fenton reaction and generates one hydroxy radical (HO·) and one hydroxyl anion (HO-):  H2O2 + Fe2+ -> HO· + HO- + Fe3+  4.   Two hydroxyl radicals (HO·) react with two molecules of glutathione (GSH) and generate two molecules of water, glutathione disulfide and two photons. This is the reaction that generates energy in the mitochondria: 2 HO· + 2 GSH -> 2 H20 + GS-SG + 2 photons   Why are photons generated in the reaction between hydroxyl radical and glutathione?  According to our hypothesis, glutathione and hydroxyl radical undergo a  HOMO/LUMO reaction, in which the thiol group (-SH) of glutathione donates a hydride ion (H-) that carries a high energy electron situated on the Highest Occupied Molecular Orbital (HOMO), that will lose energy and, on the way, emit a photon, so that it can fill the Lowest Unoccupied Molecular Orbital (LUMO) of the hydroxyl radical.   The photons emitted are then stored in the phosphorus atoms of the iPO4 present in the mitochondrial matrix, which then moves to Complex V (ATP synthase) to synthesize ATP by joining a molecule of ADP.   5.   Oxidized glutathione (GS-SG) is reduced back to two molecules of glutathione by NADPH, in a reaction mediated by glutathione reductase (GSR): GS-SG – 2 NADPH -> 2 GSH + NADP+ 6.   NADP+ is reduced back to NADPH in the Pentose Phosphate Pathway (PPP), in a reaction mediated by glucose-6-phosphate dehydrogenase, a rate-limiting enzyme. Copyright 2023 Innovatics Laboratories, Inc. Continued in the 'Comments' section.

After many years of research and data collection and integration from multiple scientific disciplines, we believe we arrived at an understanding of the redox processes that take place in biological systems that challenges to large extent the current understanding of the phenomenon and at the same time offers a new perspective of it, with far reaching implications. As it is clear that the current frame of reference in healthcare does not offer the right solutions, we aim to present the information in a new, different frame of reference that will enable the development of the right therapeutic approaches. The awareness of the central role oxidative and reductive stress play in the pathophysiology of many acute and chronic and degenerative diseases is quite high in the naturopathic and alternative medicine sectors, but it is still very limited in mainstream medicine and dentistry.  We believe this awareness must reach mainstream levels and we aim to contribute to that goal. While targeted therapies for redox homeostasis are limited, currently, to combinations of natural antioxidant extracts and vitamins, there is very limited testing of the level of oxidative stress in the subjects, leading to high risk of converting oxidative stress to reductive stress, among others. There is high value in knowing the levels of pro-oxidant and anti-oxidant status in subjects receiving antioxidant therapies and not only. It empowers the healthcare professionals to adjust and maximize the outputs of their therapeutic protocols. We will start sharing our accumulated knowledge and insights in a series of posts on the company page and will correlate the information with the importance of testing for oxidative stress in general and with our unique FRAS5 testing system in particular. #redoxhomeostasis #oxidativestress #reductivestress #FRAS5 #dROMstest #PATtest #OxidativeStressIndex #InnovaticsLaboratories