Last Updated 20 Oct 2026

Preliminary data analysis

An exploratory comparison of de-identified Oligoscan and Theriome Aristotle Plus results. The aim is to examine shared marker patterns, differences between examples, and their coherence with proposed biochemical mechanisms.

This analysis describes eighteen selected report pairs. Assessing causes, tissue deficiency or enzyme activity requires additional clinical and experimental evidence.

Patterns in data

Each example pairs one Oligoscan report with one Theriome Aristotle Plus report. Seventeen Theriome reports were supplied as PDFs. Example 4 was an early vendor-supplied CSV. The prepared data contain 42 direct Oligoscan rows and 296 direct Theriome analyte rows per example: 6,084 direct observations in total, with three unavailable numerical scores. A further 415 inferred cofactor scores are available separately.

Aluminium - all eighteen Oligoscan tool outputs flag aluminium as elevated. Scores range from 44.1 in Example 17 to 79.8 in Example 16. These values use the tool's metal scoring scale.

Silicon - all eighteen Oligoscan outputs flag silicon as Low, with deviation scores from -47.6% to -25.2%. More negative silicon scores tend to accompany higher aluminium scores: Spearman rank correlation -0.79 across these eighteen reports.

Magnesium and ATP - all eighteen Oligoscan outputs label magnesium low. Fourteen Theriome outputs flag ATP as IR-LOW, Examples 7 and 18 flag it HIGH, Example 13 flags it IR-HIGH, and Example 10 has no ATP status flag.

NAD reference profiles - in all seventeen PDF reports, NADP is closer to its reference centre than NAD, while NADPH is further below its reference centre. This pattern recurs across the seventeen sets of source scores.

Example 4 - reported use of sublingual NAD+ and NAG adds potential confounding to its NAD and sugar-metabolite profile. Its CSV scoring method also differs from the PDF reports. These factors limit interpretation of its contrast with the other examples. A replacement PDF from the same original specimen has been requested to clarify the reporting-format difference. Supplement exposure remains relevant to interpretation.

The direct-marker comparison uses reported analyte rows. The tools' inferred NAD, NADP, NADPH and ATP deficiency scores are available separately in the explorer. These derived scores depend on the observations used to calculate them.

For more on the potential causes and impact of these patterns, see remineralisation and toxic metals.

Other recurring marker patterns

The following comparisons use direct analyte rows from the seventeen Theriome PDF reports. Here, low flags combine LOW and IR-LOW; high flags combine HIGH and IR-HIGH. These are the tool's classifications. An unflagged result retains its reported score and has no source status flag.

Biotin - carries an IR-LOW flag in all seventeen PDF reports, with z-scores ranging from -0.81 to -0.53.

5-Hydroxytryptophan - carries an IR-LOW flag in all seventeen PDF reports, with z-scores ranging from -0.74 to -0.59.

Fructose - fourteen of seventeen PDF reports carry a high flag. Examples 12, 13 and 17 are unflagged. Fifteen of seventeen scores are above the reference centre; Examples 12 and 13 are slightly below it.

Fructose 1,6-bisphosphate - sixteen of seventeen PDF reports carry a low flag. Example 7 is unflagged, with a z-score of +0.40. High fructose and low fructose 1,6-bisphosphate occur together in thirteen of seventeen reports.

N-Acetyl-D-galactosamine - all seventeen PDF scores are below the reference centre, with low flags in sixteen reports. Example 14 is unflagged, with a z-score of -0.49.

Uridine diphosphate-N-acetylglucosamine - all seventeen PDF scores are below the reference centre, with low flags in fifteen reports. Examples 12 and 17 are unflagged, at -0.46 and -0.45 z respectively. This marker and N-acetyl-D-galactosamine both carry low flags in fourteen of seventeen reports.

Mannose 6-phosphate - eleven of seventeen PDF reports carry high flags. Examples 6 and 16 carry low flags; Examples 8, 12, 14 and 18 are unflagged. The predominant direction is upward, with variation between examples.

Glutamic acid - eleven of seventeen PDF reports carry high flags. The other six, Examples 1, 3, 9, 12, 14 and 18, carry low flags.

Gamma-aminobutyric acid (GABA) - all seventeen PDF scores are below the reference centre, ranging from -0.81 to -0.19 z. Fourteen reports carry an IR-LOW flag; Examples 11, 12 and 15 are unflagged. High glutamic acid and low GABA flags occur together in nine of seventeen reports.

Betaine and methionine - both carry an IR-LOW flag in all seventeen PDF reports. Betaine z-scores range from -0.81 to -0.62; methionine ranges from -0.75 to -0.54.

Creatine - all seventeen PDF scores are below the reference centre, ranging from -0.77 to -0.15 z, with a median of -0.59. Twelve reports carry an IR-LOW flag; Examples 6, 8, 11, 14 and 17 are unflagged.

Phosphocreatine - all seventeen PDF scores are below the reference centre, ranging from -0.67 to -0.49 z. Sixteen reports carry an IR-LOW flag; Example 13 is unflagged at -0.49 z. Creatine and phosphocreatine show a shared downward pattern. Establishing impaired synthesis requires additional evidence.

Example 4's CSV also flags biotin and 5-hydroxytryptophan low. It flags glutamic acid high and GABA, betaine and methionine low. Creatine is flagged HIGH at +543.1% on its CSV reference-based scale; phosphocreatine is IR-LOW at -69.6%. Its low fructose and high N-acetyl-D-galactosamine contrast with the predominant PDF directions. Its different scoring method and reported supplement exposure need to be considered together.

Urea-cycle-related markers

Arginine, citrulline, ornithine, aspartate and fumarate participate in the urea cycle. Their direct scores in the seventeen PDF reports show the following patterns.

Arginine and citrulline - each carries low flags in nine of seventeen PDF reports. Arginine scores are below the reference centre in fifteen reports; Example 15 is HIGH at +1.40 z, and seven reports are unflagged, including Example 18 at +0.45 z. Citrulline scores are below the reference centre in fourteen reports; Examples 14 and 18 are IR-HIGH at +0.77 and +0.79 z respectively. Six reports are unflagged, including Example 6 at +0.20 z.

Ornithine and aspartate - ornithine carries an IR-LOW flag in nine of seventeen PDF reports, with eight unflagged results and no high flags. Eleven scores are below the reference centre, five above it and one at zero. Aspartate carries an IR-LOW flag in eight reports. It has twelve scores below the reference centre, eight unflagged results and an IR-HIGH flag in Example 16 at +0.89 z.

Fumaric acid - fourteen PDF reports are unflagged. Examples 6 and 16 carry IR-HIGH flags and Example 7 is HIGH. Eleven scores are below the reference centre, with no low flags.

Example 4's CSV flags arginine, citrulline, ornithine and aspartate low; fumaric acid is unflagged. Urea, ammonia and argininosuccinate are absent from the prepared dataset. Assessing urea-cycle flux or ammonia clearance requires additional measurements and clinical context.

Explore the paired data

Select an example to follow its NAD pathway profile, inspect individual markers and their peaks or troughs, or compare grouped heatmaps. Marker summaries are calculated within their source scoring group, with separate summaries for CSV and PDF direct scores.

Data analysis controls

Direct report values and inferred scores stay separate. Example 4 uses a different Theriome scoring method.

Loading de-identified example data...

NADP formation and redox handling

NAD kinase phosphorylates NAD+ to NADP+ using ATP. Magnesium-dependent activity has been demonstrated in human NAD kinase preparations. A distinct mitochondrial NAD kinase also exists. These findings provide a biochemical basis for examining magnesium, ATP, NAD and NADP together. Williams and Jones, 1985; Ohashi and colleagues, 2012.

Two questions guide this comparison: whether the marker pattern is compatible with constraints on NADP formation, and whether NADP/NADPH findings are compatible with altered reduction or consumption. Both require several markers and relevant clinical context. NADPH metabolism is compartmentalised; resolving cytosolic and mitochondrial processes requires compartment-specific methods. Lewis and colleagues, 2014.

Examples 7 and 18 combine high ATP with low Oligoscan magnesium. Example 8 combines IR-LOW ATP with a NADP score of -0.11 z. These combinations help assess how well a proposed explanation accommodates variation across the examples.

Estimating NADPH:NADP concentration ratios, total NADP/NADPH pool size or NAD kinase activity requires suitable measurements, units and assay calibration. These inputs are unavailable in the prepared data.

Silicon and aluminium

Low silicon and elevated aluminium co-occur across all eighteen Oligoscan reports. The inverse rank association describes variation within this selected set of tool scores. Exposure history, diet, assay behaviour and other shared influences could contribute to it.

Silica chemistry provides a reason to examine these markers together. In a tracer study of three healthy volunteers, oligomeric silica reduced gastrointestinal aluminium uptake. The monomeric form showed no significant effect under the study conditions. Jugdaohsingh and colleagues, 2000.

Those findings concern chemical form and gut absorption. Applying this mechanism to the report pattern requires information about exposure, silicon form and independent measurements of aluminium handling. Both scores here come from the same Oligoscan report, so their association also requires validation with independent measurements.

Common patterns described in MPO

The following proposed patterns and mechanisms form the working interpretation framework for the Metabolic Pathway Overlay examples. Each requires assessment against the individual reports and supporting evidence. Mechanism-specific scoring is a future analysis step.

  1. Elevated toxic metals, with mineral and electrolyte deficiencies.
  2. A reduced total NADP/NADPH pool and impaired conversion of NADP to NADPH (low NADPH), with potentially broad effects. The reported exception is where aluminium is not elevated.
  3. Constraints on glycolysis and glycogen synthesis.
  4. Altered pyruvate and/or lactate levels.
  5. Constraints on the pentose phosphate pathway and downstream PRPP synthesis.
  6. Impairments in purine and pyrimidine metabolism.
  7. Oxidative stress.
  8. TCA cycle alterations and constraints.
  9. Issues with NAD+ synthesis and redox balance.
  10. Reduced carnitine synthesis and associated carnitine-shuttle constraints.
  11. Elevated glutamate relative to low GABA, except where severe malnutrition constrains glutamate synthesis.
  12. Issues with glutathione metabolism.
  13. Issues involving magnesium and Mg-ATP.
  14. Urea-cycle and nitric oxide synthase impairments, with potential effects on vascular tone.
  15. Dysregulated neurotransmitter synthesis and degradation.
  16. Impaired conversion of choline to betaine.
  17. Impaired methylation and associated SAMe, glycine, and cysteine metabolism.
  18. Impaired creatine synthesis.

Testing the exception in pattern 2 requires a comparison group with non-elevated aluminium; all eighteen examples here have elevated aluminium scores. Assessing the malnutrition exception in pattern 11 requires clinical nutritional information, which is unavailable. The proposed changes in pool size, pathway flux and enzyme activity also require further investigation.

How to read the scores

Theriome PDFs - the displayed score is the source z-score multiplied by 100, consistent with MPO's numerical convention. For example, -168 means z = -1.68, or 1.68 standard deviations below the reference centre. This explorer has its own labelled heatmap colour scale.

Example 4 CSV - the original tool-derived relative-abundance/reference score is retained and summarised separately from the PDF scores.

Oligoscan - mineral deviation scores, vitamin percentages and metal scores retain their source meanings. Each scale requires its own interpretation.

Inferred cofactors - rules applied to report observations generate these visual scores. Their clinical significance requires independent validation. Missing inferred rows remain unreported.

Missing data - unavailable scores appear as NA. Blank source status flags remain blank. Both retain an unspecified status in the comparison.

Limits of this comparison

The dataset contains eighteen selected pairs and lacks a healthy control group. Its population relevance is unknown. Collection dates, the interval between paired tests, specimen details, units and most clinical context are unavailable in the prepared data. Example 4 has reported exposure to sublingual NAD+ and NAG; doses and timing are not recorded here. Supplement use in the other examples is unspecified. Tissue and biochemical compartment differences remain unresolved.

Examples 1-10 retain their original prepared results. Examples 11-17 were processed with newer parser versions: Oligoscan 1.6.9 and Universal OAT 0.1.43, compared with 1.6.6 and 0.1.40 for the original batch. Example 18 uses Oligoscan 1.6.9 and Universal OAT 0.1.44. Parser versions are retained in the downloads; rule-derived comparisons may be affected by changes between versions.

There are many markers relative to the number of examples. Apparent correlations may be sensitive to individual cases, scoring conventions, selection and shared reference distributions. Analysis is limited to descriptive summaries. A direct-row label records the parser's classification; assay validity requires separate evaluation.

Pending

Resolve Example 4 - compare the replacement PDF with the retained vendor CSV from the same specimen, and review the timing and doses of the reported NAD+ and NAG use when interpreting its profile.

Describe marker coherence - compare direction, magnitude and variability within compatible scoring groups, retaining all exceptions and denominators.

Mechanism map and references - MPO already provides much of the mechanism mapping. A supporting table of references is in progress.

Assess concordance - show supporting observations, contradictions and missing evidence. Report mechanistic evidence, measurement relevance, coverage and sensitivity to individual examples separately.

The downloadable CSV and JSON contain the prepared de-identified observations, source scores and marker identities for future analyses.

Last Updated 5 Feb 2026


by
Joshua Leisk | (@joshual_tm on X/Twitter)
[Please note, this is a large webpage (>250 printable pages) with dynamic content. A quick-start guide can be found here. Please allow a few moments for all database tables to load.]


What's new?

Here are the 10 most recent changes / updates:

For more detailed information, please see Changelog
Please report any bugs / errors.

NB. Whilst efforts have been made to make this information accessible on mobile devices, it is best viewed on a computer.

Last Updated 3 May 2026

This is an experimental protocol based on the disease model by Leisk et al., (preview here - https://bornfree.life/learn/) which may be helpful for supporting the unique nutritional and wellness / lifestyle requirements of teenagers and adults with chronic illnesses that share various common features, including mitochondrial dysfunction, hormone dysregulation, ferritin and other mineral deficiencies, microbiome dysbiosis, biofilms, neurotransmitter dysregulation and immune dysregulation, etc., such as:

Myalgic Encephalomyelitis / Chronic Fatigue Syndrome (ME/CFS) Long COVID, Post-Vaccine Syndrome Viral / Post-Viral Chronic Fatigue
Post-Exertional Malaise (PEM) “Floxed” Guillain-Barre Syndrome (GBS)
Chronic Inflammatory Response Syndrome (CIRS) Breathlessness Hypoxia
Acidosis / Alkalosis / pH Dysregulation Dysregulated Cortisol Dyslipidemia
Post Finasteride Syndrome (PFS) Post SSRI Sexual Dysfunction (PSSD) Post Accutane Syndrome
Small Fibre Peripheral Neuropathy (SFPN) Mast Cell Activation Syndrome (MCAS) Diamine Oxidase (DAO) Deficiency
Stiff Person Syndrome Gut Fermentation Syndrome Glycogen Storage Disease (functional)
Histamine intolerance Brain Fog Sleep disorders
Anxiety Postural Orthostatic Tachycardia Syndrome (POTS) Autism Spectrum Disorders (ASD)
Fibromyalgia Dysautonomia Frequent Urination
GI Disorders “Small Intestinal Bacterial Overgrowth” (SIBO) Vitamin B6 Toxicity
Vitamin A Toxicity Candidiasis Aspergillosis
Ehlers Danlos Syndrome (EDS) Joint Pain Connective Tissue Disorders
Macular Degeneration Multiple Sclerosis (MS) Parkinson's Disease (PD)
Tinnitus Polycystic Ovary Syndrome (PCOS) Erectile Dysfunction
Elevated Cholesterols / Fasting Glucose / D-Dimer Hashimotos Alopecia
Major and other types of Depression Cataracts Migraines
Last Updated 11 Feb 2026

Please read the entire protocol documentation before taking any action based on its contents.

Medical Disclaimer:

This protocol is for research and educational purposes only. Always consult your doctor to determine if this information is appropriate for you, especially if you are taking medications like SNRIs/SSRIs, beta-blockers, MAO inhibitors, or drugs metabolized by CYP2D6 enzymes.


Important Notice:

While we strive to minimize adverse effects, you may experience some temporary symptoms when starting this protocol -

  • Paradoxical Effects: As deficiencies are corrected and neurotransmitter balance is restored, you might notice short-term increases in adrenaline and heart rate.
  • Copper Deficiency Correction: Addressing a severe copper deficiency may cause temporary kidney pain and nausea.
  • Immune Activity Increase: An initial boost in energy availability can heighten immune activity and symptom severity. This might be psychologically challenging and may cause hesitation in continuing the process.
  • Herxheimer Reactions: You may experience effects from microbial die-off, such as increased "burning" sensations and headaches.
  • Hormonal Changes: Temporary changes in libido or menstrual cycles can occur.
  • Inflammation and Flu-like Symptoms: Improvements in immune function might lead to brief periods of inflammation, fever, flu-like symptoms, nausea, and fatigue.

Support:

We host an online discussion and support group on Discord. Feel free to join us:

https://discord.gg/CAMXV78


Monitoring Recommendations:

In addition to the tests used in the protocol, we suggest regular blood tests to monitor:

  • D-dimer Levels
  • Vitamin B6 Levels
  • Cholesterol Levels
  • Fasting Glucose Levels
Last Updated 4 Feb 2026

Warning: This documentation contains a significant amount of information and it is expected that you will need multiple read-throughs and a number of hours to absorb and / or action the information.

How to Navigate This Information

There are multiple ways to approach this material:

  1. First Reading: We suggest that you read everything from start to finish in a linear fashion. This will help you become acquainted with all of the concepts.

  2. Guided Navigation: If you decide to act on the information, return here and use the visual navigation tool below to step through the protocol in a logical and structured manner.

  3. AI-supported (experimental): If you would like AI assistance in understanding the model and/or protocol, Google's NotebookLM is able to perform this function for free. A setup guide is available here - https://bornfree.life/learn/notebooklm-ai/ 


Last Updated 3 Oct 2026



Long-term illness can affect energy, immunity, mineral balance and the signals between the nervous system and the gut. The Born Free model asks how problems in these systems could reinforce one another and make recovery difficult.

It proposes several possible routes into a persistent defensive state, rather than one cause for everyone. The explanation below brings those ideas together. It is a working model, not an established explanation for every illness or a guide to individual treatment.

The proposed model

Cells need to produce energy, move minerals, manage chemical stress and respond to danger. These tasks are connected. An infection, for example, can change immune activity and the resources available for energy production.

The model proposes that several disruptions could become self-reinforcing. Less available energy may make maintenance and recovery harder. Continued inflammation, altered mineral handling and stress signals could then place further demands on the same systems.

Four areas that may interact

The proposed interaction model links energy, mineral handling, immune activity and nervous system and gut signalling. The arrows show possible feedback, not a universal sequence or proven causation.

A defensive response can be useful during short-term danger. The question here is whether some of the responses could continue after the initiating event, or be renewed by ongoing threats. This proposed persistence is the central idea of the model.

How the cycle could begin

There may be more than one starting point. The model considers infection, immune activation, toxic metal exposure, poor nutrient availability, gut disturbances, connective tissue problems and sustained physiological stress.

Sleep loss, heat, surgery, pregnancy, overexertion or trauma could add to an existing burden. It also considers the effects of neurodevelopmental stress and the demands associated with neurodivergence. These are possible contributors, not an explanation of an individual person's illness.

Some people may lose capacity gradually before a major event pushes them beyond what they can tolerate. Others may become unwell more abruptly. A prenatal contribution is one possibility among these routes.

Optional detail: the prenatal example

The model proposes that conditions during pregnancy could influence a child's later vulnerability. Its examples include maternal metal exposure, mineral status, immune activation, inflammation, stress hormones and glucose handling.

It also asks whether gestational diabetes could sometimes involve difficulty storing and processing glucose, alongside high blood glucose itself. Magnesium, phosphate, zinc, selenium, folate and choline appear in this proposal because of their roles in metabolism and nutrient-dependent processes.

This example does not establish prenatal conditions as the usual origin of chronic illness. It cannot identify the cause of a person's illness or attribute responsibility to a parent.

Why it could persist

Energy production and chemical balance

Cells transfer electrons when they process fuel and maintain antioxidant defences. This is called redox. Antioxidants help manage reactive chemicals that can damage cells when their production exceeds the body's capacity to handle them.

Two important redox systems are NAD⁺/NADH and NADP⁺/NADPH. These chemical carriers accept and donate electrons. NAD⁺ supports fuel processing; NADPH helps recycle antioxidants, including glutathione. Glutathione contributes to antioxidant defence and the handling of some metals and reactive chemicals.

The model proposes that impaired redox function could make energy production, chemical clearance and repair less reliable. Continued stress on these processes could then make redox function harder to restore.

ATP inside and outside cells

ATP, or adenosine triphosphate, transfers energy for work inside cells. Outside cells, it can also act as a signal. ATP released during tissue stress or damage can activate receptors on immune cells and other cells.

ATP can do work or carry a signal

Proposed persistence: the model asks whether repeated release or altered signal breakdown could sustain immune activity.

Inside cells, ATP transfers energy for cellular work. ATP released outside cells can act as a stress or damage signal through receptors.

Extracellular ATP is broken down through ADP and AMP to adenosine; adenosine can then be converted to inosine. These molecules have their own signalling effects. The model proposes that changes in this sequence could affect how an inflammatory response resolves.

Mineral amount and mineral location

A mineral's total amount and its availability in a particular tissue are different questions. Minerals move between the gut, blood, cells and storage sites. Inflammation can affect some of this movement.

Iron provides a specific example. Hepcidin is a hormone that regulates iron movement through ferroportin, a protein that exports iron from cells. This does not prove that all minerals or toxic metals share the same regulation.

Amount is not the same as location

AmountHow much is present?

Location and availabilityWhere is it, and can it be used?

Conceptual mineral movement between circulation and tissues. The central symbol represents transport pathways, not a single transporter shared by all minerals. No quantities or diagnostic interpretation are shown.

The model extends this question to essential minerals and toxic metals: could altered transport, storage, loss or chemical binding leave some processes short of usable resources? That broader proposal needs evidence for each mineral, tissue and condition.

Immune activity, histamine and stress responses

Mast cells are immune cells found in tissues. They can release histamine and other signalling molecules. Histamine affects blood vessels, the gut, nerves and immune responses, as well as participating in allergic reactions.

Histamine release and histamine breakdown are separate processes. The model considers both increased release, following immune or tissue stress, and reduced clearance. It proposes that these changes could interact with glucose storage, inflammation and nervous system activity.

The sympathetic nervous system helps mobilise energy and maintain alertness during stress. The model asks whether sustained activity in this system, together with disturbed glycogen storage, could contribute to fatigue alongside agitation, tremor or a racing heart. Glycogen is the body's stored form of glucose.

Post-exertional malaise (PEM) is a worsening of symptoms after physical or mental activity. The model seeks to explain why exertion could place demands on several already strained systems. This is not evidence that every post-exertional symptom has the same mechanism.

Gut microbes, tissue injury or unresolved immune triggers could also keep stimulating these responses. Once several loops are active, the initial trigger may no longer explain the whole picture.

Optional mechanism detail

The optional sections explore the proposed biochemical connections in more detail.

Microbial products, acetaldehyde and nutrient function

The model proposes that altered gut movement, bile flow, tissue oxygen availability, immune activity and glucose handling could favour different microbial activity. Its examples include fermentation products such as ethanol, acetaldehyde, D-lactate, hydrogen sulphide and ammonia. Their effects depend on amount, location and context.

Acetaldehyde is a reactive chemical produced during alcohol metabolism and by some microbes. Aldehyde dehydrogenase enzymes, abbreviated to ALDH, help clear it. These reactions use NAD⁺. The model proposes a feedback loop in which impaired NAD⁺ recycling limits clearance, while acetaldehyde adds further chemical stress.

Biotin (vitamin B7) enters intestinal cells through the sodium-dependent multivitamin transporter, or SMVT. Microbial products may reduce this uptake in different ways. In human colon cells and biopsy-derived cultures, prolonged acetaldehyde exposure reduced biotin uptake and the amount of SMVT produced (Ramamoorthy et al., 2021). In separate intestinal-cell experiments, bacterial lipopolysaccharide (LPS) reduced uptake by decreasing the amount of SMVT at the cell surface, without reducing the total amount of SMVT in the cells; reduced uptake was also observed in mice (Lakhan & Said, 2017). Within this model, these findings provide a possible link between microbial activity and reduced biotin availability. The size of this effect in people with chronic illness has not yet been established.

The model also proposes effects on zinc availability, magnesium-dependent enzymes, thiamine binding and active vitamin B6 handling. It includes altered riboflavin conversion into FMN and FAD, folate and vitamin B12 metabolism, SAMe, retinoic acid signalling, CoQ10 production and BH4 oxidation.

FMN and FAD are forms of riboflavin used by enzymes. SAMe participates in methyl-group transfer. BH4 is a helper molecule for enzymes involved in nitric oxide and neurotransmitter production.

The term functional deficiency is used here for a proposed situation in which a nutrient is present but its use is impaired. This cannot be inferred from symptoms or from a normal or abnormal blood value alone.

For histamine, the model distinguishes mast-cell release from breakdown through pathways involving diamine oxidase (DAO) and other enzymes. It proposes that acetaldehyde, nutrient availability and redox changes could affect clearance. ALDH's role in aldehyde handling should not be treated as the same process as DAO's action on histamine.

In the proposed cycle, histamine-related changes in liver glycogen use and responsiveness to adrenaline could contribute to unstable glucose availability, orthostatic symptoms and PEM. The model also considers stronger histamine or inflammatory responses to exercise. These links remain hypotheses within the model.

Nitric oxide, BH4 and mitochondrial redox

Nitric oxide helps regulate blood-vessel relaxation. BH4 supports the enzymes that make it. Under some conditions, impaired BH4 function can contribute to nitric oxide synthase becoming uncoupled, producing superoxide instead of normal nitric oxide output.

Nitric oxide and superoxide can react to form peroxynitrite. The model proposes that this chemical stress could inhibit mitochondrial complex I, affect NAD⁺/NADH balance and make acetaldehyde clearance harder. Complex I is part of the mitochondria's electron transport chain.

Endogenous morphine, cAMP-PKA and CREB

The model proposes that morphine produced within the body, described as endogenous morphine, and sympathetic activity could pull a cell-signalling pathway in opposite directions. That pathway involves cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA).

In the proposed mechanism, activation of µ-opioid receptors engages inhibitory G proteins (Gi), reduces adenylyl cyclase activity and lowers cAMP. β-adrenergic signalling engages stimulatory G proteins (Gs) and can increase cAMP. Adenylyl cyclase is an enzyme that produces cAMP.

The model proposes that prolonged inhibition could produce compensatory changes in neurons and liver cells, including increased cyclase, PKA and CREB activity. CREB, the cAMP response-element binding protein, helps control gene transcription. Its proposed role is a persistent change in the cell's response to later signals.

The model links these changes to tyrosine hydroxylase and catecholamine synthesis in neurons, and to glycogen phosphorylase, glycogen synthase and glucose release in liver cells. It proposes that reduced opioid restraint, alongside high sympathetic drive, could lead to larger swings in cAMP, adrenaline and glucose availability.

It further links glutamate, calcium stress and extra-synaptic NMDA receptors to increased nervous system excitability. NMDA receptors respond to the neurotransmitter glutamate. These proposals are used to explain why relatively small changes might produce a disproportionate response.

The protocol aims to reduce excessive cAMP-PKA fluctuations while supporting glycogen storage and redox function. The proposed role of endogenous morphine, and the safety and benefit of modifying these pathways, remain to be tested.

Mineral handling, acidaemia and the longer proposed loop

The model links mitochondrial dysfunction, greater reliance on glycolysis, lactate accumulation, acid-base changes, inflammation and kidney losses. Glycolysis is the initial breakdown of glucose; mitochondria process fuel through the TCA cycle and electron transport chain.

Acidaemia means an abnormally low blood pH. It should not be assumed from fatigue, muscle discomfort, exertion or a subjective feeling of internal “sourness”. Greater glycolytic activity or lactate production does not by itself establish persistent acidaemia.

The model proposes that inflammatory signals, including TNF-α, IL-1β, IL-6, IL-10 and IL-22, could participate in altered mineral handling. It discusses hepcidin, ferroportin and divalent metal transporter 1 (DMT1). Their involvement in iron transport does not validate a single mechanism for all essential minerals or toxic metals.

It also considers reduced intestinal absorption, tissue sequestration, chemical binding and urinary loss. It names phosphate, calcium, magnesium, sodium, potassium and zinc in relation to kidney handling and energy chemistry. It also proposes acetaldehyde-related changes involving zinc and silicon, and effects involving lithium and rubidium. These are elements of the model, not established individual deficiencies or indications for supplementation.

The model proposes altered homeostasis across at least 12 minerals or metals, without a demonstrated common regulatory pathway. Blood and red-cell measurements address particular compartments and cannot confirm the model's proposed tissue pattern on their own.

The longer loop can be summarised as follows:

  1. Acid-base changes and mineral losses could impair energy chemistry.
  2. Impaired energy and redox function could reduce glutathione capacity and chemical or metal handling.
  3. Acetaldehyde burden and altered histamine clearance could affect glycogen stability and stress signalling.
  4. Stress signalling and extracellular ATP could contribute to continued inflammation.
  5. Inflammation could further affect resource availability, including iron handling.

This is a set of proposed feedback links, not a demonstrated chain that must occur in that order. An ongoing infection, tissue injury or immune trigger could also contribute. Low circulating iron or phosphate, altered ferritin, temperature instability, poor concentration and orthostatic intolerance do not uniquely identify this model.

Connective tissue and persistent immune triggers

The model includes connective tissue maintenance as part of the proposed cycle. It considers collagen production in relation to oxygen, iron, vitamin C, copper, silica, glycine, proline, lysine, ATP, redox function and enzymes such as prolyl hydroxylases.

It proposes that poor resource availability, hypoxia signalling, acid-base disturbance, oxidative stress or inflammation could affect tissue maintenance. Tissue fragility or mechanical stress could, in turn, release extracellular ATP and stimulate mast cells.

Similar symptoms or tissue features do not establish hypermobile Ehlers-Danlos syndrome (hEDS) or inherited vascular Ehlers-Danlos syndrome (vEDS).

The model proposes that inflammation could coexist with impaired pathogen clearance. It considers poor sleep, low usable energy, mineral misallocation, cortisol dysregulation, weak interferon responses and impaired nutrient activation as possible contributors.

Viral, bacterial, fungal or other reservoirs could then renew inflammatory signals. The proposed wider effects include neurotransmitters, stress hormones, vitamin metabolism, cholesterol, histamine, kidney function and temperature regulation. These possibilities require evidence in the relevant condition and cannot be inferred from a shared symptom pattern.

The protocol's proposed rationale for changing the trajectory

Changing one input may be insufficient if several feedback loops remain active. The protocol aims to restore coordination across mineral availability, glutathione and redox function, chemical clearance, glycogen storage, immune activity, nervous system regulation, connective tissue maintenance and the microbiome.

Its “Reversing the Trajectory” section groups the proposed work into four areas: nutrient and metal handling; microbiome disturbances and persistent infections; structural and lymphatic issues; and the physical and psychological consequences of chronic illness. These aims are not evidence that the proposed interventions work.

The model proposes that improving nutrient availability and addressing toxic metal burdens could support energy metabolism, redox function and immune responses. The proposed link from mitochondrial dysfunction to immunodeficiency and chronic herpesvirus reactivation is a modelling inference.

The model also considers chronic Lyme disease and T. gondii among possible persistent infectious burdens. Naming these examples does not establish their presence in a particular person, or show that restoring metabolism clears an infection.

The microbiome proposal discusses mucosal surfaces, biofilms, pathogen reservoirs and microbial products such as mycotoxins and endotoxins. The structural proposal concerns posture, lymphatic function, blood flow and tissue oxygen availability. Their inclusion in the model does not demonstrate that treating them resolves the wider cycle.

Living with chronic illness can be traumatic, especially after medical missteps or dismissive care. The protocol considers metabolic and infectious burdens alongside the need to recover a sense of safety. Stellate ganglion blocks are included as an example of altering autonomic activity; whether such changes can provide durable benefit remains to be tested.

Illness is not evidence of psychological weakness or lack of effort. This model does not establish that trauma support must wait until physical problems are resolved, or guarantee that either type of intervention will restore normal metabolism.

Evidence and uncertainty

The model combines recognised biological processes, findings from particular conditions and proposed links between systems. Evidence for one process, such as hepcidin's role in iron regulation, is not evidence for every connection in the larger model.

Patterns of deficiencies and biomarkers may help characterise research subgroups and generate hypotheses about disease features and severity. They do not provide reliable individual prediction on their own. See references [121], [122], [123] and [124] for research on these patterns.

Early reports from people using the experimental Born Free protocol include significant improvement and long-term remission in some individuals. Formal clinical trials are planned to evaluate the protocol systematically. These reports are not trial results.

Reported experiences

Severe fatigue, post-exertional crashes, disturbed sleep, cognitive difficulty and a mixture of agitation and exhaustion can be distressing and disabling.

Some people report brief periods of clearer thinking or easier movement, changes in appetite or sleep, and later periods of steadier energy. Others describe plateaus, relapses, anxiety, palpitations, gut symptoms or renewed fatigue. Responses vary.

Why recovery may feel uneven

The model proposes a period of adjustment as sympathetic drive eases and parasympathetic activity becomes more prominent. The sympathetic system helps mobilise energy and maintain alertness; the parasympathetic system supports rest and digestion. The systems supporting steadier energy and recovery may lag during this proposed transition, and the balance of changes may differ between people.

People report periods of lower energy, reduced motivation or less enjoyment during recovery. The model anticipates that these experiences may sometimes accompany the adjustment. Anhedonic tone means a reduced capacity to feel pleasure or enjoyment.

Immunological debt is the model's term for unresolved immune burdens. It proposes that, as energy systems recover, renewed immune activity may begin to address some of these burdens. The model interprets some reported changes in immune-related symptoms as possible signs of this renewed activity.

The model proposes that some of these changes could reflect greater flexibility in using glucose and fats. Changes in symptoms do not, by themselves, show whether an intervention is helping or explain the mechanism.

The protocol favours a gradual approach, with room to pause or adjust when symptoms change. Its purpose is to avoid repeated bursts of activity or intervention followed by a crash.

Recovery patterns and proposed mechanisms

The model considers how starting several antioxidants, NAD⁺-related products, acetaldehyde-directed products, dietary changes or antimicrobials together could be followed by brief improvement and then anxiety, irritability, insomnia, pain, sweats, fatigue, tremor, palpitations, bowel symptoms or cravings.

It proposes microbial alcohol and acetaldehyde changes, and endogenous morphine signalling, as possible explanations. Those descriptions do not establish withdrawal or exclude an adverse effect or another cause of worsening.

The proposed staged approach includes exogenous NAD⁺, active B vitamins, minerals, antioxidant support, carbohydrate intake and later NAD⁺ precursors or cAMP-PKA modulation. Proposed acetaldehyde-directed approaches include NAC, DHM, Acetium, Z-Biotics and L-carnosine, followed by targeted antimicrobials and prebiotics or probiotics.

Light exposure, sleep, gentle movement and the ability to pause are also part of the proposed approach. Reported changes include sugar cravings, appetite, emotional responsiveness, sleep depth and dreams, along with easier activity or shorter post-exertional symptoms.

The model proposes a gradual return of flexibility in using glucose and fats. It considers whether glycolysis could improve before fatty acid oxidation, alongside possible “keto flu”-like sensations, changing carbohydrate needs and fluctuations after stress, infection or poor sleep.

Some people also describe stronger emotional responsiveness or old memories returning. Support and a sense of agency matter without requiring a particular mechanistic explanation.

Further illustrations

These illustrations explore proposed pathways and stages of recovery. The stages remain to be tested.

Coloured inward spiral linking mineral depletion, pathogens, trauma, microbiome changes, metabolism, oxidative stress and immune activity in a proposed worsening cycle.
Proposed worsening cycle
Coloured outward spiral showing proposed connections among mineral restoration, metabolism, immune activity, circulation, microbiomes and nervous-system recovery.
Proposed recovery cycle
Flowchart linking proposed work on deficiencies, oxidative stress, immune responses, microbiomes, structural problems and trauma.
Proposed trajectory diagram
Flowchart contrasting rapid reversal with a proposed phased recovery approach, with reported experiences beside each phase.
Proposed recovery stages
Open image
Last Updated 21 Oct 2026

This protocol involves several steps that require multiple tests and a wide range of important supplements. While the expected results are unlikely to be realized without completing all these steps, efforts have been made to simplify them as much as possible. The steps are as follows:

  1. 1) Collect your data
  2. 2) Protocol onboarding: Mineral and electrolyte repletion, toxic metals chelation, mitochondrial remediation and support, diet and lifestyle changes, glycogen and collagen synthesis improvements, nasal dysbiosis remediation, structural assessment, cardiovascular assessment, initial immune activity phase, plus ongoing transition from sympathetic overdrive -> parasympathetic balance
  3. 3) Stage 1: Microbiome re-seeding and performing a controlled initial “die-off” (at least two weeks)
  4. 4) Stage 2: Enhanced gastrointestinal and mucosal interventions (at least two weeks)
  5. 5) Stage 3: Stimulate and support a strong immune response while breaking (circulating) biofilms (at least four weeks)
  6. 6) Optional follow-up
  7. 7) Ongoing preventative maintenance


"Be thankful for your triggers, for they show where you are not free."


We fully anticipate that individuals exploring this protocol may be highly sensitive to supplements. The reasons for this sensitivity are detailed in the disease model and have been accounted for within the protocol.
The sensitivity to supplements and foods is expected to resolve as part of the protocol.

One of the most important lessons learned through this process is that ultimately, whenever a basic, essential nutrient triggers an initial reaction - either good or bad - you need it, although you may also unmask other deficiencies / compensations in the process of correcting one.

That stated, blindly charging your way through barriers is not a sensible way to achieve success, and there is a structured process to follow which makes nutrient repletion much more tolerable.


This protocol is designed to stimulate immune activity and may produce a “J-curve” trajectory, meaning you might feel progressively "quite sick" at the beginning of each stage. Increasing energy availability allows for heightened immune activity, which prioritizes energy metabolism toward these goals and can temporarily intensify symptoms. Maintaining electrolytes, selected minerals, glycogen and active B-vitamin status may help reduce risk of severe symptoms or PEM in this protocol model, but should not be presented as guaranteed prevention.


trajectory


Your individual experience may vary based on factors such as the pace of protocol implementation, the pathogens involved, your initial baseline condition, and the consistency and accuracy of your daily participation. Avoiding or delaying parts of the protocol can hinder progress and potentially lead to unnecessary discomfort. Support is available through our Discord discussion group.

Before beginning, assess your lymphatic system as described in the “2.2.2 Lymphatic Maintenance” section. Note any tissues that have been or continue to be sore, stiff, or inflamed—these areas are expected to flare during increased immune activity and may be adjacent to other infected tissues.

To help reduce expected symptoms when correcting deficiencies and experiencing Herxheimer reactions (microbial die-off effects) due to improved immune activity, a recommended starting sequence has been provided.

The "die-off effect" can cause a range of temporary but highly debilitating symptoms, including headaches, nausea, additional fatigue, dizziness, swollen glands, bloating, gas, constipation or diarrhoea, joint or muscle pain, tachycardia, chills, cold hands or feet, itchiness, rashes, sweating, and fever resembling the "disulfiram effect."

A schedule of specific products is available, with vendor website links provided for different regions (see “5. Ordering Products”). These are not affiliate marketing links. With the exception of the Oxalobacter.com website (which was created out of necessity to address worldwide supply issues for "Oxalobacter formigenes" probiotics and is not intended as a for-profit endeavor), there are no financial incentives or benefits from providing links to products or tests. If you encounter supply shortages, please try to find the same product from another vendor or ask for help in the online Discord discussion group.

Deviating from the recommended starting order or substituting products may lead to avoidable and unpleasant symptoms or may simply be ineffective. You can space out the stages further if needed. Despite the extensive list of supplements—aside from one or two items included to make your experience more tolerable—all are necessary. Attempts to create a "lite" version of this protocol by swapping or omitting items have consistently resulted in failure.

Onboarding and Stage 1 focuses on delivering missing minerals and other nutrients to the body, sometimes via non-oral routes, to resolve measurable deficiencies caused by inhibited oral absorption of these micronutrients at levels typically found in foods. While this helps restore normal metabolism, these nutrients are also accessible to the microorganisms inhabiting your body. A further protocol goal is selectivity: reducing unwanted organisms while preserving or rebuilding beneficial species through diet, probiotics and carefully chosen antimicrobial strategies. Some of these interventions are included in Stage 1, while others are introduced in Stages 2 and 3.


It is not advisable to take the Stage 1 metabolic support supplements for an extended period without progressing to the parts of the protocol that address microbiome dysbiosis, such as dietary optimisation. Otherwise, you may feel increasingly better at first, but over time this may lead to amplification of existing microbiome issues. Nutrients that benefit your cells also benefit microorganisms that depend on you, and vice versa.

Fulvic acid is used extensively in Stage 3 of the protocol. Due to its potent ability to "liberate" and recirculate metals sequestered during chronic inflammation; it is recommended to test your response to a single drop of Good State Ionic Man (fulvic acid multi-mineral) diluted in a glass of water during Stage 2. Increase the dosage slowly. Fulvic acid is also a potent biofilm breaker. If you experience any unpleasant effects, take a few days to gradually increase from a smaller dose—for example, place a drop in a bottle of water, mix it, and then transfer a drop from this first dilution into another glass of water—until you can consume a standard serving without adverse effects.


If you have nasal inflammation or notice any airway resistance or restriction when breathing, it may be appropriate to perform a nasal microbiome test and investigate for environmental mould issues. Unwanted microorganisms such as Candida, Aspergillus, Streptococcus, Staphylococcus, and Klebsiella species are sometimes found in the nasopharynx. These tests may also indicate the susceptibility of detected strains to interventions like fluconazole, itraconazole, voriconazole, or amoxicillin.

If your levels of magnesium, zinc, molybdenum, selenium, iron, iodine, or calcium are low, you may experience additional symptoms related to aldehyde metabolism insufficiency. These symptoms can be exacerbated by consuming excess alcohol, histamine, polyethylene glycol (PEG), vitamin A, or vitamin B6.

Issues with neurotransmitter metabolism, histamine regulation, and gut function are common and to be expected.


Correcting severe electrolyte deficiencies can temporarily create paradoxical effects, including increased metabolism rates and heart rate. Consuming certain minerals like copper or zinc on an empty stomach can easily induce nausea.


If taurine or magnesium worsens sleep, palpitations or tremor, treat this as a protocol signal to evaluate thyroid status and temporarily adjust electrolyte dosing. If calcium deficiency is suspected, calcium, vitamin D3 and vitamin K2 may be reviewed, and 1-2g/day elemental calcium (a high supplemental range) could be individualised against diet, labs, kidney-stone risk and medications. Addressing calcium status may change oxalate handling in the protocol model and can coincide with muscle or tissue symptoms. This is a possible transitional issue, not a necessary process. Low calcium levels may coincide with strontium and phosphorus deficiencies. The tests included in this protocol will help identify these issues. Low strontium levels in hair testing may serve as a proxy indication for calcium deficiency.


Correcting an iodine deficiency is likely to temporarily increase anti-thyroid peroxidase antibodies (anti-TPO) and Thyroid Stimulating Hormone (TSH), enhancing the activity of the sodium-iodide symporter (NIS) while improving levels of triiodothyronine (T3) and thyroxine (T4).


We recognise that the extensive list of individual supplements required for this protocol presents a new challenge. We are collaborating with compounding pharmacies to simplify this process - transforming most of Stage 1 into sublingual lozenges (troches) or DMSO-based multivitamin, and a tub or bag of premixed powder. A DIY powder recipe is also included for those who prefer to mix their own.

Mixed inositols may influence insulin/glucose signalling and glycogen handling. Inositol inhibits catalase, which may decrease your capacity to handle oxidative stress during immune activity, necessitating additional antioxidant support. If your liver and muscle glycogen stores are low - which is expected - you may initially feel extremely hungry or experience hypoglycemic symptoms such as dizziness, depression, and extreme fatigue. 

During the protocol onboard phase, there is expected to be a transition from sympathetic dominance and lipolysis to parasympathetic balance and glycogen / glycolysis preference. It is expected that consuming potentially >400 grams of net carbohydrates and water, per day, for typically two to three days, to replenish glycogen stores should help you feel significantly better, and continuing at >250g net carbohydrates per day should keep you that way. Glycogen binds with water at a 1:3 ratio, so your total body mass may increase by around 2 kilograms. Note that inositol can be problematic if you are following a ketogenic diet. D-chiro-inositol should be started at approximately 2.5 mg and very slowly increased as tolerated.

Pre-protocol support


Remineralisation Will Alter (Restore) Neurotransmitter Homeostasis or "Managing Transitional Neuropsychiatric Symptoms"

In complex chronic illness, neurotransmitter imbalances are almost never occurring in isolation. They emerge from a landscape riddled with microbiome dysbiosis, nutrient insufficiencies, redox imbalance, enzyme inhibition, and compensatory feedback loops. Restoration of neurotransmitter function therefore requires much more than supplementing amino acids or throwing SSRIs at the wall. It requires understanding why key cofactors, especially active B vitamins and minerals, are almost always functionally low.

It's critical to note that most of the enzymes responsible for synthesising, degrading, or recycling neurotransmitters are B-vitamin dependent, and their active forms, such as FMN, FAD, TPP, P5P, and methylcobalamin, are functionally deficient in the data of people with chronic illness. This is not always due to dietary inadequacy, even relative to general population requirements. The applied data shows this is due to cofactor bottlenecks, impaired enzymatic activation, oxidative degradation, and zinc- / magnesium-dependent enzyme instability.

This is evident, for example, in the metabolism of vitamin B6. As shown in this detailed diagram of B6 metabolism, converting dietary B6 into its active form P5P involves a cascade of steps: alkaline phosphatase (ALP) activity outside the cell, membrane transport, and intracellular phosphorylation via pyridoxal kinase (PK). Each of these steps is dependent on magnesium and zinc, and many are redox-sensitive. Acetaldehyde, chronic inflammation, and low ATP all interfere. It's no surprise, then, that functional P5P levels are low, even when blood levels of pyridoxine or pyridoxal appear normal or highly elevated.

p5p_v3


However, this problem isn’t limited to Vitamin B6.

eg. Riboflavin must be converted to FMN and FAD, a process impaired by low ATP, oxidative stress, zinc or magnesium loss. Thyroid synthesis and related minerals, substrates also influence this. 

fad-fmn


Thiamine must become TPP via thiamine pyrophosphokinase. B12 must be converted to methylcobalamin, a zinc- and glutathione-sensitive step. These are often affected by chronic acetaldehyde, either directly or indirectly.

Acetaldehyde burden affects not only ALDH directly, but also indirectly interferes with zinc-dependent enzymes, magnesium availability, and redox-sensitive pathways throughout this entire cascade. It also stimulates mast cell activation, promoting histamine release, while simultaneously impairing histamine degradation through inhibition of ALDH and depletion of cofactors like P5P and copper. This can disrupt the balance of neurotransmitters by overactivating histamine receptors, and indirectly suppressing dopamine synthesis via H₃-mediated inhibition of tyrosine hydroxylase. It acts as both a primary disruptor and a multiplier of vulnerability in neurotransmitter and cofactor systems.

catecholamines_v2


The result is that small efforts to stimulate or correct neurotransmitter production can trigger paradoxical responses, including overshooting, agitation, anxiety, "brain on fire", insomnia, sedation, getting "stoned" / "high", or what many patients call a “weird” or “off” feeling.


Onboarding

The fix isn’t to push harder, it’s to intervene smarter - a staged approach works better.

For severe cases and/or people with elevated susceptibility to (neuropsychiatric) symptoms from supplements which normally improve methylation and gluathione metabolism, the standard protocol onboarding sequence now actively targets issues which comprehensive data collection has shown creates that sensitivity.


A schedule of "essential" products and dosing can be found below, in "Getting started".

Use non-oral routes to bypass known bottlenecks. Transdermal, sublingual, or parenteral delivery helps sidestep transporter saturation, liver metabolism, or dysregulated GI environments. Dissolving nutrients in DMSO is another useful strategy.

P5P in DMSO is one example of an advanced strategy - dissolving P5P in a DMSO solution bypasses multiple upstream steps: it does not rely on ALP, avoids compromised membrane transporters, and skips over PK activation. This makes it ideal in states where P5P is known to be low despite B6 repletion. It may also enter intracellular compartments more efficiently, including mitochondria. The same effect applies to most compounds needing to be absorbed and distributed systemically.

In this context, neurotransmitter restoration is more about kinetics, cofactor availability, and redox context. A patient may be taking all the right nutrients, but if those nutrients are stuck upstream, degraded by peroxynitrite, or unable to bind to their target enzymes due to low zinc, the net effect is failure. Worse, poorly timed repletion can destabilise the system further.

The half-life of each vitamin is frequently overlooked. Dosing vitamins 2-4 times per day is often necessary to achieve sufficient benefit and this is reflected in the schedule.
eg. Vitamins B1, B2, B3, B5, B6, B7/H, B8, B9, B12 and C are taken 2-3x/day, often sublingually.

In the context of restoring severe deficiencies, strategic sequencing matters. B vitamins are powerful, especially in active forms, however they also act as signals in a complex control system. If introduced out of order or in isolation, they may trigger compensatory reactions that mimic toxicity, when in fact they are just unmasking hidden dysfunction.

Many of these mechanisms may also help explain why certain neuropsychiatric conditions appear during chronic illness and how they can act as precursor states to systemic collapse.


ADHD, for instance, is often characterised by signs of noradrenaline and dopamine dysregulation, impaired impulse control, and chronic restlessness, traits which may reflect an early-stage compensation for failing neurotransmitter recycling or low-grade redox and metabolic dysfunction + glycogen insufficiency, along with chronic endogenous acetaldehyde and opioid synthesis. If these compensations begin to fail, especially under stress, infection, or prolonged nutritional depletion downstream of chronic lactic acid metabolism and mineral transporter alterations, the system may tip into a more energy-conserving, downregulated state, as observed in ME/CFS, long COVID, post vaccination syndrome, etc.

Bipolar disorder may reflect a different kind of instability, one in which neurotransmitter tone swings between overcompensation and collapse, and this too may be compounded by aldehyde load, especially acetaldehyde, which inhibits ALDH enzymes responsible for clearing dopamine breakdown products like DOPAL.

Dietary contributions to acetaldehyde burden, such as excessive intake of alcohol, sugar, or fermentable carbohydrates, can further amplify this inhibition by fuelling microbial overgrowth and endogenous ethanol production in the gut. When ALDH function is compromised, neurotoxic intermediates accumulate, further impairing redox balance and downstream dopamine synthesis. Over time, this may shift the system toward an oscillatory or collapsing state, where any attempt to stabilise mood or energy is undermined by bottlenecks in detoxification and neurotransmitter recycling These cycles could, in part, emerge from fluctuating availability of critical cofactors like P5P, B2, or SAMe, or episodic redox crashes that disrupt normal mood-regulating feedback.

Last Updated 4 Feb 2026

This protocol begins with a data collection process that helps you to identify specific deficiencies and microbial influences relevant to your metabolism.

There are multiple tests required to collect this data and the protocol is unlikely to succeed without them.

protocol-testing2
Last Updated 8 Aug 2026

Testing blood is a helpful way to understand specific aspects of immune activity and metabolism. Minimally, it’s useful to have markers for blood group (needed for Oligoscan), red blood cell (RBC) and white blood cell (WBC) counts, comprehensive metabolic panel (CMP) and lipids, LDH isoenzymes, iron studies (iron, transferrin, transferrin saturation %, ferritin), thyroid markers (TSH, fT3, fT4), vitamin B6 and vitamin D (inactive form). Your doctor is usually the best person to assist you with these, however private lab services are also available in many countries, should these be helpful.

In the disease model, elevated serum vitamin B6, vitamin A and LDL are all potential proxy markers for hepatic ALDH2 inhibition, via NAD+:NADH redox impairment, downstream of oxidative stress and mitochondrial Complex I inhibition, indicating mitochondrial dysfunction. LDL is the most reliable, as it's less impacted by dietary input patterns. Serum vitamin B6 elevation is also influenced by Zn and Mg-ATP insufficiency, inhibiting pyridoxal kinase.


Impaired hepatic mitochondrial function can be expected to affect gluconeogenesis, retinol -> retinal -> retinoic acid metabolism and a range of other important pathways. 

Similarly, insufficient thyroid vs insulin signalling can also inhibit gluconeogenesis at PEPCK. Thyroid hormone synthesis is influenced by tyrosine, iodine, selenium and electrolytes / ATPase activity. Iodide transporters (NIS) require retinoic acid signalling. Selenium status is affected by silver and mercury toxicity.


USA 1

DirtCheapLabs.xyz have kindly created a custom panel which includes all of the markers above, currently via Quest -
https://dirtcheaplabs.xyz/c/joshua-leisk/bornfree-life-standard-set
(again, a clear reminder that I intentionally do NOT receive any income or benefits from any lab test or supplement vendors)
 


USA 2


Australia


Lactate dehydrogenase (LDH) isoenzymes

Lactate dehydrogenase (LDH) is an enzyme primarily involved in converting pyruvate to lactate (and vice versa). It has five isoenzymes (LDH-1 to LDH-5), each consisting of different combinations of two subunits: H (heart) and M (muscle). Further, there are individual LDH isoforms - LDH-A/B/C which make up these isoenzymes. These isoforms have specific roles / functions. LDH is also a metalloenzyme, affected by zinc status and NAD+:NADH ratio. LDH isoenzymes are distributed across various tissues based on their metabolic needs. 

The terms "H" (heart) and "M" (muscle) subunits were historically named based on the tissues where these subunits were first observed in high concentrations. The H (heart) subunit was first identified in cardiac tissue, which relies on aerobic metabolism. The M (muscle) subunit was discovered in skeletal muscle, which often utilises anaerobic glycolysis, especially during high activity.

This test allows you to see if an elevation of a specific isoenzyme or pattern of elevations is present, further suggesting if a particular tissue is experiencing energy metabolism dysregulation, and/or upstream  influences affecting energy metabolism.

eg. If LDH-4 and LDH-5 (which contain more M subunits, characteristic of anaerobic glycolysis) are elevated, it can be indicative of reduced oxygen availability or prolyl hydroxylase inhibition for other reasons (oxidative stress, low Fe/Si, low Zn, low alpha-ketoglutarate, elevated succinate). These isoenzymes are more prevalent in tissues that rely on anaerobic metabolism, such as skeletal muscle and liver, and their elevation suggests a shift towards lactate production due to reduced oxygen, a common feature of hypoxic conditions and / or various issues which inhibit prolyl hydroxylase activity (see Figure 1).


Standard LDH testing only measures TOTAL LDH. The individual isoenzymes are combined into a single figure, easily obscuring any anomalies in an individual tissue. This makes standard LDH testing highly unreliable and easily provides another false negative result, upsetting diagnostics efforts by clinicians.
eg.

LDH_total
Last Updated 15 Sep 2026

After several years working with most of the commercially available approaches to mineral and metal testing, I have found Oligoscan to be one of the more useful tools for pattern recognition, provided that several apparent algorithmic errors are corrected before interpreting the results.

In my dataset, the uncorrected output appears in some cases to classify depleted minerals as elevated. The iron and calcium outputs also behave differently from corresponding measurements across other specimen types and should not be interpreted in the same way as the remaining markers. I have therefore developed a tool on the Born Free website that applies these corrections automatically before the data are analysed, which is linked below.

The company has been unusually open to reviewing these observations and responsive to the analysis I have shared with them, including discussing the work with me publicly in a recent podcast.

This should not be taken to mean that Oligoscan provides a direct measurement of intracellular mineral concentrations throughout the body. It does not. Rather, its value has to be understood in the context of a broader problem: virtually every available method for assessing human mineral and metal status measures a different compartment, and every compartment has important limitations.

Serum
primarily tells us what is circulating outside cells at the time of collection. For many minerals, serum concentrations are tightly regulated and can remain apparently normal despite substantial intracellular depletion or tissue redistribution. During inflammatory or stress responses, circulating concentrations may also move in a direction that does not reflect the concentration within metabolically active tissues.

Red blood 
cells provide an intracellular measurement, but erythrocytes are highly specialised cells. Mature red blood cells contain no mitochondria and participate extensively in buffering and transporting circulating elements. Their mineral content can therefore diverge from that of tissues such as liver, brain, kidney, muscle and bone marrow.

White blood cells
are metabolically more representative cells and can sometimes provide a more useful intracellular proxy. However, they still represent a specific circulating cell population rather than the organs we ultimately want to understand. Commercial intracellular assays also tend to provide little or no useful information about toxic-metal accumulation.

Whole blood
combines plasma, red blood cells, white blood cells and platelets. This can be useful for certain questions, particularly exposure assessment, but it produces a composite signal that can obscure redistribution between extracellular and intracellular compartments.

Urine
primarily tells us what is being excreted. A low urinary concentration may indicate low body burden, poor mobilisation, low exposure, altered renal handling or impaired excretion. It cannot by itself distinguish between these possibilities.

Hair
provides a longer-term record of material incorporated into growing tissue. It can be useful for identifying patterns over time, but it is still an excretory and peripheral tissue measurement. Hair-forming cells do not participate in inflammatory redistribution in the same manner as liver, brain, kidney, spleen or bone marrow. Conceptually, hair can sometimes be thought of as providing a longer-duration average of what has been available to peripheral tissue via serum, rather than a contemporaneous measurement of metabolically active organs.

This leaves us with an important problem. Many tests described as "validated" are analytically valid for the specimen they measure, but that does not necessarily make them valid measures of total-body status or of concentrations inside a particular organ. A normal result can therefore be falsely reassuring when the biological question is whether an element has been depleted from, redistributed into, or accumulated within another tissue.


Oligoscan has the same fundamental limitation. Its measurements are derived from the skin and should be treated as an inference about systemic mineral and metal status, not as a direct measurement of the brain, liver, kidneys, bone marrow or other organs. Toxic-metal accumulation may differ substantially between these tissues and the skin.

This distinction becomes especially important during depletion and repletion. Minerals and metals do not distribute uniformly across the body. Different tissues accumulate, release and retain individual elements at different rates. During repletion, the concentration in blood, skin or hair may therefore rise, fall or remain unchanged while the concentration in another tissue is moving in the opposite direction.

For this reason, the protocol does not treat any single mineral or metal test as definitive. Results are interpreted as compartment-specific observations and are compared with other measurements, clinical context, metabolic pathway data from OAT or Theriome Aristotle Plus reports and, where available, repeated measurements over time.


The objective is not to identify a supposedly perfect test. At present, one does not exist. The objective is to understand what each test is actually measuring, recognise what it cannot tell us, and use the combined pattern to make a more defensible inference about intracellular and tissue mineral status.

To perform testing


Use at least one of the following two testing methods (hair mineral testing no longer supported for measuring most deficiencies due to unavoidable limitations in the methodology). An Oligoscan or SO/Check test is potentially the most useful and includes toxic metal data.

There are limitations to any testing methodology and potential ways for errors in sampling to occur. Some of these problems can be identified and avoided. Others can relate to eg. unpredictable sample transport delays.

White blood cells (WBC)


Currently, there are 3 competing laboratories which specialise in measuring the intracellular nutrient status inside WBCs - Cell Science Systems CMA (part of their CNA offering), SpectraCell Micronutrients and Vibrant America Micronutrients.

WBCs are used as a proxy to infer the nutrient status of other cell types, eg. brain, muscle, liver, etc. This data is very different to what is reported in serum (data sourced from outside the cells and any/all related mineral transporter function) or red blood cells, which lack mitochondria and therefore have very different metabolism. Vibrant America Micronutrients test does an excellent job of highlighting this issue in their report. It's frankly astonishing that serum testing is still considered the "gold standard" by public health systems, in light of these issues. However, this also highlights why chronic diseases have been difficult to research and solve.

Although all of these laboratories measure WBC nutrients, each of these tests has a different methodology and different array of included markers. Therefore, the advantages and disadvantages for each have been carefully considered. Primarily due to the expansive array of markers more closely matching our requirements, Cell Science Systems CMA is used by this protocol. However, there are limitations to accommodate. In a more perfect world, we would have a test that uses the SpectraCell or Vibrant America methodology, with the CMA scope of markers.

NB. Due to the preparation methodology used by the CMA assay, it's possible under certain circumstances for the report results to appear almost "normal" in the more severe patients who also have more SEVERE deficiencies. These deficiencies affect how the cells respond in the CMA assay, even when other deficient nutrients are added. Severity of these deficiencies can be identified by cross-referencing OAT metabolites and other data. Usually these are eg. phosphate and/or sulphur, however a severe enough deficiency of eg. B1 or magnesium can also limit cellular activity sufficiently to mask the effect of correcting other nutrient deficiencies.

By checking OAT results before ordering the CMA test, you can help identify if you have the hypoxia / acidemia patterns and are likely to have issues:
1. If you see elevated lactic acid (22), relative to pyruvic acid (23), elevated oxalic acid (21), and/or low phosphorus (76), this pattern is suggestive of excessive anaerobic glycolysis / lactic acid metabolism, metabolic acidemia and renal phosphate dumping.
2. A relative drop from citric acid (29) to aconitic acid (28), with similarly low 2-oxoglutaric acid (27) may indicate severe deficiency of magnesium.

The Getting started section may be helpful in stabilsing the baseline metabolism enough to perform the CMA test without issues. Alternatively, you could use the Oligoscan test discussed below, without this limitation.


However, if you receive a CMA report which is verifiably affected by this issue, you may be able to partially salvage the results by reinterpreting the markers as follows:
1. Any marker not all the way to the left, yet still green is re-interpreted as yellow.
2. Any marker that is yellow is re-interpreted as red.
3. Any marker that is red is re-interpreted as severely deficient.

Advantages:

1. Directly actionable data from an accredited laboratory.
2. Significant array of markers included.
3. Data is considered a snapshot of "current status", without any lag.
4. Blood sample can be collected at home, using a mobile phlebotomist.

Limitations:

1. 8–12 hours fasting is required.
2. Does not include fluorine, sulphur, silicon, various toxic metals, and some of the electrolytes – phosphorus, sodium and potassium
3. Does not indicate excess nutrient data, although this hasn't been a concern.
4. Transport of the sample needs to be rapid, to accommodate eg. cytokine half-life and nutrient transporter behaviour in the cellular membrane, otherwise report sensitivity can be lost. For non-US citizens, an express FedEx courier service is supplied by the laboratory (with an additional cost), along with any import/export clearance paperwork needed for getting blood samples through border security.
5. Sensitivity in the reporting appears to be significantly impacted by hypoxia, phosphate, sulphur and other deficiencies.
6. Intracellular iron status does not capture systemic iron status data, as 75% of iron is circulating in RBC and serum. A separate test is required.
7. Does not capture toxic metals data.

This comprehensive test is available internationally from a US laboratory. The test normally costs USD$239 - 264, plus express FedEx shipping (usually quoted / invoiced separately, shortly after purchase) and any phlebotomist costs.
Ordering: (International, via USA)

1) https://www.dhalab.com/shop/cma-cellular-micronutrient-assay (USD$264)

The reports track well with the other mineral testing methodology, Oligoscan, where Oligoscan is interpreted using our upper and lower thresholds, plus algorithmic corrections. However, CMA has a better evidence base and includes a number of additional nutrients. As it also lacks some markers, some workarounds may apply:  
Phosphorus data from the Organic Acids Test (OAT) can infer phosphorus status, although there are some differences, being urine / excretory data.
The protocol uses daily targets for electrolytes, however you won’t have an understanding of the baselines for these markers.
image-1


Skin minerals and metals

Research suggests that an Oligoscan, So/Check or Zell-Check report can provide an approximation of your systemic mineral status, using the skin of your hand as a data source. Note that “OligoLab” / “OligoHealth” / “Scantest” branded reports are not supported. This is normally an in-clinic test and a local practitioner will be required. For severe patients, a home-testing kit may be available from some vendors - use the contact form if you'd like help obtaining this.

Your blood type, age, height and weight affect the sample calibration used to generate this report, so this information will be requested when you visit a clinic. Any inaccuracies can dramatically alter your results. 


NB. Spectrophotometry cannot directly measure minerals / metals in the skin - that's impossible. Instead, their "black box" methodology captures patterns in skin colouring data from your hand, which is then algorithmically matched against a large database of training data. 


While not happy with the level of published evidence currently supporting this emerging methodology, our testing showed a clear pattern of deficiencies in ME/CFS, Long Covid and Post Vaccine Syndrome, with minor variability, which was not observed in controls.

Follow up Oligoscan reports have also matched supplements taken and previous results. The array of elemental markers collected is almost the same as the Mosaic DX (Doctors Data) HTMA reports, however it also includes silicon and usually fluorine. Unfortunately, it does not show rubidium or strontium markers.
image-2
Notes and limitations:
1. Because iron distribution is compartmentalised and clinically important, skin/mineral scan data should not be used to infer systemic iron status; serum iron studies remain the appropriate confirmation route.
Low manganese may also provide a hint for low iron, unless IV iron has recently been administered.
2. Skin/mineral scan calcium should be interpreted cautiously or discarded, as most calcium is stored in bone and serum calcium is tightly regulated. This also makes serum calcium insensitive. As strontium and calcium can potentially substitute for each other in biological processes, using the “strontium” marker from the hair test as a proxy to indicate systemic calcium may be possible, but needs further study. Tremors and/or heart palpitations, especially in response to taurine or magnesium could indicate low calcium, also. Low phosphorus may indicate low calcium.
3. Copper and zinc appear to be over-estimated in some cases.
4. Due to an unusual quirk in methodology / reporting, highly elevated minerals need to be re-interpreted as highly deficient.


Oligoscan and CMA reports are not known to be adversely affected by the inflammatory cascade, meaning that these reports can provide an actionable set of objective markers to use when choosing most mineral supplements to address deficiencies. (HTMA data can be referenced to help supplementation planning for rubidium and strontium, also inferring calcium).

For more information about the effects of specific mineral deficiencies, see the “Rationale for protocol inclusions and general notes” section. The Oligoscan and SO/Check reports include a number of vitamin markers, which appear to track relatively well with other data. A good general target for these is to exceed 55%.

Toxic metals are also reported. These are also targeted in the protocol. Any markers in the “High -/+” ranges should be monitored. Due to redistribution, heavy metals in skin often go up in the first stage of the protocol, then decline.

More information can be found in 4 Calculating supplement doses


If you have a good (or bad) experience with an Oligoscan practitioner, please share details with our community Discord group in the #oligoscan-providers channel.

For Oligoscan, you can also find local practitioners here: https://www.theoligoscan.com/nearest-provider
or try using Google and run individual searches for each company / technology and your suburb (4 separate searches, rather than 1):
eg.
Oligoscan | Oligocheck | So-Check | ZellCheck <your suburb>


Broadly speaking, Oligocheck, So-Check and ZellCheck are more commonly found in and around Europe. Oligoscan is more commonly found in USA, Australia and UK.


Hair minerals (deprecated / limited use)


Research suggests that a Hair Toxin Mineral Analysis (HTMA) may provide an approximation of your functional mineral status, averaged over the period of follicle growth, sampled adjacent to the transporter alterations from inflammation – where circulating minerals can be sequestered inside various brain, liver and kidney cells and consequently low in other cells. It cannot accurately report on the intracellular levels or systemic levels, as sequestered minerals are not shown in this excretory data.

The results obtained from the HTMA data may be used to infer averaged intracellular mineral availability, downstream of any chronic inflammation, for the period of time in which the hair follicles grew, in a similar way to read HbA1C for glucose metabolism. This mechanism is largely relevant to the eleven elements transported by DMT-1 and ferroportin.

A list of vendors offering a compatible list of markers and reference ranges is provided below. Doctors Data and other compatible laboratories
US https://www.walkinlab.com/products/view/dd-hair-elements-profile-doctors-data (not available in NY/ NYC and other areas.)
AU https://www.toxno.com.au/articles/heavy-metals/hair-tissue-mineral-analysis-hma-or-htma-in-australia/ (can manually add rubidium)
EU / UK https://regeneruslabs.com/products/hair-toxic-essential-elements-1
EU https://www.biocoherence.eu/en/shop/htma/ (doesn't include rubidium)
EU https://www.lifelinediag.eu/en/product/eha-standard-en/#products (doesn't include rubidium)
NZ https://www.houseofhealth.co.nz/product/hair-toxic-mineral-analysis/
PL https://analizawlosa.com.pl/produkt/analiza-wlosa/
UK / some EU https://www.lauristonlabs.co.uk/product/hair-elements-doctors-data/
Hair collection guide:
  1. Ignore any conflicting vendor-specific instructions.
  2. The hair needs to be clean, well-rinsed, dry, untreated and uncoloured. Unwashed hair may provide false (elevated) data for sodium and potassium, from dried sweat. Use gloves.
  3. Select suitable areas of sideburn, scalp and/or neck hair. If the hair is longer than 2.5cm / 1", first trim the sample area to this maximum length with clean stainless steel scissors. By using shorter hair, the report data relates to a more recent period of time.
  4. Cut the (remaining) hair to be sampled as close to the skin as possible. Do not use clippers or an electric razor – metal contamination from tungsten blades may occur. Collect at least two heaped tablespoons worth of hair, or approximately 250mg, or the sample may be rejected.
  5. Place the hair sample in the sample kit envelope provided, or in a clean, clearly labelled envelope or sachet. Whilst Trace Elements Inc says not to use resealable plastic bags as apparently hair “sweats” and the sample may be rejected, whereas Doctors Data supplies plastic bags in their kit.