Falsification-First Validation of the Fructose Model

Purpose and Overview

This appendix defines an open, reproducible framework to test—and, where appropriate, falsify—the core predictions of the Fructose Model:

• Fragile Energy Hypothesis: mitochondrial dysfunction accumulates into fragile cells → fragile systems, explaining clustered chronic disease.
• Fructose Amplifier Hypothesis: among many stressors, fructose → fructokinase (KHK-C) activation is the most significant, modifiable amplifier in modern life.

Near-term goals:

• Demonstrate (or falsify) human KHK inhibition by natural compounds.
• Resolve the physiological significance of endogenous fructose (polyol pathway).
• Track whether pathway modulation improves a shared, multi-system “fragile energy” signature.

Open Science & License: This protocol is released under Creative Commons CC BY-SA 4.0. You may copy, adapt, and build upon it—even commercially—provided you give appropriate credit and share derivatives under the same license. Pre-register deviations and report all results (including null) for transparency.

Strategy: Falsification Before Confirmation

Claim	How It’s Tested	Falsifying Outcome
KHK inhibition attenuates uric-acid generation after fructose load	Exogenous Fructose Challenge (EFC)	No measurable attenuation
Endogenous fructose contributes materially to urate rise	Glucose + Sodium Challenge (G+NaC)	No baseline rise or no effect of inhibitor
Modulating the pathway improves a multi-system signature	4–8 week endpoint panel	No correlation between KHK attenuation and systemic improvement

Compounds and Arms

The appendix remains platform-agnostic. Example arms for independent labs:

Arm	Description	Primary Target
A. Luteolin (liposomal)	Natural polyphenol; strongest preclinical signals for KHK inhibition; liposomal improves exposure	KHK-C
B. Osthole	Coumarin derivative; exploratory KHK/urate modulation; useful as a falsification foil	KHK ± urate signaling
C. Luteolin + Tart Cherry	“Block spark + lower smoke”: KHK inhibition plus urate lowering	KHK + urate amplification
D. D-Mannose	Exploratory sugar; potential competitive interaction at early hexose handling; hypothesis-generating	Upstream/competitive modulation
E. Placebo	Matched excipient	—

Independent laboratories may request access to a reference formulation containing a defined liposomal blend of luteolin and tart cherry extract, supplied by LIV3 Health for use in non-commercial, open-science research. The formulation is made available to enable independent replication of the KHK-inhibition and uric-acid attenuation studies described in this appendix. Participating researchers are encouraged to compare its effects with other candidate modulators under their own supervision, with all findings shared under a Creative Commons open-license framework.

Phased Experimental Design

Stage 1 — Immediate (Acute) Validation

Purpose: demonstrate same-day attenuation of the uric-acid/ATP response—direct evidence of human KHK modulation.

Day	Condition	Compound	Purpose
Day 1	Baseline challenge	None	Reference uric-acid and ATP response
Day 2	Repeat challenge	Single pre-dose (30–60 min before load)	Test acute inhibition
Day 3 (optional)	Placebo challenge	Placebo	Reproducibility / expectancy control

Primary readouts (0 / 60 / 90 min): urinary uric acid (UA); optional PBMC ATP/ADP ratio (luciferase).
Success threshold: ≥ 30% median reduction in UA AUC(0–90) vs baseline.

Stage 2 — Short Wash-In (Steady-State) Validation

Purpose: determine whether continued exposure maintains or amplifies inhibition and whether endogenous fructose generation is affected.

Duration: 7–14 days of daily compound dosing (no daily sugar or glucose loads).

Protocol: After the wash-in period, perform the following two challenge days on separate dates:

1. Exogenous Fructose Challenge (EFC) — reassess sustained KHK inhibition.
2. Glucose + Sodium Challenge (G+NaC) — assess endogenous (polyol-pathway) fructose generation.

Interpretation: attenuation in both supports KHK hit and endogenous relevance; EFC-only attenuation pressures breadth of the amplifier claim.

Stage 3 — Short-Term Endpoint Tracking (4–8 Weeks)

Purpose: test whether biochemical modulation aligns with multi-system improvement.

Baseline collection: All metabolic and subjective endpoints should be measured prior to the first compound exposure (typically at the same visit as the initial fructose challenge). These baseline values serve as the reference for 4–8 week comparisons and for computing changes in the Fragile Energy Composite (FEC).

Pre-specify 4–5 endpoints:

• Liver/metabolic: ALT (±AST), TG/HDL ratio, HOMA-IR.
• Vascular: mean arterial pressure (home AM/PM average).
• Subjective: Energy VAS, Craving VAS.
• Optional: CGM time-in-range and variability.

Detailed Challenge Protocols

1) Exogenous Fructose Challenge (EFC)

Load: 50 g D-fructose in 350 mL water, consumed in ≤ 5 min (pure fructose for KHK specificity).

• −30 min: administer modulator (modulator arm only)
• 0 min: Void #1 (baseline UA; optional blood/ATP)
• 60 min: Void #2 (UA; optional ATP)
• 90 min: Void #3 (UA; optional ATP)

Hydration: 250 mL water 30 min pre; ≤ 250 mL during 0–90 min. Keep timing and hydration identical across visits.

2) Glucose + Sodium Challenge (G+NaC)

Load: 75 g glucose + 3.0 g NaCl in 500 mL water, ≤ 5 min (raises glycemia and osmolality to trigger polyol-pathway fructose).

Sampling: same schedule as EFC (0 / 60 / 90 min UA; optional ATP).

Controls and Exclusions

• Exclude: gout, CKD, nephrolithiasis, pregnancy, uncontrolled diabetes, active infection; recent alcohol (< 48 h); diuretics, allopurinol/febuxostat (unless supervised).
• Standardize: prior-day purines, caffeine, exercise; same clock timing (± 30 min) and hydration across visits.

Analytical Notes

• AUUAC (0–90): trapezoidal area under UA curve (mg·min/dL) from 0/60/90 min values.
• ΔATP%: ((post / pre) − 1) × 100% (if PBMC assay used).
• Statistics: paired t-test or Wilcoxon; report Cohen’s dz, median %, and 95% CI.
• Pilot n: 10–20 participants (~80% power for dz ≈ 0.6).

Fragile Energy Composite (FEC)

Create a compact composite to track the shared signature. Z-score each item; invert Energy VAS sign before averaging; then compute mean z as the FEC:

• AUUAC attenuation (lower = better)
• Mean arterial pressure (lower)
• TG/HDL ratio (lower)
• ALT (lower)
• Energy VAS (higher; invert sign)

Correlation between AUUAC attenuation and ΔFEC supports the “common signature” thesis; discordance motivates refinement (dose, duration, cohort).

Falsification Logic

Observation	Interpretation
No UA rise at baseline	Assay/timing mismatch—revise protocol
UA rise present, no attenuation	Compound/dose does not inhibit human KHK
EFC attenuates; G+NaC does not	Dietary-gate specificity; endogenous significance unproven
Both attenuate	KHK hit + endogenous relevance supported
Acute attenuation without ΔFEC gain	Questions clinical relevance at this dose/duration; iterate
Attenuation correlates with ΔFEC	Supports shared “fragile energy” signature

Ethical and Practical Notes

• Mechanistic, non-diagnostic testing; follow local IRB/ethics where applicable.
• Do not alter prescribed medications without clinician oversight.
• Record and share anonymized raw data and analysis code under CC BY-SA 4.0.

Summary Table

Stage	Timeframe	Primary Question	Key Metric	Falsifies If …
1	Day 1 → Day 2	Can naturals acutely inhibit KHK?	Δ AUUAC (0–90)	No attenuation
2	0–14 days	Is endogenous fructose significant and modifiable?	Δ AUUAC (EFC & G+NaC)	No rise / no inhibition
3	4–8 weeks	Does KHK modulation improve systemic signature?	Δ FEC	No correlation / no improvement

Suggested Citation

Mearns C., Gross P., et al. (2025). “The Fructose Model: Appendix A — Falsification-First Experimental Test Protocols.” LIV3 Health Whitepaper Series. Licensed CC BY-SA 4.0.