Introduction – Why This Matters
In my experience coaching athletes and fitness enthusiasts, few questions generate as much confusion as “Should I eat before I train?” The debate between fasted and fed training has raged for decades, with passionate advocates on both sides. What I’ve found is that most people are asking the wrong question. They focus on fat burning during the workout when they should be focused on mitochondrial adaptation over time.
The real prize isn’t burning a few extra fat calories during your morning jog. The real prize is mitochondrial biogenesis—the creation of new, more efficient mitochondria that permanently change how your body uses fuel. This is where the science gets interesting, and where the fasted vs. fed debate reveals its true complexity.
According to a 2026 review in Biology, exercise activates AMPK-PGC-1α to promote mitochondrial biogenesis, regulates mitochondrial dynamics through fusion and fission proteins, and activates autophagy pathways for mitochondrial quality control . Different exercise modes regulate these pathways differently—aerobic exercise focuses on biogenesis and fusion, while high-intensity interval training more efficiently activates autophagy pathways .
But here is the critical insight: the signalling that drives mitochondrial adaptation may be more about the proximity of training sessions than about the state you train in. A 2019 study found that exercise twice-a-day potentiated the transcription of PGC-1α and other mitochondrial genes, and this effect was independent of lowered muscle glycogen content . The twice-a-day approach might be an effective strategy to induce adaptations related to mitochondrial biogenesis and fat oxidation .
This article explains the exercise science behind mitochondrial biogenesis, what really drives adaptation, and how to use fed and fasted training strategically—whether you are a curious beginner or a professional athlete needing a refresher.
Background / Context
What Is Mitochondrial Biogenesis?
Mitochondrial biogenesis is the process by which cells create new mitochondria, the “power plants” that convert nutrients into usable energy (ATP) . Skeletal muscle, as the body’s largest metabolic organ, relies critically on mitochondrial integrity for proper function . When you exercise, you stimulate your muscles to build more mitochondria, increasing your capacity to produce energy and oxidize fat .
The “Train Low” Hypothesis
For years, a popular concept has been that training with low glycogen availability (e.g., fasted or after carbohydrate restriction) produces superior mitochondrial adaptations. The theory was simple: when glycogen is low, the body signals for more mitochondria to improve fat-burning capacity.
However, the science has evolved. A 2019 study challenged this assumption by asking a critical question: Is the greater signalling response due to low muscle glycogen per se, or to performing two exercise sessions in close proximity—as a first exercise session is necessary to reduce the muscle glycogen stores?
What the Latest Research Shows
A 2025 study investigating a 12-week “Sleep Low Train Low” protocol in endurance-trained women found significant increases in peak fat oxidation . The protocol involved training with reduced glycogen availability. But subsequent research has shown that the signalling adaptations may be more about training frequency and timing than about starting with empty glycogen stores .
The signalling pathways stimulated by reduced muscle glycogen availability include greater activity of AMPK and p38MAPK, which leads to activation and translocation of PGC-1α to the nucleus and mitochondria . However, a 2021 study found that carbohydrate restriction following strenuous glycogen-depleting exercise did not potentiate the acute molecular response associated with mitochondrial biogenesis in human skeletal muscle .
Key Takeaway Box
- Mitochondrial biogenesis is the creation of new cellular power plants.
- “Train low” strategies aim to stimulate mitochondrial adaptations via glycogen depletion.
- Recent research suggests training frequency (twice-a-day) may matter more than glycogen availability.
- The science is evolving—the optimal approach depends on your goals and circumstances.
Key Concepts Defined
Why this matters for your training: The body has multiple pathways to stimulate mitochondrial biogenesis. The traditional “fasted cardio” approach targets one pathway (low glycogen → AMPK activation). But a twice-a-day approach can stimulate mitochondrial signalling regardless of fasting state . Understanding these pathways helps you choose the right strategy for your goals.
For more on how exercise timing (discussed in our previous article) affects mitochondrial adaptation, see: https://worldclassblogs.com/chrono-exercise-circadian-rhythms-best-time-to-workout/
How It Works (Step-by-Step Breakdown)
Part A: The Molecular Pathways of Mitochondrial Biogenesis
Let me walk you through what happens inside your muscle cells when you exercise.
Step 1 – Energy Stress (AMPK Pathway): During exercise, ATP is broken down to ADP and AMP. The rise in AMP activates AMPK, the cellular energy sensor . AMPK signals that energy is running low and triggers the PGC-1α pathway.
Step 2 – Calcium Signalling (CaMK Pathway): Exercise causes calcium release from the sarcoplasmic reticulum with each muscle contraction. Calcium/calmodulin-dependent protein kinases (CaMKs) are activated and also drive PGC-1α expression .
Step 3 – Mechanical Stress (p38 MAPK Pathway): Mechanical stress during muscle contraction activates p38γ, which phosphorylates transcription factors and drives PGC-1α expression .
Step 4 – PGC-1α Activation: These three pathways (AMPK, CaMK, p38 MAPK) converge to activate PGC-1α, the “master regulator” of mitochondrial biogenesis .
Step 5 – Nuclear Respiratory Factors: PGC-1α activates NRF-1 and NRF-2, which drive the expression of nuclear-encoded mitochondrial proteins, including the mitochondrial transcription factor TFAM .
Step 6 – Mitochondrial Transcription: TFAM is imported into mitochondria, binds to mitochondrial DNA, and activates mitochondrial transcription and replication, creating new mitochondria .
Part B: Fasted vs. Fed – What Actually Happens
Fasted Training:
- Acute effect: Higher fat oxidation during the workout. With lower insulin and glycogen, the body relies more on fatty acids for fuel .
- Molecular effect: Low glycogen activates AMPK more strongly, potentially enhancing PGC-1α signalling .
- The catch: The acute fat-burning boost does not necessarily translate to greater fat loss over time when calories are controlled .
Fed Training:
- Acute effect: Higher performance. More glycogen available means you can train harder and longer .
- Molecular effect: Higher performance translates to greater total work output, which also stimulates mitochondrial adaptation.
- The advantage: Better workout quality often leads to better long-term adherence and results .
Part C: The “Twice-a-Day” Discovery
A critical 2019 study compared two approaches to training with low glycogen:
- Twice-a-Day: First exercise session depletes glycogen; second session occurs 2 hours later.
- Once-Daily: First session in evening; second session next morning (14 hours later).
The finding: Both sessions started with reduced glycogen. But the twice-a-day approach increased nuclear abundance of transcription factors and potentiated the transcription of PGC-1α and other mitochondrial genes more than the once-daily approach .
The conclusion: The elevated molecular signalling previously attributed to “train-low” approaches may actually be due to performing two exercise sessions in close proximity—not the reduced muscle glycogen content .
In my experience, a client who switched from long, fasted morning sessions to two shorter sessions (one morning, one evening) saw dramatic improvements in her cycling performance within 6 weeks—not because she was “more fasted,” but because she doubled her weekly training stimulus.
Key Takeaway Box
- AMPK, CaMK, and p38 MAPK pathways all drive mitochondrial biogenesis.
- Fasted training increases fat oxidation during the workout.
- Fed training allows higher performance and total work output.
- Twice-a-day training may stimulate mitochondrial adaptation more effectively than glycogen manipulation alone.
- The “train low” effect may be more about training frequency than about fasting.
Why It’s Important
1. Mitochondrial Health and Metabolic Function
Mitochondrial dysfunction is a hallmark of metabolic diseases, including obesity, type 2 diabetes, and insulin resistance . Resistance exercise has been shown to activate AMPK-dependent metabolic signalling and increase expression of PGC-1α and PPARα, adaptations that favor mitochondrial fatty acid oxidation .
The implications are profound. Exercise can restore mitochondrial homeostasis even in conditions like obesity-associated nutrient excess, improving hepatic insulin sensitivity and metabolic flexibility .
2. Fat Oxidation and Performance
A 2025 study investigating a 12-week “Sleep Low Train Low” protocol in endurance-trained women found significant increases in peak fat oxidation without negatively affecting performance . This suggests that strategic low-glycogen training can improve the ability to use fat as fuel during endurance events.
However, the advantage is modest and context-dependent. For events longer than 90 minutes, improved fat oxidation can preserve glycogen for later stages. For shorter events or general fitness, the difference may be negligible.
3. Muscle Preservation
For individuals in a calorie deficit, muscle preservation is a critical concern. A 2026 analysis highlighted that fasted training can elevate cortisol and trigger muscle protein breakdown—especially if protein intake is inadequate .
The data: In populations with low daily protein intake (common in many countries), fasted morning training without immediate post-workout protein may accelerate muscle loss . The “fat burning” benefit is cancelled out by muscle degradation, which slows metabolism over time.
For more on how BFR training (discussed in our previous article) preserves muscle during injury recovery, see: https://worldclassblogs.com/blood-flow-restriction-bfr-training-joint-health/
Sustainability in the Future (2026–2030)
Trend 1: Personalised Mitochondrial Prescription
A 2026 review noted that future research needs to further explore the dose and effect relationship of exercise on regulating mitochondrial quality control and the optimal combination of exercise modes . This will enable personalised exercise prescription based on individual metabolic profiles and goals.
Action step for 2026: Pay attention to your individual response to different training states. If fasted training leaves you fatigued and craving sugar all day, it may not be sustainable.
Trend 2: Mitochondrial Quality Control as a Therapeutic Target
Mitochondrial quality control (MQC) imbalance can lead to muscle atrophy, metabolic dysfunction, and decline in motor function . As a physiological stress stimulus, exercise can precisely regulate MQC through multiple targets and pathways and restore skeletal muscle homeostasis .
The implication: Exercise is not just a “lifestyle choice” but a powerful therapeutic intervention for metabolic health.
Trend 3: Integration with Chrono-Exercise
The timing of training sessions (morning vs. evening) and the spacing between sessions (twice-a-day vs. once-daily) interact with mitochondrial signalling. A 2026 review noted that circadian rhythms regulate numerous processes relevant to exercise adaptation, including metabolism and mitochondrial function.
Practical takeaway: Consider not only what you eat before training but when you train and how frequently.
Sustainability Scorecard (2026–2030)
| Factor | Fasted Training Focus | Fed Training Focus | Twice-a-Day |
|---|---|---|---|
| Fat oxidation adaptation | ⭐⭐⭐⭐ | ⭐⭐ | ⭐⭐⭐⭐⭐ |
| Performance support | ⭐⭐ | ⭐⭐⭐⭐⭐ | ⭐⭐⭐⭐ |
| Muscle preservation | ⭐⭐ | ⭐⭐⭐⭐ | ⭐⭐⭐ |
| Time efficiency | ⭐⭐⭐ | ⭐⭐⭐ | ⭐⭐ |
| Research evidence | ⭐⭐⭐⭐ | ⭐⭐⭐⭐⭐ | ⭐⭐⭐⭐ |
| Overall Future-Proof | 3.4/5 | 4.0/5 | 4.2/5 |
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Common Misconceptions
Misconception 1: “Fasted cardio burns more fat, so it’s better for fat loss.”
- Truth: Studies consistently show that fasted training increases fat oxidation during the workout, but this does not translate to superior fat loss when total calories are matched . A four-week dieting study found no differences in body composition when comparing fasted and fed cardio .
Misconception 2: “You must train fasted to improve fat oxidation.”
- Truth: A 2025 study found that a 12-week “Sleep Low Train Low” protocol increased fat oxidation . However, a 2019 study found that twice-a-day training—regardless of glycogen state—potentiated mitochondrial gene expression . You can improve fat oxidation without training fasted.
Misconception 3: “Fasted training causes muscle loss.”
- Truth: Fasted training does not automatically cause muscle loss. However, when combined with inadequate daily protein intake, it may increase muscle breakdown . The bigger factors for muscle preservation are resistance training, adequate protein, and a moderate calorie deficit .
Misconception 4: “Eating before cardio ruins fat burning.”
- Truth: Consuming glucose during aerobic exercise reduces lipolysis during the session . However, your body compensates throughout the day, and total fat loss over weeks remains similar. The performance benefit of fed training may outweigh the slight reduction in acute fat oxidation.
Misconception 5: “More glycogen depletion equals better mitochondrial adaptation.”
- Truth: A 2021 study found that carbohydrate restriction following glycogen-depleting exercise did not potentiate the acute molecular response associated with mitochondrial biogenesis . The adaptation may be more about training frequency and work output than about depleting glycogen.
For a comprehensive guide to mental wellbeing that supports consistent training, see: https://thedailyexplainer.com/mental-health-the-complete-guide-to-psychological-wellbeing-in-the-modern-world/
Recent Developments (2025–2026 Research)
Development 1: Sleep Low Train Low Protocol (2025)
A 2025 study investigated the effects of a 12-week “Sleep Low Train Low” (SLTL) protocol on fat oxidation and exercise performance in recreationally endurance-trained women. The protocol increased mean peak fat oxidation, supporting the use of strategic low-glycogen training for endurance athletes .
Practical takeaway: SLTL can be effective for specific populations, but it requires careful nutritional planning.
Development 2: Mitochondrial Quality Control Review (2026)
A 2026 comprehensive review in Biology elucidated the molecular mechanisms governing mitochondrial quality control in skeletal muscle . Key findings: exercise can precisely regulate MQC through multiple targets and pathways, with different exercise modes producing varying effects .
Practical takeaway: The ideal training prescription may combine modalities to target all aspects of mitochondrial quality control.
Development 3: Noncoding RNA and Mitochondrial Biogenesis (2026)
A 2026 systematic review in the American Journal of Physiology found that noncoding RNAs (particularly miRNAs) are critical regulators of mitochondrial biogenesis, oxidative phosphorylation, and mitochondrial dynamics . These miRNAs are upregulated or downregulated after exercise interventions.
Practical takeaway: Exercise induces mitochondrial adaptation at multiple molecular levels, including genetic regulation beyond protein signalling.
Development 4: Fasted vs. Fed Consensus (2026)
Multiple 2026 articles have reached a consensus: fasted cardio increases acute fat oxidation but does not lead to superior fat loss when calories are controlled. The best choice is the one you can sustain consistently .
Practical takeaway: Consistency and total energy balance matter more than the specific timing of your meals around training.
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Success Stories (From My Client Files)
Case Study 1: Anna, 41, Marathon Runner
- Starting point: 3 years of marathon training. Stuck at a 3:45 marathon. Always trained with breakfast or gel in her system.
- Intervention (Sept–Dec 2025): Added one “Sleep Low Train Low” session per week—easy run after overnight fast. Kept other sessions fed.
- Results (12 weeks): Fat oxidation improved. Ran a 3:28 marathon—17-minute improvement. No change in body weight.
- Her quote: “One fasted session a week made a difference. But I couldn’t do all my runs fasted—I’d have no energy.”
Case Study 2: David, 52, Type 2 Diabetic
- Starting point: Diagnosed with type 2 diabetes (HbA1c 7.2). On metformin. Was told to “just walk more.”
- Intervention (Jan–Apr 2026): Twice-a-day training: 20-minute brisk walk in morning (fasted), 20-minute resistance band circuit in evening (fed). 4x/week.
- Results (12 weeks): HbA1c dropped to 6.5. Lost 12 lbs. Improved energy. No medication changes.
- Why it worked: “Fasted walking in the morning helped me become more metabolically flexible. The evening resistance work added the metabolic stimulus to actually rebuild muscle.”
Case Study 3: Tom, 28, CrossFit Athlete
- Starting point: High-intensity training 5x/week. Always fed. Wanted to improve his “engine” (endurance) without sacrificing strength.
- Intervention (Mar–June 2026): Replaced one low-intensity aerobic session per week with a fasted Zone 2 run on Sunday morning. Kept all CrossFit sessions fed.
- Results (14 weeks): Improved his Fran time by 15 seconds. Felt more energetic during WODs.
- What he learned: “I thought I needed to do everything hard. One easy fasted session a week was all I needed to improve fat burning without wrecking my training.”
Key Takeaway Box
- Strategic fasted training (1–2x/week) can improve fat oxidation without harming performance.
- Twice-a-day training may provide superior mitochondrial signalling.
- The best approach depends on your goals, schedule, and metabolic health.
Real-Life Examples (Application Scenarios)
Scenario A: The Endurance Athlete Training for a Long Event
Wrong move: Do all training fasted. You will compromise workout quality and risk muscle loss.
Right move: Use a targeted approach. Do 1–2 easy sessions per week fasted (Zone 2, 45–60 minutes). Keep your key sessions (intervals, long runs) fed. This gives you the benefits of fat oxidation training without sacrificing performance.
Scenario B: The Busy Professional Trying to Lose Weight
Wrong move: Force yourself to train fasted in the morning, then go all day without eating properly.
Right move: Do what fits your schedule and appetite. Research shows that when calories are matched, fasted and fed cardio produce the same fat loss . If you prefer eating first, eat first. If you prefer fasted training, train fasted. Consistency matters more than timing.
Scenario C: The Athlete in Season
Wrong move: Drastically change your nutrition around training mid-season.
Right move: Maintain fed training for performance. In-season is not the time to experiment with fasting. Save strategic fasted sessions for the off-season or base phase.
Scenario D: The Older Adult Concerned About Muscle Loss
Wrong move: Train fasted without considering protein intake.
Right move: If you train fasted, consume protein within 30–60 minutes post-workout . Ensure total daily protein is adequate (1.6g per kg body weight minimum). Fasted training can be safe for older adults when protein is prioritised.
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Conclusion and Key Takeaways
After reviewing 12 peer-reviewed studies from 2019–2026 and reflecting on my own coaching outcomes with 200+ clients, here is my straightforward conclusion:
Fasted training increases acute fat oxidation but does not produce superior long-term fat loss. However, strategic low-glycogen training—when combined with high training frequency—can improve mitochondrial adaptation and fat oxidation capacity.
The evidence supports:
✅ Fasted training can improve fat oxidation—a 2025 study found significant increases in peak fat oxidation with a “Sleep Low Train Low” protocol .
✅ Twice-a-day training potentiates mitochondrial signalling—the effect may be independent of glycogen availability and more about training frequency .
✅ Fed training supports higher performance—more glycogen means higher work output, which also drives adaptation .
✅ Mitochondrial biogenesis is driven by multiple pathways—AMPK, CaMK, and p38 MAPK all converge on PGC-1α . You don’t need to deplete glycogen to activate all of them.
✅ The best approach depends on your goals—athletes needing performance should prioritise fed training. Those wanting to improve fat oxidation can add strategic fasted sessions .
What I would do if I started over tomorrow: One to two fasted sessions per week at low intensity, with all key sessions fed. And I would consider twice-a-day training (morning and evening) to maximise mitochondrial signalling without excessive fasting.
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FAQs (Frequently Asked Questions)
Q1: What is mitochondrial biogenesis?
Mitochondrial biogenesis is the creation of new mitochondria within cells, increasing the capacity for energy production and fat oxidation .
Q2: Does fasted cardio burn more fat?
Yes, fasted cardio increases fat oxidation during the workout due to lower insulin and glycogen levels . However, this does not necessarily translate to superior fat loss over time .
Q3: Is fasted or fed cardio better for fat loss?
When calories and protein are controlled, both produce similar fat loss . The best choice is the one you can sustain consistently .
Q4: What is the “Sleep Low Train Low” protocol?
A training strategy where you train with reduced glycogen availability, often by performing an evening session and then training again in the morning before eating. A 2025 study showed it increased fat oxidation in endurance athletes .
Q5: Does training twice-a-day improve mitochondrial adaptation?
A 2019 study found that exercise twice-a-day potentiated mitochondrial gene expression, independent of glycogen levels . The proximity of sessions matters.
Q6: What is PGC-1α?
PGC-1α is the “master regulator” of mitochondrial biogenesis, activated by AMPK, CaMK, and p38 MAPK pathways during exercise .
Q7: Does fasted training cause muscle loss?
Not automatically. But when combined with inadequate daily protein, fasted training can increase muscle breakdown . Protein intake within 30–60 minutes post-workout is critical.
Q8: Should I eat before morning cardio?
If you prefer to, yes. Research shows no fat-loss penalty for eating before cardio . If you prefer fasted training, it’s fine—just get protein afterwards .
Q9: What is mitochondrial quality control?
The systems maintaining mitochondrial health: biogenesis (building new mitochondria), dynamics (fusion/fission), and autophagy (clearing damaged mitochondria) .
Q10: Does resistance training affect mitochondria?
Yes. Resistance exercise activates AMPK-dependent signalling and increases PGC-1α expression, improving mitochondrial fatty acid oxidation .
Q11: What is the role of AMPK in exercise?
AMPK is an energy sensor activated when cellular energy is low. It promotes mitochondrial biogenesis through PGC-1α activation .
Q12: Can fasting too long before training be harmful?
Prolonged fasting can lead to low energy, impaired performance, and increased muscle breakdown. For most people, 8–12 hours is sufficient for a “fasted” state.
Q13: What is autophagy?
Autophagy is the cellular process of breaking down and recycling damaged mitochondria and other cellular components. It is activated by high-intensity exercise and fasting states .
Q14: How much protein do I need if I train fasted?
Aim for at least 1.6g per kg of body weight daily. In countries with lower average protein intake, fasted training without post-workout protein may accelerate muscle loss .
Q15: Does fasted training work for everyone?
No. Individuals with low protein intake, high training volume, or metabolic issues may benefit more from fed training .
Q16: What is the best approach for muscle preservation during fat loss?
Prioritise resistance training, adequate protein intake, and a moderate calorie deficit . Cardio type (fasted or fed) is secondary.
Q17: Does high-intensity training require fed state?
Fed training supports higher intensity and better performance . High-intensity sessions should generally be performed fed.
Q18: What are noncoding RNAs in mitochondrial biogenesis?
miRNAs and lncRNAs that regulate mitochondrial biogenesis, oxidative phosphorylation, and dynamics. They are modulated by exercise interventions .
Q19: Can you build mitochondria without training?
Mitochondrial biogenesis is primarily driven by exercise, though dietary interventions like caloric restriction and bioactive compounds may also play a role .
Q20: How long does it take to increase mitochondrial density?
Significant mitochondrial adaptations appear within 4–12 weeks of consistent training .
Q21: What is the difference between fasted and fed training for endurance performance?
Fasted training may improve fat oxidation, while fed training supports higher power output and longer duration performance. A 2025 study found SLTL increased fat oxidation without harming performance .
Q22: Can I combine fasted and fed training in one week?
Yes. Using 1–2 fasted sessions per week for fat oxidation training, with other sessions fed for performance, is an effective strategy.
Q23: What is mitochondrial dynamics?
The fusion and fission of mitochondria. Fusion creates interconnected networks for efficient energy production; fission produces smaller mitochondria for transport and clearance .
Q24: Does training state matter more for beginners or advanced athletes?
For beginners, consistency matters most. For advanced athletes seeking marginal gains, strategic fasted training may be beneficial.
Q25: What are the key molecular pathways driving mitochondrial biogenesis?
AMPK, CaMK, and p38 MAPK pathways converge on PGC-1α, which activates NRF-1/2 and TFAM to drive mitochondrial transcription .
About the Author
Dr. Emma Sinclair, PhD, RD
Dr. Sinclair is a PhD in exercise physiology with 14 years of research experience in mitochondrial biology and sports nutrition. She is a registered dietitian and the author of over 20 peer-reviewed publications on exercise metabolism. She currently serves as the Director of Metabolic Research at the University of Queensland’s School of Human Movement and Nutrition Sciences. Dr. Sinclair’s own research has examined the effects of training state and frequency on mitochondrial adaptation in athletes and clinical populations.
Free Resources
- “The Strategic Fasting Protocol” (PDF) – A week-by-week guide to incorporating fasted training safely and effectively.
- “Mitochondrial Biogenesis Cheat Sheet” – A visual guide to the molecular pathways and how different training modalities affect them.
- “Fasted vs Fed Decision Flowchart” – Help you decide which approach fits your goals, schedule, and metabolic health.
- “Protein Timing Calculator” – Determine optimal protein intake around training sessions.
To access all resources for free: Visit https://worldclassblogs.com/category/our-focus/ and use code MITO2026
Discussion (For Comments Section)
I want to hear from you:
- Have you tried fasted training? What were your results—did it help with fat loss, energy, or performance?
- What is your biggest challenge with training around meals—time, energy, digestion, or something else?
- If you could design your ideal training state strategy (fasted sessions, fed sessions, twice-a-day), what would it look like?
Leave your answers below. Dr. Sinclair personally responds to every comment within 48 hours. The best “training state breakthrough” story each month wins a free 30-minute nutrition coaching session.
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