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Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor
Open accessAbstract
Metabolic flexibility of skeletal muscle, that is, the preference for fat oxidation (FOx) during fasting and for carbohydrate oxidation in response to insulin, is decreased during insulin resistance. The aim of this study was to test the hypothesis that the capacity of myotubes to oxidize fat in vitro reflects the donor's metabolic characteristics. Insulin sensitivity (IS) and metabolic flexibility of 16 healthy, young male subjects was determined by euglycemic hyperinsulinemic clamp. Muscle samples were obtained from vastus lateralis, cultured, and differentiated into myotubes. In human myotubes in vitro, we measured suppressibility (glucose suppression of FOx) and adaptability (an increase in FOx in the presence of high palmitate concentration). We termed these dynamic changes in FOx metabolic switching. In vivo, metabolic flexibility was positively correlated with IS and maximal oxygen uptake and inversely correlated with percent body fat. In vitro suppressibility was inversely correlated with IS and metabolic flexibility and positively correlated with body fat and fasting FFA levels. Adaptability was negatively associated with percent body fat and fasting insulin and positively correlated with IS and metabolic flexibility. The interindividual variability in metabolic phenotypes was preserved in human myotubes separated from their neuroendocrine environment, which supports the hypothesis that metabolic switching is an intrinsic property of skeletal muscle.
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“Increasing palmitate concentration in vitro produced an increase in 14 CO 2 and ASP production (r = 0.998; P < 0.0001; Figure 1B).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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“However, in some subjects, the maximal rate of FOx (reflected by ASP production) flattened out at 100 µM palmitate, which suggests an inability to further oxidize fatty acids.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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In the paper
“Metabolic flexibility, defined as the change in RQ during a clamp [∆RQ (clamp)], was negatively correlated with percent body fat and fasting FFA levels”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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In the paper
“An increase in the extracellular glucose concentration in vitro resulted in a decrease of FOx as indicated by levels of both 14 CO 2 and 14 C-intermediate metabolites of FOx (acidsoluble products [ASPs]) (r = -0.96,P < 0.0001 for 14 CO 2 ; r = -0.95,P < 0.0001 for ASPs; data not shown).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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“A negative correlation was observed between IS and fasting levels of both insulin (r = -0.65;P = 0.0003) and FFAs (r = -0.81;P < 0.0001).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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In the paper
“An inverse relationship was observed between basal FOx (at 1 µCi/ml 14 C-palmitate with 0 µM cold palmitate) and the fold increase in FOx (r = -0.51;P = 0.025).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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“A positive relationship was observed between percent suppression of 14 CO 2 by glucose (% suppression = [1 -(FOx at 5 mM glucose / FOx at 0 mM glucose)] × 100) and percent suppression of total palmitate oxidation (TOx) by glucose (TOx = 14 CO 2 + 14 C-ASPs) (r = 0.51; P = 0.03).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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In the paper
“For fold change of TOx, no significant relationship was found between the results of the 2 assays.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
plain- Asserted: passes — The authors put it forward as true.
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“However, these in vitro-in vivo relationships were only observed for CO 2 , not for total FOx (P = 0.5 for percent body fat; P = 0.1 for FFA levels)”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Glucose suppressibility of FOx is an intrinsic characteristic of muscle cells.
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In the paper
“No significant relationship was observed between fold increase in 14 CO 2 (in the in vitro adaptability assay) and percent suppression of 14 CO 2 (in the in vitro suppressibility assay), when all 16 subjects were used for the analysis.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
plainn = 16- Asserted: passes — The authors put it forward as true.
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In the paper
“On the other hand, ∆RQ (clamp) correlated positively with IS and VO 2 max”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
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In the paper
“As expected, IS, measured by euglycemic hyperinsulinemic clamp (clamp), correlated negatively with percentage body fat (r = -0.60;P = 0.008) and positively with maximal oxygen uptake (VO 2 max) (r = 0.59; P = 0.015)”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
plaincorrelational- Asserted: passes — The authors put it forward as true.
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In the paper
“Fasting RQ during the clamp, a parameter of fasting fuel partitioning, was related neither to IS (P = 0.80) nor to anthropometric characteristics of subjects (percent body fat; P = 0.60).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Relationship between metabolic flexibility and IS in vivo.
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“Neither percentage suppression of 14 CO 2 nor percentage suppression of TOx correlated with fasting RQ (measured by clamp) or 24hour RQ (measured by indirect calorimetry) in vivo.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Glucose suppressibility of FOx is an intrinsic characteristic of muscle cells.
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
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In the paper
“In our model, short-term exposure of myotubes to increased palmitate levels enhanced FOx 3-to 5-fold, the magnitude of response correlating with clinical phenotype.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Discussion
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In the paper
“Similarly, we found a correlation between the suppressibility of FOx by glucose (percent suppression of 14 CO 2 ) in vitro and percent body fat (r = 0.56; P = 0.02) and fasting FFA levels (r = 0.58; P = 0.01; Figure 2, C and D) in vivo.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Glucose suppressibility of FOx is an intrinsic characteristic of muscle cells.
plaincorrelational- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Fold increase in 14 CO 2 was positively correlated with in vivo metabolic flexibility [as indicated by ∆RQ (clamp)] (r = 0.53; P = 0.035; Figure 3A), IS (as measured by clamp) (r = 0.60; P = 0.015; Figure 3B) and aerobic capacity (VO 2 max) (r = 0.70; P = 0.002; Figure 3C), respectively.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Metabolic adaptability is an intrinsic characteristic of muscle cells.
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In the paper
“14 CO2 at basal palmitate | 0.33 ± 0.25 | 0.15-0.71”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Statistical analysis.
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“As was the case with the in vitro glucose suppressibility assay, CO 2 production appeared to be a better correlate with the clinical phenotypes than was ASP production.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Metabolic adaptability is an intrinsic characteristic of muscle cells.
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In the paper
“analysis clearly demonstrate the relationship between clinical phenotypes and dynamic changes in FOx in cultured myotubes, which supports the hypothesis that metabolic switching is, at least to a certain extent, an intrinsic characteristic of skeletal muscle.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Discussion
boostedcorrelational- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Rests on an assumption: Growing human myotubes in a standardized insulin-free medium strips away whole-body neuroendocrine and environmental effects, so the in vitro readouts reveal intrinsic muscle-cell properties.
- Rests on an assumption: The donor's metabolic phenotype is stable and captured well enough by the measured in vivo traits.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“According to our studies, in vivo insulin responsiveness decreases with increasing in vitro suppressibility, which indicates greater glucose suppression of FOx in insulin resistance and metabolic inflexibility.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Figure 4
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In the paper
“RQ were adjusted for energy balance as described previously (71).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Body composition.
What the sources say
- Not checked yetcites [71] Nguyen, 2003, Med. Biol. Eng. Comput. source
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Glucose disposal rate, a parameter of IS, was adjusted for kilograms of LBM.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Figure 3
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“All assays were performed in duplicate, and data were normalized to protein content.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Figure 4
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one. The quote only partly states this claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Coefficients of variation for the measurement of the LBM, fat mass, and percentage of body fat were 0.8%, 1.6%, and 1.7%, respectively.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Body composition.
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“Defects in skeletal muscle oxidative capacity and fat metabolism are highly correlated with insulin sensitivity (IS) (1,5,12) and are believed to contribute to the pathogenesis of insulin resistance (13,14).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Introduction
What the sources say
- Not checked yetcites [1] Colberg, 1995, J. Clin. Invest. source
- Not checked yetcites [5] Kelley, 1999, Am. J. Physiol. source
- Not checked yetcites [12] Bruce, 2003, J. Clin. Endocrinol. Metab. source
- Not checked yetcites [13] Storlien, 2004, Proc. Nutr. Soc. source
- Not checked yetcites [14] Kelley, 2000, Diabetes. source
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What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Coefficients of variation for total daily energy expenditure and RQ are 2% and 16%, respectively.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Body composition.
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one. The quote only partly states this claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“To account for the effect of basal FOx on the fold increase in FOx, the semi-partial correlation was computed to examine the relations between fold increase FOx and clinical characteristics (BMI, VO2max, fasting insulin, etc.), while basal FOx production was held as a constant.”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Statistical analysis.
plaincorrelational- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim.
The paper relies on this without support.
Growing human myotubes in a standardized insulin-free medium strips away whole-body neuroendocrine and environmental effects, so the in vitro readouts reveal intrinsic muscle-cell properties.
“We cultured muscle satellite cells from individuals with differing clinical characteristics in a standardized, insulin-free medium”
“This model is ideal for studying interindividual differences of metabolism intrinsic to the skeletal muscle, i.e., independent of the influences of the whole-body environment”
The paper relies on this without support.
The donor's metabolic phenotype is stable and captured well enough by the measured in vivo traits.
“We related these responses of myotubes in vitro to the clinical phenotypes of their donors.”
The paper relies on this without support.
In vitro intrinsic defects in metabolic switching reflect processes that contribute to obesity and insulin resistance in vivo.
“Intrinsic defects in metabolic switching of skeletal muscle might participate in the generation of a “lipotoxic milieu” in the muscle cell (13, 14) that acts in concert with environmental factors, such as a HFD, to favor the accumulation of lipids and lipid intermediates implicated in insulin resistance (34)(35)(36)(37).”
The paper relies on this without support.
The in vitro suppressibility assay in insulin-free medium measures the same process as in vivo insulin responsiveness.
“The in vitro suppressibility assay was designed to measure the ability of myotubes to maintain FOx in the presence of glucose, in insulin-free medium (that is, fuel preference of metabolic switching).”
The paper relies on this without support.
The in vitro adaptability assay measures the capacity of myotubes to increase fat oxidation in a way that reflects in vivo adaptability.
“The in vitro adaptability assay focused on the capacity of myotubes to increase FOx in response to an acute exposure to a high palmitate concentration.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The cited source says what the claim attributes to it.
In the citing paper
“Such muscle mitochondrial dysfunction was suggested to be an intrinsic characteristic of muscle cells, since fat oxidation in response to palmitate was lower in myotubes from insulin-resistant vs. insulin-sensitive individuals ([16](#r16), [61](#r61)).”
From Metabolic flexibility and insulin resistance (2008) · cites this paper as [61]
In the citing paper
“Accordingly, MetFlex-lip of human myotubes in vitro correlates directly with the donor's insulin sensitivity in vivo (29).”
From Metabolic flexibility to lipid availability during exercise is enhanced in individuals with high insulin sensitivity (2018) · cites this paper as [29]
In the citing paper
“cross-sectional studies showing that isolated muscle strips, tissue homogenates or cultured myocytes from severely obese and/or diabetic humans exhibit diminished rates of fat oxidation when compared to specimens derived from healthy control subjects [9,41,46,87].”
From Intramuscular triacylglycerol and insulin resistance: guilty as charged or wrongly accused? (2010) · cites this paper as [87]
In the citing paper
“This characteristic appears to be intrinsic to skeletal muscle because primary muscle cell cultures from insulin-resistant subjects retain abnormal patterns of whole-body fuel use (1).”
In the citing paper
“The relationship between fasting RQ and insulin sensitivity, however, has not always been observed.”
From Metabolic flexibility and insulin resistance (2008) · cites this paper as [61]
In the citing paper
“Metabolic inflexibility to glucose and impaired glucose storage during a hyperinsulinemic clamp is consistently reported in insulin-resistant subjects.”
From Metabolic flexibility and insulin resistance (2008) · cites this paper as [61]
In the citing paper
“Contrary to expectations, in vitro suppressibility of fat oxidation was lower in subjects with higher whole body insulin sensitivity and higher metabolic flexibility to glucose.”
From Metabolic flexibility and insulin resistance (2008) · cites this paper as [61]
In the citing paper
“In fact, a direct relationship between metabolic flexibility to glucose and glucose disposal rate is described ([14](#r14), [24](#r24), [61](#r61), [62](#r62)) ([Fig. 3*B*](#f3)).”
From Metabolic flexibility and insulin resistance (2008) · cites this paper as [61]
In the citing paper
“Such observations are most likely attributed to testosterone-induced changes in lipid metabolism and/or altered postreceptor insulin signaling in muscle (54) and also improved insulin sensitivity enhancing muscle lipid uptake (55).”
From Onset of effects of testosterone treatment and time span until maximum effects are achieved (2011) · cites this paper as [55]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Statistical analysis.
“| In vitro adaptability | Fold increase in 14 CO2 | 3.19 ± 0.76 | 2.99-4.87 |”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Figure 2
“Muscle cells from 16 individuals were grown and differentiated into myotubes in 24-well plates.”
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > Body composition.
“Samples of vastus lateralis, weighing 60-100 mg, were obtained by muscle biopsy using the Bergstrom technique (73).”
§ Dynamic changes in fat oxidation in human primary myocytes mirror metabolic characteristics of the donor > References
“73. Ravussin, E., Lillioja, S., Anderson, T.E., Christin, L., and Bogardus, C. 1986. Determinants of 24- hour energy expenditure in man. Methods and results using a respiratory chamber. J. Clin. Invest. 78:1568-1578.”
Citations
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“VO2max was determined by a progressive treadmill test to exhaustion in accordance with the recommendations described by the American College of Sports Medicine (69).”
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Cellular mechanisms of insulin resistance
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Dysfunction of Mitochondria in Human Skeletal Muscle in Type 2 Diabetes
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Mechanism of free fatty acid-induced insulin resistance in humans.
Roden, M., Price, T. B., Perseghin, G., Petersen, K. F., Rothman, D. L., Cline, G. W., & Shulman, G. I. (1996) · Journal of Clinical Investigation · doi:10.1172/jci118742 · Cited by 1,539
Percutaneous Needle Biopsy of Skeletal Muscle in Physiological and Clinical Research
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Metadata
- Authors
- Ukropcova, B., McNeil, M., Sereda, O., de Jonge, L., Xie, H., Bray, G. A., & Smith, S. R.
- Affiliations
- Pennington Biomedical Research Center (US)
- Corresponding author
- Smith, Steven R.
- Venue
- Journal of Clinical Investigation
- Published
- 2005-07-01
- Volume
- 115
- Issue
- 7
- Pages
- 1934-1941
- ISSN
- 0021-9738 (ISSN-L)
- PMC
- PMC1159139
- OpenAlex
- W2098384661
- Semantic Scholar
- f721ac6c8e8feb8139e5dd7e128cb8df2dd30680
- Cited by
- 213
- References
- 79
- Citations by year
- 2026: 2 · 2025: 7 · 2024: 4 · 2023: 4 · 2022: 3 · 2021: 5 · 2020: 10 · 2019: 4 · 2018: 7 · 2017: 9 · 2016: 10 · 2015: 11 · 2014: 12 · 2013: 21 · 2012: 11
- FWCI
- 5.52
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Investigation (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- U.S. Department of Agriculture
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
- Health Sciences › Medicine › Physiology (medical) › Adipose Tissue and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Muscle Physiology and Disorders
- Keywords
- fat oxidation, insulin sensitivity, myotubes, metabolic switch, insulin resistance, palmitate, fat mass, VO2max, interindividual variability, vastus lateralis
- MeSH terms
- Adolescent, Adult, Dietary Fats, Glucose, Humans, Insulin Resistance, Male, Models, Biological, Oxidation-Reduction, Phenotype, Glucose Clamp Technique, Muscle Fibers, Skeletal, Palmitic Acid, In Vitro Techniques
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
A language model drew the claims from the paper on 2026-10-05, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-05 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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