Acute effects of a ketone monoester, whey protein, or their coingestion on mTOR trafficking and protein-protein colocalization in human skeletal muscle.

We recently demonstrated that acute oral ketone monoester intake induces a stimulation of postprandial myofibrillar protein synthesis rates comparable to that elicited following the ingestion of 10 g whey protein or their coingestion. The present investigation aimed to determine the acute effects of ingesting a ketone monoester, whey protein, or their coingestion on mechanistic target of rapamycin (mTOR)-related protein-protein colocalization and intracellular trafficking in human skeletal muscle. In a randomized, double-blind, parallel group design, 36 healthy recreationally active young males (age: 24.2 ± 4.1 yr) ingested either: 1) 0.36 g·kg-1 bodyweight of the ketone monoester (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (KET), 2) 10 g whey protein (PRO), or 3) the combination of both (KET + PRO). Muscle biopsies were obtained in the overnight postabsorptive state (basal conditions), and at 120 and 300 min in the postprandial period for immunofluorescence assessment of protein translocation and colocalization of mTOR-related signaling molecules. All treatments resulted in a significant (Interaction: P < 0.0001) decrease in tuberous sclerosis complex 2 (TSC2)-Ras homolog enriched in brain (Rheb) colocalization at 120 min versus basal; however, the decrease was sustained at 300 min versus basal (P < 0.0001) only in KET + PRO. PRO and KET + PRO increased (Interaction: P < 0.0001) mTOR-Rheb colocalization at 120 min versus basal; however, KET + PRO resulted in a sustained increase in mTOR-Rheb colocalization at 300 min that was greater than KET and PRO. Treatment intake increased mTOR-wheat germ agglutinin (WGA) colocalization at 120 and 300 min (Time: P = 0.0031), suggesting translocation toward the fiber periphery. These findings demonstrate that ketone monoester intake can influence the spatial mechanisms involved in the regulation of mTORC1 in human skeletal muscle.NEW & NOTEWORTHY We explored the effects of a ketone monoester (KET), whey protein (PRO), or their coingestion (KET + PRO) on mTOR-related protein-protein colocalization and intracellular trafficking in human muscle. All treatments decreased TSC2-Rheb colocalization at 120 minutes; however, KET + PRO sustained the decrease at 300 min. Only PRO and KET + PRO increased mTOR-Rheb colocalization; however, the increase at 300 min was greater in KET + PRO. Treatment intake increased mTOR-WGA colocalization, suggesting translocation to the fiber periphery. Ketone bodies influence the spatial regulation of mTOR.

Acute ingestion of a ketone monoester, whey protein, or their co-ingestion in the overnight postabsorptive state elicit a similar stimulation of myofibrillar protein synthesis rates in young males: a double-blind randomized trial.

BACKGROUND: Ketone bodies may have anabolic effects in skeletal muscle via their capacity to stimulate protein synthesis. Whether orally ingested exogenous ketones can stimulate postprandial myofibrillar protein synthesis (MyoPS) rates with and without dietary protein co-ingestion is unknown. OBJECTIVES: This study aimed to evaluate the effects of ketone monoester intake and elevated blood ß-hydroxybutyrate (ß-OHB) concentration, with and without dietary protein co-ingestion, on postprandial MyoPS rates and mechanistic target of rapamycin complex 1 (mTORC1) pathway signaling. METHODS: In a randomized, double-blind, parallel group design, 36 recreationally active healthy young males (age: 24.2 ± 4.1 y; body fat: 20.9% ± 5.8%; body mass index: 23.4 ± 2 kg/m2) received a primed continuous infusion of L-[ring-2H5]-phenylalanine and ingested one of the following: 1) the ketone monoester (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (KET), 2) 10 g whey protein (PRO), or 3) the combination of both (KET+PRO). Blood and muscle biopsy samples were collected during basal and postprandial (300 min) conditions to assess ß-OHB, glucose, insulin, and amino acid concentrations, MyoPS rates, and mTORC1 pathway signaling. RESULTS: Capillary blood ß-OHB concentration increased similarly during postprandial conditions in KET and KET+PRO, with both being greater than PRO from 30 to 180 min (treatment × time interaction: P < 0.001). Postprandial plasma leucine and essential amino acid (EAA) incremental area under the curve (iAUC) over 300 min was greater (treatment: both P < 0.001) in KET+PRO compared with PRO and KET. KET, PRO, and KET+PRO stimulated postprandial MyoPS rates (0-300 min) higher than basal conditions [absolute change: 0.020%/h; (95% CI: 0.013, 0.027%/h), 0.014%/h (95% CI: 0.009, 0.019%/h), 0.019%/h (95% CI: 0.014, 0.024%/h), respectively (time: P < 0.001)], with no difference between treatments (treatment: P = 0.383) or treatment × time interaction (interaction: P = 0.245). mTORC1 pathway signaling responses did not differ between treatments (all P > 0.05). CONCLUSIONS: Acute oral intake of a ketone monoester, 10 g whey protein, or their co-ingestion in the overnight postabsorptive state elicit a similar stimulation of postprandial MyoPS rates in healthy young males. This trial was registered at clinicaltrials.gov as NCT04565444 (https://clinicaltrials.gov/study/NCT04565444).

The acute effects of insect vs. beef-derived protein on postprandial plasma aminoacidemia, appetite hormones, appetite sensations, and energy intake in healthy young men.

BACKGROUND/OBJECTIVES: The purpose of this study was to evaluate the acute effects of ingesting beef- and insect-derived protein on postprandial plasma amino acid and appetite hormone concentrations, appetite sensations, and ad libitum energy intake. SUBJECTS/METHODS: In a randomized, double-blind, crossover study, 20 young men (23 (SD: 4) y) completed two trials during which arterialized blood samples and VAS questionnaires were collected at baseline, and over 300-min after ingestion of beverages with similar energy and macronutrient content containing 25 g beef- or insect-derived (cricket) protein. Blood samples were analyzed for plasma amino acid and appetite hormone concentrations, while VAS questionnaires were applied to assess appetite sensations. After each trial, an ad libitum meal was immediately provided to assess energy intake. RESULTS: Adjusted mean postprandial incremental area under the curve (iAUC) was greater for cricket vs. beef-derived protein for plasma leucine, branched-chain amino acid, and essential amino acid concentrations (all P < 0.0001). Adjusted mean postprandial iAUC for hunger was lower following beef (-3030 (SE: 860)) vs. cricket-derived (-1197 (SE: 525)) protein (Difference: -1833 (95% CI: -3358, -308); P = 0.02), but was not different for other appetite sensations or appetite hormones (all P > 0.05). Adjusted mean ad libitum energy intake was 4072 (SE: 292) and 4408 (SE: 316) kJ following beef- and cricket-derived protein (Difference: -336 (95% CI: -992, 320); P = 0.30). CONCLUSION: Acute ingestion of cricket and beef-derived protein leads to differences in postprandial plasma amino acid concentrations, but elicits similar effects on appetite hormones, appetite sensations, and ad libitum energy intake in young men.

Ketone Monoester Supplementation Does Not Expedite the Recovery of Indices of Muscle Damage After Eccentric Exercise.

Purpose: The purpose of this study was to evaluate the effects of a ketone monoester supplement on indices of muscle damage during recovery after eccentric exercise. Methods: In a randomized, double-blind, independent group design, 20 moderately active healthy young adults consumed 360 mg per kg-1 bodyweight of a ketone monoester (KET) or energy-matched carbohydrate (CON) supplement twice daily following eccentric exercise (drop jumps). Maximal isometric voluntary contraction (MIVC) torque, counter-movement jump (CMJ) height, and muscle soreness were measured before (PRE), and immediately (POST), 24 h and 48 h post-exercise. Blood samples were collected for analysis of ß-hydroxybutyrate (ß-OHB), creatine kinase (CK), and select pro- and anti-inflammatory cytokines. Results: Peak blood ß-OHB concentration after supplement intake was greater (P < 0.001) in KET (4.4 ± 0.8 mM) vs. CON (0.4 ± 0.3 mM). Exercise increased CK concentration at 24 h and 48 h vs. PRE (time: P < 0.001) with no difference between KET and CON. Exercise reduced MIVC (KET: -19.9 ± 14.6; CON: -22.6 ± 11.1%) and CMJ (KET: -11.0 ± 7.5; CON: -13.0 ± 8.7%) at POST relative PRE; however, there was no difference between KET and CON on the recovery of MIVC at 24 h (KET: -15.4 ± 20.4; CON: -18.7 ± 20.1%) or 48 h (KET: -7.2 ± 21.2; CON: -11.8 ± 20.2%), or CMJ at 24 h (KET: -9.2 ± 11.5; CON: -13.4 ± 10.8) or 48 h (KET: -12.5 ± 12.4; CON: -9.1 ± 11.7). Muscle soreness was increased during post-exercise recovery (time: P < 0.001) with no differences between KET and CON. Monocyte chemoattractant protein-1 was greater (group: P = 0.007) in CON (236 ± 11 pg/mL) vs. KET (187 ± 11 pg/mL). Conclusion: In conclusion, twice daily ingestion of a ketone monoester supplement that acutely elevates blood ß-OHB concentration does not enhance the recovery of muscle performance or reduce muscle soreness following eccentric exercise in moderately active, healthy young adults.