Systemic proteome adaptions to 7-day complete caloric restriction in humans.

Surviving long periods without food has shaped human evolution. In ancient and modern societies, prolonged fasting was/is practiced by billions of people globally for religious purposes, used to treat diseases such as epilepsy, and recently gained popularity as weight loss intervention, but we still have a very limited understanding of the systemic adaptions in humans to extreme caloric restriction of different durations. Here we show that a 7-day water-only fast leads to an average weight loss of 5.7 kg (±0.8 kg) among 12 volunteers (5 women, 7 men). We demonstrate nine distinct proteomic response profiles, with systemic changes evident only after 3 days of complete calorie restriction based on in-depth characterization of the temporal trajectories of ~3,000 plasma proteins measured before, daily during, and after fasting. The multi-organ response to complete caloric restriction shows distinct effects of fasting duration and weight loss and is remarkably conserved across volunteers with >1,000 significantly responding proteins. The fasting signature is strongly enriched for extracellular matrix proteins from various body sites, demonstrating profound non-metabolic adaptions, including extreme changes in the brain-specific extracellular matrix protein tenascin-R. Using proteogenomic approaches, we estimate the health consequences for 212 proteins that change during fasting across ~500 outcomes and identified putative beneficial (SWAP70 and rheumatoid arthritis or HYOU1 and heart disease), as well as adverse effects. Our results advance our understanding of prolonged fasting in humans beyond a merely energy-centric adaptions towards a systemic response that can inform targeted therapeutic modulation.

Carbohydrate Ingestion during Prolonged Cycling Improves Next-Day Time Trial Performance and Alters Amino Acid Concentrations.

INTRODUCTION: Exercise with low carbohydrate availability increases protein degradation, which may reduce subsequent performance considerably. The present study aimed to investigate the effect of carbohydrate ingestion during standardized exercise with and without exhaustion on protein degradation and next-day performance. METHODS: Seven trained male cyclists (V̇O 2max 66.8 ± 1.9 mL·kg -1 ·min -1 ; mean ± SEM) cycled to exhaustion (~2.5 h) at a power output eliciting 68% of V̇O 2max (W 68% ). This was followed by repeating 1-min work/1-min recovery intervals at 90% of V̇O 2max (W 90% ) until exhaustion. During W 68% , cyclists consumed a placebo water drink (PLA) the first time and a carbohydrate drink (CHO), 1 g carbohydrate·kg -1 ·h -1 , the second time. The participants performed the same amount of work under the two conditions, separated by at least 1 wk. A standardized diet was provided to the participants so that the two conditions were isoenergetic. To test the impact of carbohydrates on recovery, participants completed a time trial (TT) the next day. RESULTS: Carbohydrate ingestion maintained carbohydrate availability during W 68% and W 90% : total carbohydrate oxidation was significantly higher in CHO ( P = 0.022), and plasma glucose concentration was maintained compared with PLA ( P = 0.025). Next-day performance during TT was better after CHO ingestion (CHO, 41:49 ± 1:38 min; PLA, 42:50 ± 1:46 min; P = 0.020; effect size d = 0.23, small), as was gross efficiency (CHO, 18.6% ± 0.3%; PLA, 17.9% ± 0.3%; P = 0.019). Urinary nitrogen excretion ( P = 0.897) and urinary 3-methylhistidine excretion ( P = 0.673) did not significantly differ during the study period. Finally, tyrosine and phenylalanine plasma concentrations increased in PLA but not in CHO ( P = 0.018). CONCLUSIONS: Carbohydrate ingestion during exhaustive exercise reduced deterioration in next-day performance through reduced metabolic stress and development of fatigue. In addition, some parameters point toward less protein degradation, which would preserve muscle function.

Similar performance after intake of carbohydrate plus whey protein and carbohydrate only in the early phase after non-exhaustive cycling.

AIM: The aim of the present study was to compare performance 5 h after a 90-min endurance training session when either carbohydrate only or carbohydrate with added whey hydrolysate or whey isolate was ingested during the first 2 h of the recovery period. METHODS: Thirteen highly trained competitive male cyclists completed three exercise and diet interventions (double-blinded, randomized, crossover design) separated by 1 week. The 90-min morning session (EX1) included a 60 min time-trial (TT60 ). Immediately and 1 h after exercise, participants ingested either (1) 1.2 g carbohydrate∙kg-1 ∙h-1 (CHO), (2) 0.8 g carbohydrate∙kg-1 ∙h-1 + 0.4 g isolate whey protein∙kg-1 ∙h-1 (ISO) or (3) 0.8 g carbohydrate∙kg-1 ∙h-1 + 0.4 g hydrolysate whey protein∙kg-1 ∙h-1 (HYD). Additional intakes were identical between interventions. After 5 h of recovery, participants completed a time-trial performance (TTP ) during which a specific amount of work was performed. Blood and urine were collected throughout the day. RESULTS: TTP did not differ significantly between dietary interventions (CHO: 43:54 ± 1:36, ISO: 46:55 ± 2:32, HYD: 44:31 ± 2:01 min). Nitrogen balance during CHO was lower than ISO (p < 0.0001) and HYD (p < 0.0001), with no difference between ISO and HYD (p = 0.317). In recovery, the area under the curve for blood glucose was higher in CHO compared to ISO and HYD. HR, VO2 , RER, glucose, and lactate during EX2 were similar between interventions. CONCLUSION: Performance did not differ after 5 h of recovery whether carbohydrate only or isocaloric carbohydrate plus protein was ingested during the first 2 h. Correspondingly, participants were not in negative nitrogen balance in any dietary intervention.

An Acute Bout of Endurance Exercise Does Not Prevent the Inhibitory Effect of Caffeine on Glucose Tolerance the following Morning.

Caffeine reduces glucose tolerance, whereas exercise training improves glucose homeostasis. The aim of the present study was to investigate the effect of caffeine on glucose tolerance the morning after an acute bout of aerobic exercise. Methods: The study had a 2 × 2 factorial design. Oral glucose tolerance tests (OGTT) were performed after overnight fasting with/without caffeine and with/without exercise the evening before. Eight healthy young active males were included (Age 25.5 ± 1.5 years; 83.9 ± 9.0 kg; VO2max: 54.3 ± 7.0 mL·kg-1·min-1). The exercise session consisted of 30 min cycling at 71% of VO2max followed by four 5 min intervals at 84% with 3 min of cycling at 40% of VO2max between intervals. The exercise was performed at 17:00 h. Energy expenditure at each session was ~976 kcal. Lactate increased to ~8 mM during the exercise sessions. Participants arrived at the laboratory the following morning at 7.00 AM after an overnight fast. Resting blood samples were taken before blood pressure and heart rate variability (HRV) were measured. Caffeine (3 mg/kg bodyweight) or placebo (similar taste/flavor) was ingested, and blood samples, blood pressure and HRV were measured after 30 min. Next, the OGTTs were initiated (75 g glucose dissolved in 3 dL water) and blood was sampled. Blood pressure and HRV were measured during the OGTT. Caffeine increased the area under curve (AUC) for glucose independently of whether exercise was done the evening before (p = 0.03; Two-way ANOVA; Interaction: p = 0.835). Caffeine did not significantly increase AUC for C-peptides compared to placebo (p = 0.096), and C-peptide response was not influenced by exercise. The acute bout of exercise did not significantly improve glucose tolerance the following morning. Diastolic blood pressure during the OGTT was slightly higher after intake of caffeine, independent of whether exercise was performed the evening before or not. Neither caffeine nor exercise the evening before significantly influenced HRV. In conclusion, caffeine reduced glucose tolerance independently of whether endurance exercise was performed the evening before. The low dose of caffeine did not influence heart rate variability but increased diastolic blood pressure slightly.

Effect of aerobic exercise and low-carbohydrate high-fat diet on glucose tolerance and android/gynoid fat in overweight/obese women: A randomized controlled trial.

The study was designed to compare the effects of weight loss induced by a low-carbohydrate-high-fat diet or a normal diet, with and without exercise, on glucose tolerance measured as area under the curve (AUC), and android (A) and gynoid (G) fat distribution. The study was registered at clinicaltrials.gov; NCT04100356. In total, 57 women classified as overweight or obese (age 40 ± 3.5 years, body mass index 31.1 ± 2.6 kg/m2) were randomly assigned and completed a 10-week intervention using a low-carbohydrate high-fat diet or a normal diet, with or without aerobic interval exercise. An equal deficit of 700 kcal/day was prescribed, either restricting the diet only, or moderately restricting diet and including three 50-min high-intensity bicycle sessions per week. There were thus four groups: normal diet (NORM); low-carbohydrate-high-fat diet (LCHF); normal diet with exercise (NORM-EX); and low-carbohydrate-high-fat diet with exercise (LCHF-EX). Linear mixed models was used to assess differences between groups. With all groups pooled, the intervention resulted in a weight loss of 6.7 ± 2.5% (p < 0.001). The intervention did not result in differences between groups in AUC glucose, nor in fasting glucose or indicis for insulin resistance such as Homeostatic Model Assessment, Matsuda Insulin Sensitivity Index, insulinogenic index and disposition index. Post-intervention android fat was lower in LCHF than NORM (3,223 ± 727 vs. 2,533 ± 535 g, p = 0.041). LCHF reached a lower A/G ratio than NORM (0.94 ± 0.12 vs. 1.04 ± 0.09, p = 0.011) and LCHF-EX (0.94 ± 0.12 vs. 1.09 ± 0.09, p < 0.001) after the intervention. LCHF resulted in lower android fat mass compared to NORM and the lowest A/G ratio compared to the other matched groups, but with no accompanying improvement in AUC glucose. In conclusion, although all groups achieved improvements in glucose tolerance, no superior effect was observed with the LCHF diet, neither with nor without exercise.

Increased Mass-Specific Maximal Fat Oxidation Rate with Small versus Large Muscle Mass Exercise.

INTRODUCTION: Skeletal muscle perfusion and oxygen (O2) delivery are restricted during whole-body exercise because of a limited cardiac output (Q˙). This study investigated the role of reducing central limitations to exercise on the maximal fat oxidation rate (MFO) by comparing mass-specific MFO (per kilogram of active lean mass) during one-legged (1L) and two-legged (2L) cycling. We hypothesized that the mass-specific MFO would be higher during 1L than 2L cycling. METHODS: Twelve male subjects (V̇O2peak, 59.3 ± 8.4 mL·kg-1·min-1; mean ± SD) performed step-incremental 2L- (30%-80% of V̇O2peak) and 1L (50% of 2L power output, i.e., equal power output per leg) cycling (counterbalanced) while steady-state pulmonary gas exchanges, Q˙ (pulse-contour analysis), and skeletal muscle (vastus lateralis) oxygenation (near-infrared spectroscopy) were determined. MFO and the associated power output (FatMax) were calculated from pulmonary gas exchanges and stoichiometric equations. A counterweight (10.9 kg) was added to the contralateral pedal arm during 1L cycling. Leg lean mass was determined by DEXA. RESULTS: The absolute MFO was 24% lower (0.31 ± 0.12 vs 0.44 ± 0.20 g·min-1, P = 0.018), whereas mass-specific MFO was 52% higher (28 ± 11 vs 20 ± 10 mg·min-1·kg-1, P = 0.009) during 1L than 2L cycling. FatMax was similar expressed as power output per leg (60 ± 28 vs 58 ± 22 W, P = 0.649). Q˙ increased more from rest to exercise during 1L than 2L cycling when expressed per active leg (ANOVA main effect: P = 0.003). Tissue oxygenation index and Δ[deoxy(Hb + Mb)] were not different between exercise modes (ANOVA main effects: P ≥ 0.587), indicating similar skeletal muscle fractional O2 extraction. CONCLUSIONS: Mass-specific MFO is increased by exercising a small muscle mass, potentially explained by increased perfusion and more favorable conditions for O2 delivery than during whole-body exercise.

Differential Effects of One Meal per Day in the Evening on Metabolic Health and Physical Performance in Lean Individuals.

Background: Generally, food intake occurs in a three-meal per 24 h fashion with in-between meal snacking. As such, most humans spend more than ∼ 12-16 h per day in the postprandial state. It may be reasoned from an evolutionary point of view, that the human body is physiologically habituated to less frequent meals. Metabolic flexibility (i.e., reciprocal changes in carbohydrate and fatty acid oxidation) is a characteristic of metabolic health and is reduced by semi-continuous feeding. The effects of time-restricted feeding (TRF) on metabolic parameters and physical performance in humans are equivocal. Methods: To investigate the effect of TRF on metabolism and physical performance in free-living healthy lean individuals, we compared the effects of eucaloric feeding provided by a single meal (22/2) vs. three meals per day in a randomized crossover study. We included 13 participants of which 11 (5 males/6 females) completed the study: age 31.0 ± 1.7 years, BMI 24.0 ± 0.6 kg/m2 and fat mass (%) 24.0 ± 0.6 (mean ± SEM). Participants consumed all the calories needed for a stable weight in either three meals (breakfast, lunch and dinner) or one meal per day between 17:00 and 19:00 for 11 days per study period. Results: Eucaloric meal reduction to a single meal per day lowered total body mass (3 meals/day -0.5 ± 0.3 vs. 1 meal/day -1.4 ± 0.3 kg, p = 0.03), fat mass (3 meals/day -0.1 ± 0.2 vs. 1 meal/day -0.7 ± 0.2, p = 0.049) and increased exercise fatty acid oxidation (p < 0.001) without impairment of aerobic capacity or strength (p > 0.05). Furthermore, we found lower plasma glucose concentrations during the second half of the day during the one meal per day intervention (p < 0.05). Conclusion: A single meal per day in the evening lowers body weight and adapts metabolic flexibility during exercise via increased fat oxidation whereas physical performance was not affected.

Eight sessions of endurance training decrease fasting glucose and improve glucose tolerance in middle-aged overweight males.

Exercise improves metabolic regulation and reduces the risk of developing type 2 diabetes and other metabolic diseases. The recommendations for exercise are rather general and the health benefits of controlled training studies are important to make better recommendations. In the present study, we report that eight endurance training sessions over 3 weeks reduced fasting glucose, and improved glucose tolerance and plasma lipids in sedentary middle-aged males (44-64 years) with overweight or obesity (BMI: 27-38). The decrease in fasting glucose was substantial (from 5.3 ± 0.3 to 4.8 ± 0.2 mM; p < .001). The training sessions consisted of 60-min indoor-cycling at ∼83% of peak heart rate divided in four blocks of 15 min cycling, with 2-min rest between blocks. Maximal oxygen uptake did not increase (38.8 ± 1.8 vs. 39.0 ± 1.6 ml kg-1 min-1). In conclusion, 3-weekly sessions of moderate-/high-intensity endurance training can be recommended for untrained males with overweight or obesity to improve glucose homeostasis.

Coingestion of protein and carbohydrate in the early recovery phase, compared with carbohydrate only, improves endurance performance despite similar glycogen degradation and AMPK phosphorylation.

The present study compared the effects of postexercise carbohydrate plus protein (CHO+PROT) and carbohydrate (CHO)-only supplementation on muscle glycogen metabolism, anabolic cell signaling, and subsequent exercise performance. Nine endurance-trained males cycled twice to exhaustion (muscle glycogen decreased from ~495 to ~125 mmol/kg dry wt) and received either CHO only (1.2 g·kg-1·h-1) or CHO+PROT (0.8/0.4 g·kg-1·h-1) during the first 90 min of recovery. Glycogen content was similar before the performance test after 5 h of recovery. Glycogen synthase (GS) fractional activity increased after exhaustive exercise and remained activated 5 h after, despite substantial glycogen synthesis (176.1 ± 19.1 and 204.6 ± 27.0 mmol/kg dry wt in CHO and CHO+PROT, respectively; P = 0.15). Phosphorylation of GS at site 3 and site 2+2a remained low during recovery. After the 5-h recovery, cycling time to exhaustion was improved by CHO+PROT supplementation compared with CHO supplementation (54.6 ± 11.0 vs. 46.1 ± 9.8 min; P = 0.009). After the performance test, muscle glycogen was equally reduced in CHO+PROT and CHO. Akt Ser473 and p70s6k Thr389 phosphorylation was elevated after 5 h of recovery. There were no differences in Akt Ser473, p70s6k Thr389, or TSC2 Thr1462 phosphorylation between treatments. Nitrogen balance was positive in CHO+PROT (19.6 ± 7.6 mg nitrogen/kg; P = 0.04) and higher than CHO (-10.7 ± 6.3 mg nitrogen/kg; P = 0.009). CHO+PROT supplementation during exercise recovery improved subsequent endurance performance relative to consuming CHO only. This improved performance after CHO+PROT supplementation could not be accounted for by differences in glycogen metabolism or anabolic cell signaling, but may have been related to differences in nitrogen balance.NEW & NOTEWORTHY Endurance athletes competing consecutive days need optimal dietary intake during the recovery period. We report that coingestion of protein and carbohydrate soon after exhaustive exercise, compared with carbohydrate only, resulted in better performance the following day. The better performance after coingestion of protein and carbohydrate was not associated with a higher rate of glycogen synthesis or activation of anabolic signaling compared with carbohydrate only. Importantly, nitrogen balance was positive after coingestion of protein and carbohydrate, which was not the case after intake of carbohydrate only, suggesting that protein synthesis contributes to the better performance the following day.

Sprint Interval Running and Continuous Running Produce Training Specific Adaptations, Despite a Similar Improvement of Aerobic Endurance Capacity-A Randomized Trial of Healthy Adults.

The purpose of the present study was to investigate training-specific adaptations to eight weeks of moderate intensity continuous training (CT) and sprint interval training (SIT). Young healthy subjects (n = 25; 9 males and 16 females) performed either continuous training (30-60 min, 70-80% peak heart rate) or sprint interval training (5-10 near maximal 30 s sprints, 3 min recovery) three times per week for eight weeks. Maximal oxygen consumption, 20 m shuttle run test and 5·60 m sprint test were performed before and after the intervention. Furthermore, heart rate, oxygen pulse, respiratory exchange ratio, lactate and running economy were assessed at five submaximal intensities, before and after the training interventions. Maximal oxygen uptake increased after CT (before: 47.9 ± 1.5; after: 49.7 ± 1.5 mL·kg-1·min-1, p < 0.05) and SIT (before: 50.5 ± 1.6; after: 53.3 ± 1.5 mL·kg-1·min-1, p < 0.01), with no statistically significant differences between groups. Both groups increased 20 m shuttle run performance and 60 m sprint performance, but SIT performed better than CT at the 4th and 5th 60 m sprint after the intervention (p < 0.05). At submaximal intensities, CT, but not SIT, reduced heart rate (p < 0.05), whereas lactate decreased in both groups. In conclusion, both groups demonstrated similar improvements of several performance measures including VO2max, but sprint performance was better after SIT, and CT caused training-specific adaptations at submaximal intensities.

Low-Carbohydrate High-Fat Diet and Exercise: Effect of a 10-Week Intervention on Body Composition and CVD Risk Factors in Overweight and Obese Women-A Randomized Controlled Trial.

We assessed the effect of weight-loss induced with a low-carbohydrate-high-fat diet with and without exercise, on body-composition, cardiorespiratory fitness and cardiovascular risk factors. A total of 57 overweight and obese women (age 40 ± 3.5 years, body mass index 31.1 ± 2.6 kg∙m-2) completed a 10-week intervention using a low-carbohydrate-high-fat diet, with or without interval exercise. An equal deficit of 700 kcal∙day-1 was prescribed, restricting diet only, or moderately restricting diet and adding exercise, producing four groups; normal diet (NORM); low-carbohydrate-high-fat diet (LCHF); normal diet and exercise (NORM-EX); and low-carbohydrate-high-fat diet and exercise (LCHF-EX). Linear Mixed Models were used to assess between-group differences. The intervention resulted in an average 6.7 ± 2.5% weight-loss (p < 0.001). Post-intervention % fat was lower in NORM-EX than NORM (40.0 ± 4.2 vs. 43.5 ± 3.5%, p = 0.024). NORM-EX reached lower values in total cholesterol than NORM (3.9 ± 0.6 vs. 4.7 ± 0.7 mmol/L, p = 0.003), and LCHF-EX (3.9 ± 0.6 vs. 4.9 ± 1.1 mmol/L, p = 0.004). Post intervention triglycerides levels were lower in NORM-EX than NORM (0.87 ± 0.21 vs. 1.11 ± 0.34 mmol/L, p = 0.030). The low-carbohydrate-high-fat diet had no superior effect on body composition, V˙O2peak or cardiovascular risk factors compared to a normal diet, with or without exercise. In conclusion, the intervention decreased fat mass, but exercise improved body composition and caused the most favorable changes in total cholesterol and triglycerides in the NORM-EX. Exercise increased cardiorespiratory fitness, regardless of diet.

Exercise in vivo marks human myotubes in vitro: Training-induced increase in lipid metabolism.

BACKGROUND AND AIMS: Physical activity has preventive as well as therapeutic benefits for overweight subjects. In this study we aimed to examine effects of in vivo exercise on in vitro metabolic adaptations by studying energy metabolism in cultured myotubes isolated from biopsies taken before and after 12 weeks of extensive endurance and strength training, from healthy sedentary normal weight and overweight men. METHODS: Healthy sedentary men, aged 40-62 years, with normal weight (body mass index (BMI) < 25 kg/m2) or overweight (BMI ≥ 25 kg/m2) were included. Fatty acid and glucose metabolism were studied in myotubes using [14C]oleic acid and [14C]glucose, respectively. Gene and protein expressions, as well as DNA methylation were measured for selected genes. RESULTS: The 12-week training intervention improved endurance, strength and insulin sensitivity in vivo, and reduced the participants' body weight. Biopsy-derived cultured human myotubes after exercise showed increased total cellular oleic acid uptake (30%), oxidation (46%) and lipid accumulation (34%), as well as increased fractional glucose oxidation (14%) compared to cultures established prior to exercise. Most of these exercise-induced increases were significant in the overweight group, whereas the normal weight group showed no change in oleic acid or glucose metabolism. CONCLUSIONS: 12 weeks of combined endurance and strength training promoted increased lipid and glucose metabolism in biopsy-derived cultured human myotubes, showing that training in vivo are able to induce changes in human myotubes that are discernible in vitro.

Insulin sensitivity, body composition and adipose depots following 12 w combined endurance and strength training in dysglycemic and normoglycemic sedentary men.

CONTEXT: Insulin resistance and dysglycemia are associated with physical inactivity and adiposity, and may be improved by exercise. OBJECTIVE: Investigate the effect of exercise on insulin sensitivity, body composition and adipose depots in sedentary men with (n = 11) or without (n = 11) overweight and dysglycemia. MATERIAL AND METHODS: Euglycemic-hyperinsulinemic clamp, ankle-to-neck MRI, MRS, muscle and adipose tissue biopsies before and after 12 weeks combined strength and endurance exercise. RESULTS: Insulin sensitivity, VO2max, strength, whole-body and muscle fat content, and abdominal adipose depots were improved without obvious differences between normo- and dysglycemic men. Hepatic fat, waist circumference and subcutaneous adipose tissue were reduced in the dysglycemic group. For both groups plasma adiponectin was reduced, whereas IL-6 was unchanged. Visceral fat was preferentially lost compared with other adipose depots. DISCUSSION AND CONCLUSION: Body composition, fat distribution and insulin sensitivity improved following training in sedentary middle-aged men with and without dysglycemia.

Sprint interval running increases insulin sensitivity in young healthy subjects.

High intensity cycling training increases oxidative capacity in skeletal muscles and improves insulin sensitivity. The present study compared the effect of eight weeks of sprint interval running (SIT) and continuous running at moderate intensity (CT) on insulin sensitivity and cholesterol profile in young healthy subjects (age 25.2 ± 0.7; VO(2max) 49.3 ± 1.2 ml·kg(-1)·min(-1)). SIT and CT increased maximal oxygen uptake by 5.3 ± 1.8 and 3.8 ± 1.6%, respectively (p < 0.05 for both). Oral glucose tolerance test (OGTT) was performed before and 60 h after the last training session. SIT, but not CT, reduced glucose area under curve and improved HOMA ß-cell index (p < 0.05). Insulin area under curve did not decrease significantly in any group. SIT, but not CT, reduced LDL and total cholesterol. In conclusion, sprint interval running improves insulin sensitivity and cholesterol profile in healthy subjects, and sprint interval running may be more effective to improve insulin sensitivity than continuous running at moderate intensity.