Potential New Methods to Analyze Basal and Total Endogenous Protein Losses of Host and Bacterial Origin in Pigs.

BACKGROUND: Current systems for assessing protein quality such as the Digestible Indispensable Amino Acid Score correct apparent amino acid (AA) digestibility for basal endogenous protein losses (bEPL), ignoring the potential influence of the diet on these losses. However, the quantification of total endogenous protein losses (tEPL) poses a challenge. OBJECTIVES: To evaluate different methods for quantifying tEPL and bEPL, and to assess their potential in discriminating between tEPL originating from bacteria and host. METHODS: Using an incomplete Youden square design, 12 ileal cannulated pigs received 10 different protein sources, and a nitrogen-free (NF) diet. Ileal digesta were collected on days 6 and 7 of each 1-wk feeding period, to quantify endogenous protein losses (EPL) and analyze apparent ileal digestibility. Ileal EPL were estimated based on 1) 16S-+18S gene copy quantitative polymerase chain reaction, 2) diaminopimelic acid (DAPA)+18S, 3) differential AA profiles in digesta, EPL, and bacteria, equaling tEPL, and 4) an NF diet and 5) whey protein isolate (WPI), equaling bEPL. RESULTS: Ileal bEPL based on the NF and WPI method correlated moderately highly (r = 0.69, P < 0.05), but the NF method probably underestimated bEPL. In pigs fed the WPI diet, EPL based on the WPI and AA profile method were highly correlated (r = 0.88, P < 0.01). Overall, tEPL based on the AA profile method were moderately correlated with the 16S+18S method (r = 0.58, P < 0.001), and DAPA+18S (r = 0.57, P < 0.001). Low correlations were observed between bacterial tEPL based on the AA profile method and 16S or DAPA. Host tEPL based on the AA profile method and 18S were weakly correlated (r = 0.39, P < 0.001). CONCLUSIONS: The AA profile method seems the most appropriate method for tEPL quantification, whereas the WPI method is preferred for bEPL quantification. Despite challenges in distinguishing between bacterial and host EPL, it is evident that bacterial proteins substantially (on average 37%-83%, depending on method) contribute to the EPL.

Protein fermentation in the gut; implications for intestinal dysfunction in humans, pigs, and poultry.

The amount of dietary protein is associated with intestinal disease in different vertebrate species. In humans, this is exemplified by the association between high-protein intake and fermentation metabolite concentrations in patients with inflammatory bowel disease. In production animals, dietary protein intake is associated with postweaning diarrhea in piglets and with the occurrence of wet litter in poultry. The underlying mechanisms by which dietary protein contributes to intestinal problems remain largely unknown. Fermentation of undigested protein in the hindgut results in formation of fermentation products including short-chain fatty acids, branched-chain fatty acids, ammonia, phenolic and indolic compounds, biogenic amines, hydrogen sulfide, and nitric oxide. Here, we review the mechanisms by which these metabolites may cause intestinal disease. Studies addressing how different metabolites induce epithelial damage rely mainly on cell culture studies and occasionally on mice or rat models. Often, contrasting results were reported. The direct relevance of such studies for human, pig, and poultry gut health is therefore questionable and does not suffice for the development of interventions to improve gut health. We discuss a roadmap to improve our understanding of gut metabolites and microbial species associated with intestinal health in humans and production animals and to determine whether these metabolite/bacterial networks cause epithelial damage. The outcomes of these studies will dictate proof-of-principle studies to eliminate specific metabolites and or bacterial strains and will provide the basis for interventions aiming to improve gut health.

Protein Ingestion before Sleep Increases Muscle Mass and Strength Gains during Prolonged Resistance-Type Exercise Training in Healthy Young Men.

BACKGROUND: It has been demonstrated that protein ingestion before sleep increases muscle protein synthesis rates during overnight recovery from an exercise bout. However, it remains to be established whether dietary protein ingestion before sleep can effectively augment the muscle adaptive response to resistance-type exercise training. OBJECTIVE: Here we assessed the impact of dietary protein supplementation before sleep on muscle mass and strength gains during resistance-type exercise training. METHODS: Forty-four young men (22 ± 1 y) were randomly assigned to a progressive, 12-wk resistance exercise training program. One group consumed a protein supplement containing 27.5 g of protein, 15 g of carbohydrate, and 0.1 g of fat every night before sleep. The other group received a noncaloric placebo. Muscle hypertrophy was assessed on a whole-body (dual-energy X-ray absorptiometry), limb (computed tomography scan), and muscle fiber (muscle biopsy specimen) level before and after exercise training. Strength was assessed regularly by 1-repetition maximum strength testing. RESULTS: Muscle strength increased after resistance exercise training to a significantly greater extent in the protein-supplemented (PRO) group than in the placebo-supplemented (PLA) group (+164 ± 11 kg and +130 ± 9 kg, respectively; P < 0.001). In addition, quadriceps muscle cross-sectional area increased in both groups over time (P < 0.001), with a greater increase in the PRO group than in the PLA group (+8.4 ± 1.1 cm(2) vs. +4.8 ± 0.8 cm(2), respectively; P < 0.05). Both type I and type II muscle fiber size increased after exercise training (P < 0.001), with a greater increase in type II muscle fiber size in the PRO group (+2319 ± 368 µm(2)) than in the PLA group (+1017 ± 353 µm(2); P < 0.05). CONCLUSION: Protein ingestion before sleep represents an effective dietary strategy to augment muscle mass and strength gains during resistance exercise training in young men. This trial was registered at clinicaltrials.gov as NCT02222415.

High dietary protein restores overreaching induced impairments in leukocyte trafficking and reduces the incidence of upper respiratory tract infection in elite cyclists.

The present study examined whether a high protein diet prevents the impaired leukocyte redistribution in response to acute exercise caused by a large volume of high-intensity exercise training. Eight cyclists (VO2max: 64.2±6.5mLkg(-1)min(-1)) undertook two separate weeks of high-intensity training while consuming either a high protein diet (3gkg(-1)proteinBM(-1)day(-1)) or an energy and carbohydrate-matched control diet (1.5gkg(-1)proteinBM(-1)day(-1)). High-intensity training weeks were preceded by a week of normal-intensity training under the control diet. Leukocyte and lymphocyte sub-population responses to acute exercise were determined at the end of each training week. Self-reported symptoms of upper-respiratory tract infections (URTI) were monitored daily by questionnaire. Undertaking high-intensity training with a high protein diet restored leukocyte kinetics to similar levels observed during normal-intensity training: CD8(+) TL mobilization (normal-intensity: 29,319±13,130cells/µL×∼165min vs. high-intensity with protein: 26,031±17,474cells/µL×∼165min, P>0.05), CD8(+) TL egress (normal-intensity: 624±264cells/µL vs. high-intensity with protein: 597±478cells/µL, P>0.05). This pattern was driven by effector-memory populations mobilizing (normal-intensity: 6,145±6,227cells/µL×∼165min vs. high-intensity with protein: 6,783±8,203cells/µL×∼165min, P>0.05) and extravastating from blood (normal-intensity: 147±129cells/µL vs. high-intensity with protein: 165±192cells/µL, P>0.05). High-intensity training while consuming a high protein diet was associated with fewer symptoms of URTI compared to performing high-intensity training with a normal diet (P<0.05). To conclude, a high protein diet might reduce the incidence of URTI in athletes potentially mediated by preventing training-induced impairments in immune-surveillance.

In vitro fermentation potential of gut endogenous protein losses of growing pigs.

Fermentation of dietary and endogenous protein in the hindgut is generally considered detrimental to the health of pigs. We investigated the in vitro fermentation potential of porcine endogenous protein in ileal digesta and colonic mucus, using a N-free buffer with an excess of fermentable carbohydrates. Urea, whey protein isolate (WPI, positive control), WPI hydrolysate (WPIH), and combinations of the latter two were used to validate the assay. A new biphasic model, including a linear end simulation, fitted to the gas production data over a 48-h period identified the time point when substrate fermentation ended. A higher degree of hydrolysis of WPI resulted in a higher maximum gas production rate (Rmax, P < 0.01). Differences in Rmax and the time required to reach Rmax were observed among ileal digesta samples, with Rmax increasing with the insoluble protein content, and the highest Rmax occurring with colonic mucus samples (P < 0.05). The endogenous proteins entering the large intestine of pigs can ferment more rapidly compared to highly soluble and digestible protein sources, with Rmax positively correlated with decreasing solubility of endogenous nitrogenous components.

Protein fermentation in the hindgut of pigs can impact their health, affecting factors like growth rates and feed efficiency. Besides dietary protein, up to 50% of the protein entering the large intestine of growing pigs may be of endogenous origin. Therefore, we explored the fermentation potential of endogenous proteins compared to a well-known protein source, whey protein isolate (WPI). In developing and validating an in vitro gas production technique, we employed urea, WPI, WPI hydrolysate, and various combinations as substrates. The study introduces a new biphasic model for in vitro gas production, offering a detailed analysis of the fermentation process over a 48-h period. Our results revealed that porcine endogenous proteins can undergo rapid fermentation because the maximum gas production rate was higher compared to WPI. This insight is crucial for understanding the dynamics of protein fermentation in pigs. Additionally, we explored the solubility and molecular size of proteins, providing a comprehensive understanding of their fermentation characteristics. We found that endogenous proteins were less soluble compared to WPI but contained more smaller peptides. Unraveling the complexities of protein fermentation in pigs contributes to improvement of feed formulation for optimal gut health.

Over-toasting dehulled rapeseed meal and soybean meal, but not sunflower seed meal, increases prececal nitrogen and amino acid digesta flows in broilers.

Poorly digestible proteins may lead to increased protein fermentation in the ceca of broilers and hence, the production of potentially harmful metabolites. To evaluate effects of protein fermentation on gut health, an experimental contrast in ileal nitrogen (N) and amino acid (AA) flow is required. Therefore, our objective was to develop a model that creates a contrast in protein fermentation by increasing the prececal flow of protein within ingredients. To this end, we used additional toasting of protein sources and evaluated the effect on prececal N and AA flows. One-day-old Ross 308 male broilers (n = 480) were divided over 6 dietary treatments, with 8 replicate pens with 10 broilers each. Diets contained 20% of a regular soybean meal (SBM), high protein sunflower seed meal (SFM) or a dehulled rapeseed meal (dRSM) as is, or heat damaged by secondary toasting at 136°C for 20 min (tSBM, tSFM, or tdRSM). Ileal and total tract digesta flows of N and AA were determined with 5 birds per pen in their third week of life using an inert marker (TiO2) in the feed. Additional toasting increased the feed conversion ratio (FCR) only in birds fed dRSM (1.39 vs. 1.31), but not SBM and SFM (interaction P = 0.047). In SBM, additional toasting increased the flow of histidine, lysine, and aspartate through the distal ileum and excreted, while in SFM it had no effect on flows of N and AA. Toasting dRSM increased the prececal flows and excretion of N (862 vs 665 and 999 vs 761 mg/d, respectively) and of the AA. Of the ingredients tested, toasting dRSM is a suitable model to increase protein flows into the hind-gut, permitting the assessment of effects of protein fermentation.

Dietary protein digestion and absorption rates and the subsequent postprandial muscle protein synthetic response do not differ between young and elderly men.

Impaired digestion and/or absorption of dietary protein lowers postprandial plasma amino acid availability and, as such, could reduce the postprandial muscle protein synthetic response in the elderly. We aimed to compare in vivo dietary protein digestion and absorption and the subsequent postprandial muscle protein synthetic response between young and elderly men. Ten elderly (64 +/- 1 y) and 10 young (23 +/- 1 y) healthy males consumed a single bolus of 35 g specifically produced, intrinsically l-[1-(13)C]phenylalanine-labeled micellar casein (CAS) protein. Furthermore, primed continuous infusions with l-[ring-(2)H(5)]phenylalanine, l-[1-(13)C]leucine, and l-[ring-(2)H(2)]tyrosine were applied and blood and muscle tissue samples were collected to assess the appearance rate of dietary protein-derived phenylalanine in the circulation and the subsequent muscle protein fractional synthetic rate over a 6-h postprandial period. Protein ingestion resulted in a rapid increase in exogenous phenylalanine appearance in both the young and elderly men. Total exogenous phenylalanine appearance rates (expressed as area under the curve) were 39 +/- 3 mumol.6 h.kg(-1) in the young men and 38 +/- 2 mumol.6 h.kg(-1) in the elderly men (P = 0.73). In accordance, splanchnic amino acid extraction did not differ between young (72 +/- 2%) and elderly (73 +/- 1%) volunteers (P = 0.74). Muscle protein synthesis rates, calculated from the oral tracer, were 0.063 +/- 0.006 and 0.054 +/- 0.004%/h in the young and elderly men, respectively, and did not differ between groups (P = 0.27). We conclude that protein digestion and absorption kinetics and the subsequent muscle protein synthetic response following the ingestion of a large bolus of intact CAS are not substantially impaired in healthy, elderly men.

Coingestion of carbohydrate and protein hydrolysate stimulates muscle protein synthesis during exercise in young men, with no further increase during subsequent overnight recovery.

We investigated the effect of carbohydrate and protein hydrolysate ingestion on whole-body and muscle protein synthesis during a combined endurance and resistance exercise session and subsequent overnight recovery. Twenty healthy men were studied in the evening after consuming a standardized diet throughout the day. Subjects participated in a 2-h exercise session during which beverages containing both carbohydrate (0.15 g x kg(-1) x h(-1)) and a protein hydrolysate (0.15 g x kg(-1) x h(-1)) (C+P, n = 10) or water only (W, n = 10) were ingested. Participants consumed 2 additional beverages during early recovery and remained overnight at the hospital. Continuous i.v. infusions with L-[ring-(13)C(6)]-phenylalanine and L-[ring-(2)H(2)]-tyrosine were applied and blood and muscle samples were collected to assess whole-body and muscle protein synthesis rates. During exercise, whole-body and muscle protein synthesis rates increased by 29 and 48% with protein and carbohydrate coingestion (P < 0.05). Fractional synthetic rates during exercise were 0.083 +/- 0.011%/h in the C+P group and 0.056 +/- 0.003%/h in the W group, (P < 0.05). During subsequent overnight recovery, whole-body protein synthesis was 19% greater in the C+P group than in the W group (P < 0.05). However, mean muscle protein synthesis rates during 9 h of overnight recovery did not differ between groups and were 0.056 +/- 0.004%/h in the C+P group and 0.057 +/- 0.004%/h in the W group (P = 0.89). We conclude that, even in a fed state, protein and carbohydrate supplementation stimulates muscle protein synthesis during exercise. Ingestion of protein with carbohydrate during and immediately after exercise improves whole-body protein synthesis but does not further augment muscle protein synthesis rates during 9 h of subsequent overnight recovery.

Pharmacokinetics of proline-rich tripeptides in the pig.

Tripeptides may possess bioactive properties. For instance, blood pressure lowering is attributed to the proline-rich tripeptides Ile-Pro-Pro (IPP), Leu-Pro-Pro (LPP), and Val-Pro-Pro (VPP). However, little is known about their absorption, distribution, and elimination characteristics. The aim of this study was to characterize the pharmacokinetic behavior of IPP, LPP, and VPP in a conscious pig model. Synthetic IPP, LPP, and VPP were administered intravenously or intragastrically (4.0 mg kg(-1) BW in saline) to 10 piglets (approximately 25 kg body weight) in the postabsorptive state. After intravenous dosing, the elimination half-life for IPP was significantly higher (P<0.001) than for LPP and VPP (2.5+/-0.1, 1.9+/-0.1, and 2.0+/-0.1 min, respectively). After intragastric dosing, however, the elimination half-lives were not significantly different between the peptides (9+/-1, 15+/-4, and 12+/-6 min, respectively). Maximum plasma concentrations were about 10 nmol l(-1) for the three tripeptides. The fraction dose absorbed was 0.077+/-0.010, 0.059+/-0.009, and 0.073+/-0.015%, for IPP, LPP, and VPP, respectively. Proline-rich tripeptides reach the blood circulation intact, with an absolute bioavailability of about 0.1% when administered via a saline solution. Because half-lives of absorption and elimination were maximally about 5 and 15 min, respectively, this suggests that under these conditions a bioactive effect of these tripeptides would be rather acute.

Protein and protein hydrolysates in sports nutrition.

With the increasing knowledge about the role of nutrition in increasing exercise performance, it has become clear over the last 2 decades that amino acids, protein, and protein hydrolysates can play an important role. Most of the attention has been focused on their effects at a muscular level. As these nutrients are ingested, however, it also means that gastrointestinal digestibility and absorption can modulate their efficacy significantly. Therefore, discussing the role of amino acids, protein, and protein hydrolysates in sports nutrition entails holding a discussion on all levels of the metabolic route. On May 28-29, 2007, a small group of researchers active in the field of exercise science and protein metabolism presented an overview of the different aspects of the application of protein and protein hydrolysates in sports nutrition. In addition, they were asked to share their opinions on the future progress in their fields of research. In this overview, an introduction to the workshop and a short summary of its outcome is provided.

Food-derived bioactive peptides influence gut function.

Bioactive peptides either present in foods or released from food proteins during digestion have a wide range of physiological effects, including on gut function. Many of the bioactive peptides characterized to date that influence gut motility, secretion, and absorption are opioid agonists or antagonists. The authors review a body of experimental evidence that demonstrates an effect of peptides from food proteins on endogenous (nondietary) protein flow at the terminal ileum of simple-stomached mammals, including adult humans. At least some dietary peptides (1000-5000 Da) significantly enhance the loss of protein from the small intestine, causing an increased amount of protein to enter the colon. Food-derived peptides appear to either stimulate protein secretion into the gut lumen or inhibit amino acid reabsorption or influence both processes simultaneously. The effect of dietary peptides on small-intestine secretory-protein dynamics is discussed in the context of the major components of gut endogenous protein, sloughed cells, enzymatic secretions, mucin, and bacterial protein.

Interaction between protein, phytate, and microbial phytase. In vitro studies.

The interaction between protein and phytate was investigated in vitro using proteins extracted from five common feedstuffs and from casein. The appearance of naturally present soluble protein-phytate complexes in the feedstuffs, the formation of complexes at different pHs, and the degradation of these complexes by pepsin and/or phytase were studied. Complexes of soluble proteins and phytate in the extracts appeared in small amounts only, with the possible exception of rice pollards. Most proteins dissolved almost completely at pH 2, but not after addition of phytate. Phytase prevented precipitation of protein with phytate. Pepsin could release protein from a precipitate, but the rate of release was increased by phytase. Protein was released faster from a protein-phytate complex when phytase was added, but phytase did not hydrolyze protein. Protein was released from the complex and degraded when both pepsin and phytase were added. It appears that protein-phytate complexes are mainly formed at low pH, as occurs in the stomach of animals. Phytase prevented the formation of the complexes and aided in dissolving them at a faster rate. This might positively affect protein digestibility in animals.

Effect of Intensive Training on Mood With No Effect on Brain-Derived Neurotrophic Factor.

CONTEXT: Monitoring mood state is a useful tool for avoiding nonfunctional overreaching. Brain-derived neurotrophic factor (BDNF) is implicated in stress-related mood disorders. PURPOSE: To investigate the impact of intensified training-induced mood disturbance on plasma BDNF concentrations at rest and in response to exercise. METHODS: Eight cyclists performed 1 wk of normal (NT), 1 wk of intensified (INT), and 1 wk of recovery (REC) training. Fasted blood samples were collected before and after exercise on day 7 of each training week and analyzed for plasma BDNF and cortisol concentrations. A 24-item Profile of Mood State questionnaire was administered on day 7 of each training week, and global mood score (GMS) was calculated. RESULTS: Time-trial performance was impaired during INT (P = .01) and REC (P = .02) compared with NT. Basal plasma cortisol (NT = 153 ± 16 ng/mL, INT = 130 ± 11 ng/mL, REC = 150 ± 14 ng/ml) and BDNF (NT = 484 ± 122 pg/mL, INT = 488 ± 122 pg/mL, REC = 383 ± 56 pg/mL) concentrations were similar between training conditions. Likewise, similar exercise-induced increases in cortisol and BDNF concentrations were observed between training conditions. GMS was 32% greater during INT vs NT (P < .001). CONCLUSIONS: Consistent with a state of functional overreaching (FOR), impairments in performance and mood state with INT were restored after 1 wk of REC. These results support evidence for mood changes before plasma BDNF concentrations as a biochemical marker of FOR and that cortisol is not a useful marker for predicting FOR.

Enhanced Lacto-Tri-Peptide Bio-Availability by Co-Ingestion of Macronutrients.

Some food-derived peptides possess bioactive properties, and may affect health positively. For example, the C-terminal lacto-tri-peptides Ile-Pro-Pro (IPP), Leu-Pro-Pro (LPP) and Val-Pro-Pro (VPP) (together named here XPP) are described to lower blood pressure. The bioactivity depends on their availability at the site of action. Quantitative trans-organ availability/kinetic measurements will provide more insight in C-terminal tri-peptides behavior in the body. We hypothesize that the composition of the meal will modify their systemic availability. We studied trans-organ XPP fluxes in catheterized pigs (25 kg; n=10) to determine systemic and portal availability, as well as renal and hepatic uptake of a water-based single dose of synthetic XPP and a XPP containing protein matrix (casein hydrolyte, CasH). In a second experiment (n=10), we compared the CasH-containing protein matrix with a CasH-containing meal matrix and the modifying effects of macronutrients in a meal on the availability (high carbohydrates, low quality protein, high fat, and fiber). Portal availability of synthetic XPP was 0.08 ± 0.01% of intake and increased when a protein matrix was present (respectively 3.1, 1.8 and 83 times for IPP, LPP and VPP). Difference between individual XPP was probably due to release from longer peptides. CasH prolonged portal bioavailability with 18 min (absorption half-life, synthetic XPP: 15 ± 2 min, CasH: 33 ± 3 min, p<0.0001) and increased systemic elimination with 20 min (synthetic XPP: 12 ± 2 min; CasH: 32 ± 3 min, p<0.0001). Subsequent renal and hepatic uptake is about 75% of the portal release. A meal containing CasH, increased portal 1.8 and systemic bioavailability 1.2 times. Low protein quality and fiber increased XPP systemic bioavailability further (respectively 1.5 and 1.4 times). We conclude that the amount and quality of the protein, and the presence of fiber in a meal, are the main factors that increase the systemic bioavailability of food-derived XPP.

High-intensity training reduces CD8+ T-cell redistribution in response to exercise.

PURPOSE: We examined whether exercise-induced lymphocytosis and lymphocytopenia are impaired with high-intensity training. METHODS: Eight trained cyclists (VO(2max) = 64.2 ± 6.5 mL · kg(-1) · min(-1)) undertook 1 wk of normal-intensity training and a second week of high-intensity training. On day 7 of each week, participants performed a cycling task, consisting of 120 min of submaximal exercise followed by a 45-min time trial. Blood was collected before, during, and after exercise. CD8(+) T lymphocytes (CD8(+)TLs) were identified, as well as CD8(+)TL subpopulations on the basis of CD45RA and CD27 expression. RESULTS: High-intensity training (18,577 ± 10,984 cells per microliter × ~165 min) was associated with a smaller exercise-induced mobilization of CD8(+)TLs compared with normal-intensity training (28,473 ± 16,163 cells per microliter × ~165 min, P = 0.09). The response of highly cytotoxic CD8(+)TLs (CD45RA(+)CD27(-)) to exercise was smaller after 1 wk of high-intensity training (3144 ± 924 cells per microliter × ~165 min) compared with normal-intensity training (6417 ± 2143 cells per microliter × ~165 min, P < 0.05). High-intensity training reduced postexercise CD8(+)TL lymphocytopenia (-436 ± 234 cells per microliter) compared with normal-intensity training (-630 ± 320 cells per microliter, P < 0.05). This was driven by a reduced egress of naive CD8(+)TLs (CD27(+)CD45RA(+)). High-intensity training was associated with reduced plasma epinephrine (-37%) and cortisol (-15%) responses (P < 0.05). CONCLUSIONS: High-intensity training impaired CD8(+)TL mobilization and egress in response to exercise. Highly cytotoxic CD8(+)TLs were primarily responsible for the reduced mobilization of CD8(+)TLs, which occurred in parallel with smaller neuroendocrine responses. The reduced capacity for CD8(+)TLs to leave blood after exercise with high-intensity training was accounted for primarily by naive, and also, highly cytotoxic CD8(+)TLs. This impaired CD8(+)TL redistribution in athletes undertaking intensified training may imply reduced immune surveillance.

Effect of increased dietary protein on tolerance to intensified training.

PURPOSE: The purpose of the present study was to examine the effect of increased protein intake on short-term decrements in endurance performance during a block of high-intensity training. METHODS: Trained male cyclists (VO(2max) = 64.2 ± 6.5 mL·kg(-1)·min(-1)) completed two 3-wk trials both divided equally into normal (NOR), intensified (INT), and recovery (REC) training. In a counterbalanced crossover experimental design, cyclists received either a high-protein (PRO; 3 g protein·kg(-1) body mass (BM)·d(-1)) or a normal diet (CON; 1.5 g protein·kg(-1) BM·d(-1)) during INT and REC. Dietary carbohydrate content remained constant at 6 g·kg(-1) BM·d(-1). Energy balance was maintained during each training week. Endurance performance was assessed with a VO(2max) test and a preloaded time trial. Alterations in blood metabolite responses to exercise were measured at rest, during, and after exercise. Cyclists completed the Daily Analysis of Life Demands for Athletes (DALDA) questionnaire each day. RESULTS: Increased dietary protein intake led to a possible attenuation (4.3%; 90% confidence limits ×/÷5.4%) in the decrement in time trial performance after a block of high-intensity training compared with NOR (PRO = 2639 ± 350 s; CON = 2555 ± 313 s). Restoration of endurance performance during recovery training possibly benefited (2.0%; ×/÷4.9%) from additional protein intake. Frequency of symptoms of stress described as "worse than normal" reported after a block of high-intensity training was very likely (97%) attenuated (17; ±11 AUC of "a" scores part B, DALDA for INT + REC) by increasing the protein content of the diet. No discernable changes in blood metabolite concentrations were observed in PRO. CONCLUSIONS: Additional protein intake reduced symptoms of psychological stress and may result in a worthwhile amelioration of the performance decline experienced during a block of high-intensity training.

Carbohydrate and protein hydrolysate coingestions improvement of late-exercise time-trial performance.

This study examined whether a carbohydrate + casein hydrolysate (CHO+ProH) beverage improved time-trial performance vs. a CHO beverage delivering approximately 60 g CHO/hr. Markers of muscle disruption and recovery were also assessed. Thirteen male cyclists (VO2peak = 60.8 +/- 1.6 ml . kg-1 . min-1) completed 2 computer-simulated 60-km time trials consisting of 3 laps of a 20-km course concluding with a 5-km climb (approximately 5% grade). Participants consumed 200 ml of CHO (6%) or CHO+ProH beverage (6% + 1.8% protein hydrolysate) every 5 km and 500 ml of beverage immediately postexercise. Beverage treatments were administered using a randomly counterbalanced, double-blind design. Plasma creatine phosphokinase (CK) and muscle-soreness ratings were assessed immediately before and 24 hr after cycling. Mean 60-km times were 134.4 +/- 4.6 and 135.0 +/- 4.0 min for CHO+ProH and CHO beverages, respectively. All time differences between treatments occurred during the final lap, with protein hydrolysate ingestion explaining a significant (p < .05) proportion of between-trials differences over the final 20 km (44.3 +/- 1.6, 45.0 +/- 1.6 min) and final 5 km (16.5 +/- 0.6, 16.9 +/- 0.6 min). Plasma CK levels and muscle-soreness ratings increased significantly after the CHO trial (161 +/- 53, 399 +/- 175 U/L; 15.8 +/- 5.1, 37.6 +/- 5.7 mm) but not the CHO+ProH trial (115 +/- 21, 262 +/- 88 U/L; 20.9 +/- 5.3, 32.2 +/- 7.1 mm). Late-exercise time-trial performance was enhanced with CHO+ProH beverage ingestion compared with a beverage containing CHO provided at maximal exogenous oxidation rates during exercise. CHO+ProH ingestion also prevented increases in plasma CK and muscle soreness after exercise.

Ingestion of a protein hydrolysate is accompanied by an accelerated in vivo digestion and absorption rate when compared with its intact protein.

BACKGROUND: It has been suggested that a protein hydrolysate, as opposed to its intact protein, is more easily digested and absorbed from the gut, which results in greater plasma amino acid availability and a greater muscle protein synthetic response. OBJECTIVE: We aimed to compare dietary protein digestion and absorption kinetics and the subsequent muscle protein synthetic response to the ingestion of a single bolus of protein hydrolysate compared with its intact protein in vivo in humans. DESIGN: Ten elderly men (mean +/- SEM age: 64 +/- 1 y) were randomly assigned to a crossover experiment that involved 2 treatments in which the subjects consumed a 35-g bolus of specifically produced L-[1-(13)C]phenylalanine-labeled intact casein (CAS) or hydrolyzed casein (CASH). Blood and muscle-tissue samples were collected to assess the appearance rate of dietary protein-derived phenylalanine in the circulation and subsequent muscle protein fractional synthetic rate over a 6-h postprandial period. RESULTS: The mean (+/-SEM) exogenous phenylalanine appearance rate was 27 +/- 6% higher after ingestion of CASH than after ingestion of CAS (P < 0.001). Splanchnic extraction was significantly lower in CASH compared with CAS treatment (P < 0.01). Plasma amino acid concentrations increased to a greater extent (25-50%) after the ingestion of CASH than after the ingestion of CAS (P < 0.01). Muscle protein synthesis rates averaged 0.054 +/- 0.004% and 0.068 +/- 0.006%/h in the CAS and CASH treatments, respectively (P = 0.10). CONCLUSIONS: Ingestion of a protein hydrolysate, as opposed to its intact protein, accelerates protein digestion and absorption from the gut, augments postprandial amino acid availability, and tends to increase the incorporation rate of dietary amino acids into skeletal muscle protein.

Effects of emulsified policosanols with different chain lengths on cholesterol metabolism in heterozygous LDL receptor-deficient mice.

Policosanol is a mixture of long-chain primary aliphatic saturated alcohols. Previous studies in humans and animals have shown that these compounds improved lipoprotein profiles. However, more-recent placebo-controlled studies could not confirm these promising effects. Octacosanol (C28), the main component of sugarcane-derived policosanol, is assumed to be the bioactive component. This has, however, never been tested in an in vivo study that compared individual policosanol components side by side. Here we present that neither the individual policosanol components (C24, C26, C28, or C30) nor the natural policosanol mixture (all 30 mg/100 g diet) lowered serum cholesterol concentrations in LDL receptor knock-out (LDLr(+/-)) mice. Moreover, there was no effect on gene expression profiles of LDLr, ABCA1, HMG-CoA synthase 1, and apolipoprotein A-I (apoA-I) in hepatic and small intestinal tissue of female LDLr(+/-) mice after the 7 week intervention period. Finally, none of the individual policosanols or their respective long-chain fatty acids or aldehydes affected de novo apoA-I protein production in vitro in HepG2 and CaCo-2 cells. Therefore, we conclude that the evaluated individual policosanols, as well as the natural policosanol mixture, have no potential for reducing coronary heart disease risk through effects on serum lipoprotein concentrations.

Protein coingestion stimulates muscle protein synthesis during resistance-type exercise.

In contrast to the effect of nutritional intervention on postexercise muscle protein synthesis, little is known about the potential to modulate protein synthesis during exercise. This study investigates the effect of protein coingestion with carbohydrate on muscle protein synthesis during resistance-type exercise. Ten healthy males were studied in the evening after they consumed a standardized diet throughout the day. Subjects participated in two experiments in which they ingested either carbohydrate or carbohydrate with protein during a 2-h resistance exercise session. Subjects received a bolus of test drink before and every 15 min during exercise, providing 0.15 g x kg(-1) x h(-1) carbohydrate with (CHO + PRO) or without (CHO) 0.15 g x kg(-1) x h(-1) protein hydrolysate. Continuous intravenous infusions with l-[ring-(13)C(6)]phenylalanine and l-[ring-(2)H(2)]tyrosine were applied, and blood and muscle biopsies were collected to assess whole body and muscle protein synthesis rates during exercise. Protein coingestion lowered whole body protein breakdown rates by 8.4 +/- 3.6% (P = 0.066), compared with the ingestion of carbohydrate only, and augmented protein oxidation and synthesis rates by 77 +/- 17 and 33 +/- 3%, respectively (P < 0.01). As a consequence, whole body net protein balance was negative in CHO, whereas a positive net balance was achieved after the CHO + PRO treatment (-4.4 +/- 0.3 vs. 16.3 +/- 0.4 micromol phenylalanine x kg(-1) x h(-1), respectively; P < 0.01). In accordance, mixed muscle protein fractional synthetic rate was 49 +/- 22% higher after protein coingestion (0.088 +/- 0.012 and 0.060 +/- 0.004%/h in CHO + PRO vs. CHO treatment, respectively; P < 0.05). We conclude that, even in a fed state, protein coingestion stimulates whole body and muscle protein synthesis rates during resistance-type exercise.