Pharmacokinetics of soy-derived lysophosphatidylcholine compared with that of glycerophosphocholine: a randomized controlled trial.

Lysophosphatidylcholine (LPC) is present in various foods and contains a choline moiety such as in glycerophosphocholine (GPC). However, the potential of LPC as a choline source remains unclear. This study investigated the single-dose pharmacokinetics of 480 mg soy-derived LPC in 12 healthy men compared with that of either soy oil with the same lipid amount (placebo) or GPC with the same choline amount. Both LPC and GPC supplementation increased plasma choline, serum phospholipid, and serum triglyceride concentrations, but neither of them significantly elevated plasma trimethylamine N-oxide concentration. In addition, although the intake of LPC slightly increased plasma LPC16:0, LPC18:2, and total LPC concentrations, their concentrations remained within physiological ranges. No adverse events were attributed to the LPC supplementation. To the best of our knowledge, this study is the first to compare LPC and GPC pharmacokinetics in humans and shows that LPC can be a source of choline.

Re-evaluation of the role of autophagy in thyroid cancer treatment.

Numerous studies have examined the role of autophagy in thyroid cancer treatment; however there are discrepancies among the reported data, with some showing the pro-survival and others the anti-survival effects of autophagy. These discrepant results appear to be at least in part due to insufficient analyses or data misinterpretation as well as improper assessments of autophagic activity. Therefore, the present study re-evaluated the regulation of autophagic activity by various anticancer modalities and examined the role of autophagy in thyroid cancer treatment in three thyroid cancer cell lines (TPC1, ACT1 and KTC1). The immunofluorescence and DalGreen findings demonstrated that cisplatin, irradiation and sorafenib were all autophagy inducers as previously reported, but, unlike previous studies using thyroid cancer cells, doxorubicin acted as an inhibitor. KTC1 cells are unique because they only responded to cisplatin. The efficacy of anticancer therapeutics was significantly higher in chloroquine or 3-methyladenine-treated autophagy-defective cells than in autophagy-competent cells, thereby indicating the pro-survival effect of autophagy induced by anticancer therapeutics, which is partly due to inhibition of apoptosis. Thus, the present findings relating to several anticancer therapeutics and three thyroid cancer cell lines demonstrate the pro-survival effect of autophagy in thyroid cancer treatment. Although the present study only involved cell lines, it provides evidence for the beneficial combination of the anticancer therapeutic modalities with autophagy inhibitors, and proposes that autophagy inhibitors may serve as a possible adjunctive therapy for thyroid cancer.

Salivary Immunoglobulin A Secretion Rate During Peak Period Conditioning Regimens in Triathletes.

ABSTRACT: Matsumoto M, Satoh, K, Kushi, H, Hamuro, K, Sakurai, M, Saito, H, Tanaka, R, Saito, T, Kohda, N, and Hamada, K. Salivary immunoglobulin A secretion rate during peak period conditioning regimens in triathletes. J Strength Cond Res 35(5): 1389-1396, 2021-Triathletes often feel unwell during the conditioning period (peak period) leading up to a race. The aim of this study was to evaluate the factors relevant to the condition of athletes and their impact on mucosal immune responses and the salivary immunoglobulin A (IgA) secretion rate. This study recruited college student triathletes (33 men and 7 women) who participated in an Olympic distance race. For each subject, the salivary IgA rate was measured continuously for 1 month before the race (peak period). Data on physical activity during the peak period were calculated in metabolic equivalents, and the relationships between these factors and the salivary IgA secretion rate were evaluated. The average amount of physical activity was highest during the 2- to 3-week period before the race, at 744.7 ± 51.5 kcal expended per day. In subjects who, on average, expended more than 1,000 kcal·d-1 in physical activity between 12 and 14 days before the race, the salivary IgA secretion rate was significantly reduced compared with the value at 1 week before the race (p < 0.05). On the day before the race, a further reduction was observed (p < 0.1). The salivary IgA secretion rate was decreased by high-intensity exercise during the peak period in advance of a race; this was associated with a loss of optimal condition just before the race.

Oral Administration of Egg- and Soy-Derived Lysophosphatidylcholine Mitigated Acetylcholine Depletion in the Brain of Scopolamine-Treated Rats.

Enzyme-modified lecithin that contains lysophosphatidylcholine (LPC) is generally recognized as safe. However, its potential as a functional ingredient has been less investigated than other choline (Ch)-containing compounds, such as glycerophosphocholine (GPC). Reports on the possibility of LPC functioning as a cholinergic precursor in vivo and on its kinetics are limited to docosahexaenoic acid-bound LPC. Herein, three experiments were performed to investigate these processes in scopolamine (SCO)-treated rats. First, an egg-derived LPC reagent was orally administered to rats, and brain acetylcholine (ACh), Ch, plasma Ch, and LPC were measured. Second, soy- and rapeseed-derived enzyme-modified lecithins and GPC were administered for comparison. Third, soy-derived enzyme-modified lecithins with different fat contents were administered for comparison. The LPC reagent mitigated SCO-induced ACh depletion at 500 mg/kg body weight and increased plasma Ch, but not LPC, concentrations. Additionally, soy-derived LPC-containing food additive counteracted brain ACh depletion similarly to GPC. Interestingly, plasma Ch and linoleoyl-LPC levels were higher when soy-derived LPC with a higher fat content was administered, whereas the plasma levels of palmitoyl-LPC decreased and those of total LPC remained constant. In conclusion, egg- and soy-derived LPC species function as cholinergic precursors in vivo, and future studies should explore this potential.

Characterization of metabolic reprogramming by metabolomics in the oncocytic thyroid cancer cell line XTC.UC1.

Oncocytic thyroid cancer is characterized by the aberrant accumulation of abnormal mitochondria in the cytoplasm and a defect in oxidative phosphorylation. We performed metabolomics analysis to compare metabolic reprogramming among the oncocytic and non-oncocytic thyroid cancer cell lines XTC.UC1 and TPC1, respectively, and a normal thyroid cell line Nthy-ori 3-1. We found that although XTC.UC1 cells exhibit higher glucose uptake than TPC1 cells, the glycolytic intermediates are not only utilized to generate end-products of glycolysis, but also diverted to branching pathways such as lipid metabolism and the serine synthesis pathway. Glutamine is preferentially used to produce glutathione to reduce oxidative stress in XTC.UC1 cells, rather than to generate α-ketoglutarate for anaplerotic flux into the TCA cycle. Thus, growth, survival and redox homeostasis of XTC.UC1 cells rely more on both glucose and glutamine than do TPC1 cells. Furthermore, XTC.UC1 cells contained higher amounts of intracellular amino acids which is due to higher expression of the amino acid transporter ASCT2 and enhanced autophagy, thus providing the building blocks for macromolecules and energy production. These metabolic alterations are required for oncocytic cancer cells to compensate their defective mitochondrial function and to alleviate excess oxidative stress.

Energy expenditure during 2-day trail walking in the mountains (2,857 m) and the effects of amino acid supplementation in older men and women.

We compared relative exercise intensity and active energy expenditure (AEE) on trail walking in the mountains, with those of daily exercise training, and whether branched-chain amino acid (BCAA) and arginine supplementation attenuated the release of markers indicating muscle damage and declines in physical performance. Twenty-one subjects (~63 years) were divided into two groups: amino acid (AA, 51 g of amino acids and 40 g of carbohydrate, male/female = 6/4) or placebo (PL, 91 g of carbohydrate, male/female = 6/5) supplementation during 2 days of trail walking in the mountains. We measured heart rate (HR), AEE, fatigue sensation, water and food intake, and sweat loss during walking. In addition, we measured peak aerobic capacity [Formula: see text] and heart rate (HR(peak)) with graded-intensity walking, vertical jumping height (VJ) before and after walking. We found that average HR and AEE during uphill walking were ~100% HR(peak) and ~60% [Formula: see text], while they were ~80 and ~20% during downhill walking, respectively. Moreover, average total AEE per day was sevenfold that of their daily walking training. VJ after walking remained unchanged compared with the baseline in AA (P > 0.2), while it was reduced by ~10% in PL (P < 0.01), although with no significant difference in the reduction between the groups (P > 0.4). The responses of other variables were not significantly different between groups (all, P > 0.2). Thus, trail walking in the mountains required a high-intensity effort for older people, while the effects of BCAA and arginine supplementation were modest in this condition.

Effects of a soybean nutrition bar on the postprandial blood glucose and lipid levels in patients with diabetes mellitus.

We investigated the influence of a soybean nutrition bar made from whole soy powder on the blood glucose, insulin and lipid levels in comparison with a test cookie with the same amount of energy in patients with diabetes mellitus. In the cross-over designed study, meal tolerance tests using the soybean nutrition bar and test cookie were performed. Two kinds of test meals were used: Study 1 80 kcal, Study 2 592 kcal. The blood glucose response was significantly lower in the soybean nutrition bar trial than in the cookie trial (Studies 1 and 2, p < 0.001). The blood insulin response was also significantly lower in the soybean nutrition bar trial than in the cookie trial (Study 2, p < 0.001). The blood triglyceride and non-esterified fatty acid responses were not significantly different between the two trials, nor were the changes in breath H2 enrichment (Study 2). The soybean nutrition bar did not induce postprandial hyperglycaemia in diabetic patients unlike the isoenergetic test cookies.

Reprogramming of Cellular Metabolism and Its Therapeutic Applications in Thyroid Cancer.

Metabolism is a series of life-sustaining chemical reactions in organisms, providing energy required for cellular processes and building blocks for cellular constituents of proteins, lipids, carbohydrates and nucleic acids. Cancer cells frequently reprogram their metabolic behaviors to adapt their rapid proliferation and altered tumor microenvironments. Not only aerobic glycolysis (also termed the Warburg effect) but also altered mitochondrial metabolism, amino acid metabolism and lipid metabolism play important roles for cancer growth and aggressiveness. Thus, the mechanistic elucidation of these metabolic changes is invaluable for understanding the pathogenesis of cancers and developing novel metabolism-targeted therapies. In this review article, we first provide an overview of essential metabolic mechanisms, and then summarize the recent findings of metabolic reprogramming and the recent reports of metabolism-targeted therapies for thyroid cancer.

MIEAP and ATG5 are tumor suppressors in a mouse model of BRAFV600E-positive thyroid cancer.

Mitochondria-eating protein (MIEAP) is a molecule important for non-canonical mitophagy and thought to be a tumor suppressor. Our previous study found that MIEAP expression is defective in thyroid oncocytomas, irrespective of being benign or malignant, and also in non-oncocytic thyroid cancers. Thyroid oncocytomas are composed of large polygonal cells with eosinophilic cytoplasm that is rich in abnormal mitochondria. Thus, our data indicate that, together with increased mitochondrial biogenesis that compensates for the dysfunction of the mitochondria, MIEAP plays a critical role in the accumulation of mitochondria in thyroid oncocytic tumors, whereas a defective MIEAP expression alone is not sufficient for mitochondrial accumulation in non-oncocytic cancers with normal mitochondria. To clarify whether MIEAP is a tumor suppressor in the thyroids and whether MIEAP knockout (KO) alone is sufficient for the oncocytic phenotype and also to extend our effort toward canonical mitophagy (a selective autophagy), we here conducted mouse studies using genetically engineered mice. BrafCA/wt mice developed thyroid cancers 1 year after intrathyroidal injection of adenovirus expressing Cre, while cancer development was observed at 6 months in adenovirus-Cre-injected BrafCA/wt;MieapKO/KO and BrafCA/wt;Atg5flox/flox mice [where autophagy-related 5 (ATG5) is a component of autophagic machinery], although KO of either molecule alone was not sufficient for cancer development. These data demonstrate that MIEAP or ATG5 KO accelerated thyroid cancer development. However, cancers in adenovirus-Cre-injected BrafCA/wt ;MieapKO/KO and BrafCA/wt ;Atg5flox/flox mice were not oncocytic. In conclusion, we here show that MIEAP and ATG5 are both tumor suppressors in thyroid carcinogenesis, but as we have anticipated from our previous data, KO of either molecule does not confer the oncocytic phenotype to BRAFV600E-positive thyroid cancers. The combination of disruptive mitochondrial function and impaired mitochondrial quality control may be necessary to establish a mouse model of thyroid oncocytoma.

A novel in vitro assay model developed to measure both extracellular and intracellular acetylcholine levels for screening cholinergic agents.

BACKGROUND: Cholinergic neurons utilize choline (Ch) to synthetize acetylcholine (ACh) and contain a high-affinity Ch transporter, Ch acetyltransferase (ChAT), ACh receptors, and acetylcholinesterase (AChE). As the depletion or malfunction of each component of the cholinergic system has been reported in patients with dementia, many studies have sought to evaluate whether treatment candidates affect each of the cholinergic components. The associated changes in the cholinergic components may be reflected by intra- or extra-cellular ACh levels, with an increase in extracellular ACh levels occurring following AChE inhibition. We hypothesized that increases in intracellular ACh levels can be more sensitively detected than those in extracellular ACh levels, thereby capturing subtle effects in the cholinergic components other than AChE. The objective of this study was to test this hypothesis. METHODS: We developed an in vitro model to measure both extracellular and intracellular ACh levels using the human cholinergic neuroblastoma cell line, LA-N-2, which have been reported to express Ch transporter, ChAT, muscarinic ACh receptor (mAChR), and AChE. With this model, we evaluated several drug compounds and food constituents reported to improve cholinergic function through various mechanisms. In addition, we conducted western blotting to identify the subtype of mAChR that is expressed on the cell line. RESULTS: Our cell-based assay system was capable of detecting increases in extracellular ACh levels induced by an AChE inhibitor at relatively high doses, as well as increases in intracellular ACh levels following the administration of lower AChE-inhibitor doses and an mAChR agonist. Moreover, increases in intracellular ACh levels were observed even after treatment with food constituents that have different mechanisms of action, such as Ch provision and ChAT activation. In addition, we revealed that LA-N-2 cells expressed mAChR M2. CONCLUSION: The findings support our hypothesis and indicate that the developed assay model can broadly screen compounds from drugs to food ingredients, with varying strengths and mechanisms of action, to develop treatments for ACh-relevant phenomena, including dementia and aging-related cognitive decline.

Efficacy of ice slurry and carbohydrate-electrolyte solutions for firefighters.

OBJECTIVES: To examine the thermoregulatory and fluid-electrolyte responses of firefighters ingesting ice slurry and carbohydrate-electrolyte solutions before and after firefighting operations. METHODS: Twelve volunteer firefighters put on fireproof clothing and ingested 5 g/kg of beverage in an anteroom at 25°C and 50% relative humidity (RH; pre-ingestion), and then performed 30 minutes of exercise on a cycle ergometer (at 125 W for 10 minutes and then 75 W for 20 minutes) in a room at 35℃ and 50% RH. The participants then returned to the anteroom, removed their fireproof clothing, ingested 20 g/kg of beverage (post-ingestion), and rested for 90 minutes. Three combinations of pre-ingestion and post-ingestion beverages were provided: a 25℃ carbohydrate-electrolyte solution for both (CH condition); 25℃ water for both (W condition); and a -1.7℃ ice slurry pre-exercise and 25℃ carbohydrate-electrolyte solution post-exercise (ICE condition). RESULTS: The elevation of body temperature during exercise was lower in the ICE condition than in the other conditions. The sweat volume during exercise was lower in the ICE condition than in the other conditions. The serum sodium concentration and serum osmolality were lower in the W condition than in the CH condition. CONCLUSIONS: The ingestion of ice slurry while firefighters were wearing fireproof clothing before exercise suppressed the elevation of body temperature during exercise. Moreover, the ingestion of carbohydrate-electrolyte solution by firefighters after exercise was useful for recovery from dehydration.

The effects of endotoxin on plasma free amino acid concentrations in rats.

We examined the effects of lipopolysaccharide (LPS) injection on body temperature and plasma free amino acid concentrations in rats. A catheter was placed in the jugular vein of the rats in order to draw blood from and to inject LPS into awake animals. On the day of the experiment, body temperature was recorded during the experiment (330 min) and blood was drawn before and at several time points after injection of LPS (10 microg/kg body weight). Body temperature in LPS-treated rats began to rise approximately 30 min after injection with a peak at 120 min, and afterward remained approximately 1 degree C higher than that in control rats through the end of the experiment. Concentrations of many plasma free amino acids were decreased by LPS treatment, with a nadir at approximately 120 min, and then were increased to the level of or over the control. It appears that thermoregulatory responses induced by LPS treatment may be related to alterations in plasma free amino acid concentrations. Effects of LPS treatment on the dynamics of plasma free branched-chain amino acid (BCAA) concentrations in rats with peroral or intravenous administration of BCAAs were also examined. The results showed that the rise in plasma BCAA concentrations after peroral BCAA administration was significantly suppressed by LPS treatment, but the dynamics of plasma BCAAs after intravenous administration was not affected by LPS, suggesting that LPS treatment inhibited the intestinal absorption of BCAAs into the circulation. These results suggest that the availability of administered BCAAs to the body tissues during sepsis is higher following parenteral than peroral administration.

Interval Walking Training and Nutritional Intake to Increase Plasma Volume in Elderly.

PURPOSE: Aerobic training-induced plasma volume (PV) expansion improves thermoregulation, and carbohydrate (CHO) + whey protein supplementation enhanced the effects in older people; however, these were suggested by studies on gym-based cycling training but not on home-based interval walking training (IWT). Moreover, long-term walking training effects on PV remain unknown. METHODS: Seventeen male and 10 female subjects (~69 yr), having performed IWT for ≥24 months before the study, were used. After pre-intervention measurement (PRE) of PV, plasma albumin content (Albcont), fasting glucose concentration ([Glc]f), and HbA1c, the subjects were randomly divided into two groups: CHO and Pro-CHO, either consuming CHO (22.5 g) alone or CHO (15 g) + whey protein (10 g), respectively, during additional 5-month IWT from May to November, 2009. After the additional IWT, we measured the same variables again (postintervention measurement). RESULTS: The baseline PV and Albcont were significantly correlated with the number of IWT days for the 12 months preceding PRE (r = 0.716, P < 0.001 and r = 0.671, P < 0.001, respectively). In postintervention, PV and Albcont marginally decreased in CHO from the baselines (P = 0.081 and P = 0.130, respectively) with increased HbA1c (P < 0.001) after correction for the baseline [Glc]f by ANCOVA, but these values remained unchanged in Pro-CHO (both, P > 0.74), with significant differences in the changes between groups (P = 0.020, P = 0.041, and P = 0.018 respectively). CONCLUSIONS: PV was proportional to the number of IWT days for 12 months and a CHO + whey protein supplementation during the 5-month IWT prevented PV reduction for the period of no supplementation, which might be partially linked with blood glucose control mechanisms.

Senescence of human skeletal muscle impairs the local inflammatory cytokine response to acute eccentric exercise.

The impact of aging on the cytokine response of human skeletal muscle to exercise-induced injury remains poorly understood. We enrolled physically active, young (23-35 years old, n=15) and old (66-78 years old, n=15) men to perform 45 min of downhill running (16% descent) at 75% VO2max. Biopsies of vastus lateralis were obtained 24 h before and 72 h after acute eccentric exercise. Transcripts for inflammatory (TNF-alpha, IL-1beta) and anti-inflammatory cytokines (IL-6, TGF-beta1) were quantified by real-time PCR. Before exercise, cytokine transcripts did not differ with age. At old age, exercise induced a blunted accumulation of transcripts encoding the pan-leukocyte surface marker CD18 (young: 10.1-fold increase, P<0.005; old: 4.7-fold increase, P=0.02; young vs. old: P<0.05). In both age groups, CD18 transcript accumulation strongly correlated with TNF-alpha (young, r=0.87, P<0.001; old, r=0.72, P=0.002) and TGF-beta1 transcript accumulation (young, r=0.80, P<0.001; old, r=0.64, P=0.008). At old age, there was no correlation between IL-1beta and CD18 transcript accumulation. Furthermore, exercise induced IL-6 transcript accumulation in young (3.6-fold, P=0.057) but not in old men. Our results suggest that aging impairs the adaptive response of human skeletal muscle to eccentric exercise by differential modulation of a discrete set of inflammatory and anti-inflammatory cytokine genes.

Effect of vitamin E supplementation on vitamin K status in adults with normal coagulation status.

BACKGROUND: Cases of enhanced anticoagulant effect in response to high-dose vitamin E supplementation have been reported among patients taking oral anticoagulants. Although a vitamin E-vitamin K interaction was proposed to underlie this effect, it has not been systematically investigated in adults with normal baseline coagulation status. OBJECTIVE: The objective was to study the effect of 12 wk of supplementation with 1000 IU RRR-alpha-tocopherol/d on biochemical measures of vitamin K status in men and women not taking oral anticoagulants. DESIGN: Vitamin K status, which was assessed with the use of plasma phylloquinone concentrations, the degree of under-gamma-carboxylation of prothrombin (proteins induced by vitamin K absence-factor II, PIVKA-II), and the percentage of undercarboxylated osteocalcin (ucOC), was determined in 38 men and women with rheumatoid arthritis (study A) and in 32 healthy men (study B) participating in 2 independent, 12-wk randomized clinical trials of vitamin E supplementation (1000 IU/d). RESULTS: Mean (+/- SD) PIVKA-II increased from 1.7 +/- 1.7 to 11.9 +/- 16.1 ng/mL (P < 0.001) in study A and from 1.8 +/- 0.6 to 5.3 +/- 3.9 ng/mL (P < 0.001) in study B in response to 12 wk of vitamin E supplementation. An increase in PIVKA-II is indicative of poor vitamin K status. In contrast, the other measures of vitamin K status (ie, plasma phylloquinone concentration and percentage of ucOC) did not change significantly in response to the supplementation. CONCLUSIONS: High-dose vitamin E supplementation increased PIVKA-II in adults not receiving oral anticoagulant therapy. The clinical significance of these changes warrants further investigation, but high doses of vitamin E may antagonize vitamin K. Whether such an interaction is potentially beneficial or harmful remains to be determined.

Plasma volume and blood viscosity during 4 h sitting in a dry environment: effect of prehydration.

BACKGROUND: Deep vein thrombosis and pulmonary thromboembolism are potential problems for travelers, including those who fly. We hypothesized that prehydration with an electrolyte-glucose beverage (EGB) would be better than water for maintaining body fluid balance and preventing increased blood viscosity in immobilized men. METHODS: There were 12 healthy men (24-38 yr) who participated in crossover trials of prehydration using EGB and H2O as well as a control condition (Con) with no prehydration. Fluid intake was set at 6 ml x kg(-1) body weight (mean 418 ml). For each trial, subjects sat for 4 h at a dry-bulb temperature of 23.0-23.5 degrees C and a relative humidity of 18-36%. Plasma volume (PV) and whole blood viscosity (Bvis) were determined every hour; routine laboratory hematological tests, urine volume, and body weight were recorded at 2 h and 4 h. RESULTS: For Con, subjects lost approximately 110 ml h(-1); at 2 h, PV had decreased significantly by 3.4%, and Bvis had increased significantly by 9.3%, with no further change at 4 h. For prehydration, retention of the consumed fluid at 2 h was significantly higher for EGB (57%) than for H2O (38%), while both drinks prevented significant change in PV and Bvis. There were no significant differences between trials in coagulation variables, but Bvis measured at higher shear rates for EGB were significantly attenuated compared with Con. CONCLUSION: EGB and water prevented the increase of blood viscosity that occurred without prehydration. EGB was better than water for maintaining body fluid balance and preventing hypovolemia.

Effect of prehydration on nasal mucociliary clearance in low relative humidity.

OBJECTIVE: Nasal mucociliary clearance, which plays an important role in defending the respiratory system, tends to decrease under conditions of low relative humidity (RH). The purpose of this randomized cross-over study was to investigate the preventive effect of prehydration on nasal mucociliary clearance under low RH conditions. METHODS: Fourteen young healthy subjects were assigned to three interventions: prehydration (6ml/kg body weight) using water (W), prehydration using a carbohydrate-electrolyte beverage (CE), and control conditions (Cont) with no prehydration. For each intervention trial, subjects sat for 4h in an environmental chamber (23°C, 10% RH). Nasal mucociliary clearance was measured by the saccharin transit time (ST). Furthermore, a dry sensation of the mucosa, urine volume, and body weight were measured at the baseline, 2h and 4h after entering the environmental chamber, respectively. RESULTS: ST was prolonged under the Cont conditions after entering the environmental chamber. Prehydration with CE led to a significantly lower ST compared to the Cont pretreatment at 2h, whereas the W trial showed no significance difference in comparison to the Cont values. CONCLUSION: These results suggest that prehydration with a carbohydrate-electrolyte beverage is therefore beneficial for maintaining better nasal mucociliary clearance at least for 2h under low RH conditions.

Branched-chain amino acid supplementation increases the lactate threshold during an incremental exercise test in trained individuals.

The effects of branched-chain amino acid (BCAA) supplementation on the lactate threshold (LT) were investigated as an index of endurance exercise capacity. Eight trained male subjects (21+/-2 y) participated in a double-blind crossover placebo-controlled study. The subjects were randomly assigned to two groups and were provided either a BCAA drink (0.4% BCAA, 4% carbohydrate; 1,500 mL/d) or an iso-caloric placebo drink for 6 d. On the 7th day, the subjects performed an incremental loading exercise test with a cycle ergometer until exhaustion in order to measure the LT. The test drink (500 mL) was ingested 15-min before the test. Oxygen consumption VO2 and the respiratory exchange ratio (RER) during the exercise test were measured with the breath-by-breath method. Blood samples were taken before and during the exercise test to measure the blood lactate and plasma BCAA concentrations. The same exercise test was performed again 1 wk later. BCAA supplementation increased the plasma BCAA concentration during the exercise test, while plasma BCAA concentration decreased in the placebo trial. The RER during the exercise test in the BCAA trial was lower than that in the placebo trial (p<0.05). The VO2 and workload levels at LT point in the BCAA trial were higher than those in the placebo trial (VO2: 29.8+/-6.8 vs. 26.4+/-5.4 mL/kg/min; workload: 175+/-42 vs. 165+/-38 W, p<0.05, respectively). The VO2max in the BCAA trial was higher than that in the placebo trial (47.1+/-5.7 vs. 45.2+/-5.0 mL/kg/min, p<0.05). These results suggest that BCAA supplementation may be effective to increase the endurance exercise capacity.

Leucine-induced activation of translational initiation is partly regulated by the branched-chain alpha-keto acid dehydrogenase complex in C2C12 cells.

Branched-chain amino acid leucine has been shown to activate the translational regulators through the mammalian target of rapamycin. However, the leucine's effects are self-limiting because leucine promotes its own disposal by an oxidative pathway. The irreversible and rate-limiting step in the leucine oxidation pathway is catalyzed by the branched-chain alpha-keto acid dehydrogenase (BCKDH) complex. The complex contains E1 (alpha2beta2), E2, and E3 subunits, and its activity is abolished by phosphorylation of the E1alpha subunit by BCKDH kinase. The relationship between the activity of BCKDH complex and leucine-mediated activation of the protein translation was investigated using the technique of RNA interference. The activity of BCKDH complex in C2C12 cell was modulated by transfection of small interfering RNA (siRNA) for BCKDH E2 subunit or BCKDH kinase. Transfection of siRNAs decreased the mRNA expression and protein amount of corresponding gene. Suppression of either E2 subunit or kinase produced opposite effects on the cell proliferation and the activation of translational regulators by leucine. Suppression of BCKDH kinase for 48h resulted in decreasing cell proliferation. In contrast, E2 suppression led to increased amount of total cellular protein. The phosphorylation of p70 S6 kinase by leucine was increased in E2-siRNA transfected C2C12 cells, whereas the leucine's effect was diminished in kinase-siRNA transfected cells. These results suggest that the activation of the translational regulators by leucine was partly regulated by the activity of BCKDH complex.

Age-related loss of associations between acute exercise-induced IL-6 and oxidative stress.

IL-6 mediates many aspects of the exercise-induced acute-phase response, including upregulation of antioxidant defenses. Moreover, IL-6 synthesis is regulated in part by oxidative stress. This investigation tested the hypothesis that an IL-6-mediated acute-phase response after exercise provides negative-feedback protection against exercise-induced oxidative stress. Healthy young (n = 16, 26.4 +/- 1.8 yr) and older men (n = 16, 71.1 +/- 2.0 yr) ran downhill for 45 min at 75% maximal oxygen consumption before and after a 12-wk period of supplementation with vitamin E (1,000 IU/day) or placebo. Circulating IL-6 and soluble IL-6 receptors, peripheral mononuclear cell production of IL-6, and IL-6 transcripts in muscle were measured before and within a 72-h time window after each acute exercise bout. At all time points plasma IL-6, IL-6 bioavailability, and C-reactive protein were higher in the older men; yet in response to exercise, young and older subjects experienced similar increases in these factors. Although the magnitude of postexercise changes in acute-phase variables was independent of age, correlations among plasma, mononuclear cell, and muscle IL-6 and oxidative stress were evident only in young men (R2 = 0.64, 0.35, and 0.33, respectively). These changes in circulating IL-6 were closely associated with a prooxidant state (R2 = 0.47), whereas muscle IL-6 mRNA correlated with an antioxidant state (R2 = 0.65). Supplementation with vitamin E did not affect exercise-induced responses or differences between the young and old men in a consistent manner. Therefore, oxidative stress is linked to the acute-phase response after exercise in young men, but not in older men who had elevated acute-phase reactants, suggesting that further research is warranted to determine the basis for these differences.