3-methylhistidine and clinical outcomes in maintenance haemodialysis patients.

BACKGROUND: Chronic kidney disease is an important contributor to morbidity and mortality. 3-methylhistidine (3-MH) is the by-product of actin and myosin degradation reflecting skeletal muscle turnover. Markedly elevated 3-MH levels have been documented in uraemic patients, but the interpretation of high 3-MH concentration in maintenance haemodialysis (MHD) patients remains unclear. Indeed, it is not known whether elevated serum 3-MH levels are a marker of excessive muscle catabolism or a better lean tissue mass. Here, we evaluated the association between serum 3-MH levels and clinical outcomes in these patients. METHODS: Serum 3-MH concentration was measured by reverse-phase liquid chromatography/tandem mass spectrometry in a cohort of MHD patients. We analysed the relationships between various clinical/laboratory indices, lean tissue mass measured by bioimpedance spectroscopy, mortality and cardiovascular (CV) events. RESULTS: Serum 3-MH concentration was positively correlated with serum albumin, normalized protein catabolic rate (nPCR), simplified creatinine index (SCI) and lean tissue mass. Of 291 MHD patients, during a mean follow-up of 847 days, 91 patients died and 101 patients experienced a CV event. Survival was significantly better in patients with high 3-MH concentrations (P = .002). A higher level of 3-MH was also associated with a lower CV mortality and lower incidence of CV events (P = .015 and P < .001, respectively). Low serum 3-MH levels remained significantly associated with CV events but not with mortality after adjustment for demographic, metabolic and CV risk factors. CONCLUSION: Elevated serum 3-MH concentration appears to be a marker of better lean tissue mass and nutritional status. Low serum 3-MH is a robust and independent predictor of CV events in the MHD population.

Low Serum 3-Methylhistidine Levels Are Associated With First Hospitalization in Kidney Transplantation Recipients.

BACKGROUND: Low protein intake and increased muscle breakdown are associated with increased mortality risk in patients with kidney transplantation (KT). 3-methylhistidine (3-MH), a nonproteinogenic amino acid residue, is an index of muscle breakdown. the present study investigated the association between serum 3-MH levels and subsequent first hospitalization events in patients with KT. METHODS: A total of 64 KT patients were enrolled and 43 first hospitalization events occurred. Fasting blood samples were obtained and serum 3-MH level was performed with high-performance liquid chromatography and mass spectrometry. Associations between serum 3-MH levels and first hospitalization over a 5-year follow-up period were examined. RESULTS: Compared with patients without hospitalization, the 64 patients with KT revealed higher diabetes (P = .012) and hypertension (P = .006) prevalence, higher body fat mass (P = .012) and systolic blood pressure (P = .002), higher serum blood urea nitrogen (BUN) levels (P = .003), and lower serum 3-MH levels (P = .001). Statistical analysis revealed that serum 3-MH (95% confidence interval [CI]: 0.902-0.986, P = .010) and serum BUN (95% CI: 1.003-1.040, P = .022) levels were independently associated with first hospitalization events in patients with KT. Kaplan-Meier analysis showed a greater cumulative incidence of first hospitalization events in the patients with lower 3-MH levels (≤5.91 ng/mL) than that in those with higher 3-MH levels (P = .014; log-rank test). CONCLUSIONS: Low serum 3-MH levels are associated with increased first hospitalization risk in KT recipients.

Race modifies the association between animal protein metabolite 1-methylhistidine and blood pressure in middle-aged adults: the Bogalusa Heart Study.

OBJECTIVE: Dietary factors mediate racial disparities in hypertension. However, the physiological mechanisms underlying this relationship are incompletely understood. We sought to assess the association between 1-methylhistidine (1-MH), a metabolite marker of animal protein consumption, and blood pressure (BP) in a community-based cohort of black and white middle-aged adults. METHODS: This analysis consisted of 655 participants of the Bogalusa Heart Study (25% black, 61% women, aged 34-58 years) who were not taking antihypertensive medication. Fasting serum 1-MH was measured using liquid chromatography-tandem mass spectroscopy. Animal food intakes were quantified by food-frequency questionnaires. Multivariable linear regression assessed the association between 1-MH and BP in combined and race-stratified analyses, adjusting for demographic, dietary, and cardiometabolic factors. RESULTS: A significant dose--response relationship was observed for the association of red meat (P-trend <0.01) and poultry (P-trend = 0.03) intake with serum 1-MH among all individuals. Serum 1-MH, per standard deviation increase, had a significant positive association with SBP (ß=3.4 ±â€Š1.6 mmHg, P = 0.04) and DBP (ß=2.0 ±â€Š1.1 mmHg, P = 0.05) in black participants, whereas no appreciable association was observed in white participants. Among a subgroup of black participants with repeat outcome measures (median follow-up = 3.0 years), one standard deviation increase in 1-MH conferred a 3.1 and 2.2 mmHg higher annual increase in SBP (P = 0.03) and DBP (P = 0.03), respectively. CONCLUSION: Serum 1-MH associates with higher SBP and DBP in blacks, but not whites. These results suggest a utility for further assessing the role of dietary 1-MH among individuals with hypertension to help minimize racial disparities in cardiovascular health.

Effects of rumen-protected lysine and histidine on milk production and energy and nitrogen utilization in diets containing hydrolyzed feather meal fed to lactating Jersey cows.

Hydrolyzed feather meal (HFM) is high in crude protein, most of which bypasses rumen degradation when fed to lactating dairy cows, allowing direct supply of AA to the small intestine. Compared with other feeds that are high in bypass protein, such as blood meal or heat-treated soybean meal, HFM is low in His and Lys. The objectives of this study were to determine the effects of supplementing rumen-protected (RP) Lys and His individually or in combination in a diet containing 5% HFM on milk production and composition as well as energy and N partitioning. Twelve multiparous Jersey cows (mean ± SD: 91 ± 18 d in milk) were used in a triplicated 4 × 4 Latin square with 4 periods of 28 d (24-d adaptation and 4-d collection). Throughout the experiment, all cows were fed the same TMR, with HFM included at 5% of diet DM. Cows were grouped by dry matter intake and milk yield, and cows within a group were randomly assigned to 1 of 4 treatments: no RP Lys or RP His; RP Lys only [70 g/d of Ajipro-L (24 g/d of digestible Lys), Ajinomoto Co. Inc., Tokyo, Japan]; RP His only [32 g/d of experimental product (7 g/d of digestible His), Balchem Corp., New Hampton, NY]; or both RP Lys and His. Plasma Lys concentration increased when RP Lys was supplemented without RP His (77.7 vs. 66.0 ± 4.69 µM) but decreased when RP Lys was supplemented with RP His (71.4 vs. 75.0 ± 4.69 µM). Plasma concentration of 3-methylhistidine decreased with RP Lys (3.19 vs. 3.40 ± 0.31 µM). With RP His, plasma concentration of His increased (21.8 vs. 18.7 ± 2.95 µM). For milk production and milk composition, no effects of Lys were observed. Supplementing RP His increased milk yield (22.5 vs. 21.6 ± 2.04 kg/d) and tended to increase milk protein yield (0.801 vs. 0.772 ± 0.051 kg/d). Across treatments, dry matter intake (18.5 ± 0.83 kg/d) and energy supply (32.2 ± 2.24 Mcal of net energy for lactation) were not different. Supplementing RP His did not affect N utilization; however, supplementing RP Lys increased N balance (25 vs. 16 ± 9 g/d). The lack of production responses to RP Lys suggests that Lys was not limiting or that the increase in Lys supply was not large enough to cause an increase in milk protein yield. However, increased N balance and decreased 3-methylhistidine with RP Lys suggest that increased Lys supply increased protein accretion and decreased protein mobilization. Furthermore, His may be a limiting AA in diets containing HFM.

Acute Effect of the Timing of Resistance Exercise and Nutrient Intake on Muscle Protein Breakdown.

BACKGROUND: Combining resistance exercise (RE) with nutrient intake stimulates muscle protein net balance. However, it is still unclear whether the optimal timing of nutrient intake is before or after RE, especially on muscle protein breakdown (MPB) for an augmented muscle anabolic response. The aim of this study was to investigate the effect of a substantial mixed meal (i.e., nutrient- and protein-dense whole foods) before or after RE, compared with RE without a meal on the acute response of MPB in a crossover-design study. METHODS: Eight healthy young men performed three trials: (1) meal intake before RE (Pre), (2) meal intake after RE (Post), and (3) RE without meal intake (No). Plasma insulin and 3-methylhistidine (3-MH), an MPB marker, were measured. RESULTS: Time course change in plasma insulin level after RE was significantly higher in the Post condition than in the Pre and No conditions. The area under the curve of 3-MH concentration was significantly lower in the Post condition than in the Pre and No conditions. CONCLUSIONS: These results suggest that a substantial mixed meal immediately after RE may effectively suppress MPB in the morning.

Mammalian target of rapamycin signaling and ubiquitin-proteasome-related gene expression in skeletal muscle of dairy cows with high or normal body condition score around calving.

The objective of the current study was to investigate the effects of overconditioning around calving on gene expression of key components of the mammalian target of rapamycin (mTOR) pathway and ubiquitin-proteasome system (UPS) in skeletal muscle as well as the AA profiles in both serum and muscle of periparturient cows. Fifteen weeks antepartum, 38 multiparous Holstein cows were allocated to either a high body condition group (HBCS; n = 19) or a normal body condition group (NBCS; n = 19) and were fed different diets until dry-off (d -49 relative to calving) to amplify the difference. The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg). At dry-off, the NBCS cows (parity: 2.42 ± 1.84; body weight: 665 ± 64 kg) had a body condition score (BCS) <3.5 and backfat thickness (BFT) <1.2 cm, whereas the HBCS cows (parity: 3.37 ± 1.67; body weight: 720 ± 57 kg) had a BCS >3.75 and BFT >1.4 cm. During the dry period and the subsequent lactation, both groups were fed identical diets but maintained the BCS and BFT differences. Blood samples and skeletal muscle biopsies (semitendinosus) were repeatedly (d -49, +3, +21, and +84 relative to calving) collected for assessing the concentrations of free AA and the mRNA abundance of various components of mTOR and UPS. The differences in BCS and BFT were maintained throughout the study. The circulating concentrations of most AA with the exception of Gly, Gln, Met, and Phe increased in early lactation in both groups. The serum concentrations of Ala (d +21 and +84) and Orn (d +84) were lower in HBCS cows than in NBCS cows, but those of Gly, His, Leu, Val, Lys, Met, and Orn on d -49 and Ile on d +21 were greater in HBCS cows than in NBCS cows. The serum concentrations of 3-methylhistidine, creatinine, and 3-methylhistidine:creatinine ratio increased after calving (d +3) but did not differ between the groups. The muscle concentrations of all AA (except for Cys) remained unchanged over time and did not differ between groups. The muscle concentrations of Cys were greater on d -49 but tended to be lower on d +21 in HBCS cows than in NBCS cows. On d +21, mTOR and eukaryotic translation initiation factor 4E binding protein 1 mRNA abundance was greater in HBCS cows than in NBCS cows, whereas ribosomal protein S6 kinase 1 was not different between the groups. The mRNA abundance of ubiquitin-activating enzyme 1 (d +21), ubiquitin-conjugating enzyme 1 (d +21), atrogin-1 (d +21), and ring finger protein-1 (d +3) enzymes was greater in HBCS cows than in NBCS cows, whereas ubiquitin-conjugating enzyme 2 was not different between the groups. The increased mRNA abundance of key components of mTOR signaling and of muscle-specific ligases of HBCS cows may indicate a simultaneous activation of anabolic and catabolic processes and thus increased muscle protein turnover, likely as a part of the adaptive response to prevent excessive loss of skeletal muscle mass during early lactation.

Plasma concentrations of anserine, carnosine and pi-methylhistidine as biomarkers of habitual meat consumption.

BACKGROUND/OBJECTIVES: Dietary intake of red and processed meat has been associated with disease risk. Since dietary intake assessment methods are prone to measurement errors, identifying biomarkers of meat intake in bio-samples could provide more valid intake estimates. We examined associations of habitual red and processed meat, poultry, fish, and dairy products consumption with plasma concentrations of anserine, carnosine, pi-methylhistidine (Π-MH), tau-methylhistidine (T-MH), and the ratio of T-MH to Π-MH in a cross-sectional study. SUBJECTS/METHODS: Plasma anserine, carnosine, Π-MH, and T-MH concentrations were measured using ion-pair LC-MS/MS in 294 participants in the second Bavarian Food Consumption Survey (BVS II). Habitual food consumption was assessed using three 24-h dietary recalls. Associations between plasma metabolites concentrations and meat, fish, eggs, and dairy products consumption were assessed by fitting generalized linear model, adjusted for age, sex, and BMI. RESULTS: Total meat intake was associated with plasma concentrations of anserine, carnosine, Π-MH and, the ratio of T-MH to Π-MH. Red meat intake was related to carnosine (p-trend = 0.0028) and Π-MH plasma levels (p-trend = 0.0493). Poultry (p-trend = 0.0006) and chicken (p-trend = 0.0003) intake were associated with Π-MH. The highest anserine concentrations were observed in individuals consuming processed meat or turkey. For T-MH we did not observe any association with meat intake. CONCLUSIONS: Our results indicate an association between habitual meat consumption and plasma concentrations of anserine, carnosine, Π-MH and the ratio of T-MH to Π-MH. Intervention studies should clarify whether the analyzed plasma metabolites are indicative for a specific type of meat before proposing them as biomarkers of habitual meat intake in epidemiologic studies.

Untargeted 1H NMR-Based Metabolomics Analysis of Urine and Serum Profiles after Consumption of Lentils, Chickpeas, and Beans: An Extended Meal Study To Discover Dietary Biomarkers of Pulses.

High legume intake has been shown to have beneficial effects on the health of humans. The use of nutritional biomarkers, as a complement to self-reported questionnaires, could assist in evaluating dietary intake and downstream effects on human health. The aim of this study was to investigate potential biomarkers of the consumption of pulses (i.e., white beans, chickpeas, and lentils) by using untargeted NMR-based metabolomics. Meals rich in pulses were consumed by a total of 11 participants in a randomized crossover study and multilevel partial least-squares regression was employed for paired comparisons. Metabolomics analysis indicated that trigonelline, 3-methylhistidine, dimethylglycine, trimethylamine, and lysine were potential, though not highly specific, biomarkers of pulse intake. Furthermore, monitoring of these metabolites for a period of 48 h after intake revealed a range of different excretion patterns among pulses. Following the consumption of pulses, a metabolomic profiling revealed that the concentration ratios of trigonelline, choline, lysine, and histidine were similar to those found in urine. In conclusion, this study identified potential urinary biomarkers of exposure to dietary pulses and provided valuable information about the time-response effect of these putative biomarkers.

The Influence of Dietary Habits and Meat Consumption on Plasma 3-Methylhistidine-A Potential Marker for Muscle Protein Turnover.

SCOPE: 3-Methylhistidine (3-MH) as a potential biomarker for muscle protein turnover is influenced by meat intake but data on the impact of meat on plasma 3-MH are scarce. We determined the association of plasma 3-MH, 1-methylhistidine (1-MH), and creatinine with dietary habits and assessed the impact of a single white meat intervention during a meat-free period. METHODS AND RESULTS: Plasma 3-MH, 1-MH, and creatinine concentrations of healthy young omnivores (n = 19) and vegetarians (n = 16) were analyzed together with data on anthropometry, body composition, grip strength, and nutrition. After baseline measurements omnivores adhered to a meat-free diet for 6 days and received a defined administration of chicken breast on day four. At baseline, omnivores had higher plasma 3-MH and 1-MH concentrations than vegetarians. White meat administration led to a slight increase in plasma 3-MH in omnivores. The elevated 3-MH concentrations significantly declined within 24 h after white meat intake. CONCLUSION: 1-MH concentrations in plasma seem to be suitable to display (white) meat consumption and its influence on 3-MH plasma concentration. 3-MH in plasma may be used as a biomarker for muscle protein turnover if subjects have not consumed meat in the previous 24 h.

Estimation of Chicken Intake by Adults Using Metabolomics-Derived Markers.

BACKGROUND: Improved assessment of meat intake with the use of metabolomics-derived markers can provide objective data and could be helpful in clarifying proposed associations between meat intake and health. OBJECTIVE: The objective of this study was to identify novel markers of chicken intake using a metabolomics approach and use markers to determine intake in an independent cohort. METHODS: Ten participants [age: 62 y; body mass index (in kg/m2): 28.25] in the NutriTech food intake study consumed increasing amounts of chicken, from 88 to 290 g/d, in a 3-wk span. Urine and blood samples were analyzed by nuclear magnetic resonance and mass spectrometry, respectively. A multivariate data analysis was performed to identify markers associated with chicken intake. A calibration curve was built based on dose-response association using NutriTech data. A Bland-Altman analysis evaluated the agreement between reported and calculated chicken intake in a National Adult Nutrition Survey cohort. RESULTS: Multivariate data analysis of postprandial and fasting urine samples collected in participants in the NutriTech study revealed good discrimination between high (290 g/d) and low (88 g/d) chicken intakes. Urinary metabolite profiles showed differences in metabolite levels between low and high chicken intakes. Examining metabolite profiles revealed that guanidoacetate increased from 1.47 to 3.66 mmol/L following increasing chicken intakes from 88 to 290 g/d (P < 0.01). Using a calibration curve developed from the NutriTech study, chicken intake was calculated through the use of data from the National Adult Nutrition Survey, in which consumers of chicken had a higher guanidoacetate excretion (0.70 mmol/L) than did nonconsumers (0.47 mmol/L; P < 0.01). A Bland-Altman analysis revealed good agreement between reported and calculated intakes, with a bias of -30.2 g/d. Plasma metabolite analysis demonstrated that 3-methylhistidine was a more suitable indicator of chicken intake than 1-methylhistidine. CONCLUSIONS: Guanidoacetate was successfully identified and confirmed as a marker of chicken intake, and its measurement in fasting urine samples could be used to determine chicken intake in a free-living population. This trial was registered at clinicaltrials.gov as NCT01684917.

Application of a new procedure for liquid chromatography/mass spectrometry profiling of plasma amino acid-related metabolites and untargeted shotgun proteomics to identify mechanisms and biomarkers of calcific aortic stenosis.

Calcific aortic valve stenosis (CAS) increasingly affects our ageing population, but the mechanisms of the disease and its biomarkers are not well established. Recently, plasma amino acid-related metabolite (AA) profiling has attracted attention in studies on pathology and development of biomarkers of cardiovascular diseases, but has not been studied in CAS. To evaluate the potential relationship between CAS and AA metabolome, a new ion-pairing reversed-phase liquid chromatography-tandem mass spectrometry (IP-RPLC-MS/MS) method has been developed and validated for simultaneous determination of 43 AAs in plasma of stenotic patients and age-matched control subjects. Furthermore, untargeted mass spectrometry-based proteomic analysis and confirmatory ELISA assays were performed. The method developed offered high accuracy (intra-assay imprecision averaged 4.4% for all compounds) and sensitivity (LOQ within 0.01-0.5µM). We found that 22 AAs and three AA ratios significantly changed in the CAS group as compared to control. The most pronounced differences were observed in urea cycle-related AAs and branched-chain AA (BCAA)-related AAs. The contents of asymmetric dimethylarginine (ADMA) and its monomethylated derivative (NMMA) were increased by 30-64% with CAS. The arginine/ADMA and Fischer's ratios as well as arginine, homoarginine, ADMA, symmetric dimethylarginine, hydroxyproline, betaine and 3-methylhistidine correlated with cardiac function-related parameters and concomitant systemic factors in the CAS patients. The results of proteomic analysis were consistent with involvement of inflammation, lipid abnormalities, hemostasis and extracellular matrix remodeling in CAS. In conclusion, changes in plasma AA profile and protein pattern that we identified in CAS provide information relevant to pathomechanisms and may deliver new biomarkers of the disease.

Effects of short-term fasting on the Akt-mediated pathway involved in protein metabolism in chicken skeletal muscle.

In the present study, we show that short-term (4 h) fasting significantly decreased the levels of protein synthesis-related factors such as the plasma insulin concentration, skeletal muscle pAkt, and pS6 levels in 2-wk-old chickens (P < 0.05). An intravenous injection of insulin significantly elevated the contents of pAkt and p-S6 in the skeletal muscle (P < 0.01). These findings suggest that decreasing the plasma insulin causes the downregulation of the Akt/S6 pathway in chicken skeletal muscle under short-term fasting conditions. However, protein synthesis was not significantly affected by short-term fasting. In addition, no significant change was observed in the levels of proteolysis-related factors such as plasma Nτ-methylhistidine, phosphorylated forkhead box class O, and muscle ring finger-1 during 4-h fasting, indicating that short-term fasting does not induce skeletal muscle proteolysis in chickens. Interestingly, atrogin-1 expression significantly increased after 2-h fasting (P < 0.05), and insulin injection significantly reversed the fasting-induced atrogin-1 expression in chicken skeletal muscle (P < 0.01). Collectively, these findings suggest that short-term fasting downregulates the insulin-stimulated Akt/S6 pathway but does not significantly affect protein synthesis and proteolysis in chicken skeletal muscle, and that atrogin-1 expression is upregulated in a FOXO1-independent manners.

Short- and longer-term effects of feeding increased metabolizable protein with or without an altered amino acid profile to dairy cows immediately postpartum.

The first few weeks after parturition is marked by low, but increasing feed intake and sharply increasing milk production by dairy cows. Because of low intake, the nutrient density of the diet may need to be higher during this period to support increasing milk yields. We hypothesized that feeding higher levels of metabolizable protein (MP) or a protein supplement with rumen-protected lysine and methionine during the immediate postpartum period would increase yields of milk and milk components. Fifty-six Holstein cows (21 primiparous and 35 multiparous) starting at 3 d in milk were used in a randomized block design. In phase 1 (3 through 23 d in milk), cows were fed 1 of 3 diets that differed in supply of MP and AA profile. At 23 d in milk, all cows were moved to a common freestall pen and fed the control diet used in phase 1 for an additional 63 d (phase 2). Diets were formulated using the National Research Council model and were control [16.5% crude protein (CP), 10.9% rumen-degradable protein (RDP), and 5.6% rumen-undegradable protein (RUP)], high MP (HMP; 18.5% CP, 11.6% RDP, 6.9% RUP), and AA (MPAA; 17.5% CP, 10.5% RDP, 7.0% RUP 29.7). The MPAA diet included a proprietary spray-dried blood meal product (Perdue Agribusiness, Salisbury, MD) and contained a model-estimated 7.2 and 2.6% of digestible lysine and methionine (% of MP). The HMP and control diets contained 6.3 and 6.7% digestible lysine and both had 1.8% digestible methionine. In phase 1, diet did not affect milk yield (33.6, 34.7, and 33.2 kg for control, HMP, and MPAA, respectively), dry matter intake (17.8, 18.0, and 18.5 kg/d for control, HMP, and MPAA), or milk protein yield (1.07 kg/d). Feeding additional protein (HMP or MPAA) increased both the concentration and yield of milk fat, and milk protein concentration was greater (3.30 vs. 3.17%) for MPAA compared with the HMP diet. Energy-corrected milk was greater (38.4 and 38.6 vs. 35.3 kg/d, respectively) for MPAA and HP than for the control. Cows fed MPAA had the greatest plasma concentrations of Met and the lowest concentrations of isoleucine, but lysine was not affected by treatment. Feeding additional MP (HMP or MPAA) reduced the concentrations of 3-methylhistidine in plasma, indicating reduced muscle breakdown. Diet effects on milk composition continued after cows were changed to a common diet in that cows fed MPAA the first 3 wk of lactation had greater concentration of milk protein for the entire experiment than cows fed HMP, and cows fed additional MP (HMP and MPAA) during phase 1 had greater concentrations of milk fat for the entire experiment. Increasing dietary protein and AA supply in early lactation had short-term effects on yield of energy-corrected milk and long-term effects on milk composition.

Dietary supplementation of branched-chain amino acids increases muscle net amino acid fluxes through elevating their substrate availability and intramuscular catabolism in young pigs.

Branched-chain amino acids (BCAA) have been clearly demonstrated to have anabolic effects on muscle protein synthesis. However, little is known about their roles in the regulation of net AA fluxes across skeletal muscle in vivo. This study was aimed to investigate the effect and related mechanisms of dietary supplementation of BCAA on muscle net amino acid (AA) fluxes using the hindlimb flux model. In all fourteen 4-week-old barrows were fed reduced-protein diets with or without supplemental BCAA for 28 d. Pigs were implanted with carotid arterial, femoral arterial and venous catheters, and fed once hourly with intraarterial infusion of p-amino hippurate. Arterial and venous plasma and muscle samples were obtained for the measurement of AA, branched-chain α-keto acids (BCKA) and 3-methylhistidine (3-MH). Metabolomes of venous plasma were determined by HPLC-quadrupole time-of-flight-MS. BCAA-supplemented group showed elevated muscle net fluxes of total essential AA, non-essential AA and AA. As for individual AA, muscle net fluxes of each BCAA and their metabolites (alanine, glutamate and glutamine), along with those of histidine, methionine and several functional non-essential AA (glycine, proline and serine), were increased by BCAA supplementation. The elevated muscle net AA fluxes were associated with the increase in arterial and intramuscular concentrations of BCAA and venous metabolites including BCKA and free fatty acids, and were also related to the decrease in the intramuscular concentration of 3-MH. Correlation analysis indicated that muscle net AA fluxes are highly and positively correlated with arterial BCAA concentrations and muscle net BCKA production. In conclusion, supplementing BCAA to reduced-protein diet increases the arterial concentrations and intramuscular catabolism of BCAA, both of which would contribute to an increase of muscle net AA fluxes in young pigs.

Methionine and Choline Supply during the Periparturient Period Alter Plasma Amino Acid and One-Carbon Metabolism Profiles to Various Extents: Potential Role in Hepatic Metabolism and Antioxidant Status.

The objective of this study was to profile plasma amino acids (AA) and derivatives of their metabolism during the periparturient period in response to supplemental rumen-protected methionine (MET) or rumen-protected choline (CHOL). Forty cows were fed from -21 through 30 days around parturition in a 2 × 2 factorial design a diet containing MET or CHOL. MET supply led to greater circulating methionine and proportion of methionine in the essential AA pool, total AA, and total sulfur-containing compounds. Lysine in total AA also was greater in these cows, indicating a better overall AA profile. Sulfur-containing compounds (cystathionine, cystine, homocystine, and taurine) were greater in MET-fed cows, indicating an enriched sulfur-containing compound pool due to enhanced transsulfuration activity. Circulating essential AA and total AA concentrations were greater in cows supplied MET due to greater lysine, arginine, tryptophan, threonine, proline, asparagine, alanine, and citrulline. In contrast, CHOL supply had no effect on essential AA or total AA, and only tryptophan and cystine were greater. Plasma 3-methylhistidine concentration was lower in response to CHOL supply, suggesting less tissue protein mobilization in these cows. Overall, the data revealed that enhanced periparturient supply of MET has positive effects on plasma AA profiles and overall antioxidant status.

Clenbuterol changes phosphorylated FOXO1 localization and decreases protein degradation in the sartorius muscle of neonatal chicks.

To investigate the intracellular signaling mechanisms by which clenbuterol reduces muscle protein degradation, we examined the phosphorylation level and intracellular localization of FOXO1 in the sartorius muscle of neonatal chicks. One-day-old chicks were given a single intraperitoneal injection of clenbuterol (0.1 mg/kg body weight). Three hours after injection, AKT protein was phosphorylated in the sartorius muscle by clenbuterol injection. Coincidentally, clenbuterol increased cytosolic level of phosphorylated FOXO1 protein, while it decreased nuclear level of FOXO1 protein in the sartorius muscle. Furthermore, clenbuterol decreased the expression of mRNAs for muscle-specific ubiquitin ligases (atrogin-1/MAFbx and MuRF1) in the sartorius muscle accompanied by decreased plasma 3-methylhistidine concentration, an index of muscle protein degradation, at 3 h after injection. These results suggested that, in the sartorius muscle of the chicks, clenbuterol changed the intracellular localization of phosphorylated FOXO1, and consequently decreased protein degradation via suppressing the expression of genes encoding muscle-specific ubiquitin ligases.

Possible relationship between amino acids, aggression and psychopathy.

OBJECTIVE: Aggressive behaviour is associated with reduced serotonin metabolism in the brain, but there is not enough knowledge on potential changes of the serotonin precursor levels among violent offenders. In this study, we aimed to evaluate the relationships among the tendency of psychopathy, anger and the basic amino acids. METHODS: Fifty-two young adult male patients with antisocial personality disorder (APD) and 30 healthy men included the study. Serum amino acid levels were measured by HPLC method. Aggression questionnaire and Hare Psychopathology Scale were used for all participants. RESULTS: Blood levels of phosphoserine, aspartic acid, glutamic acid, aminoadipic acid and 1-methylhistidine in group of patients with APD were significantly higher than the control group. Blood levels of TRP, asparagine, citrulline, cystine, isoleucine, tyrosine, histidine, hydroxylysine, lysine, ethanolamine and arginine in the group of patients were found lower than the control group. A significant positive correlation between anger scores and histidine, methionine and GABA was found. GABA and methionine showed a significant correlation with the indirect aggression score. CONCLUSION: Our study showed a relationship between serum amino acid levels and the scores of aggression and psychopathy. We think that this is a productive research area for understanding the relationship among biochemical factors, aggression and psychopathy.

Time-course changes in muscle protein degradation in heat-stressed chickens: Possible involvement of corticosterone and mitochondrial reactive oxygen species generation in induction of the ubiquitin-proteasome system.

Heat stress (HS) induces muscle protein degradation as well as production of mitochondrial reactive oxygen species (ROS). In the present study, to improve our understanding of how protein degradation is induced by HS treatment in birds, a time course analysis of changes in the circulating levels of glucocorticoid and N(τ)-methylhistidine, muscle proteolysis-related gene expression, and mitochondrial ROS generation, was conducted. At 25 days of age, chickens were exposed to HS conditions (33 °C) for 0, 0.5, 1 or 3 days. While no alteration in plasma N(τ)-methylhistidine concentration relative to that of the control group was observed in the 0.5 day HS group, the concentration was significantly higher in the 3-d HS treatment group. Plasma corticosterone concentrations increased in response to 0.5-d HS treatment, but subsequently returned to near-normal values. HS treatment for 0.5 days did not change the levels of µ-calpain, cathepsin B, or proteasome C2 subunit mRNA, but increased the levels of mRNA encoding atrogin-1 (P<0.05) and its transcription factor, forkhead box O3 (P=0.09). Under these hyperthermic conditions, mitochondrial superoxide production was significantly increased than that of thermoneutral control. Here, we show that HS-induced muscle protein degradation may be due to the activation of ubiquitination by atrogin-1, and that this process may involve mitochondrial ROS production as well as corticosterone secretion.

Serum amine-based metabolites and their association with outcomes in primary prevention implantable cardioverter-defibrillator patients.

AIMS: Heart failure patients are at increased risk of ventricular arrhythmias and all-cause mortality. However, existing clinical and serum markers only modestly predict these adverse events. We sought to use metabolic profiling to identify novel biomarkers in two independent prospective cohorts of patients with implantable cardioverter-defibrillators (ICDs) for primary prevention of sudden cardiac death (SCD). METHODS AND RESULTS: Baseline serum was quantitatively profiled for 42 known biologically relevant amine-based metabolites among 402 patients from the Prospective Observational Study of Implantable Cardioverter-Defibrillators (PROSE-ICD) Study (derivation group) and 240 patients from the Genetic Risk Assessment of Defibrillator Events (GRADE) Study (validation group) for ventricular arrhythmia-induced ICD shocks and all-cause mortality. Three amines, N-methyl-l-histidine, symmetric dimethylarginine (SDMA), and l-kynurenine, were derived and validated to be associated with all-cause mortality. The hazard ratios of mortality in PROSE-ICD and GRADE were 1.48 (95% confidence interval 1.14-1.92) and 1.67 (1.22-2.27) for N-methyl-l-histidine, 1.49 (1.17-1.91) and 1.77 (1.27-2.45) for SDMA, 1.31 (1.06-1.63) and 1.73 (1.32-2.27) for l-kynurenine, respectively. l-Histidine, SDMA, and l-kynurenine were associated with ventricular arrhythmia-induced ICD shocks in PROSE-ICD, but they did not reach statistical significance in the GRADE cohort. CONCLUSION: Utilizing metabolic profiling in two independent prospective cohorts of patients undergoing ICD implantation for primary prevention of SCD, we identified several novel amine markers that were associated with appropriate shock and mortality. These findings shed insight into the potential biologic pathways leading to adverse events in ICD patients. Further studies are needed to confirm the prognostic value of these findings.

Glucose supplementation stimulates peripheral branched-chain amino acid catabolism in lactating dairy cows during essential amino acid infusions.

To determine how glucose modulates protein synthesis when essential AA are in abundant supply, 5 early-lactation, rumen-fistulated Holstein dairy cows were fed a diet containing 6.95 MJ/kg of net energy for lactation and 12.4% crude protein and abomasally infused for 5 d with saline, 844 or 1,126 g/d of a complete essential AA mix, with and without the inclusion of 1,000 g/d of glucose, in a 5×5 Latin square design. Infusion of essential AA increased milk yield by 4.1 kg/d, milk protein by 256 g/d, milk fat by 95 g/d, and milk urea nitrogen by 70% compared with saline, with no differences between the level of essential AA infusion. The addition of glucose to essential AA infusate did not stimulate milk protein yield or concentration, but reduced milk urea nitrogen by 17% and decreased milk fat yield. Arterial concentrations of total essential AA increased 3- to 4-fold, mammary clearance decreased 61%, and mammary uptake of essential AA increased 65% in response to essential AA infusion. Arterial branched-chain AA concentrations declined 29% in response to glucose and mammary clearance increased 48%, but mammary AA uptake was unchanged. Essential AA infusion increased plasma 3-methylhistidine by 50% and reduced muscle branched-chain α-keto acid dehydrogenase kinase abundance by 14%, indicating stimulation of muscle protein turnover and branched-chain AA catabolism, respectively. Glucose had no further effect on muscle branched-chain α-keto acid dehydrogenase kinase abundance but decreased mRNA expression of branched chain aminotransferase 1. Lack of further increases in plasma 3-methylhistidine or greater stimulation of muscle branched-chain AA catabolism indicates that muscle protein degradation was unchanged with glucose but that accretion may have been stimulated. The decrease in circulating branched-chain AA concentrations and nitrogen excretion in response to glucose suggests that surplus essential AA were redirected to peripheral, extra-mammary tissues.