Pre- and postpartum metabolizable protein supply: II. Effects on plasma amino acids and markers of tissue mobilization in transition Holstein dairy cows.

The influence of diet composition on the degree of adipose and lean muscle mobilization and concentrations of circulating AA has been demonstrated during the transition period. Altering the MP supply might offer a strategy to control tissue mobilization and increase circulating AA availability, but the optimum supply of MP fed pre- and postpartum remains unknown. We investigated the effect of increasing the MP supply in the prepartum, postpartum, or both diets on plasma AA concentrations and ultrasound and circulating indicators of tissue mobilization. Multiparous Holstein cows (n = 96) were assigned to 1 of 4 treatment groups at 28 d before expected calving following a randomized block design. Prepartum diets were formulated to contain either a control (C; 85 g of MP/kg DM; 1,175 g of MP/d) or high (H; 113 g of MP/kg DM; 1,603 g of MP/d) level of estimated MP. From calving to 21 DIM, fresh diets were formulated to contain either a control (C; 104 g of MP/kg DM; 2,044 g of MP/d) or high (H; 131 g of MP/kg DM; 2,685 g of MP/d) level of estimated MP. To control the potential confounding effect of Met and Lys supply, diets were formulated to supply an equal amount at 1.24 and 3.84 g/Mcal of ME in both prepartum diets and 1.15 and 3.16 g/Mcal of ME in both postpartum diets, respectively. The combination of a pre- and postpartum diet resulted in 4 treatment groups: 1) CC (n = 23), 2) CH (n = 24), 3) HC (n = 22), and 4) HH (n = 23). A common lactation diet (113 g of MP/kg DM; 2,956 g of MP/d) was fed from 22 DIM to the end of the observation period at 42 DIM. Transcutaneous ultrasonography was used to determine the longissimus dorsi muscle diameter and backfat thickness. Concentrations of plasma AA, 3-methylhistidine (3MH), and creatinine were determined on a subset of cows (n = 60) using ultra-high-performance liquid chromatography and mass spectrometry. Treatment did not affect the longissimus dorsi muscle diameter from -14 to 21 d relative to calving, but the diameter was greater in CH compared with HH at 40 DIM. Backfat thickness and the ratio of 3MH to creatinine did not differ by treatment. Concentrations of EAA were greater at -13 d relative to calving in HH compared with CC and CH and at -6 d relative to calving EAA concentrations were higher in HC compared with CC. Cows fed the H diet postpartum had elevated EAA concentrations at 6 and 20 DIM compared with cows fed the C postpartum diet but EAA concentration did not differ at 40 DIM. Total NEAA concentrations were higher in CH compared with HC and HH at -6 d relative to calving, but NEAA concentration did not differ by treatment at -13, 6, 20, or 40 d relative to calving. In conclusion, increasing the supply of MP fed prepartum, postpartum, or both had minimal effects on tissue mobilization but influenced concentrations of plasma AA.

Changes in Plasma Concentration of Free Proteinogenic and Non-Proteinogenic Amino Acids in High-Performance Sprinters over a 6-Month Training Cycle.

Background/Objectives: Free amino acids substantially contribute to energy metabolism. Also, their profile may identify (over)training status and effectiveness. The long-term effects of speed-power training on plasma free amino acid (PFAA) profiles are not known. We aimed to observe variations in PFAA levels in high-performance sprinters in a six-month training cycle. Methods: Ten male athletes (24.6 ± 3.3 years) were examined during four training phases: transition (1 month), general preparation (2 months), specific preparation (1 month), and pre-competition/competition (2 months). Venous blood was collected at rest, after exhaustive exercise, and recovery. Forty-two PFAAs were analyzed by the LC-ESI-MS/MS method. Results: Significant decreases in resting concentrations were observed between the transition and competition phases for glutamine (762 ± 117 vs. 623 ± 53 µmol∙L-1; p < 0.001, η2 = 0.47) and histidine (89 ± 15 vs. 75 ± 10 µmol∙L-1; p = 0.010, η2 = 0.27), whereas ß-alanine (30 ± 7 vs. 41 ± 9 µmol∙L-1; p = 0.024, η2 = 016) and sarcosine (3.6 ± 0.4 vs. 4.8 ± 0.6 µmol∙L-1; p = 0.006, η2 = 0.188) levels increased. Between the specific and competition phases, significant decreases in the resting levels of 1-methylhistidine (22.1 ± 19.4 vs. 9.6 ± 8.8 µmol∙L-1; p = 0.14, η2 = 0.19), 3-methylhistidine (7.1 ± 1.5 vs. 6.5 ± 1.6 µmol∙L-1; p = 0.009, η2 = 0.18), citrulline (40 ± 10 vs. 29 ± 4 µmol∙L-1; p = 0.05, η2 = 0.29), and ornithine (74 ± 15 vs. 56 ± 10 µmol∙L-1; p = 0.015, η2 = 185) were noticed. Also, for ß-alanine and sarcosine, the pattern of response to exercise strongly changed between the training phases. Blood ammonia levels at exhaustion decreased between the transition and competition phases (32 ± 4 vs. 23 ± 5 µmol∙L-1; p < 0.001, η2 = 0.67), while lactate, the phenylalanine-tyrosine ratio, the glutamine-glutamate ratio, hematological parameters, and cardiorespiratory indices remained at similar levels. Conclusions: Speed-power training seems to affect PFAAs involved in skeletal muscle metabolic pathways responsible for neutralizing toxic ammonia (glutamine, arginine, citrulline, ornithine), attenuating the deleterious effects of H+ ions (histidine, ß-alanine), and reducing exercise-induced protein breakdown (1- and 3-methylhistidine). Our findings suggest that sprint-oriented training supports metabolic pathways that are responsible for the removal of harmful metabolites produced during exercise.

Timing of feeding a protein supplement on nitrogen balance and plasma amino acids during exercise recovery in horses.

Eight geldings weighing 544 ± 16 kg were used to evaluate feeding a postexercise protein meal on plasma amino acids during recovery. Horses were fed sweet feed, corn, grass hay and equal amounts of a protein pellet (32% CP) with meals (MP group) or postexercise (EP group). Horses exercised 1-2 h/day, 5 days/week, for 12 weeks. A pre and poststudy 4 days total urine and feces collection was conducted. Urine and fecal samples were analyzed for nitrogen (N) to calculate N balance. Blood samples were drawn immediately after and at 1 and 3 h postexercise at the start and end of the study for plasma amino acid concentrations. Absorbed N and N retention were greater for the MP group compared to the EP group (p = 0.038, 0.033 respectively). An interaction revealed an increase in fecal N (p = 0.01) and decreased N digestibility for the MP group compared to the EP group at the end of the study. Plasma concentrations for 8 out of 14 amino acids were less for the EP group immediately after exercise compared to the MP group (p < 0.02). Plasma concentrations of lysine and arginine were greater for the EP group compared to the MP group at 1 and 3 h after exercise (p < 0.05 and 0.04 respectively). Changes were different for 8 out of the 14 amino acids immediately post exercise, 7 out of 14 amino acids at 1 h postexercise and 10 out of 14 amino acids at 3 h postexercise with positive changes for the EP group and negative changes for the MP group. The EP group had improved supply of plasma amino acids in the recovery period that sustained for 3 h postexercise and are indicative of better amino acid supply supporting muscle development.

Plasma amino acids in major depressive disorder: between pathology to pharmacology.

Addressing the formidable challenge posed by the development of effective and personalized interventions for major depressive disorder (MDD) necessitates a comprehensive comprehension of the intricate role that plasma amino acids play and their implications in MDD pathology and pharmacology. Amino acids, owing to their indispensable functions in neurotransmission, metabolism, and immune regulation, emerge as pivotal entities in this intricate disorder. Our primary objective entails unraveling the underlying mechanisms and unveiling tailored treatments through a meticulous investigation into the interplay between plasma amino acids, MDD, and pharmacological strategies. By conducting a thorough and exhaustive review of the existing literature, we have identified pertinent studies on plasma amino acids in MDD, thereby uncovering noteworthy disturbances in the profiles of amino acids among individuals afflicted by MDD when compared to their healthy counterparts. Specifically, disruptions in the metabolism of tryptophan, phenylalanine, and tyrosine, which serve as precursors to essential neurotransmitters, have emerged as prospective biomarkers and critical contributors to the pathophysiology of depression. Amnio acids play an essential role in MDD and could represent an attractive pharmacological target, more studies are further required to fully reveal their underlying mechanisms.

Elevated Plasma Tryptophan in Patients with Anorexia Nervosa Compared with Hypotrophic Controls.

Introduction: We hypothesized that anorexia nervosa (AN) is associated with pathological amino acid metabolism. This study aimed to identify amino acids exhibiting abnormal metabolism in patients with AN compared with those in low-nutrient controls. Methods: This was a single-center, retrospective, observational study that compared patients with AN with a low-nutrient control group. All participants were admitted to the Kitasato University Hospital Emergency Center between January 1, 2018, and January 31, 2021. Both the AN and low-nutrient control groups had five patients each. Plasma amino acid category testing was conducted at the same institution for both groups. Patient sex, age, height, weight, and comorbidities were retrospectively extracted. Plasma amino acid fractions, total amino acids, total essential amino acids, total nonessential amino acids, branched-chain amino acids (sum of valine, isoleucine, and leucine), and amino acid concentrations and ratios were compared between the two groups. Data were analyzed using the Mann-Whitney U test. Results: Body mass index was lower in the AN group (p = 0.00794). Tryptophan levels were significantly higher in the AN group (p = 0.00794). Other amino acid values, the sum of amino acid values, and amino acid ratios were not significantly different between both groups. Conclusions: Serum tryptophan levels were higher in the AN group than in the low-nutrient group, and AN may be associated with abnormal amino acid metabolism.

Enzymatically modified isoquercitrin in soy protein temporarily enhanced the plasma amino-acid concentrations, antioxidant index, and plasma hormone levels: a randomized, double-blind cross-over trial.

This study investigated the effects of a dietary protein supplement containing enzymatically modified isoquercitrin (EMIQ) on plasma amino-acid levels in healthy people. A randomized double-blind cross-over trial (UMIN000044791) was conducted with a sample of nine healthy individuals. These participants ingested soy protein with or without 42 mg EMIQ for 7 days after performing mild exercise. Plasma amino-acid levels were measured before ingestion and at 15, 30, 45, 60, 90, 120, 180, and 240 min after ingestion on the last day. The concentrations of total amino acids at 0 and 120 min and easily oxidized amino acids at 120 min were significantly higher in the plasma of individuals who consumed 42 mg EMIQ. Oxidative stress levels were lower and plasma testosterone levels were higher in participants who ingested soy protein with 42 mg EMIQ than in those who did not. These results suggest that daily ingestion of soy protein with 42 mg EMIQ can be useful for effective protein absorption.

Changes in Faecal and Plasma Amino Acid Profile in Dogs with Food-Responsive Enteropathy as Indicators of Gut Homeostasis Disruption: A Pilot Study.

Dogs suffering from food-responsive enteropathy (FRE) respond to an elimination diet based on hydrolysed protein or novel protein; however, studies regarding the amino acid profile in FRE dogs are lacking. The aim of this pilot study was to evaluate whether the plasma and faecal amino acid profiles differed between control and FRE dogs and whether these could serve as indicators of severity of illness. Blood, faecal samples, body condition score, and severity of clinical signs based on the canine inflammatory bowel disease activity index were collected before starting the elimination diet. FRE dogs had lower proportions of plasma Asparagine, Histidine, Glycine, Cystine, Leucine, and branched-chain/aromatic amino acids; however, Phenylalanine increased. In faecal samples, Cystine was greater whereas Phenylalanine was lesser in sick dogs compared to control. Leucine correlated negatively with faecal humidity (r = -0.66), and Leucine and Phenylalanine with faecal fat (r = -0.57 and r = -0.62, respectively). Faecal Phenylalanine (r = 0.80), Isoleucine (r = 0.75), and Leucine (r = 0.92) also correlated positively with total short-chain fatty acids, whereas a negative correlation was found with Glycine (r = -0.85) and Cystine (r = -0.61). This study demonstrates the importance of Leucine and Phenylalanine amino acids as indicators of the disease severity in FRE dogs.

Plasma Amino Acids in Horses Suffering from Pituitary Pars Intermedia Dysfunction.

Pituitary pars intermedia dysfunction is one of the most common diseases of aged horses and ponies. In Parkinson's disease, which is, similar to PPID, a disease that involves oxidative damage to dopaminergic pathways but with different clinical signs, alterations to the serum amino acid profile have been reported. To examine changes in the plasma amino acid profile in horses with PPID, EDTA plasma of horses that were presented for various reasons that required laboratory examinations of blood anticoagulated with EDTA was collected. With this plasma, the basal ACTH concentration as well as the amino acid profile was determined. Horses were considered PPID patients if the ACTH concentration was ≥ 100 pg/mL, i.e., they would be considered affected at any time. Horses were defined as non-PPID (nPPID) patients if the ACTH concentration was below 30 pg/mL. Horses receiving pergolide with ACTH ≤ 30 pg/mL were allocated to the group PPIDrr (PPID, ACTH in reference range) and horses receiving pergolide with ACTH ≥ 100 pg/mL to the group PPIDarr (PPID, ACTH above reference range). In total, 93 horses were examined, including 88 horses at the clinic and 5 horses at a private practice. Of these, 53 horses fulfilled the inclusion criteria (ACTH ≤ 30 pg/mL or ACTH ≥ 100 pg/mL). A total of 25 horses were diagnosed as nPPID, 20 as PPID, 5 as PPIDrr, and 3 as PPIDarr. Arginine was significantly higher in PPIDrr than in PPID and nPPID, asparagine was significantly higher in PPID, PPIDrr, and PPIDarr than in nPPID, citrulline was significantly higher in PPIDrr than in nPPID and PPID, cysteine was significantly lower in PPIDrr than in PPID, nPPID, and PPIDarr, and glutamine was significantly higher in PPID and PPIDarr than in nPPID. Especially, asparagine, citrulline, and glutamine may be potential diagnostic markers and may offer interesting approaches for research regarding amino supplementation in PPID.

[Plasma amino acids levels in 1859 physical examination subjects in Hangzhou City in 2020].

OBJECTIVE: To study the relationship between fasting plasma amino acids and obesity in the physical examination population. METHODS: A total of 1859 persons who underwent physical examination in the Second Affiliated Hospital Zhejiang University School of Medicine from January to December 2020 were enrolled. Fasting plasma amino acids concentrations and body fat ratio(BFR) were respectively determined by ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) and Inbody instrument. According to body mass index and gender grouping, Python language was used for data analysis and graphic visualization to explore the relationship between body mass index, gender and plasma amino acids levels. RESULTS: The plasma levels of glycine, serine and asparagine in the overweight and obesity groups were significantly lower than those in the normal group, especially the levels in the obesity group were lower than those in the overweight group in males(P<0.05). There was no significant difference in plasma levels of lysine, tryptophan, arginine and hydroxyproline among normal, overweight and obesity groups(P>0.05). The plasma levels of leucine, valine, tyrosine, phenylalanine(in males), isoleucine(in females) and alanine in the overweight and obesity groups were significantly higher than those in the normal group, and the levels in the obesity group were higher than those in the overweight group except alanine(P<0.05). BFR was positively correlated with body mass index. BFR in females increased significantly with age after 50 years old(P<0.05). CONCLUTION: The rate of overweight and obesity among the health screening population is high.

Recent tPA administration can cause pseudo-hyperargininemia and may mimic arginase deficiency or arginine supplementation.

Individuals suspected of or diagnosed with a rare disorder, including inherited metabolic disorders (IMD), often need frequent and/or urgent vascular access for blood draws and treatment, making central indwelling catheters commonly used devices in this patient population. These indwelling catheters are prone to thrombosis, limiting vascular access. This complication is frequently resolved with the use of altepase, a recombinant tissue plasminogen activator (tPA). This report describes two individuals, one with a known IMD and one undergoing evaluation for an IMD, who were found to have hyperargininemia (>500 µM; reference 10-140 µM) by plasma amino acid (PAA) analysis of a specimen collected ~1.5-3 h after clearance of an indwelling catheter with tPA. In both cases, hyperargininemia resolved with repeat testing, suggesting pseudo-hyperargininemia secondary to tPA administration. Quantitative amino acid analysis of the administered tPA demonstrated an arginine level of ~200 mM, supporting tPA as the cause of pseudo-hyperargininemia. Certain formulations of tPA contain high concentrations of arginine, which if not cleared properly can result in marked elevations of arginine, mimicking arginase deficiency or suggesting arginine supplementation. Thus, the possibility of pseudohyperargininemia due to tPA administration should be considered when obtaining PAAs from an indwelling catheter in any individual being evaluated or managed for an IMD.

Plasma Amino Acids May Improve Prediction Accuracy of Cerebral Vasospasm after Aneurysmal Subarachnoid Haemorrhage.

Aneurysmal subarachnoid haemorrhages (aSAH) account for 5% of strokes and continues to place a great burden on patients and their families. Cerebral vasospasm (CVS) is one of the main causes of death after aSAH, and is usually diagnosed between day 3 and 14 after bleeding. Its pathogenesis remains poorly understood. To verify whether plasma concentration of amino acids have prognostic value in predicting CVS, we analysed data from 35 patients after aSAH (median age 55 years, IQR 39-62; 20 females, 57.1%), and 37 healthy volunteers (median age 50 years, IQR 38-56; 19 females, 51.4%). Fasting peripheral blood samples were collected on postoperative day one and seven. High performance liquid chromatography-mass spectrometry (HPLC-MS) analysis was performed. The results showed that plasma from patients after aSAH featured a distinctive amino acids concentration which was presented in both principal component analysis and direct comparison. No significant differences were noted between postoperative day one and seven. A total of 18 patients from the study group (51.4%) developed CVS. Hydroxyproline (AUC = 0.7042, 95%CI 0.5259-0.8826, p = 0.0248) and phenylalanine (AUC = 0.6944, 95%CI 0.5119-0.877, p = 0.0368) presented significant CVS prediction potential. Combining the Hunt-Hess Scale and plasma levels of hydroxyproline and phenylalanine provided the model with the best predictive performance and the lowest leave-one-out cross-validation of performance error. Our results suggest that plasma amino acids may improve sensitivity and specificity of Hunt-Hess scale in predicting CVS.

Association of Plasma Branched-Chain and Aromatic Amino Acids with Reduction in Kidney Function Evaluated in Apparently Healthy Adults.

The published literature on the association of circulatory branched-chain amino acids (BCAAs) and aromatic amino acids (AAAs) with reduced kidney function is inconsistent or conflicting. Clarification of it might help to better understand the underlying pathophysiology and to determine potential biomarkers for early detection and evaluation of kidney function decline. Our main purpose was to explore and clarify the potential relationships of individual BCAAs and AAAs with estimated glomerular filtration rate (eGFR) decline. We included the data from 2804 healthy subjects and categorized them into three groups based on eGFR tertiles. The associations between individual amino acids and eGFR were explored by covariate-adjusted logistic regression models. There was a progressive increase in the concentrations of BCAAs and AAAs from the upper to the lower tertiles. We revealed significant positive associations of isoleucine, leucine, and phenylalanine with lower tertiles of eGFR in the adjusted models (p < 0.01-0.001). The findings hold a promising potential of using plasma isoleucine, leucine, and phenylalanine levels for evaluation of kidney function decline. Future longitudinal studies should investigate the causal association between altered levels of these amino acids and impaired kidney function and also the utility of the former as potential biomarkers for evaluating the risk and early detection of the latter.

Correlations of blood and brain biochemistry in phenylketonuria: Results from the Pah-enu2 PKU mouse.

BACKGROUND: In phenylketonuria (PKU), treatment monitoring is based on frequent blood phenylalanine (Phe) measurements, as this is the predictor of neurocognitive and behavioural outcome by reflecting brain Phe concentrations and brain biochemical changes. Despite clinical studies describing the relevance of blood Phe to outcome in PKU patients, blood Phe does not explain the variance in neurocognitive and behavioural outcome completely. METHODS: In a PKU mouse model we investigated 1) the relationship between plasma Phe and brain biochemistry (Brain Phe and monoaminergic neurotransmitter concentrations), and 2) whether blood non-Phe Large Neutral Amino Acids (LNAA) would be of additional value to blood Phe concentrations to explain brain biochemistry. To this purpose, we assessed blood amino acid concentrations and brain Phe as well as monoaminergic neurotransmitter levels in in 114 Pah-Enu2 mice on both B6 and BTBR backgrounds using (multiple) linear regression analyses. RESULTS: Plasma Phe concentrations were strongly correlated to brain Phe concentrations, significantly negatively correlated to brain serotonin and norepinephrine concentrations and only weakly correlated to brain dopamine concentrations. From all blood markers, Phe showed the strongest correlation to brain biochemistry in PKU mice. Including non-Phe LNAA concentrations to the multiple regression model, in addition to plasma Phe, did not help explain brain biochemistry. CONCLUSION: This study showed that blood Phe is still the best amino acid predictor of brain biochemistry in PKU. Nevertheless, neurocognitive and behavioural outcome cannot fully be explained by blood or brain Phe concentrations, necessitating a search for other additional parameters. TAKE-HOME MESSAGE: Blood Phe is still the best amino acid predictor of brain biochemistry in PKU. Nevertheless, neurocognitive and behavioural outcome cannot fully be explained by blood or brain Phe concentrations, necessitating a search for other additional parameters.

Plasma amino acid levels in individuals with bacterial pneumonia and healthy controls.

BACKGROUND & AIMS: Amino acids play an important role in immune responses and as neurotransmitters. During the course of a bacterial pneumonia episode, from the onset to the recovery phase, immune responses dramatically change, as does the metabolism of amino acids, a concept referred to as immuno-nutrition. We investigated the differences in plasma amino acid levels (PAA) between the acute and recovery phases in individuals with community-acquired pneumonia (CAP) and healthy controls. METHODS: Two groups of participants were recruited: Healthy adults aged over 60 years and patients hospitalized with CAP. Samples were collected on Day 0 (the day of admission) and Day 7 (after 6-8 days treatment). RESULTS: A total of 93 healthy adults and 60 patients with CAP participated in the study. Of those with CAP, 43 had their amino acids measured on Day 7. Patients with CAP had markedly decreased PAA of 12 amino acids on Day 0. Citrulline, histidine, and tryptophan remained low in male, while aspartic acid, asparagine, ornithine, proline, and threonine were higher on Day 7 in both males and females. Phenylalanine increased at Day 0 and Day7. CONCLUSIONS: The findings suggest that the host response against bacterial infection changed the plasma amino acid levels. PAA on Day 7 (representing convalescence) continued to display an amino acid profile distinct from that observed in healthy individuals. Based on these findings, reconsideration for providing amino acids to patients with bacterial pneumonia should be needed depending on stage of the pneumonia from the perspective of immuno-nutrition.

Altered Amino Acid Metabolism in Patients with Cardiorenal Syndrome Type 2: Is It a Problem for Protein and Exercise Prescriptions?

The goal of this retrospective study was to document any alterations in plasma amino acids (AAs) in subjects with cardiorenal syndrome type 2 (CRS 2). We analyzed data from sixteen patients with CRS 2 and eight healthy subjects (control group, C), whose plasma arterial (A) and venous (V) AA concentrations had been measured. Compared to C, the group of CRS 2 patients showed significant reductions by more than 90% in A (p < 0.01) and V (p < 0.01) individual AAs, whereas negative A-V differences that indicated a net muscle AA release (muscle hypercatabolism) were found in 59% of CRS 2 patients (p < 0.03). No significant differences in plasma A and V AA concentrations nor in A-V differences were found between patients with mild kidney damage (N = 5; estimated glomerular filtration rate, eGFR ≥ 60 mL/min/1.73 m2) and patients with moderate-severe kidney damage (N = 11; eGFR < 60 mL/min/1.73 m2). Several plasma arterial AAs correlated with hemodynamic variables, but not with GFR. The study showed that patients with CRS 2 had very low concentrations of circulating AAs, independent of the degree of GFR damage.

Comparison of 24-Hour Recall and 3-Day Food Records during the Complementary Feeding Period in Thai Infants and Evaluation of Plasma Amino Acids as Markers of Protein Intake.

BACKGROUND: An accurate and reliable measurement of nutrient intake is the first and foremost step in order to optimise infant nutrition and evaluate its impact on health outcomes. However, research on the validity of dietary assessment tools used during the weaning period is limited, especially in lower-middle income countries. The primary aim of this study was to evaluate relative validity of a 24-h recall method (24-HR) using a 3-day food record (3-DFR). A secondary aim was to investigate association between protein intake from 3-DFR and plasma amino acids as a potential protein biomarker. Methods A multicentre, prospective cohort study was conducted in Chiang Mai, Thailand from June 2018 to May 2019. Food consumption data were collected in healthy infants using 24-HR and 3-DFR at 9 and 12 months of age. Blood samples were obtained at 12 months (M). Plasma amino acids were analysed using high performance liquid chromatography. Results Of 145 infants, 49% were female. At group level, paired t-tests/Wilcoxon signed rank tests did not show significant differences between average nutrient intakes from the 2 dietary assessment methods, except for vitamin A and vitamin C. Weighted kappa (Kw) was acceptable for all nutrients, except for vitamin A intake at 9 M (Kw = 0.15). The Bland-Altman analyses were unbiased for most nutrients with variable limits of agreement. At individual level, correlation coefficients (r) ranged from acceptable to excellent (r = 0.37-0.87) while cross-classifications showed acceptable outcomes, except for vitamin A. Multivariate analyses showed significant associations between protein intake at 12 M from the 3-DFR and plasma concentrations of branched-chain amino acids (BCAA) and essential amino acids (EAA), even after adjusting for gender, milk feeding type and energy intake. Conclusions For infants aged 9-12 M, a 24-HR can be used as a more practical alternative to a 3-DFR for most nutrients although caution is required for some micronutrients, especially vitamin A. A repeated interview might further improve the accuracy. Furthermore, protein intake, particularly animal-based protein, significantly predicted plasma BCAA and EAA concentrations regardless of gender, type of milk feeding and energy consumption.

Concurrent and carryover effects of feeding blends of protein and amino acids in high-protein diets with different concentrations of forage fiber to fresh cows. 1. Production and blood metabolites.

Because of low feed intake during the first weeks of lactation, dietary concentration of metabolizable protein (MP) must be elevated. We evaluated effects of providing additional rumen-undegradable protein (RUP) from a single source or a blend of protein and AA sources during the first 3 wk of lactation. We also evaluated whether replacing forage fiber (fNDF) or nonforage fiber with the blend affected responses. In a randomized block design, at approximately 2 wk prepartum, 40 primigravid (664 ± 44 kg of body weight) and 40 multigravid (797 ± 81 kg of body weight) Holsteins were blocked by calving date and fed a common diet (11.5% crude protein, CP). After calving to 25 d in milk (DIM), cows were fed 1 of 4 diets formulated to be (1) 20% deficient in metabolizable protein (MP) based on predicted milk production (17% CP, 24% fNDF), (2) adequate in MP using primarily RUP from soy to increase MP concentration (AMP; 20% CP, 24% fNDF), (3) adequate in MP using a blend of RUP and rumen-protected AA sources to increase MP concentration (Blend; 20% CP, 24% fNDF), or (4) similar to Blend but substituting fNDF with added RUP rather than nonforage neutral detergent fiber (Blend-fNDF; 20% CP, 19% fNDF). The blend was formulated to have a RUP supply with an AA profile similar to that of casein. A common diet (17% CP) was fed from 26 to 92 DIM, and milk production and composition were measured from 26 to 92 DIM, but individual dry matter intake (DMI) was measured only until 50 DIM. During the treatment period for both parities, AMP and Blend increased energy-corrected milk (ECM) yields compared with the diet deficient in MP based on predicted milk production (40.7 vs. 37.8 kg/d) and reduced concentrations of plasma 3-methyl-His (4.1 vs. 5.3 µmol/L) and growth hormone (9.0 vs. 11.9 ng/mL). Blend had greater DMI than AMP (17.4 vs. 16.1 kg/d), but ECM yields were similar. Blend had greater plasma Met (42.0 vs. 26.4 µmol/L) and altered metabolites associated with antioxidant production and methyl donation compared with AMP. Conversely, the concentration of total essential AA in plasma was less in Blend versus AMP (837 vs. 935 µmol/L). In multiparous cows, Blend-fNDF decreased DMI and ECM yield compared with Blend (19.2 vs. 20.1 kg/d of DMI, 45.3 vs. 51.1 kg/d of ECM), whereas primiparous cows showed the opposite response (15.3 vs. 14.6 kg/d of DMI, 32.9 vs. 31.4 kg/d of ECM). Greater DMI for multiparous cows fed Blend carried over from 26 to 50 DIM and was greater compared with AMP (23.1 vs. 21.2 kg /d) and Blend-fNDF (21.3 kg/d). Blend also increased ECM yield compared with AMP (49.2 vs. 43.5 kg/d) and Blend-fNDF (45.4 kg/d) from 26 to 92 DIM. Few carryover effects of fresh cow treatments on production were found in primiparous cows. Overall, feeding blends of RUP and AA may improve the balance of AA for fresh cows fed high MP diets and improve concurrent and longer-term milk production in multiparous cows. However, with high MP diets, multiparous fresh cows require greater concentrations of fNDF than primiparous cows.

Plasma Branched-Chain and Aromatic Amino Acids in Relation to Hypertension.

Findings of the available studies regarding the roles of branched-chain amino acids (BCAAs) and aromatic amino acids (AAAs) in hypertension are inconsistent, conflicting and inconclusive. The purpose of this study was to explore and clarify the existence of any relationships of individual BCAAs and AAAs with hypertension with adjustments for potential relevant confounders. A total of 2805 healthy controls and 2736 hypertensive patients were included in the current analysis. The associations between individual amino acids and hypertension were explored by logistic regression analyses adjusted for potential confounding variables. Among the investigated amino acids, only the BCAAs showed consistently significant positive associations with hypertension in the adjusted models (p-trend < 0.05 to 0.001). However, compared with the corresponding lowest quartile of individual BCAAs, the positive association with hypertension remained significant only in the highest quartile (p < 0.01 to 0.001). We confirmed in a relatively large cohort of subjects that BCAAs, not AAAs, demonstrated consistent positive associations with hypertension. The results display the promising potential for the use of BCAAs as relevant and accessible biomarkers, and provide perspectives on interventions directed towards the reduction in plasma BCAA levels in the prevention and management of hypertension.

Role of Amino Acids in Blood Glucose Changes in Young Adults Consuming Cereal with Milks Varying in Casein and Whey Concentrations and Their Ratio.

BACKGROUND: Increasing the total protein content and reducing the casein to whey ratio in milks consumed with breakfast cereal reduce postprandial blood glucose (BG). OBJECTIVES: We aimed to explore associations between plasma amino acids (AAs), BG, and glucoregulatory hormones. METHODS: In this repeated-measures design, 12 healthy adults consumed cereal (58 g) and milks (250 mL) with 3.1 wt% or high 9.3 wt% protein concentrations and with casein to whey ratios of either 80:20 or 40:60. Blood was collected at 0, 30, 60, 120, 140, 170, and 200 min for measurement of the primary outcome, BG, and for the exploratory outcomes such as plasma AA, gastric emptying, insulin (INS), and glucoregulatory hormones. Measures were made prior to and after an ad libitum lunch at 120 min. Exploratory correlations were conducted to determine associations between outcomes. RESULTS: Pre-lunch plasma AA groups [total (TAA), essential (EAA), BCAA, and nonessential (NEAA)] were higher after 9.3 wt% than 3.1 wt% milks by 12.7%, 21.4%, 20.9%, and 7.6%, respectively (P ≤ 0.05), while post-lunch AA groups were higher by 10.9%, 19.8%, 18.8%, and 6.0%, respectively (P ≤ 0.05). Except for NEAA, pre-lunch AAs were higher after 40:60 than 80:20 ratio milks by 4.5%, 8.3%, and 9.3% (P ≤ 0.05). When pooled by all treatments, pre-lunch AA groups associated negatively with BG (r/ρ ≥ -0.45, P ≤ 0.05), but post-lunch only TAA and NEAA correlated (r ≥ -0.37, P < 0.05). Pre-lunch BG was inversely associated with Leu, Ile, Lys, Met, Thr, Cys-Cys, Asn, and Gln (r/ρ ≥ -0.46, P ≤ 0.05), but post-lunch, only with Thr, Ala, and Gly (r ≥ -0.50, P ≤ 0.05). Pre-lunch associations between AA groups and INS were not found. CONCLUSIONS: Protein concentration and the ratio of casein to whey in milks consumed at breakfast with cereal affect plasma AA concentrations and their associations with decreased BG. The decrease in BG could be explained by INS-independent mechanisms. This trial was registered at www.clinicaltrials.gov as NCT02471092.

A Machine Learning Approach for the Automated Interpretation of Plasma Amino Acid Profiles.

BACKGROUND: Plasma amino acid (PAA) profiles are used in routine clinical practice for the diagnosis and monitoring of inherited disorders of amino acid metabolism, organic acidemias, and urea cycle defects. Interpretation of PAA profiles is complex and requires substantial training and expertise to perform. Given previous demonstrations of the ability of machine learning (ML) algorithms to interpret complex clinical biochemistry data, we sought to determine if ML-derived classifiers could interpret PAA profiles with high predictive performance. METHODS: We collected PAA profiling data routinely performed within a clinical biochemistry laboratory (2084 profiles) and developed decision support classifiers with several ML algorithms. We tested the generalization performance of each classifier using a nested cross-validation (CV) procedure and examined the effect of various subsampling, feature selection, and ensemble learning strategies. RESULTS: The classifiers demonstrated excellent predictive performance, with the 3 ML algorithms tested producing comparable results. The best-performing ensemble binary classifier achieved a mean precision-recall (PR) AUC of 0.957 (95% CI 0.952, 0.962) and the best-performing ensemble multiclass classifier achieved a mean F4 score of 0.788 (0.773, 0.803). CONCLUSIONS: This work builds upon previous demonstrations of the utility of ML-derived decision support tools in clinical biochemistry laboratories. Our findings suggest that, pending additional validation studies, such tools could potentially be used in routine clinical practice to streamline and aid the interpretation of PAA profiles. This would be particularly useful in laboratories with limited resources and large workloads. We provide the necessary code for other laboratories to develop their own decision support tools.