Acute Dissociation and Ketamine's Antidepressant and Anti-Suicidal Ideation Effects in a Midazolam-Controlled Trial.

OBJECTIVE: We sought to explore relationships of acute dissociative effects of intravenous ketamine with change in depression and suicidal ideation and with plasma metabolite levels in a randomized, midazolam-controlled trial. METHODS: Data from a completed trial in suicidal, depressed participants (n = 40) randomly assigned to ketamine was used to examine relationships between ketamine treatment-emergent dissociative and psychotomimetic symptoms with pre/post-infusion changes in suicidal ideation and depression severity. Nonparametric correlational statistics were used. These methods were also used to explore associations between dissociative or psychotomimetic symptoms and blood levels of ketamine and metabolites in a subset of participants (n = 28) who provided blood samples immediately post-infusion. RESULTS: Neither acute dissociative nor psychotomimetic effects of ketamine were associated with changes in suicidal ideation or depressive symptoms from pre- to post-infusion. Norketamine had a trend-level, moderate inverse correlation with dissociative symptoms on Day 1 post-injection (P = .064; P =.013 removing 1 outlier). Dehydronorketamine correlated with Clinician-Administered Dissociative States Scale scores at 40 minutes (P = .034), 230 minutes (P = .014), and Day 1 (P = .012). CONCLUSION: We did not find evidence that ketamine's acute, transient dissociative, or psychotomimetic effects are associated with its antidepressant or anti-suicidal ideation actions. The correlation of higher plasma norketamine with lower dissociative symptoms on Day 1 post-treatment suggests dissociation may be more an effect of the parent drug.

Effects of direct-fed microbial supplement on ruminal and plasma metabolome of early-lactation dairy cows: Untargeted metabolomics approach.

We examined the effects of 2 multispecies direct-fed microbial (DFM) supplements on ruminal and plasma metabolome of early-lactation dairy cows using a high-coverage untargeted metabolomics approach. A total of 45 multiparous Holstein cows (41 ± 7 DIM) were enrolled for the 14-d pre-experimental and 91-d experimental period and were a subset from a lactation performance study, which used 114 cows. Cows were blocked using pre-experimental energy-corrected milk yield and randomly assigned within each block to 1 of 3 treatments: (1) corn silage-based diet with no DFM supplement (control; CON), (2) basal diet top-dressed with a mixture of Lactobacillus animalis and Propionibacterium freudenreichii at 3 × 109 cfu/d (PRO-A), or (3) basal diet top-dressed with a mixture of L. animalis, P. freudenreichii, Bacillus subtilis, and Bacillus licheniformis at 11.8 × 109 cfu/d (PRO-B). The basal diet was fed ad libitum daily as a TMR at 0600 and 1200 h for a duration of 91 d. Rumen fluid and blood samples were taken on d -3, 28, 49, 70, and 91 and immediately stored at -80°C. Before analysis, ruminal and plasma samples from d 28, 49, 70, and 91 were composited. An in-depth, untargeted metabolome profile of the composite rumen and plasma samples and the d -3 samples was developed by using a chemical isotope labeling/liquid chromatography-mass spectrometry (LC-MS)-based technique. Differentially abundant metabolites (taking into account fold change [FC] values and false discovery rates [FDR]) were identified with a volcano plot. In the rumen, compared with the CON diet, supplemental PRO-A increased (FC ≥1.2; FDR ≤0.05) the relative concentrations of 9 metabolites, including 2-hydroxy-2,4-pentadienoic acid, glutaric acid, quinolinic acid, and shikimic acid, and PRO-B increased relative concentrations of 16 metabolites, including 2-hydroxy-2,4-pentadienoic acid, glutaric acid, 16-hydroxypalmitic acid, and 2 propionate precursors (succinic and methylsuccinic acids). Relative to PRO-A, supplemental PRO-B increased (FC ≥1.2; FDR ≤0.05) relative rumen concentrations of 3 metabolites, 16-hydroxypalmitic acid, indole-3-carboxylic acid, and 5-aminopentanoic acid, but reduced relative rumen concentrations of 13 metabolites, including carnitine, threonic acid, and shikimic acid. Compared with the CON diet, relative concentrations of 13 plasma metabolites, including myxochelin A and glyceraldehyde, were increased (FC ≥1.2; FDR ≤0.05) by PRO-A supplementation, whereas those of 9 plasma metabolites, including 4-(2-aminophenyl)-2,4-dioxobutanoic acid, N-acetylornithine, and S-norlaudanosolin, were reduced (FC ≤0.83; FDR ≤0.05). Supplemental PRO-B increased (FC ≥1.2; FDR ≤0.05) relative concentrations of 9 plasma metabolites, including trans-o-hydroxybenzylidenepyruvic acid and 3-methylsalicylaldehyde, and reduced relative concentrations of 4 plasma metabolites, including ß-ethynylserine and kynurenine. Pathway analysis of the differentially abundant metabolites in both rumen and plasma revealed that these metabolites are involved in AA and fatty acid metabolism and have antimicrobial and immune-stimulating properties. The results of this study demonstrated that dietary supplementation with either PRO-A or PRO-B altered the plasma and ruminal metabolome. Notably, ruminal and plasma metabolites involved in the metabolism of AA and fatty acids and those with immunomodulatory properties were altered by either or both of the 2 microbial additives.

Metabolic signatures of insulin resistance in non-diabetic individuals.

BACKGROUND: Insulin resistance (IR) evolved from excessive energy intake and poor energy expenditure, affecting the patient's quality of life. Amino acid and acylcarnitine metabolomic profiles have identified consistent patterns associated with metabolic disease and insulin sensitivity. Here, we have measured a wide array of metabolites (30 acylcarnitines and 20 amino acids) with the MS/MS and investigated the association of metabolic profile with insulin resistance. METHODS: The study population (n = 403) was randomly chosen from non-diabetic participants of the Surveillance of Risk Factors of NCDs in Iran Study (STEPS 2016). STEPS 2016 is a population-based cross-sectional study conducted periodically on adults aged 18-75 years in 30 provinces of Iran. Participants were divided into two groups according to the optimal cut-off point determined by the Youden index of HOMA-IR for the diagnosis of metabolic syndrome. Associations were investigated using regression models adjusted for age, sex, and body mass index (BMI). RESULTS: People with high IR were significantly younger, and had higher education level, BMI, waist circumference, FPG, HbA1c, ALT, triglyceride, cholesterol, non-HDL cholesterol, uric acid, and a lower HDL-C level. We observed a strong positive association of serum BCAA (valine and leucine), AAA (tyrosine, tryptophan, and phenylalanine), alanine, and C0 (free carnitine) with IR (HOMA-IR); while C18:1 (oleoyl L-carnitine) was inversely correlated with IR. CONCLUSIONS: In the present study, we identified specific metabolites linked to HOMA-IR that improved IR prediction. In summary, our study adds more evidence that a particular metabolomic profile perturbation is associated with metabolic disease and reemphasizes the significance of understanding the biochemistry and physiology which lead to these associations.

Stopover refuelling, movement and departure decisions in the white-throated sparrow: The influence of intrinsic and extrinsic factors during spring migration.

Differential migration timing between sex or age classes is an example of how migratory movement strategies can differ among subgroups within a population. However, in songbirds, evidence for intrinsic differences in en route migratory behaviour is often mixed, suggesting that the local environmental context may play a role in accentuating or diminishing patterns. We evaluated how multiple intrinsic and extrinsic variables influenced refuelling rates, local movement behaviour and departure decisions in the white-throated sparrow Zonotrichia albicollis during spring migration. This species exhibits a unique genetically based plumage dimorphism, providing a unique class of individual in which to evaluate patterns and processes of differential migration, in addition to sex, age and migration distance. At a migratory stopover site, plasma metabolite analysis was used to quantify individual variation in stopover refuelling rate. In after second year adults, automated and manual radio telemetry was used to quantify daily activity timing, daily movement distances, stopover duration and departure time. Arrival timing to the stopover site was determined using capture data. Non-breeding and previous breeding/natal latitude were determined using analysis of hydrogen isotopes in claws and feathers. Males arrived at the stopover site 11 days on average before females, but no difference in migration timing was observed between plumage morph or age classes. After second year, adults with more southern previous breeding latitudes arrived at stopover earlier, whereas second year birds making their first return migration arrived at stopover in an inverse relationship to non-breeding latitude. Stopover refuelling rate did not differ between ages, sexes or plumage morphs, and daily departure probability of adults was higher under warmer temperatures and favourable tailwinds. White-striped morphs moved greater distances during stopover, initiated daily activity earlier in the morning and departed for migration earlier in the evening than tan-striped morphs. Our results show that while individual phenotype can influence some aspects of local stopover-scale movement behaviour, evidence for differential stopover behaviour was weak. Differential migration timing is unlikely to result from intrinsic differences in en route refuelling rate and departure decisions, especially because the latter is strongly influenced by meteorological conditions.

Illuminating the physiological implications of artificial light on an insectivorous bat community.

Global light pollution threatens to disturb numerous wildlife species, but impacts of artificial light will likely vary among species within a community. Thus, artificial lights may change the environment in such a way as to create winners and losers as some species benefit while others do not. Insectivorous bats are nocturnal and a good model to test for differential effects of light pollution on a single community. We used a physiological technique to address this community-level question by measuring plasma ß-hydroxybutyrate (a blood metabolite) concentrations from six species of insectivorous bats in lit and unlit conditions. We also recorded bat calls acoustically to measure activity levels between experimental conditions. Blood metabolite level and acoustic activity data suggest species-specific changes in foraging around lights. In red bats (Lasiurus borealis), ß-hydroxybutyrate levels at lit sites were highest early in the night before decreasing. Acoustic data indicate pronounced peaks in activity at lit sites early in the night. In red bats on dark nights and in the other species in this community, which seem to avoid lights, ß-hydroxybutyrate remained relatively constant. Our results suggest red bats are more willing to expend energy to actively forage around lights despite potential negative impacts, while other, generally rarer species avoid lit areas. Artificial light appears to have a bifurcating effect on bat communities, whereby some species take advantage of concentrated prey resources, yet most do not. Further, this may concentrate light-intolerant species into limited dark refugia, thereby increasing competition for depauperate, phototactic insect communities.

Plasma metabolite changes in dairy cows during parturition identified using untargeted metabolomics.

The metabolic responses of cows undergo substantial changes during the transition from late pregnancy to early lactation. However, the molecular mechanisms associated with these changes in physiological metabolism have not been clearly elucidated. The objective of this study was to investigate metabolic changes in transition cows from the perspective of plasma metabolites. Plasma samples collected from 24 multiparous dairy cows on approximately d 21 prepartum and immediately postpartum were analyzed using ultra-high-performance liquid chromatography/time-of-flight mass spectrometry in positive and negative ion modes. In conjunction with multidimensional statistical methods (principal component analysis and orthogonal partial least squares discriminant analysis), differences in plasma metabolites were identified using the t-test and fold change analysis. Sixty-seven differential metabolites were identified consisting of AA, lipids, saccharides, and nucleotides. The levels of 32 plasma metabolites were significantly higher and those of 35 metabolites significantly lower after parturition than on d 21 prepartum. Pathway analysis indicated that the metabolites that increased from late pregnancy to early lactation were primarily involved in lipid metabolism and energy metabolism, whereas decreased metabolites were related to AA metabolism.

Gut microbiome-related metabolic changes in plasma of antibiotic-treated rats.

The intestinal microbiota contributes to the metabolism of its host. Adequate identification of the microbiota's impact on the host plasma metabolites is lacking. As antibiotics have a profound effect on the microbial composition and hence on the mammalian-microbiota co-metabolism, we studied the effects of antibiotics on the "functionality of the microbiome"-defined as the production of metabolites absorbed by the host. This metabolomics study presents insights into the mammalian-microbiome co-metabolism of endogenous metabolites. To identify plasma metabolites related to microbiome changes due to antibiotic treatment, we have applied broad-spectrum antibiotics belonging to the class of aminoglycosides (neomycin, gentamicin), fluoroquinolones (moxifloxacin, levofloxacin) and tetracyclines (doxycycline, tetracycline). These were administered orally for 28 days to male rats including blood sampling for metabolic profiling after 7, 14 and 28 days. Fluoroquinolones and tetracyclines can be absorbed from the gut; whereas, aminoglycosides are poorly absorbed. Hippuric acid, indole-3-acetic acid and glycerol were identified as key metabolites affected by antibiotic treatment, beside changes mainly concerning amino acids and carbohydrates. Inter alia, effects on indole-3-propionic acid were found to be unique for aminoglycosides, and on 3-indoxylsulfate for tetracyclines. For each class of antibiotics, specific metabolome patterns could be established in the MetaMap®Tox data base, which contains metabolome data for more than 550 reference compounds. The results suggest that plasma-based metabolic profiling (metabolomics) could be a suitable tool to investigate the effect of antibiotics on the functionality of the microbiome and to obtain insight into the mammalian-microbiome co-metabolism.

Signals of vitellogenesis and estrus in female hawksbill turtles.

This study reports a viable means of identifying the vitellogenic cycle and limited estrus period in hawksbill turtles for the purposes of developing captive breeding program, based on the combination of blood metabolite parameters (triglyceride, total protein, and calcium levels), feeding status, and ovary condition. Follicle size of two focal captive females showed clear seasonal changes, with major development occurring between March and May (19.0-24.4 mm), and exceeding 25 mm between June and September. Triglyceride, total protein, and calcium levels dropped with follicular development and maintenance (March to October), and then began to rise when follicular retraction occurred from October onwards. The two focal turtles reduced food intake during intensive follicular development (April to May). These findings suggest that blood metabolite parameters and feeding conditions are inferred by the vitellogenic cycle. An additional 10 females exhibiting follicular development were mated with a single male for 7-day period between May and June. Follicle size was measured immediately prior to pairing, and a statistically significant difference in follicle size of 10 females was recorded between the seven failed (20.9 mm) and three successful (23.6 mm) mating events. This indicates follicle development is essential to successful mate and monitoring of vitellogenic cycle may help improve the success rates of captive hawksbill breeding programs.

Body weight, meat quality and blood metabolite responses to carbohydrate administration in the drinking water during pre-slaughter feed withdrawal in broilers.

This study was conducted to determine weights of body (BW), carcass (CW), gastrointestinal tract (GTW), meat quality and some blood metabolite responses to corn starch, saccharose or glucose administration in the drinking water during pre-slaughter feed withdrawal (FW) in broilers. On day 42 of age, 200 broilers (Ross 308) were allocated randomly to five treatments with four replicates. During a 10-h FW, control broilers (C) were provided with non-treated water and the standard finisher diet ad libitum, whereas fasted broilers provided with non-treated (NFW) or treated water, 3 g glucose (G), saccharose (S) or corn starch (CS)/L. Eight birds (four males and four females) per treatment were slaughtered. Birds receiving non-treated or treated water had lower BW and higher carcass yield than the full-fed broilers. The full-fed broilers had higher absolute and relative GTW than the fasted birds. Broilers consumed more readily treated water compared with non-treated water. While the a* value of breast meat from CS birds was higher than that from NFW, the b* value of that was higher than S and C birds. The c* values of breast meat from S birds were lower compared with that from the CS treatment. The thigh meat from NFW broilers had higher h* value than that from C and G broilers. The thigh meats of C and CS broilers had higher c* value than that of G birds. The full-fed broilers had higher plasma triglyceride concentration than NFW, S and G birds. The full-fed broilers had higher plasma uric acid and uric acid nitrogen concentrations than S birds. These results show that carbohydrate administration in the drinking water cannot be a good alternative for the FW period before slaughter due to the fact that the carbohydrates do not reduce BW losses and do not lead to increases in meat quality.

Identification of predictive biomarkers of disease state in transition dairy cows.

In dairy cows, periparturient disease states, such as metritis, mastitis, and laminitis, are leading to increasingly significant economic losses for the dairy industry. Treatments for these pathologies are often expensive, ineffective, or not cost-efficient, leading to production losses, high veterinary bills, or early culling of the cows. Early diagnosis or detection of these conditions before they manifest themselves could lower their incidence, level of morbidity, and the associated economic losses. In an effort to identify predictive biomarkers for postpartum or periparturient disease states in dairy cows, we undertook a cross-sectional and longitudinal metabolomics study to look at plasma metabolite levels of dairy cows during the transition period, before and after becoming ill with postpartum diseases. Specifically we employed a targeted quantitative metabolomics approach that uses direct flow injection mass spectrometry to track the metabolite changes in 120 different plasma metabolites. Blood plasma samples were collected from 12 dairy cows at 4 time points during the transition period (-4 and -1 wk before and 1 and 4 wk after parturition). Out of the 12 cows studied, 6 developed multiple periparturient disorders in the postcalving period, whereas the other 6 remained healthy during the entire experimental period. Multivariate data analysis (principal component analysis and partial least squares discriminant analysis) revealed a clear separation between healthy controls and diseased cows at all 4 time points. This analysis allowed us to identify several metabolites most responsible for separating the 2 groups, especially before parturition and the start of any postpartum disease. Three metabolites, carnitine, propionyl carnitine, and lysophosphatidylcholine acyl C14:0, were significantly elevated in diseased cows as compared with healthy controls as early as 4 wk before parturition, whereas 2 metabolites, phosphatidylcholine acyl-alkyl C42:4 and phosphatidylcholine diacyl C42:6, could be used to discriminate healthy controls from diseased cows 1 wk before parturition. A 3-metabolite plasma biomarker profile was developed that could predict which cows would develop periparturient diseases, up to 4 wk before clinical symptoms appearing, with a sensitivity of 87% and a specificity of 85%. This is the first report showing that periparturient diseases can be predicted in dairy cattle before their development using a multimetabolite biomarker model. Further research is warranted to validate these potential predictive biomarkers.

Impact of Chlorella vulgaris Intake Levels on Performance Parameters and Blood Health Markers in Broiler Chickens.

This systematic review examines the effects of cumulative Chlorella vulgaris intake levels on broiler chickens, focusing on growth performance and systemic health markers. The review establishes a clear relationship between cumulative C. vulgaris intake and significant outcomes in poultry nutrition and health through a detailed analysis of various studies. The correlation analysis revealed that cumulative C. vulgaris intake levels ranging from 0.8 to 718 g/bird influenced growth rates and feed efficiency, following sigmoid models. Specifically, intakes of approximately 20 g/bird maximized final body weight (R2 = 0.616, p < 0.001), cumulative body weight gain (R2 = 0.627, p < 0.001) and daily weight gain (R2 = 0.639, p < 0.001). The feed conversion ratio also improved with increasing C. vulgaris intakes up to this level, although this was non-significant (R2 = 0.289, p = 0.117). In addition, similar cumulative C. vulgaris intake levels impacted plasma health markers in broilers, leading to reductions in triacylglycerols and cholesterol and improvements in immunoglobulin levels. These findings underscore the importance of carefully calibrated C. vulgaris supplementation strategies to optimise poultry growth and health without adverse effects. Future research should focus on refining C. vulgaris dosing guidelines and further exploring its long-term effects and mechanisms of action to enhance poultry health and production sustainability.

Predicting effect of anti-PD-1/PD-L1 inhibitors therapy for hepatocellular carcinoma by detecting plasma metabolite based on UHPLC-MS.

Introduction: Anti-PD-1/PD-L1 inhibitors therapy has become a promising treatment for hepatocellular carcinoma (HCC), while the therapeutic efficacy varies significantly among effects for individual patients are significant difference. Unfortunately, specific predictive biomarkers indicating the degree of benefit for patients and thus guiding the selection of suitable candidates for immune therapy remain elusive.no specific predictive biomarkers are available indicating the degree of benefit for patients and thus screening the preferred population suitable for the immune therapy. Methods: Ultra-high-pressure liquid chromatography-mass spectrometry (UHPLC-MS) considered is an important method for analyzing biological samples, since it has the advantages of high rapid, high sensitivity, and high specificity. Ultra-high-pressure liquid chromatography-mass spectrometry (UHPLC-MS) has emerged as a pivotal method for analyzing biological samples due to its inherent advantages of rapidity, sensitivity, and specificity. In this study, potential metabolite biomarkers that can predict the therapeutic effect of HCC patients receiving immune therapy were identified by UHPLC-MS. Results: A partial least-squares discriminant analysis (PLS-DA) model was established using 14 glycerophospholipid metabolites mentioned above, and good prediction parameters (R2 = 0.823, Q2 = 0.615, prediction accuracy = 0.880 and p < 0.001) were obtained. The relative abundance of glycerophospholipid metabolite ions is closely related to the survival benefit of HCC patients who received immune therapy. Discussion: This study reveals that glycerophospholipid metabolites play a crucial role in predicting the efficacy of immune therapy for HCC.

Identification of potential biomarkers for diabetic cardiomyopathy using LC-MS-based metabolomics.

Diabetic cardiomyopathy (DCM) is a serious complication of type 2 diabetes mellitus (T2DM) that contributes to cardiovascular morbidity and mortality. However, the metabolic alterations and specific biomarkers associated with DCM in T2DM remain unclear. In this study, we conducted a comprehensive metabolomic analysis using liquid chromatography-mass spectrometry (LC-MS) to investigate the plasma metabolite profiles of T2DM patients with and without DCM. We identified significant differences in metabolite levels between the groups, highlighting the dysregulation of various metabolic pathways, including starch and sucrose metabolism, steroid hormone biosynthesis, tryptophan metabolism, purine metabolism, and pyrimidine metabolism. Although several metabolites showed altered abundance in DCM, they also shared characteristics of DCM and T2DM rather than specific to DCM. Additionally, through biomarker analyses, we identified potential biomarkers for DCM, such as cytidine triphosphate, 11-ketoetiocholanolone, saccharopine, nervonic acid, and erucic acid. These biomarkers demonstrated distinct patterns and associations with metabolic pathways related to DCM. Our findings provide insights into the metabolic changes associated with DCM in T2DM patients and highlight potential biomarkers for further validation and clinical application. Further research is needed to elucidate the underlying mechanisms and validate the diagnostic and prognostic value of these biomarkers in larger cohorts.

Postprandial plasma amino acid and metabolite kinetics of adult and growing pigs fed a diet with a balanced or unbalanced amino acid profile.

Feeding diets with an unbalanced amino acid (AA) profile can reduce the postprandial AA utilization for protein synthesis. Growing pigs use dietary AA mainly for protein accretion, whereas non-lactating and non-pregnant adult pigs use AA mainly for maintenance. The requirement for AA for growth is much larger than that for maintenance and growing pigs may therefore be more affected by a diet with an unbalanced AA profile than adult pigs. This study aimed to compare the postprandial plasma AA and metabolite concentrations of adult and growing pigs after feeding a diet with either an unbalanced (UNB) or a balanced AA profile (BAL). The postprandial plasma concentrations of AA were used to study the influence of AA balance on postprandial AA metabolism. Extensively hydrolysed feathers (EHF) were used as an AA source. Both BAL and UNB contained EHF supplemented with L-Ala, L-Asp, L-Glu, Gly, and L-Trp while BAL was also supplemented with L-His, L-Ile, L-Lys, L-Met, and L-Tyr. Four growing and four male adult pigs were fitted with a jugular catheter and received each diet as a meal test thrice. The meal test consisted of giving a small meal after an overnight fast followed by serial blood collection for 360 min. A non-linear regression model was used to describe the postprandial plasma AA kinetics. Plasma kinetics of adult and growing pigs fed BAL resulted in a higher area under the curve (AUC) for the AA that were used to balance the diet. For the other AA, feeding BAL resulted in lower AUC, suggesting faster metabolic utilization of AA for protein synthesis. The apparent quantity of dietary AA appearing in the plasma after feeding was lower in adult pigs, suggesting higher first-pass AA utilization in the intestine and liver. For adult and growing pigs, balancing the AA profile of the diet resulted in faster overall metabolic utilization of AA as seen in the generally lower AUC of BAL compared to UNB.

Analysis of the SYSDIET Healthy Nordic Diet randomized trial based on metabolic profiling reveal beneficial effects on glucose metabolism and blood lipids.

BACKGROUND & AIMS: Intake assessment in multicenter trials is challenging, yet important for accurate outcome evaluation. The present study aimed to characterize a multicenter randomized controlled trial with a healthy Nordic diet (HND) compared to a Control diet (CD) by plasma and urine metabolic profiles and to associate them with cardiometabolic markers. METHODS: During 18-24 weeks of intervention, 200 participants with metabolic syndrome were advised at six centres to eat either HND (e.g. whole-grain products, berries, rapeseed oil, fish and low-fat dairy) or CD while being weight stable. Of these 166/159 completers delivered blood/urine samples. Metabolic profiles of fasting plasma and 24 h pooled urine were analysed to identify characteristic diet-related patterns. Principal components analysis (PCA) scores (i.e. PC1 and PC2 scores) were used to test their combined effect on blood glucose response (primary endpoint), serum lipoproteins, triglycerides, and inflammatory markers. RESULTS: The profiles distinguished HND and CD with AUC of 0.96 ± 0.03 and 0.93 ± 0.02 for plasma and urine, respectively, with limited heterogeneity between centers, reflecting markers of key foods. Markers of fish, whole grain and polyunsaturated lipids characterized HND, while CD was reflected by lipids containing palmitoleic acid. The PC1 scores of plasma metabolites characterizing the intervention is associated with HDL (ß = 0.05; 95% CI: 0.02, 0.08; P = 0.001) and triglycerides (ß = -0.06; 95% CI: -0.09, -0.03; P < 0.001). PC2 scores were related with glucose metabolism (2 h Glucose, ß = 0.1; 95% CI: 0.05, 0.15; P < 0.001), LDL (ß = 0.06; 95% CI: 0.01, 0.1; P = 0.02) and triglycerides (ß = 0.11; 95% CI: 0.06, 0.15; P < 0.001). For urine, the scores were related with LDL cholesterol. CONCLUSIONS: Plasma and urine metabolite profiles from SYSDIET reflected good compliance with dietary recommendations across the region. The scores of metabolites characterizing the diets associated with outcomes related with cardio-metabolic risk. Our analysis therefore offers a novel way to approach a per protocol analysis with a balanced compliance assessment in larger multicentre dietary trials. The study was registered at clinicaltrials.gov with NCT00992641.

Metabolomics reveals mouse plasma metabolite responses to acute exercise and effects of disrupting AMPK-glycogen interactions.

Introduction: The AMP-activated protein kinase (AMPK) is a master regulator of energy homeostasis that becomes activated by exercise and binds glycogen, an important energy store required to meet exercise-induced energy demands. Disruption of AMPK-glycogen interactions in mice reduces exercise capacity and impairs whole-body metabolism. However, the mechanisms underlying these phenotypic effects at rest and following exercise are unknown. Furthermore, the plasma metabolite responses to an acute exercise challenge in mice remain largely uncharacterized. Methods: Plasma samples were collected from wild type (WT) and AMPK double knock-in (DKI) mice with disrupted AMPK-glycogen binding at rest and following 30-min submaximal treadmill running. An untargeted metabolomics approach was utilized to determine the breadth of plasma metabolite changes occurring in response to acute exercise and the effects of disrupting AMPK-glycogen binding. Results: Relative to WT mice, DKI mice had reduced maximal running speed (p < 0.0001) concomitant with increased body mass (p < 0.01) and adiposity (p < 0.001). A total of 83 plasma metabolites were identified/annotated, with 17 metabolites significantly different (p < 0.05; FDR<0.1) in exercised (↑6; ↓11) versus rested mice, including amino acids, acylcarnitines and steroid hormones. Pantothenic acid was reduced in DKI mice versus WT. Distinct plasma metabolite profiles were observed between the rest and exercise conditions and between WT and DKI mice at rest, while metabolite profiles of both genotypes converged following exercise. These differences in metabolite profiles were primarily explained by exercise-associated increases in acylcarnitines and steroid hormones as well as decreases in amino acids and derivatives following exercise. DKI plasma showed greater decreases in amino acids following exercise versus WT. Conclusion: This is the first study to map mouse plasma metabolomic changes following a bout of acute exercise in WT mice and the effects of disrupting AMPK-glycogen interactions in DKI mice. Untargeted metabolomics revealed alterations in metabolite profiles between rested and exercised mice in both genotypes, and between genotypes at rest. This study has uncovered known and previously unreported plasma metabolite responses to acute exercise in WT mice, as well as greater decreases in amino acids following exercise in DKI plasma. Reduced pantothenic acid levels may contribute to differences in fuel utilization in DKI mice.

Changes in growth performance, plasma metabolite concentrations, and myogenic gene expression in growing pigs fed a methionine-restricted diet.

Background: Methionine (Met) is usually the second or third limiting amino acid in swine diets and plays vital roles in promoting the growth, especially, the muscle growth of pigs. This research evaluated the effects of dietary Met restriction on the growth performance, plasma metabolite concentrations, and myogenic gene expression in growing pigs. Materials and methods: Eight genes in two families (myogenic regulatory factor family and myocyte enhancer factor 2 family) were selected for the analysis. Twenty individually penned barrows (crossbred, 23.6 ± 2.4 kg) were randomly allotted to two dietary treatments (n = 10). A diet based on corn and soybean meal (Diet 1, Met-restricted) was formulated to meet or exceed the energy and nutrient requirements, except for Met. Diet 2 (Met-adequate) was formulated by adding crystalline DL-Met to Diet 1 to meet the Met requirement. During the 4-week feeding trial, average daily gain (ADG), average daily feed intake (ADFI), and gain to feed ratio (G:F) were measured. Immediately before and after the feeding trial, blood was sampled via jugular venipuncture for plasma nutrient metabolite analysis, while Longissimus dorsi muscle were sampled via aseptic biopsy for gene expression analysis. Data were analyzed with Student t-test. Results: Pigs fed the Met-restricted diet had lower ADG and G:F (P < 0.01). Plasma Met, cysteine, and taurine concentrations were lower (P < 0.05), while glycine and histidine concentrations were higher (P < 0.05), in pigs fed the Met-restricted diet. Furthermore, the pigs fed the Met-restricted diet tended to express less myogenic factor 6 (Myf6) and myocyte enhancer factor 2D (Mef2D) mRNA in longissimus dorsi muscle (P < 0.09). Conclusion: Given the fact that Myf6, assisted by Mef2D, is involved in myocyte differentiation, this study suggests that the reduced growth performance in the Met-restricted pigs may be associated with a reduced muscle cell differentiation.

Production performance and plasma metabolite concentrations of broiler chickens fed low crude protein diets differing in Thr and Gly.

The aim of the study was to test the interaction between Thr and Gly in low crude protein (CP) diets in 7 to 28 d broilers on production performance and plasma metabolites. A total of 2,040 broilers were allocated to 17 treatments. A positive control (PC) diet (20.5% CP) was formulated to be adequate in dietary Thr and Gly. A negative control (NC) diet (18.5% CP, deficient in Thr and Gly) was supplemented with crystalline l-Thr and Gly to obtain a 4 Thr × 4 Gly design. Dietary Thr was tested at an apparent faecal digestibility (AFD) Thr-to-Lys ratio, which was 55%, 58%, 61% or 64%, and dietary Gly was tested at an AFD (Gly + Ser)-to-Lys ratio, which was 135%, 142%, 149% or 156%. Plasma samples were collected at 28 d. The low CP diet, formulated at 64% Thr and 156% Gly, resulted in a higher body weight gain (BWG) (P < 0.01) and similar feed conversion ratio (FCR) as the high CP treatment (PC). FCR was improved (P < 0.001) by l-Thr supplementation. Quadratic response to dietary Thr was significant for feed intake (FI), BWG and FCR (P < 0.01). A near-significant interaction for Thr × Gly was observed for FI and BWG (P linear = 0.091 and P = 0.074, respectively). Gly did not affect production performance. An interaction between Thr × Gly on plasma free AA level was observed (P < 0.05). Free AA concentration in plasma linearly decreased with increase in AFD Thr-to-Lys ratio, and increased with increase in AFD (Gly + Ser)-to-Lys ratio. Plasma uric acid concentration was higher in PC than in all of the other diets, and plasma triglyceride concentration was decreased by l-Thr supplementation, but not by Gly. In conclusion, Gly was not limiting for growth at low dietary CP level unless Thr was deficient, showing that adequate amounts of Thr in broiler diets can overcome marginal supply of Gly and Ser and allow reduction of dietary CP from 20.5% to 18.5% for broilers from 7 to 28 d of age.

Sex-Specific Relationship between the Cardiorespiratory Fitness and Plasma Metabolite Patterns in Healthy Humans-Results of the KarMeN Study.

Cardiorespiratory fitness (CRF) represents a strong predictor of all-cause mortality and is strongly influenced by regular physical activity (PA). However, the biological mechanisms involved in the body's adaptation to PA remain to be fully elucidated. The aim of this study was to systematically examine the relationship between CRF and plasma metabolite patterns in 252 healthy adults from the cross-sectional Karlsruhe Metabolomics and Nutrition (KarMeN) study. CRF was determined by measuring the peak oxygen uptake during incremental exercise. Fasting plasma samples were analyzed by nuclear magnetic resonance spectroscopy and mass spectrometry coupled to one- or two-dimensional gas chromatography or liquid chromatography. Based on this multi-platform metabolomics approach, 427 plasma analytes were detected. Bi- and multivariate association analyses, adjusted for age and menopausal status, showed that CRF was linked to specific sets of metabolites primarily indicative of lipid metabolism. However, CRF-related metabolite patterns largely differed between sexes. While several phosphatidylcholines were linked to CRF in females, single lyso-phosphatidylcholines and sphingomyelins were associated with CRF in males. When controlling for further assessed clinical and phenotypical parameters, sex-specific CRF tended to be correlated with a smaller number of metabolites linked to lipid, amino acid, or xenobiotics-related metabolism. Interestingly, sex-specific CRF explanation models could be improved when including selected plasma analytes in addition to clinical and phenotypical variables. In summary, this study revealed sex-related differences in CRF-associated plasma metabolite patterns and proved known associations between CRF and risk factors for cardiometabolic diseases such as fat mass, visceral adipose tissue mass, or blood triglycerides in metabolically healthy individuals. Our findings indicate that covariates like sex and, especially, body composition have to be considered when studying blood metabolic markers related to CRF.

Pharmacokinetics and biotransformation investigation in beagle dog of active compounds from naoxintong capsule.

Naoxintong Capsule (NXTC), a standardized herbal medicine, has been widely applied in treating cardiovascular and cerebrovascular diseases with remarkable efficacy. However, the efficacy contributing components of NXTC are unclear, and the in vivo absorption and metabolism processes of NXTC remain largely obscured. In this study, using beagle dog as model species, we have identified and tentatively characterized 25 prototype and 15 catabolites of NXTC in beagle dog plasma by ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-Q-TOF-MS/MS). We have proposed the in vivo bio-transformation pathways of these absorbed constituents. In addition, for six crucial components, we have developed a quantitative method and conducted plasma pharmacokinetic study of these six components by rapid resolution liquid chromatography tandem triple quadrupole mass spectrometry (RRLC-QQQ-MS/MS). In conclude, our study provided comprehensive insights into the understanding of the plasma absorbed components profiling of NXTC as well as their in vivo transformation behaviors, which would be of great value for identifying efficacy contributing critical components as well as mechanism related investigations of NXTC in the future.