PubMed

Bioanalysis of endogenous bile acids by LC-MS utilizing stable isotope labeled analogues: The utility of stable isotope labeled (SIL) analogues in the bioanalysis of endogenous compounds by LC-MS applied to the study of bile acids in a metabolomics assay. Anal Biochem. 2016 Mar 28; Authors: Zheng JJ, Shields EE, Snow KJ, Nelson DM, Olah TV, Reily MD, Robertson DG, Shipkova PA, Stryker SA, Xin B, Drexler DM Abstract The growing field of biomarker bioanalysis by LC-MS is challenged with the selection of suitable matrices to construct relevant and valid calibration curves resulting in not only precise but also accurate data. While surrogate matrices are often employed with the associated concerns about the accuracy of the obtained data, presented here is an assay utilizing surrogate analytes in naïve biological matrices. This approach is illustrated with the analysis of endogenous bile acids (e-BAs) in serum and plasma using stable isotope labeled (SIL)-analogues as calibration standards to address the matrix concerns. Several deuterated BAs (d-BAs) were used as standards representing respectively grouped e-BAs with structural similarity allowing for the simultaneous bioanalysis of 16 e-BA. The utility of this LC-MS assay employing d-BAs is demonstrated with the analysis of samples resultant of a controlled metabolomics study where a cohort of rats were fed/fasted to investigate the change of e-BAs dependent on food consumption and fasting time. PMID: 27033006 [PubMed - as supplied by publisher]

Effects of Perfluorooctanoic Acid on Metabolic Profiles in Brain and Liver of Mouse Revealed by a High-throughput Targeted Metabolomics Approach. Sci Rep. 2016;6:23963 Authors: Yu N, Wei S, Li M, Yang J, Li K, Jin L, Xie Y, Giesy JP, Zhang X, Yu H Abstract Perfluorooctanoic acid (PFOA), a perfluoroalkyl acid, can result in hepatotoxicity and neurobehavioral effects in animals. The metabolome, which serves as a connection among transcriptome, proteome and toxic effects, provides pathway-based insights into effects of PFOA. Since understanding of changes in the metabolic profile during hepatotoxicity and neurotoxicity were still incomplete, a high-throughput targeted metabolomics approach (278 metabolites) was used to investigate effects of exposure to PFOA for 28 d on brain and liver of male Balb/c mice. Results of multivariate statistical analysis indicated that PFOA caused alterations in metabolic pathways in exposed individuals. Pathway analysis suggested that PFOA affected metabolism of amino acids, lipids, carbohydrates and energetics. Ten and 18 metabolites were identified as potential unique biomarkers of exposure to PFOA in brain and liver, respectively. In brain, PFOA affected concentrations of neurotransmitters, including serotonin, dopamine, norepinephrine, and glutamate in brain, which provides novel insights into mechanisms of PFOA-induced neurobehavioral effects. In liver, profiles of lipids revealed involvement of β-oxidation and biosynthesis of saturated and unsaturated fatty acids in PFOA-induced hepatotoxicity, while alterations in metabolism of arachidonic acid suggesting potential of PFOA to cause inflammation response in liver. These results provide insight into the mechanism and biomarkers for PFOA-induced effects. PMID: 27032815 [PubMed - as supplied by publisher]

Dissecting lipid metabolism in meibomian glands of humans and mice: An integrative study reveals a network of metabolic reactions not duplicated in other tissues. Biochim Biophys Acta. 2016 Mar 28; Authors: Butovich IA, McMahon A, Wojtowicz JC, Lin F, Mancini R, Itani K Abstract Lipids comprise the bulk of the meibomian gland secretion (meibum) which is produced by meibocytes. Complex arrays of lipogenic reactions in meibomian glands, which we collectively call meibogenesis, have not been explored on molecular level yet. Our goals were to elucidate the possible biosynthetic pathways that underlie the generation of meibum, reveal similarities in, and differences between, lipid metabolism in meibomian glands and other organs and tissues, and integrate meibomian gland studies into the field of general metabolomics. Specifically, we have conducted detailed analyses of human and mouse specimens using genomic, immunohistochemical, and lipidomic approaches. Among equally highly expressed genes found in meibomian glands of both species were those related to fatty acid elongation, branching, desaturation, esterification, reduction of fatty acids to alcohols, and cholesterol biosynthesis. Importantly, corresponding lipid products were detected in meibum of both species using lipidomic approaches. For the first time, a cohesive, unifying biosynthetic scheme that connects genomic, lipidomic, and immunohistochemical observations is outlined and discussed. PMID: 27032494 [PubMed - as supplied by publisher]

Related Articles -omic sciences: new horizons in food allergy. Curr Opin Allergy Clin Immunol. 2015 Jun;15(3):234-6 Authors: Fiocchi A, Wang J PMID: 25899698 [PubMed - indexed for MEDLINE]

Related Articles UHPLC-PDA-ESI-TOF/MS metabolic profiling of Arctostaphylos pungens and Arctostaphylos uva-ursi. A comparative study of phenolic compounds from leaf methanolic extracts. Phytochemistry. 2015 Jul;115:79-88 Authors: Panusa A, Petrucci R, Marrosu G, Multari G, Gallo FR Abstract The aim of this study was to get a rapid metabolic fingerprinting and to gain insight into the metabolic profiling of Arctostaphylos pungens H. B. K., a plant morphologically similar to Arctostaphylos uva-ursi (L.) Spreng. (bearberry) but with a lower arbutin (Arb) content. According to the European Pharmacopoeia the Arb content in the dried leaf of A. uva-ursi (L.) Spreng. must be at least 7% (wt/wt) but other species, like A. pungens, are unintentionally or fraudulently marketed instead of it. Therefore, methanolic leaf extracts of nine A. uva-ursi and six A. pungens samples labeled and marketed as "bearberry leaf" have been analyzed. A five-minute gradient with a UHPLC-PDA-ESI-TOF/MS on an Acquity BEH C18 (50×2.1 mm i.d.) 1.7 μm analytical column has been used for the purpose. A comprehensive assignment of secondary metabolites has been carried out in a comparative study of the two species. Among twenty-nine standards of natural compounds analyzed, fourteen have been identified, while other fifty-five metabolites have been tentatively assigned. Moreover, differences in both metabolic fingerprinting and profiling have been evidenced by statistical multivariate analysis. Specifically, main variations have been observed in the relative content for Arb, as expected, and for some galloyl derivative like tetra- and pentagalloylglucose more abundant in A. uva-ursi than in A. pungens. Furthermore, differences in flavonols profile, especially in myricetin and quercetin glycosilated derivatives, were observed. Based on principal component analysis myricetrin, together with a galloyl arbutin isomer and a disaccharide are herein proposed as distinctive metabolites for A. pungens. PMID: 25702282 [PubMed - indexed for MEDLINE]

Related Articles GC-MS-based plasma metabolomic investigations of morphine dependent rats at different states of euphoria, tolerance and naloxone-precipitated withdrawal. Metab Brain Dis. 2015 Jun;30(3):767-76 Authors: Liu R, Cheng J, Yang J, Ding X, Yang S, Dong F, Guo N, Liu S Abstract Long-term or excessive application of morphine leads to tolerance and addiction, which hindered its conventional applications as a drug. Although tremendous progress has been made on the mechanisms of morphine, crucial evidence elaborating the neurobiological basis of tolerance and dependence is still lacking. To further explore the physiological adaptions during morphine's application, a systematic screening of small molecules in blood has been carried out. The plasma of morphine dependent rats was collected at different time points with or without naloxone treatment, and was analyzed by gas chromatography-mass spectrometry (GC-MS). Partial least squares discriminate analysis (PLS-DA) and the Student's t Tests with the false discovery rate (FDR) correction were conducted on the normalized data for the distinction of groups and the identification of the most contributed metabolites. Clear separation is observed between different treatments, and 29 out of 41 metabolites changes significantly compared with the corresponding controls. The concentration of threonine, glycine, serine, beta-d-glucose and oxalic acid are consistently changed in all morphine treated groups compared with controls. Through this experiment we find characteristic metabolites in different dependent states and discuss the possible compensation effects. The interpretation of these metabolites would throw light on the biological effects of morphine and reveal the possibilities to become marker of morphine addiction. PMID: 25472920 [PubMed - indexed for MEDLINE]

Discovery of potential biomarkers with dose and time dependence in cisplatin-induced nephrotoxicity using metabolomics integrated with principal component-based area calculation (PCAC) strategy. Chem Res Toxicol. 2016 Mar 31; Authors: Zhang P, Chen J, Wang Y, Huang Y, Tian Y, Zhang Z, Xu F Abstract Cisplatin is a potent chemotherapeutic agent. However, its clinical usage is restricted by serious adverse effects, especially nephrotoxicity. In order to reveal the dose- and time-dependence in cisplatin-induced nephrotoxicity, mass spectrometry (MS)-based metabolomics integrated with principal component-based area calculation (PCAC) strategy was proposed in the present study. Area plots based on the first two principal components (PCs) of principal component analysis (PCA) model were first constructed. Then, the sums of cumulative areas under PC-T curves (AUCPC-T) were calculated. Finally, fold change of AUCPC-T (FCAUC) between experimental group and control group at different time point was calculated and used as indicative parameter. With PCAC approach, dose- and time-dependence of cisplatin-induced metabolic change was quantitatively confirmed for the first time. Furthermore, twenty-seven potential biomarkers with dose- and time-dependence related to nephrotoxicity induced by cisplatin were screened out and tentatively identified. Metabolic pathways interrupted by cisplatin mainly included energy metabolism, amino acid metabolism and lipid metabolism. PMID: 27030963 [PubMed - as supplied by publisher]

Immunological off-target effects of imatinib. Nat Rev Clin Oncol. 2016 Mar 31; Authors: Zitvogel L, Rusakiewicz S, Routy B, Ayyoub M, Kroemer G Abstract Around 15 years ago, imatinib mesylate (Gleevec(®) or Glivec(®), Novartis, Switzerland) became the very first 'targeted' anticancer drug to be clinically approved. This drug constitutes the quintessential example of a successful precision medicine that has truly changed the fate of patients with Philadelphia-chromosome-positive chronic myeloid leukaemia (CML) and gastrointestinal stromal tumours by targeting the oncogenic drivers of these diseases, BCR-ABL1 and KIT and/or PDGFR, mutations in which lead to gain of function of tyrosine kinase activities. Nonetheless, the aforementioned paradigm might not fully explain the clinical success of this agent in these diseases. Growing evidence indicates that the immune system has a major role both in determining the therapeutic efficacy of imatinib (and other targeted agents) and in restraining the emergence of escape mutations. In this Review, we re-evaluate the therapeutic utility of imatinib in the context of the anticancer immunosurveillance system, and we discuss how this concept might inform on novel combination regimens that include imatinib with immunotherapies. PMID: 27030078 [PubMed - as supplied by publisher]

Plasma Biomarkers of Poor Muscle Quality in Older Men and Women from the Baltimore Longitudinal Study of Aging. J Gerontol A Biol Sci Med Sci. 2016 Mar 30; Authors: Moaddel R, Fabbri E, Khadeer MA, Carlson OD, Gonzalez-Freire M, Zhang P, Semba RD, Ferrucci L Abstract Aging is characterized by progressive decline in muscle mass, strength, and quality all of which contribute to functional impairment, falls, mobility disability, and frailty. Circulating factors may provide clues on the mechanisms for decline in muscle quality with aging. Characterizing the metabolic profile associated with reduced muscle quality in older persons could have important translational implications for the early identification of subjects at high risk of developing sarcopenia and the identification of targets for new preventive strategies and treatments. In a pilot cross-sectional, case-control study nested in the Baltimore Longitudinal Study on Aging, we compared circulating metabolites between 79 participants with low muscle quality ratio and 79 controls with high muscle quality, matched by age, sex, and height. The concentrations of 180 metabolites were determined by LC MS/MS, using the Biocrates p180 system, a targeted metabolomics approach. Participants with low muscle quality had significantly higher levels of leucine, isoleucine, tryptophan, serotonin, and methionine, while those with high muscle quality had significantly lower levels of putrescine and the selected phophatidylcholine (PCs) and lysoPCs. The results of this study open a new road for future investigations aimed at identifying new metabolic pathways involved in the decline of muscle quality with aging. PMID: 27029859 [PubMed - as supplied by publisher]

Related Articles Age at diagnosis and C-peptide level are associated with diabetic retinopathy in Chinese. PLoS One. 2014;9(3):e91174 Authors: Cai X, Han X, Zhang S, Luo Y, Chen Y, Ji L Abstract OBJECTIVE: To find the associations between diabetic retinopathy and age at diagnosis, C-peptide level and thyroid-stimulating hormone (TSH) level in Chinese type 2 diabetes mellitus. METHODS: 3100 hospitalized type 2 diabetic patients in Peking University People's Hospital were included in this retrospective study. Their medical history and the laboratory data were collected. All the patients received examination of diabetic retinopathy (DR) by professional ophthalmologist. RESULTS: Comparisons among patients with NDR, NPDR and PDR showed that with the progression of diabetic retinopathy, patients turned to have older age but younger age at diagnosis of diabetes, and have higher SBP, longer duration of diabetes, higher mean HbA1c but lower fasting and 2 hours postprandial C-peptide level. Moreover, with the progression of diabetic retinopathy, patients turned to have higher prevalence of primary hypertension, higher prevalence of peripheral vascular sclerosis, higher proportion with insulin treatment. TSH level was comparable among the three groups of patients. Association analysis showed that after adjusting for age, sex, duration of diabetes, body mass index, HbA1c, blood pressure and albuminurea creatinine ratio and insulin treatment, age at diagnosis (OR 0.888, 95%CI 0.870-0.907, p = 0.00) and postprandial C-peptide (OR 0.920, 95%CI 0.859-0.937, p = 0.00) are the independent associated factors of DR in Chinese type 2 diabetes. CONCLUSIONS: According to the results, postprandial C-peptide level and age at diabetes may be two independent associated factors with DR in Chinese type 2 diabetes. The lower level of postprandial C-peptide, the younger age at diagnosis, may indicate the higher prevalence of DR. PMID: 24614131 [PubMed - indexed for MEDLINE]

Gut microbiota profiling of pediatric NAFLD and obese patients unveiled by an integrated meta-omics based approach. Hepatology. 2016 Mar 29; Authors: Del Chierico F, Nobili V, Vernocchi P, Russo A, De Stefanis C, Gnani D, Furlanello C, Zandonà A, Paci P, Capuani G, Dallapiccola B, Miccheli A, Alisi A, Putignani L Abstract There is evidence that non-alcoholic fatty liver disease (NAFLD) is affected by gut microbiota. Therefore, we investigated its modifications in paediatric NAFLD patients using targeted-metagenomics (MG) and metabolomics (MB). Stools were collected from 61 consecutive patients diagnosed with NAFL, NASH, or obesity and 54 healthy subjects (CTRLs), matched in a case-control fashion. Operational taxonomic units were pyrosequenced targeting 16S ribosomal RNA and volatile organic compounds (VOCs) determined by solid-phase micro-extraction GC-MS. The α-diversity was highest in CTRLs followed by obese, NASH, NAFL patients and β-diversity distinguished between patients and CTRLs, but not NAFL and NASH. Compared to CTRLs, in NAFLD patients Actinobacteria were significantly increased and Bacteroidetes reduced. There were no significant differences amongst NAFL, NASH, and obese groups. Overall NAFLD patients had increased levels of Bradyrhizobium, Anaerococcus, Peptoniphilus, Propionibacterium acnes, Dorea, Ruminococcus and reduced proportions of Oscillospira and Rikenellaceae compared to CTRLs. After reducing MG and MB data dimensionality, multivariate analyses indicated Oscillospira decrease in NAFL and NASH groups, and Ruminococcus, Blautia, and Dorea increase in NASH patients compared to CTRLs. Of the 292 VOCs, 26 were up- and 2 down-regulated in NAFLD patients. Multivariate analyses found that combination of Oscillospira, Rickenellaceae, Parabacteroides, Bacteroides fragilis, Sutterella, Lachnospiraceae, 4-methyl-2-pentanone, 1-butanol, and 2-butanone could discriminate NAFLD patients from CTRLs. Univariate analyses found significantly lower levels of Oscillospira and higher levels of 1-pentanol and 2-butanone in NAFL compared to CTRLs. In NASH, lower levels of Oscillospira were associated with higher abundance of Dorea, Ruminococcus and higher levels of 2-butanone, 4-methyl-2-pentanone compared to CTRLs. CONCLUSION: Oscillospira decrease coupled to 2-butanone up-regulation and Ruminococcus, Dorea increase were identified as gut microbiota signatures of NAFL onset and NAFL-NASH progression, respectively. This article is protected by copyright. All rights reserved. PMID: 27028797 [PubMed - as supplied by publisher]

Linking field-based metabolomics and chemical analyses to prioritize contaminants of emerging concern in the Great Lakes basin. Environ Toxicol Chem. 2016 Mar 30; Authors: Davis JM, Ekman DR, Teng Q, Ankley GT, Berninger JP, Cavallin JE, Jensen KM, Kahl MD, Schroeder AL, Villeneuve DL, Jorgenson ZG, Lee KE, Collette TW Abstract The ability to focus on the most biologically relevant contaminants affecting aquatic ecosystems can be challenging because toxicity assessment programs have not kept pace with the growing number of contaminants requiring testing. Because it has proven effective in assessing biological impacts of potentially toxic contaminants, profiling of endogenous metabolites (metabolomics) may help screen out contaminants with a lower likelihood of eliciting biological impacts, thereby prioritizing the most biologically-important contaminants. We present results from a study that utilized cage-deployed fathead minnows (Pimephales promelas) at 18 sites across the Great Lakes basin. We measured water temperature and contaminant concentrations in water samples (132 contaminants targeted; 86 detected), and used (1) H-NMR spectroscopy to measure endogenous metabolites in polar extracts of livers. We used partial least-squares (PLS) regression to compare relative abundances of endogenous metabolites with contaminant concentrations and temperature. Results indicated that profiles of endogenous polar metabolites covaried with at most 49 contaminants. Thus, we identified up to 52% of detected contaminants as not significantly covarying with changes in endogenous metabolites, suggesting they likely were not eliciting measureable impacts at these sites. This represents a first step in screening for the biological-relevance of detected contaminants by shortening lists of contaminants potentially affecting these sites. Such information may allow risk assessors to prioritize contaminants and focus toxicity testing on the most biologically-relevant contaminants. This article is protected by copyright. All rights reserved. PMID: 27027868 [PubMed - as supplied by publisher]

"Omics" of Selenium Biology: A Prospective Study of Plasma Proteome Network Before and After Selenized-Yeast Supplementation in Healthy Men. OMICS. 2016 Mar 30; Authors: Sinha I, Karagoz K, Fogle RL, Hollenbeak CS, Zea AH, Arga KY, Stanley AE, Hawkes WC, Sinha R Abstract Low selenium levels have been linked to a higher incidence of cancer and other diseases, including Keshan, Chagas, and Kashin-Beck, and insulin resistance. Additionally, muscle and cardiovascular disorders, immune dysfunction, cancer, neurological disorders, and endocrine function have been associated with mutations in genes encoding for selenoproteins. Selenium biology is complex, and a systems biology approach to study global metabolomics, genomics, and/or proteomics may provide important clues to examining selenium-responsive markers in circulation. In the current investigation, we applied a global proteomics approach on plasma samples collected from a previously conducted, double-blinded placebo controlled clinical study, where men were supplemented with selenized-yeast (Se-Yeast; 300 μg/day, 3.8 μmol/day) or placebo-yeast for 48 weeks. Proteomic analysis was performed by iTRAQ on 8 plasma samples from each arm at baseline and 48 weeks. A total of 161 plasma proteins were identified in both arms. Twenty-two proteins were significantly altered following Se-Yeast supplementation and thirteen proteins were significantly changed after placebo-yeast supplementation in healthy men. The differentially expressed proteins were involved in complement and coagulation pathways, immune functions, lipid metabolism, and insulin resistance. Reconstruction and analysis of protein-protein interaction network around selected proteins revealed several hub proteins. One of the interactions suggested by our analysis, PHLD-APOA4, which is involved in insulin resistance, was subsequently validated by Western blot analysis. Our systems approach illustrates a viable platform for investigating responsive proteomic profile in 'before and after' condition following Se-Yeast supplementation. The nature of proteins identified suggests that selenium may play an important role in complement and coagulation pathways, and insulin resistance. PMID: 27027327 [PubMed - as supplied by publisher]

Real-time bilinear rotation decoupling in absorptive mode J-spectroscopy: Detecting low-intensity metabolite peak close to high-intensity metabolite peak with convenience. J Magn Reson. 2016 Mar 10;266:51-58 Authors: Verma A, Baishya B Abstract "Pure shift" NMR spectra display singlet peak per chemical site. Thus, high resolution is offered at the cost of valuable J-coupling information. In the present work, real-time BIRD (BIlinear Rotation Decoupling) is applied to the absorptive-mode 2D J-spectroscopy to provide pure shift spectrum in the direct dimension and J-coupling information in the indirect dimension. Quite often in metabolomics, proton NMR spectra from complex bio-fluids display tremendous signal overlap. Although conventional J-spectroscopy in principle overcomes this problem by separating the multiplet information from chemical shift information, however, only magnitude mode of the experiment is practical, sacrificing much of the potential high resolution that could be achieved. Few J-spectroscopy methods have been reported so far that produce high-resolution pure shift spectrum along with J-coupling information for crowded spectral regions. In the present work, high-quality J-resolved spectrum from important metabolomic mixture such as tissue extract from rat cortex is demonstrated. Many low-intensity metabolite peaks which are obscured by the broad dispersive tails from high-intensity metabolite peaks in regular magnitude mode J-spectrum can be clearly identified in real-time BIRD J-resolved spectrum. The general practice of removing such spectral overlap is tedious and time-consuming as it involves repeated sample preparation to change the pH of the tissue extract sample and subsequent spectra recording. PMID: 27026651 [PubMed - as supplied by publisher]

Diastolic Left Ventricular Function in Relation to Circulating Metabolic Biomarkers in a General Population. J Am Heart Assoc. 2016;5(3) Authors: Zhang ZY, Marrachelli VG, Thijs L, Yang WY, Wei FF, Monleon D, Jacobs L, Nawrot T, Verhamme P, Voigt JU, Kuznetsova T, Redón J, Staessen JA Abstract BACKGROUND: The metabolic signature associated with subclinical diastolic left ventricular (LV) dysfunction in the population remains ill defined. METHODS AND RESULTS: In 711 randomly recruited Flemish (50.8% women; mean age, 50.8 years), we assessed echocardiographic Doppler indexes of diastolic LV function in relation to 44 circulating metabolites determined by nuclear magnetic resonance spectroscopy. In multivariable-adjusted regression analysis with Bonferroni correction of significance levels applied, peak a' decreased (P≤0.048) and e'/a' increased (P≤0.044) with circulating tyrosine, high-density lipoprotein apolipoproteins, glucose+glutamine, and an unidentified molecule. Effect sizes expressed per 1-SD increment in the metabolite ranged from -0.277 to -0.203 cm/s for peak a' and from +0.047 to +0.054 for e'/a'. In addition, peak a' decreased (P≤0.031) with glucose+2-aminobutyrate (-0.261 cm/s) and glucose+2-phosphoglycerate (-0.209 cm/s). In partial least square discriminant analysis (PLS-DA), metabolites associated with normal diastolic LV function (n=538) included glucose+glutamine, glucose+2-aminobutyrate, and glucose+2-phosphoglycerate, whereas those siding with abnormal function encompassed 4-aminobutyrate, 4-hydroxybutyrate, creatinine, and phosphocholine. In receiver operating characteristics plots, adding 3 latent factors identified by PLS-DA to prohormone brain natriuretic peptide increased (P<0.0001) the area under the curve from 0.64 (95% CI, 0.58-0.68) to 0.73 (0.68-0.78). CONCLUSIONS: In a general population, circulating metabolites indicative of energy substrate utilization and protection against oxidative stress differentiated normal from abnormal diastolic LV function. These findings improve our understanding of the pathophysiology underlying deterioration of diastolic LV function and potentially point to new targets for prevention and treatment of this condition. PMID: 27025885 [PubMed - in process]

Plasma Metabolomics of Common Marmosets (Callithrix jacchus) to Evaluate Diet and Feeding Husbandry. J Am Assoc Lab Anim Sci. 2016;55(2):137-46 Authors: Banton SA, Soltow QA, Liu KH, Uppal K, Promislow DE, Power ML, Tardif SD, Wachtman LM, Jones DP Abstract Common marmosets (Callithrix jacchus) are an important NHP model for the study of human aging and age-related diseases. However, the full potential of marmosets as a research model has not been realized due to a lack of evidence-based, standardized procedures for their captive management, especially regarding diet and feeding husbandry. In the present study, we conducted a high-resolution metabolomics analysis of plasma from marmosets from a 3-mo dietary crossover study to determine whether significant metabolic differences occur with a semisynthetic chemically defined (purified) diet as needed for controlled nutrition research. Marmosets were fed a standard, diverse-ingredient diet, followed by a semisynthetic purified diet, and then were switched back to the standard diet. The standard diet used in this analysis was specific to the animal facility, but it is similar in content to the diets currently used for other marmoset colonies. High-resolution metabolomics of plasma with liquid chromatography-mass spectrometry and bioinformatics was used to measure metabolic differences. The concentration of the essential amino acids methionine, leucine/isoleucine, lysine, and threonine were higher when marmosets were fed the purified diet. In contrast, phenylalanine concentrations were higher during exposure to the standard diet. In addition, metabolic pathway enrichment and analysis revealed differences among metabolites associated with dopamine metabolism and the carnitine shuttle. These results show that diet-associated differences in metabolism occur in marmosets and suggest that additional nutritional studies with detailed physiologic characterization are needed to optimize standard and purified diets for common marmosets. PMID: 27025803 [PubMed - in process]

Urine metabonomic study for blood-replenishing mechanism of Angelica sinensis in a blood-deficient mouse model. Chin J Nat Med. 2016 Mar;14(3):210-9 Authors: Wang T, Sun HG, Hua YL, Li PL, Wei YM Abstract This study aimed at determining the effects of Angelica sinensis (AS) on urinary metabolites in blood deficiency mice and exploring its replenishing blood mechanism. Gas chromatography-mass spectrometry (GC-MS) was applied to detect metabolites in the urine samples in different collection periods. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to investigate the differences in metabolic profiles among control group (CG), blood deficiency model group (MG), AS groups, and Colla Corii Asini group (CCAG). The potential biomarkers were identified based on the variable importance in the projection (VIP), T-test, and National Institute of Standards and Technology (NIST) and mass spectra library. The metabolites were analyzed using metabolomics pathway analysis (MetPA) to build the metabolic pathways. Our results indicated that, on the seventh day, the levels of glucose, lactic acid, pyruvic acid, alanine, acetoacetic acid, and citric acid changed significantly in blood deficiency mice. However, these metabolic deviations came to closer to normal levels after AS intervention. The reversing blood-deficiency mechanism of AS might involve regulating synthesis and degradation of ketone bodies, Pyruvate metabolism, TCA cycle, and Glycolysis/Gluconeogenesis. In conclusion, metabonomics is a robust and promising means for the identification of biomarkers and elucidation of the mechanisms of a disease, thereby highlighting its importance in drug discovery. PMID: 27025368 [PubMed - in process]

Related Articles Comparative Metabolomic Analysis of the Neuroprotective Effects of Scutellarin and Scutellarein against Ischemic Insult. PLoS One. 2015;10(7):e0131569 Authors: Tang H, Tang Y, Li NG, Lin H, Li W, Shi Q, Zhang W, Zhang P, Dong Z, Shen M, Gu T, Duan JA Abstract For more than thirty years, scutellarin (Scu) has been used in China to clinically treat acute cerebral infarction and paralysis. Scutellarein (Scue), the major Scu metabolite in vivo, exhibits heightened neuroprotective effects when compared to Scu. To explore the neuroprotective role of these compounds, we performed ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UHPLC-QTOF/MS) coupled with a pattern recognition approach to investigate metabolomic differences in a rat model of ischemia after treatment with each compound. We examined metabolites in urine, hippocampal tissue, and plasma, and we tentatively identified 23 endogenous metabolites whose levels differed significantly between sham-operated and model groups. Upon pathway analysis, we found an additional 11 metabolic pathways in urine, 14 metabolic pathways in the hippocampal tissue, and 3 metabolic pathways in plasma. These endogenous metabolites were mainly involved in sphingolipid metabolism, lysine biosynthesis, and alanine, aspartate, and glutamate metabolism. We found that metabolic changes after ischemic injury returned to near-normal levels after Scue intervention, unlike Scu treatment, further validating the heightened protective effects exerted by Scue compared to Scu. These results demonstrate that Scue is a potential drug for treatment of ischemic insult. PMID: 26147971 [PubMed - indexed for MEDLINE]

Related Articles Urine and serum metabolomic profiling reveals that bile acids and carnitine may be potential biomarkers of primary biliary cirrhosis. Int J Mol Med. 2015 Aug;36(2):377-85 Authors: Tang YM, Wang JP, Bao WM, Yang JH, Ma LK, Yang J, Chen H, Xu Y, Yang LH, Li W, Zhu YP, Cheng JB Abstract In order to provide non-invasive, reliable and sensitive laboratory parameters for the diagnosis of primary biliary cirrhosis (PBC), metabolic technology of ultraperformance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used to compare small molecule metabolites in blood and urine from patients with PBC and healthy controls. We then screened for bio-markers in the blood and urine of the patients with PBC. Data were processed by Bruker ProfileAnalysis metabonomic software and imported to SIMCA-P software, which utilized principal component analysis (PCA) to create models of patients with PBC and healthy controls. In total, 18 urinary markers were found and the levels of 11 of these urinary markers were elevated in the patients with PBC, whereas the levels of the remaining 7 markers were lower in the PBC group compared to the control group. We also identified 20 blood-based biomarkers in the patients with PBC and the levels of 9 of these markers were higher in the PBC group, whereas the levels of the remaining 11 markers were lower in the patients with PBC compared to the controls. Among these biomarkers, the levels of bile acids increased with the progression of PBC, while the levels of carnitines, such as propionyl carnitine and butyryl carnitine, decreased with the progression of PBC. In conclusion, the findings of the present study suggest that the circulating levels of bile acids and carnitine are differentially altered in patients with PBC. PMID: 26046127 [PubMed - indexed for MEDLINE]

Related Articles Identification and mode of inheritance of quantitative trait loci for secondary metabolite abundance in tomato. Plant Cell. 2015 Mar;27(3):485-512 Authors: Alseekh S, Tohge T, Wendenberg R, Scossa F, Omranian N, Li J, Kleessen S, Giavalisco P, Pleban T, Mueller-Roeber B, Zamir D, Nikoloski Z, Fernie AR Abstract A large-scale metabolic quantitative trait loci (mQTL) analysis was performed on the well-characterized Solanum pennellii introgression lines to investigate the genomic regions associated with secondary metabolism in tomato fruit pericarp. In total, 679 mQTLs were detected across the 76 introgression lines. Heritability analyses revealed that mQTLs of secondary metabolism were less affected by environment than mQTLs of primary metabolism. Network analysis allowed us to assess the interconnectivity of primary and secondary metabolism as well as to compare and contrast their respective associations with morphological traits. Additionally, we applied a recently established real-time quantitative PCR platform to gain insight into transcriptional control mechanisms of a subset of the mQTLs, including those for hydroxycinnamates, acyl-sugar, naringenin chalcone, and a range of glycoalkaloids. Intriguingly, many of these compounds displayed a dominant-negative mode of inheritance, which is contrary to the conventional wisdom that secondary metabolite contents decreased on domestication. We additionally performed an exemplary evaluation of two candidate genes for glycolalkaloid mQTLs via the use of virus-induced gene silencing. The combined data of this study were compared with previous results on primary metabolism obtained from the same material and to other studies of natural variance of secondary metabolism. PMID: 25770107 [PubMed - indexed for MEDLINE]