Integrative Molecular Phenotyping
INTEGRATIVE MOLECULAR
PHENOTYPING
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY

PubMed

A Web Service Framework for Interactive Analysis of Metabolomics Data.

Thu, 18/05/2017 - 13:40
Related Articles A Web Service Framework for Interactive Analysis of Metabolomics Data. Anal Chem. 2017 May 17;: Authors: Lyutvinskiy Y, Watrous JD, Jain M, Nilsson R Abstract Analyzing mass spectrometry-based metabolomics data presents a major challenge to metabolism researchers, as it requires downloading and processing large data volumes through complex "pipelines", even in cases where only a single metabolite or peak is of interest. This presents a significant hurdle for data sharing, reanalysis, or meta-analysis of existing data sets, whether locally stored or available from public repositories. Here we introduce mzAccess, a software system that provides interactive, online access to primary mass spectrometry data in real-time via a Web service protocol, circumventing the need for bulk data processing. mzAccess allows querying instrument data for spectra, chromatograms, or two-dimensional MZ-RT areas in either profile or centroid modes through a simple, uniform interface that is independent of vendor or instrument type. Using a cache mechanism, mzAccess achieves response times in the millisecond range for typical liquid chromatography-mass spectrometry (LC-MS) peaks, enabling real-time browsing of large data sets with hundreds or even thousands of samples. By simplifying access to metabolite data, we hope that this system will help enable data sharing and reanalysis in the metabolomics field. PMID: 28514166 [PubMed - as supplied by publisher]

Body Composition Analysis by Using Bioelectrical Impedance in a Young Healthy Chinese Population: Methodological Considerations.

Thu, 18/05/2017 - 13:40
Related Articles Body Composition Analysis by Using Bioelectrical Impedance in a Young Healthy Chinese Population: Methodological Considerations. Food Nutr Bull. 2017 Jun;38(2):172-181 Authors: Chen W, Jiang H, Yang JX, Yang H, Liu JM, Zhen XY, Feng LJ, Yu JC Abstract OBJECTIVE: To develop a new bioelectrical impedance analysis (BIA) model for analyzing body composition by using isotope dilution, magnetic resonance imaging (MRI), and dual-energy X-ray absorptiometry (DEXA) as the reference methods in young healthy Chinese populations. METHODS: Thirty healthy participants were enrolled. Their body composition was analyzed using BIA and 3 reference methods. We established a model that uniformed data from 3 references methods (isotope, MRI, and DEXA) into 1 formula. This model was further validated with 209 participants. RESULTS: The following BIA body composition adjustment model was developed: [Formula: see text], where X represents the impedance index; when K = 1, 2, and 3, Y represents total body water, fat mass, and bone mass, respectively. The prediction accuracy of this formula was 93.3%. By incorporating the data matrix, the protein mass was calculated using BIA: [Formula: see text]. In the verification part of this study, the lean body mass measured using DEXA and BIA was 43.02 ± 8.34 kg and 45.85 ± 8.81 kg, respectively. Analysis indicated that the model fit was extremely favorable ( R(2) = .9997, P < .001). CONCLUSIONS: The accuracy of BIA measurement on body composition and protein mass is significantly improved by our work. PMID: 28513264 [PubMed - in process]

Metabolic Perturbation and Potential Markers in Patients with Esophageal Cancer.

Thu, 18/05/2017 - 13:40
Related Articles Metabolic Perturbation and Potential Markers in Patients with Esophageal Cancer. Gastroenterol Res Pract. 2017;2017:5469597 Authors: Zhu X, Wang K, Liu G, Wang Y, Xu J, Liu L, Li M, Shi J, Aa J, Yu L Abstract Clinical diagnosis of esophageal cancer (EC) at early stage is rather difficult. This study aimed to profile the molecules in serum and tissue and identify potential biomarkers in patients with EC. A total of 64 volunteers were recruited, and 83 samples (24 EC serum samples, 21 serum controls, 19 paired EC tissues, and corresponding tumor-adjacent tissues) were analyzed. The gas chromatography time-of-flight mass spectrometry (GC/TOF-MS) was employed, and principal component analysis was used to reveal the discriminatory metabolites and identify the candidate markers of EC. A total of 41 in serum and 36 identified compounds in tissues were relevant to the malignant prognosis. A marked metabolic reprogramming of EC was observed, including enhanced anaerobic glycolysis and glutaminolysis, inhibited tricarboxylic acid (TCA) cycle, and altered lipid metabolism and amino acid turnover. Based on the potential markers of glucose, glutamic acid, lactic acid, and cholesterol, the receiver operating characteristic (ROC) curves indicated good diagnosis and prognosis of EC. EC patients showed distinct reprogrammed metabolism involved in glycolysis, TCA cycle, glutaminolysis, and fatty acid metabolism. The pivotal molecules in the metabolic pathways were suggested as the potential markers to facilitate the early diagnosis of human EC. PMID: 28512469 [PubMed - in process]

GC-MS Metabolomic Analysis to Reveal the Metabolites and Biological Pathways Involved in the Developmental Stages and Tissue Response of Panax ginseng.

Thu, 18/05/2017 - 13:40
Related Articles GC-MS Metabolomic Analysis to Reveal the Metabolites and Biological Pathways Involved in the Developmental Stages and Tissue Response of Panax ginseng. Molecules. 2017 Mar 21;22(3): Authors: Liu J, Liu Y, Wang Y, Abozeid A, Zu YG, Zhang XN, Tang ZH Abstract Ginsenosides, the major compounds present in ginseng, are known to have numerous physiological and pharmacological effects. The physiological processes, enzymes and genes involved in ginsenoside synthesis in P. ginseng have been well characterized. However, relatively little information is known about the dynamic metabolic changes that occur during ginsenoside accumulation in ginseng. To explore this topic, we isolated metabolites from different tissues at different growth stages, and identified and characterized them by using gas chromatography coupled with mass spectrometry (GC-MS). The results showed that a total of 30, 16, 20, 36 and 31 metabolites were identified and involved in different developmental stages in leaf, stem, petiole, lateral root and main root, respectively. To investigate the contribution of tissue to the biosynthesis of ginsenosides, we examined the metabolic changes of leaf, stem, petiole, lateral root and main root during five development stages: 1-, 2-, 3-, 4- and 5-years. The score plots of partial least squares-discriminate analysis (PLS-DA) showed clear discrimination between growth stages and tissue samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis in the same tissue at different growth stages indicated profound biochemical changes in several pathways, including carbohydrate metabolism and pentose phosphate metabolism, in addition, the tissues displayed significant variations in amino acid metabolism, sugar metabolism and energy metabolism. These results should facilitate further dissection of the metabolic flux regulation of ginsenoside accumulation in different developmental stages or different tissues of ginseng. PMID: 28335577 [PubMed - indexed for MEDLINE]

Profiling and Preparation of Metabolites from Pyragrel in Human Urine by Online Solid-Phase Extraction Coupled with High Performance Liquid Chromatography Tandem Mass Spectrometry Followed by a Macroporous Resin-Based Purification Approach.

Thu, 18/05/2017 - 13:40
Related Articles Profiling and Preparation of Metabolites from Pyragrel in Human Urine by Online Solid-Phase Extraction Coupled with High Performance Liquid Chromatography Tandem Mass Spectrometry Followed by a Macroporous Resin-Based Purification Approach. Molecules. 2017 Mar 21;22(3): Authors: Zhao X, Jiang J, Yang G, Huang J, Yang G, He G, Chu Z, Hang T, Fan G Abstract Pyragrel, a new anticoagulant drug, is derived from the molecular combination of ligustrazine and ferulic acid. Pyragrel showed significant inhibitory activity against platelet aggregation induced by adenosine diphosphate (ADP), and had been approved for a phase I clinical trial by CFDA. To characterize the metabolites of Pyragrel in human urine after intravenous administration, a reliable online solid-phase extraction couple with high performance liquid chromatography tandem mass spectrometry (online SPE-HPLC-MS(n)) method was conceived and applied. Five metabolites were detected and tentatively identified, which suggested that the major metabolic pathways of Pyragrel in human were double-bond reduction, double-bond oxidation, and then followed by glucuronide conjugation. Two main metabolites were then prepared using β-glucuronide hydrolysis and macroporous resin purification approach followed by preparative high-performance liquid chromatography (PHPLC) method, with their structures confirmed on the basis of nuclear magnetic resonance (NMR) data. This study provided information for the further study of the metabolism and excretion of Pyragrel. PMID: 28335566 [PubMed - indexed for MEDLINE]

Monitoring of pistachio (Pistacia Vera) ripening by high field nuclear magnetic resonance spectroscopy.

Thu, 18/05/2017 - 13:40
Related Articles Monitoring of pistachio (Pistacia Vera) ripening by high field nuclear magnetic resonance spectroscopy. Nat Prod Res. 2017 Apr;31(7):765-772 Authors: Sciubba F, Avanzato D, Vaccaro A, Capuani G, Spagnoli M, Di Cocco ME, Tzareva IN, Delfini M Abstract The metabolic profiling of pistachio (Pistacia vera) aqueous extracts from two different cultivars, namely 'Bianca' and 'Gloria', was monitored over the months from May to September employing high field NMR spectroscopy. A large number of water-soluble metabolites were assigned by means of 1D and 2D NMR experiments. The change in the metabolic profiles monitored over time allowed the pistachio development to be investigated. Specific temporal trends of amino acids, sugars, organic acids and other metabolites were observed and analysed by multivariate Partial Least Squares (PLS) analysis. Statistical analysis showed that while in the period from May to September there were few differences between the two cultivars, the ripening rate was different. PMID: 27712103 [PubMed - indexed for MEDLINE]

Metabolomic and Genome-wide Association Studies Reveal Potential Endogenous Biomarkers for OATP1B1.

Thu, 18/05/2017 - 13:40
Related Articles Metabolomic and Genome-wide Association Studies Reveal Potential Endogenous Biomarkers for OATP1B1. Clin Pharmacol Ther. 2016 Nov;100(5):524-536 Authors: Yee SW, Giacomini MM, Hsueh CH, Weitz D, Liang X, Goswami S, Kinchen JM, Coelho A, Zur AA, Mertsch K, Brian W, Kroetz DL, Giacomini KM Abstract Transporter-mediated drug-drug interactions (DDIs) are a major cause of drug toxicities. Using published genome-wide association studies (GWAS) of the human metabolome, we identified 20 metabolites associated with genetic variants in organic anion transporter, OATP1B1 (P < 5 × 10(-8) ). Of these, 12 metabolites were significantly higher in plasma samples from volunteers dosed with the OATP1B1 inhibitor, cyclosporine (CSA) vs. placebo (q-value < 0.2). Conjugated bile acids and fatty acid dicarboxylates were among the metabolites discovered using both GWAS and CSA administration. In vitro studies confirmed tetradecanedioate (TDA) and hexadecanedioate (HDA) were novel substrates of OATP1B1 as well as OAT1 and OAT3. This study highlights the use of multiple datasets for the discovery of endogenous metabolites that represent potential in vivo biomarkers for transporter-mediated DDIs. Future studies are needed to determine whether these metabolites can serve as qualified biomarkers for organic anion transporters. Quantitative relationships between metabolite levels and modulation of transporters should be established. PMID: 27447836 [PubMed - indexed for MEDLINE]

Ozone Exposure Increases Circulating Stress Hormones and Lipid Metabolites in Humans.

Thu, 18/05/2017 - 13:40
Related Articles Ozone Exposure Increases Circulating Stress Hormones and Lipid Metabolites in Humans. Am J Respir Crit Care Med. 2016 Jun 15;193(12):1382-91 Authors: Miller DB, Ghio AJ, Karoly ED, Bell LN, Snow SJ, Madden MC, Soukup J, Cascio WE, Gilmour MI, Kodavanti UP Abstract RATIONALE: Air pollution has been associated with increased prevalence of type 2 diabetes; however, the mechanisms remain unknown. We have shown that acute ozone exposure in rats induces release of stress hormones, hyperglycemia, leptinemia, and glucose intolerance that are associated with global changes in peripheral glucose, lipid, and amino acid metabolism. OBJECTIVES: To examine ozone-induced metabolic derangement in humans using serum metabolomic assessment, establish human-to-rodent coherence, and identify novel nonprotein biomarkers. METHODS: Serum samples were obtained from a crossover clinical study that included two clinic visits (n = 24 each) where each subject was blindly exposed in the morning to either filtered air or 0.3 parts per million ozone for 2 hours during 15-minute on-off exercise. Serum samples collected within 1 hour after exposure were assessed for changes in metabolites using a metabolomic approach. MEASUREMENTS AND MAIN RESULTS: Metabolomic analysis revealed that ozone exposure markedly increased serum cortisol and corticosterone together with increases in monoacylglycerol, glycerol, and medium- and long-chain free fatty acids, reflective of lipid mobilization and catabolism. Additionally, ozone exposure increased serum lysolipids, potentially originating from membrane lipid breakdown. Ozone exposure also increased circulating mitochondrial β-oxidation-derived metabolites, such as acylcarnitines, together with increases in the ketone body 3-hydroxybutyrate. These changes suggested saturation of β-oxidation by ozone in exercising humans. CONCLUSIONS: As in rodents, acute ozone exposure increased stress hormones and globally altered peripheral lipid metabolism in humans, likely through activation of a neurohormonally mediated stress response pathway. The metabolomic assessment revealed new biomarkers and allowed for establishment of rodent-to-human coherence. Clinical trial registered with www.clinicaltrials.gov (NCT 01492517). PMID: 26745856 [PubMed - indexed for MEDLINE]

metabolomics; +18 new citations

Wed, 17/05/2017 - 12:57
18 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2017/05/17PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

metabolomics; +24 new citations

Tue, 16/05/2017 - 12:17
24 new pubmed citations were retrieved for your search. Click on the search hyperlink below to display the complete search results: metabolomics These pubmed results were generated on 2017/05/16PubMed comprises more than millions of citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.

Metabolomic Bioinformatic Analysis.

Mon, 15/05/2017 - 21:13
Related Articles Metabolomic Bioinformatic Analysis. Methods Mol Biol. 2017;1606:341-352 Authors: Dailey AL Abstract Metabolomics allows for the investigation of the small molecules found within living systems. Based on the design of the experiments, it is not uncommon for these analyses to include matrices of thousands of variables. In order to handle such large datasets, many have turned to multivariate statistical analyses to analyze and understand their data. Herein, we present protocols for using R to analyze metabolomic data using some of the more common multivariate statistical techniques. PMID: 28502011 [PubMed - in process]

LC-Mass Spectrometry for Metabolomics.

Mon, 15/05/2017 - 21:13
Related Articles LC-Mass Spectrometry for Metabolomics. Methods Mol Biol. 2017;1606:333-340 Authors: Dailey AL Abstract The field of metabolomics is greatly being refined by the addition of new technologies. LC-MS has allowed researchers to explore additional metabolites which were not originally captured through GC-MS. Through the customizability of the LC columns and mass spectrometer, it is now easier to tailor the instrument to your research needs. Herein, we describe a protocol for sample preparation and data acquisition for a global metabolomic analysis of tissues or feces. PMID: 28502010 [PubMed - in process]

Implementing liquid biopsies into clinical decision making for cancer immunotherapy.

Mon, 15/05/2017 - 21:13
Related Articles Implementing liquid biopsies into clinical decision making for cancer immunotherapy. Oncotarget. 2017 Apr 24;: Authors: Quandt D, Zucht HD, Amann A, Wulf-Goldenberg A, Borrebaeck C, Cannarile M, Lambrechts D, Oberacher H, Garrett J, Nayak T, Kazinski M, Massie C, Schwarzenbach H, Maio M, Prins R, Wendik B, Hockett R, Enderle D, Noerholm M, Hendriks H, Zwierzina H, Seliger B Abstract During the last decade, novel immunotherapeutic strategies, in particular antibodies directed against immune checkpoint inhibitors, have revolutionized the treatment of different malignancies leading to an improved survival of patients. Identification of immune-related biomarkers for diagnosis, prognosis, monitoring of immune responses and selection of patients for specific cancer immunotherapies is urgently required and therefore areas of intensive research. Easily accessible samples in particular liquid biopsies (body fluids), such as blood, saliva or urine, are preferred for serial tumor biopsies.Although monitoring of immune and tumor responses prior, during and post immunotherapy has led to significant advances of patients' outcome, valid and stable prognostic biomarkers are still missing. This might be due to the limited capacity of the technologies employed, reproducibility of results as well as assay stability and validation of results. Therefore solid approaches to assess immune regulation and modulation as well as to follow up the nature of the tumor in liquid biopsies are urgently required to discover valuable and relevant biomarkers including sample preparation, timing of the collection and the type of liquid samples. This article summarizes our knowledge of the well-known liquid material in a new context as liquid biopsy and focuses on collection and assay requirements for the analysis and the technical developments that allow the implementation of different high-throughput assays to detect alterations at the genetic and immunologic level, which could be used for monitoring treatment efficiency, acquired therapy resistance mechanisms and the prognostic value of the liquid biopsies. PMID: 28501851 [PubMed - as supplied by publisher]

Obesity aggravates toxic effect of BPA on spermatogenesis.

Mon, 15/05/2017 - 21:13
Related Articles Obesity aggravates toxic effect of BPA on spermatogenesis. Environ Int. 2017 May 11;105:56-65 Authors: Hu W, Dong T, Wang L, Guan Q, Song L, Chen D, Zhou Z, Chen M, Xia Y, Wang X Abstract Both bisphenol A (BPA) and obesity affect male reproductive system. However, whether there is an interaction between them remains poorly understood. The aim of the present study was to evaluate the interaction between BPA exposure and obesity on semen quality and elucidate the mechanism in humans and animals. We firstly analyzed the interaction on semen volume, sperm count per ejaculate, sperm concentration and sperm motility in 357 men, and found that urinary BPA concentration was significantly correlated with sperm count per ejaculate in obese men (β=-34.62; 95% CI: -60.75, -8.48; P=0.01). Then we validated the interaction using lean and obese mice with administration of BPA. Significant interactions between BPA exposure and obesity on sperm count and sperm concentration was observed in mice. Finally, we conducted metabolomics analyses to identify metabolites related to the interaction. Metabolites related to the interaction, including capric acid, dodecanoic acid, l-palmitoylcarnitine, niacinamide, etc., are known to play critical roles in fatty acid oxidation and tricarboxylic acid cycle indicating increased oxidative stress associated with male reproductive dysfunction. Thus, our study finds an interaction between BPA exposure and obesity on sperm count and reveals potential metabolic mechanisms. It emphasizes the importance to study interactions between endocrine disrupting chemicals and obesity, and opens avenues for the possible use of animal models in identifying the interactions. PMID: 28501790 [PubMed - as supplied by publisher]

Metabolite profile of koji amazake and its lactic acid fermentation product by Lactobacillus sakei UONUMA.

Mon, 15/05/2017 - 21:13
Related Articles Metabolite profile of koji amazake and its lactic acid fermentation product by Lactobacillus sakei UONUMA. J Biosci Bioeng. 2017 May 10;: Authors: Oguro Y, Nishiwaki T, Shinada R, Kobayashi K, Kurahashi A Abstract The koji amazake is a traditional sweet Japanese beverage. It has been consumed for over a thousand years in Japan; nonetheless, little is yet known of the ingredients in koji amazake. Therefore, this study aimed to analyze the metabolites of koji amazake using a metabolomics approach. Additionally, we reformed the flavor of koji amazake by lactic acid fermentation (LAF-amazake) using Lactobacillus sakei UONUMA, which was isolated from snow caverns. The purpose of this article is to identify the ingredients in these beverages. In LAF-amazake and koji amazake, sugars, amino acids, organic acids, and vitamin B complex were determined in the two beverages, and over 300 compounds were detected in total. Thirteen saccharides were identified including two unknown trisaccharides, and there were no differences in these between the two beverages. In LAF-amazake, lactic acid, vitamin B2 (riboflavin), B3 (nicotinic acid and nicotinamide), and B6 (pyridoxine) were significantly increased as compared to koji amazake, whereas malate and glutamine decreased. These results suggested that LAF, malolactic fermentation, and glutamine deamidation occurred simultaneously in LAF-amazake. L. sakei UONUMA strains produced these vitamins. Moreover, it was surprising that acetylcholine, a well-known neurotransmitter, was newly generated in LAF-amazake. Here, we have succeeded in reforming the flavor of koji amazake and obtained these metabolic data on the two beverages. The present study could provide useful basic information for promoting functional analyses of koji amazake and LAF-amazake for human health. PMID: 28501542 [PubMed - as supplied by publisher]

Lipase-catalyzed kinetic resolution as key step in the synthesis of enantiomerically pure σ ligands with 2-benzopyran structure.

Mon, 15/05/2017 - 21:13
Related Articles Lipase-catalyzed kinetic resolution as key step in the synthesis of enantiomerically pure σ ligands with 2-benzopyran structure. Bioorg Med Chem. 2017 Apr 30;: Authors: Knappmann I, Lehmkuhl K, Köhler J, Schepmann D, Giera M, Bracher F, Wünsch B Abstract In order to obtain enantiomerically pure σ1 receptor ligands with a 2-benzopyran scaffold an Oxa-Pictet-Spengler reaction with the enantiomerically pure 2-phenylethanol derivatives (R)-4 and (S)-4 was envisaged. The kinetic resolution of racemic alcohol (±)-4 using Amano Lipase PS-C II and isopropenyl acetate in tert-butyl methyl ether led to the (R)-configured alcohol (R)-4 in 42% yield with an enantiomeric excess of 99.6%. The (S)-configured alcohol (S)-4 was obtained by Amano Lipase PS-C II catalyzed hydrolysis of enantiomerically enriched acetate (S)-5 (76.9% ee) and provided (S)-4 in 26% yield and 99.7% ee. The absolute configuration of alcohol (R)-4 was determined by exciton coupled CD spectroscopy of the bis(bromobenzoate) (R)-7. The next important step for the synthesis of 2-benzopyrans 2 and 3 was the Oxa-Pictet-Spengler reaction of the enantiomerically pure alcohols (R)-4 and (S)-4 with piperidone ketal 8 and chloropropionaldehyde acetal 12. The conformationally restricted spirocyclic 2-benzopyrans 2 revealed higher σ1 affinity than the more flexible aminoethyl derivatives 3. The (R)- and (R,R)-configured enantiomers (R)-2 and (R,R)-3 represent the eutomers of this class of compounds with eudismic ratios of 4.8 (2b) and 4.5 (2c). High σ1/σ2 selectivity (>49) was found for the most potent σ1 ligands (R)-2b, (R)-2c, (R)-2d, and (S)-2d (Ki(σ1) 9-15nM). PMID: 28501431 [PubMed - as supplied by publisher]

Capillary electrophoresis tandem mass spectrometry determination of glutamic acid and homocysteine's metabolites: Potential biomarkers of amyotrophic lateral sclerosis.

Mon, 15/05/2017 - 21:13
Related Articles Capillary electrophoresis tandem mass spectrometry determination of glutamic acid and homocysteine's metabolites: Potential biomarkers of amyotrophic lateral sclerosis. Talanta. 2017 Aug 01;170:63-68 Authors: Cieslarova Z, Lopes FS, do Lago CL, França MC, Colnaghi Simionato AV Abstract Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease that affects both lower and upper motor neurons, leading to muscle atrophy, paralysis, and death caused by respiratory failure or infectious complications. Altered levels of homocysteine, cysteine, methionine, and glutamic acid have been observed in plasma of ALS patients. In this context, a method for determination of these potential biomarkers in plasma by capillary electrophoresis tandem mass spectrometry (CE-MS/MS) is proposed herein. Sample preparation was carefully investigated, since sulfur-containing amino acids may interact with plasma proteins. Owing to the non-thiol sulfur atom in methionine, it was necessary to split sample preparation into two methods: i) determination of homocysteine and cysteine as S-acetyl amino acids; ii) determination of glutamic acid and methionine. All amino acids were separated within 25min by CE-MS/MS using 5molL(-1) acetic acid as background electrolyte and 5mmolL(-1) acetic acid in 50% methanol/H2O (v/v) as sheath liquid. The proposed CE-MS/MS method was validated, presenting RSD values below 6% and 11% for intra- and inter-day precision, respectively, for the middle concentration level within the linear range. The limits of detection ranged from 35 (homocysteine) to 268nmolL(-1) (glutamic acid). The validated method was applied to the analysis of plasma samples from a group of healthy individuals and patients with ALS, showing the potential of glutamic acid and homocysteine metabolites as biomarkers of ALS. PMID: 28501214 [PubMed - in process]

Serum metabolomics reveals the mechanistic role of functional foods and exercise for obesity management in rats.

Sun, 14/05/2017 - 14:48
Related Articles Serum metabolomics reveals the mechanistic role of functional foods and exercise for obesity management in rats. J Pharm Biomed Anal. 2017 May 01;142:91-101 Authors: Ammar NM, Farag MA, Kholeif TE, Metwally NS, El-Sheikh NM, El Gendy AN, Abdel-Hamid AZ Abstract Obesity is one of the independent risk factors for several health problems, leading to metabolic perturbations and for which analytical approaches i.e., "metabolomics" is needed to monitor the underlying metabolic changes. In this study, obesity associated changes were assessed via serum metabolites analysis of obese rats fed on high fat diet. Obese rats were subsequently treated with different functional foods used for obesity management including pomegranate, grapefruit, and red cabbage in parallel to swimming exercise. Serum samples were analyzed using gas chromatography-mass spectrometry (GC-MS) followed by multivariate data analysis to classify samples and determine if such treatments can help revert obesity related metabolic changes back to normal status. Results led to the identification of several novel metabolites biomarkers for obesity related to lipids, amino acids and central tricarboxylic acid (TCA) pathways. Distinct variations in metabolite levels were recorded in obese rats compared to normal ones including l-aspartic, l-alanine, l-glutamine, l-glycine, phenylethanolamine, α-aminobutyric acid and β-hydroxybutyric acid. Metabolomics approach developed herein provides novel insight onto the metabolic disturbances associated with obesity, which will assist in future drug design that can help mitigate against such changes. PMID: 28500980 [PubMed - as supplied by publisher]

High throughput metabolic profiling based on small amount of hepatic cells.

Sun, 14/05/2017 - 14:48
Related Articles High throughput metabolic profiling based on small amount of hepatic cells. Electrophoresis. 2017 May 12;: Authors: Zhou L, Yin P, Luo P, Tang L, Wang Z, Gao P, Piao H, Lu X, Xu G Abstract Common metabolomics platforms require about 10(6) cells, which has a limited throughput due to the time-consuming steps of cell culture and preparation. There is a demand for metabolic profiling methods to improve analytical throughput and detection sensitivity based on small amount of cells. In this study, we proposed a high-throughput scheme, integrating 96-well plate cell cultivation, in-situ cell pretreatment and sensitive dansylation labelling coupled with liquid chromatography-mass spectrometry analysis of metabolites inside hepG2 cells (of the order of magnitude of 10(3) cells in each well). A simple and rapid cell pretreatment was performed showing good extraction efficiency and good precision (the relative standard deviations smaller than 5%) for polar metabolites. The recovery in metabolite extraction evaluated with three isotope-labeled amino acids was from 89.7% to 106.3% at low, medium and high concentrations. The suitability of the method was illustrated by exploring influences of different fatty acids on HepG2 cells. This article is protected by copyright. All rights reserved. PMID: 28500646 [PubMed - as supplied by publisher]

Comparative Metabolomic Analysis of the Green Microalga Chlorella sorokiniana Cultivated in the Single Culture and a Consortium with Bacteria for Wastewater Remediation.

Sun, 14/05/2017 - 14:48
Related Articles Comparative Metabolomic Analysis of the Green Microalga Chlorella sorokiniana Cultivated in the Single Culture and a Consortium with Bacteria for Wastewater Remediation. Appl Biochem Biotechnol. 2017 May 12;: Authors: Chen T, Zhao Q, Wang L, Xu Y, Wei W Abstract Co-culture of microalgae with many types of bacteria usually comes out with significant different treatment efficiencies for COD, nitrogen, and phosphorus in wastewater remediation, compared with the single culture. In order to understand the mechanism behind, a comparative experiment was designed in this study, using the green microalgae species Chlorella sorokiniana in the single culture and a consortium with a bacterium, Pseudomonas H4, for nutrient removal. Comparative metabolome profile analysis was conducted to reveal the Chlorella cell responses to the synergistic growth with the bacteria, and possible relations between the metabolic regulation of microalgae and the nutrient degradation were discussed. The detectable differential metabolites of Chlorella belonged to several classes, including carbohydrates, fatty acids, amino acids, phosphates, polyols, etc. The orthogonal partial least squares discriminant analysis (OPLS-DA) model of the identified metabolites suggests the metabolism in this alga was significantly affected by the bacteria, corresponding to different treatment behaviors. PMID: 28500414 [PubMed - as supplied by publisher]

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