Metabolomics Applied to Diabetes Research

Type 2 diabetes is caused by a complex set of interactions between genetic and environmental factors. Recent work has shown that human type 2 diabetes is a constellation of disorders associated with polymorphisms in a wide array of genes, with each individual gene accounting for <1% of disease risk (1). Moreover, type 2 diabetes involves dysfunction of multiple organ systems, including impaired insulin action in muscle and adipose, defective control of hepatic glucose production, and insulin deficiency caused by loss of β-cell mass and function (2). This complexity presents challenges for a full understanding of the molecular pathways that contribute to the development of this major disease. Progress in this area may be aided by the recent advent of technologies for comprehensive metabolic analysis, sometimes termed “metabolomics.” Herein, we summarize key metabolomics methodologies, including nuclear magnetic resonance (NMR) and mass spectrometry (MS)-based metabolic profiling technologies, and discuss “nontargeted” versus “targeted” approaches. Examples of the application of these tools to diabetes and metabolic disease research at the cellular, animal model, and human disease levels are summarized, with a particular focus on insights gained from the more quantitative targeted methodologies. We also provide early examples of integrated analysis of genomic, transcriptomic, and metabolomic datasets for gaining knowledge about metabolic regulatory networks and diabetes mechanisms and conclude by discussing prospects for future insights. In principal, metabolomics can provide certain advantages relative to other “omics” technologies (genomics, transcriptomics, proteomics) in diabetes research: 1 ) Estimates vary, but one current source, the Human Metabolome Database (HMDB)-Canada (3), currently lists ∼6,500 discrete small molecule metabolites, significantly less than the estimate of 25,000 genes, 100,000 transcripts, and 1,000,000 proteins. 2 ) Metabolomics measures chemical phenotypes that are the net result of genomic, transcriptomic, and proteomic variability, therefore providing the most integrated profile of biological status. 3 ) Metabolomics is in …

[1]  M. Jensen,et al.  The Mitochondrial Citrate/Isocitrate Carrier Plays a Regulatory Role in Glucose-stimulated Insulin Secretion* , 2006, Journal of Biological Chemistry.

[2]  David I. Ellis,et al.  Serum metabolomics reveals many novel metabolic markers of heart failure, including pseudouridine and 2-oxoglutarate , 2007, Metabolomics.

[3]  W. R. Morrison,et al.  PREPARATION OF FATTY ACID METHYL ESTERS AND DIMETHYLACETALS FROM LIPIDS WITH BORON FLUORIDE--METHANOL. , 1964, Journal of lipid research.

[4]  B. Wisely,et al.  Monitoring of PPAR alpha protein expression in human tissue by the use of PPAR alpha-specific MAbs. , 1998, Hybridoma.

[5]  D. Millington,et al.  The tandem mass spectrometry newborn screening experience in North Carolina: 1997–2005 , 2006, Journal of Inherited Metabolic Disease.

[6]  Xianlin Han,et al.  Global analyses of cellular lipidomes directly from crude extracts of biological samples by ESI mass spectrometry: a bridge to lipidomics. , 2003, Journal of lipid research.

[7]  W. Krone,et al.  Rapid Conversion by Insulin of Hepatic Intermediary Metabolism from Glucose Production to Glucose Utilization in the Liver of Alloxan-diabetic Rats , 1977, Diabetes.

[8]  Zugen Chen,et al.  Structural Analysis of the Regulatory Dithiol-containing Domain of the Chloroplast ATP Synthase γ Subunit* , 2006, Journal of Biological Chemistry.

[9]  W. Wahli,et al.  Peroxisome proliferator-activated receptors: nuclear control of metabolism. , 1999, Endocrine reviews.

[10]  K. Kaski,et al.  1H NMR metabonomics approach to the disease continuum of diabetic complications and premature death , 2008, Molecular systems biology.

[11]  J. Holloszy,et al.  A potential link between muscle peroxisome proliferator- activated receptor-alpha signaling and obesity-related diabetes. , 2005, Cell metabolism.

[12]  Julian L Griffin,et al.  Proton NMR analysis of plasma is a weak predictor of coronary artery disease , 2006, Nature Medicine.

[13]  A. Goodridge,et al.  Physiological and molecular mechanisms involved in nutritional regulation of fatty acid synthesis. , 1995, Physiological reviews.

[14]  D. Cory,et al.  Gradient, high‐resolution, magic‐angle spinning nuclear magnetic resonance spectroscopy of human adipocyte tissue , 1997, Magnetic resonance in medicine.

[15]  M. Sjöström,et al.  Design of experiments: an efficient strategy to identify factors influencing extraction and derivatization of Arabidopsis thaliana samples in metabolomic studies with gas chromatography/mass spectrometry. , 2004, Analytical biochemistry.

[16]  J. Bruce German,et al.  Lipid metabolome-wide effects of the PPARγ agonist rosiglitazones⃞s The online version of this article (available at http://www.jlr.org) contains an additional 4 tables. Published, JLR Papers in Press, August 16, 2002. DOI 10.1194/jlr.M200169-JLR200 , 2002, Journal of Lipid Research.

[17]  L. Svetkey,et al.  The Study of the Effects of Diet on Metabolism and Nutrition (STEDMAN) weight loss project: Rationale and design. , 2005, Contemporary clinical trials.

[18]  M. Saad,et al.  Peroxisome Proliferator-activated Receptor α (PPARα) Influences Substrate Utilization for Hepatic Glucose Production* , 2002, The Journal of Biological Chemistry.

[19]  J. Griffin.  Metabolic profiles to define the genome: can we hear the phenotypes? , 2004, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[20]  K. Petersen,et al.  Disordered lipid metabolism and the pathogenesis of insulin resistance. , 2007, Physiological reviews.

[21]  R. Kauppinen,et al.  Assignment of 1H nuclear magnetic resonance visible polyunsaturated fatty acids in BT4C gliomas undergoing ganciclovir-thymidine kinase gene therapy-induced programmed cell death. , 2003, Cancer research.

[22]  W. Kraus,et al.  Fatty Acid Homeostasis and Induction of Lipid Regulatory Genes in Skeletal Muscles of Peroxisome Proliferator-activated Receptor (PPAR) α Knock-out Mice , 2002, The Journal of Biological Chemistry.

[23]  R. Gonzalez,et al.  Quantitative neuropathology by high resolution magic angle spinning proton magnetic resonance spectroscopy. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[24]  J. Bryan,et al.  Both Triggering and Amplifying Pathways Contribute to Fuel-induced Insulin Secretion in the Absence of Sulfonylurea Receptor-1 in Pancreatic β-Cells* , 2004, Journal of Biological Chemistry.

[25]  L. Svetkey,et al.  The STEDMAN project: biophysical, biochemical and metabolic effects of a behavioral weight loss intervention during weight loss, maintenance, and regain. , 2009, Omics : a journal of integrative biology.

[26]  O. Ilkayeva,et al.  Metabolic profiling of PPARα−/− mice reveals defects in carnitine and amino acid homeostasis that are partially reversed by oral carnitine supplementation , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  I. Issemann,et al.  Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators , 1990, Nature.

[28]  J. Lehmann,et al.  The structure - Activity relationship between peroxisome proliferator-activated receptor γ agonism and the antihyperglycemic activity of thiazolidinediones , 1996 .

[29]  M. J. MacDonald,et al.  Perspective: emerging evidence for signaling roles of mitochondrial anaplerotic products in insulin secretion. , 2005, American journal of physiology. Endocrinology and metabolism.

[30]  M. Linton,et al.  Re-emergence of fibrates in the management of dyslipidemia and cardiovascular risk , 2000, Current atherosclerosis reports.

[31]  Fredrik Karpe,et al.  Rosiglitazone increases indexes of stearoyl-CoA desaturase activity in humans: link to insulin sensitization and the role of dominant-negative mutation in peroxisome proliferator-activated receptor-gamma. , 2005, Diabetes.

[32]  H. Mulder,et al.  Biochemical Mechanism of Lipid-induced Impairment of Glucose-stimulated Insulin Secretion and Reversal with a Malate Analogue* , 2004, Journal of Biological Chemistry.

[33]  Bruce D. Hammock,et al.  Metabolomics: building on a century of biochemistry to guide human health , 2005, Metabolomics.

[34]  P. Ferré,et al.  The biology of peroxisome proliferator-activated receptors: relationship with lipid metabolism and insulin sensitivity. , 2004, Diabetes.

[35]  J. Nicholson,et al.  Rapid and noninvasive diagnosis of the presence and severity of coronary heart disease using 1H-NMR-based metabonomics , 2002, Nature Medicine.

[36]  W. Willett,et al.  Biological variability dominates and influences analytical variance in HPLC-ECD studies of the human plasma metabolome , 2006, BMC clinical pathology.

[37]  J Bruce German,et al.  Lipid metabolome-wide effects of the PPARgamma agonist rosiglitazone. , 2002, Journal of lipid research.

[38]  B. Hammock,et al.  Mass spectrometry-based metabolomics. , 2007, Mass spectrometry reviews.

[39]  W. Weckwerth.  Metabolomics in systems biology. , 2003, Annual review of plant biology.

[40]  M. Febbraio,et al.  Overexpression of Carnitine Palmitoyltransferase-1 in Skeletal Muscle Is Sufficient to Enhance Fatty Acid Oxidation and Improve High-Fat Diet–Induced Insulin Resistance , 2009, Diabetes.

[41]  O. Fiehn,et al.  Metabolite profiling for plant functional genomics , 2000, Nature Biotechnology.

[42]  S. Nissen,et al.  Effect of rosiglitazone on the risk of myocardial infarction and death from cardiovascular causes. , 2007, The New England journal of medicine.

[43]  C. Newgard,et al.  Metabolic coupling factors in pancreatic beta-cell signal transduction. , 1995, Annual review of biochemistry.

[44]  K. Frayn.  Metabolic Regulation: A Human Perspective , 1996 .

[45]  Elias Chaibub Neto,et al.  Genetic Networks of Liver Metabolism Revealed by Integration of Metabolic and Transcriptional Profiling , 2008, PLoS genetics.

[46]  S. Robins,et al.  Conclusions from the VA-HIT study. , 2000, The American journal of cardiology.

[47]  Ru Wei,et al.  Metabolic profiling of the human response to a glucose challenge reveals distinct axes of insulin sensitivity , 2008, Molecular systems biology.

[48]  Robert A. Harris,et al.  Adaptive increase in pyruvate dehydrogenase kinase 4 during starvation is mediated by peroxisome proliferator-activated receptor alpha. , 2001, Biochemical and biophysical research communications.

[49]  Brian L. Frey,et al.  Alpha-Ketoisocaproate-induced hypersecretion of insulin by islets from diabetes-susceptible mice. , 2005, American journal of physiology. Endocrinology and metabolism.

[50]  Jeffrey I. Gordon,et al.  Mechanisms underlying the resistance to diet-induced obesity in germ-free mice , 2007, Proceedings of the National Academy of Sciences.

[51]  A. Häkkinen,et al.  Adipose Tissue Inflammation and Increased Ceramide Content Characterize Subjects With High Liver Fat Content Independent of Obesity , 2007, Diabetes.

[52]  Matej Oresic,et al.  Lipidomics: a new window to biomedical frontiers. , 2008, Trends in biotechnology.

[53]  D. Hachey,et al.  Validation of a new procedure to determine plasma fatty acid concentration and isotopic enrichment. , 1999, Journal of lipid research.

[54]  Xin Lu,et al.  Practical approach for the identification and isomer elucidation of biomarkers detected in a metabonomic study for the discovery of individuals at risk for diabetes by integrating the chromatographic and mass spectrometric information. , 2008, Analytical chemistry.

[55]  Christian Gieger,et al.  Genetics Meets Metabolomics: A Genome-Wide Association Study of Metabolite Profiles in Human Serum , 2008, PLoS genetics.

[56]  Leif Groop,et al.  Genetic dissection of type 2 diabetes , 2009, Molecular and Cellular Endocrinology.

[57]  D. Matern,et al.  Fetal Fatty Acid Oxidation Disorders, Their Effect on Maternal Health and Neonatal Outcome: Impact of Expanded Newborn Screening on Their Diagnosis and Management , 2005, Pediatric Research.

[58]  M. Jensen,et al.  Metabolic cycling in control of glucose-stimulated insulin secretion. , 2008, American journal of physiology. Endocrinology and metabolism.

[59]  Elaine Holmes,et al.  Metabonomics in pharmaceutical R & D , 2007, The FEBS journal.

[60]  Christopher B. Newgard,et al.  Molecular and metabolic mechanisms of insulin resistance and β-cell failure in type 2 diabetes , 2008, Nature Reviews Molecular Cell Biology.

[61]  J. Berger,et al.  The mechanisms of action of PPARs. , 2002, Annual review of medicine.

[62]  S. Eaton,et al.  Mammalian mitochondrial beta-oxidation. , 1996, The Biochemical journal.

[63]  Royston Goodacre,et al.  Metabolomics: Current technologies and future trends , 2006, Proteomics.

[64]  Kieran Clarke,et al.  A combined 1H-NMR spectroscopy- and mass spectrometry-based metabolomic study of the PPAR-alpha null mutant mouse defines profound systemic changes in metabolism linked to the metabolic syndrome. , 2006, Physiological genomics.

[65]  B. Yandell,et al.  Inferring Causal Phenotype Networks From Segregating Populations , 2008, Genetics.

[66]  B. Spiegelman,et al.  PPARγ: a Nuclear Regulator of Metabolism, Differentiation, and Cell Growth* , 2001, The Journal of Biological Chemistry.

[67]  P. Felig,et al.  Alanine: Key Role in Gluconeogenesis , 1970, Science.

[68]  John C. Lindon,et al.  Metabolomics Standards Workshop and the development of international standards for reporting metabolomics experimental results , 2006, Briefings Bioinform..

[69]  W. Wahli,et al.  Differential expression of peroxisome proliferator-activated receptors (PPARs): tissue distribution of PPAR-alpha, -beta, and -gamma in the adult rat. , 1996, Endocrinology.

[70]  Ping Li,et al.  Peroxisome Proliferator-activated Receptor-γ Co-activator 1α-mediated Metabolic Remodeling of Skeletal Myocytes Mimics Exercise Training and Reverses Lipid-induced Mitochondrial Inefficiency* , 2005, Journal of Biological Chemistry.

[71]  K. Kaski,et al.  Diagnosing diabetic nephropathy by 1H NMR metabonomics of serum , 2007, Magnetic Resonance Materials in Physics, Biology and Medicine.

[72]  S. Summers,et al.  Sphingolipids, insulin resistance, and metabolic disease: new insights from in vivo manipulation of sphingolipid metabolism. , 2008, Endocrine reviews.

[73]  Peter James,et al.  Metabolomic and proteomic analysis of a clonal insulin-producing beta-cell line (INS-1 832/13). , 2008, Journal of proteome research.

[74]  W. Dunn,et al.  Measuring the metabolome: current analytical technologies. , 2005, The Analyst.

[75]  M. Jensen,et al.  Compensatory responses to pyruvate carboxylase suppression in islet beta-cells. Preservation of glucose-stimulated insulin secretion. , 2006, The Journal of biological chemistry.

[76]  G. Gibbons,et al.  A role for PPARalpha in the control of SREBP activity and lipid synthesis in the liver. , 2005, The Biochemical journal.

[77]  Frederick P Roth,et al.  Metabolomic Identification of Novel Biomarkers of Myocardial Ischemia , 2005, Circulation.

[78]  M. Jensen,et al.  A Pyruvate Cycling Pathway Involving Cytosolic NADP-dependent Isocitrate Dehydrogenase Regulates Glucose-stimulated Insulin Secretion* , 2006, Journal of Biological Chemistry.

[79]  C. Newgard,et al.  Substrate Control of Insulin Release , 2011 .

[80]  David I. Ellis,et al.  Metabolic fingerprinting in disease diagnosis: biomedical applications of infrared and Raman spectroscopy. , 2006, The Analyst.

[81]  Nicholas J. Wareham,et al.  Genetic Factors in Type 2 Diabetes: The End of the Beginning? , 2005, Science.

[82]  E. Schleicher,et al.  Changes of the plasma metabolome during an oral glucose tolerance test: is there more than glucose to look at? , 2009, American journal of physiology. Endocrinology and metabolism.

[83]  M. Goran,et al.  Role of environmental vs. metabolic factors in the etiology of obesity: time to focus on the environment. , 2000, Obesity research.

[84]  M. McCarthy,et al.  Metabolic profiling reveals a contribution of gut microbiota to fatty liver phenotype in insulin-resistant mice , 2006, Proceedings of the National Academy of Sciences.

[85]  D. Macneil,et al.  Silencing of Cytosolic or Mitochondrial Isoforms of Malic Enzyme Has No Effect on Glucose-stimulated Insulin Secretion from Rodent Islets* , 2008, Journal of Biological Chemistry.

[86]  Michael E. Lassman,et al.  Malonyl CoenzymeA Decarboxylase Regulates Lipid and Glucose Metabolism in Human Skeletal Muscle , 2008, Diabetes.

[87]  W. Bremner,et al.  Advances in male contraception. , 2008, Endocrine reviews.

[88]  Svati H Shah,et al.  A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance. , 2009, Cell metabolism.

[89]  F. Gonzalez,et al.  The role of the peroxisome proliferator-activated receptor α (PPARα) in the control of cardiac lipid metabolism , 1999 .

[90]  G. Reaven.  Syndrome X: 6 years later. , 1994, Journal of internal medicine. Supplement.

[91]  William E. Kraus,et al.  Relationships Between Circulating Metabolic Intermediates and Insulin Action in Overweight to Obese, Inactive Men and Women , 2009, Diabetes Care.

[92]  Tamas L. Horvath,et al.  N-acylphosphatidylethanolamine, a Gut- Derived Circulating Factor Induced by Fat Ingestion, Inhibits Food Intake , 2008, Cell.

[93]  C. Newgard,et al.  Normal Flux through ATP-Citrate Lyase or Fatty Acid Synthase Is Not Required for Glucose-stimulated Insulin Secretion* , 2007, Journal of Biological Chemistry.

[94]  David Millington,et al.  Hepatic expression of malonyl-CoA decarboxylase reverses muscle, liver and whole-animal insulin resistance , 2004, Nature Medicine.

[95]  G. Barish,et al.  PPARs and the complex journey to obesity , 2004, Nature Medicine.

[96]  D. Kell,et al.  A functional genomics strategy that uses metabolome data to reveal the phenotype of silent mutations , 2001, Nature Biotechnology.

[97]  A. Vidal-Puig,et al.  PPARγ and the thiazolidinediones: molecular basis for a treatment of ‘Syndrome X’? , 2002, Diabetes, obesity & metabolism.

[98]  J. Lindon,et al.  Metabonomics: a platform for studying drug toxicity and gene function , 2002, Nature Reviews Drug Discovery.

[99]  O. Ilkayeva,et al.  Chronic Suppression of Acetyl-CoA Carboxylase 1 in β-Cells Impairs Insulin Secretion via Inhibition of Glucose Rather Than Lipid Metabolism* , 2008, Journal of Biological Chemistry.

[100]  T Hashimoto,et al.  Defect in Peroxisome Proliferator-activated Receptor α-inducible Fatty Acid Oxidation Determines the Severity of Hepatic Steatosis in Response to Fasting* , 2000, The Journal of Biological Chemistry.

[101]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[102]  Dominique Gauguier,et al.  Direct quantitative trait locus mapping of mammalian metabolic phenotypes in diabetic and normoglycemic rat models , 2007, Nature Genetics.

[103]  K. Griffin,et al.  Peroxisome proliferator activated receptor-alpha expression in human liver. , 1998, Molecular pharmacology.

[104]  Wei Jia,et al.  Metabonomic variations in the drug-treated type 2 diabetes mellitus patients and healthy volunteers. , 2009, Journal of proteome research.

[105]  Hans-Werner Mewes,et al.  Bioinformatics analysis of targeted metabolomics--uncovering old and new tales of diabetic mice under medication. , 2008, Endocrinology.

[106]  Olga Ilkayeva,et al.  Mitochondrial overload and incomplete fatty acid oxidation contribute to skeletal muscle insulin resistance. , 2008, Cell metabolism.

[107]  J. Griffin,et al.  The biochemical profile of rat testicular tissue as measured by magic angle spinning 1H NMR spectroscopy , 2000, FEBS letters.

[108]  G. Shulman,et al.  Mitochondrial GTP regulates glucose-stimulated insulin secretion. , 2007, Cell metabolism.

[109]  D. Kelly,et al.  A critical role for the peroxisome proliferator-activated receptor alpha (PPARalpha) in the cellular fasting response: the PPARalpha-null mouse as a model of fatty acid oxidation disorders. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[110]  Craig R Malloy,et al.  Analytical solutions for (13)C isotopomer analysis of complex metabolic conditions: substrate oxidation, multiple pyruvate cycles, and gluconeogenesis. , 2004, Metabolic engineering.

[111]  S. Watkins,et al.  Identification of a Lipokine, a Lipid Hormone Linking Adipose Tissue to Systemic Metabolism , 2008, Cell.

[112]  W. Wahli,et al.  The peroxisome proliferator‐activated receptor α regulates amino acid metabolism , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[113]  Christian Baumgartner,et al.  Metabolite profiling of blood from individuals undergoing planned myocardial infarction reveals early markers of myocardial injury. , 2008, The Journal of clinical investigation.

[114]  Joshua D. Knowles,et al.  Development of a robust and repeatable UPLC-MS method for the long-term metabolomic study of human serum. , 2009, Analytical chemistry.

[115]  S. Kersten,et al.  Peroxisome proliferator-activated receptor α target genes , 2004, Cellular and Molecular Life Sciences CMLS.

[116]  G. Shulman,et al.  13C NMR Isotopomer Analysis of Anaplerotic Pathways in INS-1 Cells* , 2004, Journal of Biological Chemistry.

[117]  S. Williams,et al.  A comparison of cell and tissue extraction techniques using high‐resolution 1H‐NMR spectroscopy , 2002, NMR in biomedicine.

[118]  A. Watson,et al.  Thematic Review Lipidomics: a Global Approach to Lipid Analysis in Biological Systems , 2022 .

[119]  M. Ravier,et al.  Hierarchy of the β‐cell signals controlling insulin secretion , 2003, European journal of clinical investigation.

[120]  Geoffrey S Ginsburg,et al.  High heritability of metabolomic profiles in families burdened with premature cardiovascular disease , 2009, Molecular systems biology.

[121]  Olli Simell,et al.  Dysregulation of lipid and amino acid metabolism precedes islet autoimmunity in children who later progress to type 1 diabetes , 2008, The Journal of experimental medicine.

[122]  Daniel J. Kliebenstein,et al.  Linking Metabolic QTLs with Network and cis-eQTLs Controlling Biosynthetic Pathways , 2007, PLoS genetics.

[123]  John T. Wei,et al.  Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression , 2009, Nature.

[124]  H. Mulder,et al.  13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS) , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[125]  C. Stanley,et al.  A Signaling Role of Glutamine in Insulin Secretion* , 2004, Journal of Biological Chemistry.

[126]  Thomas Hankemeier,et al.  Analytical strategies in lipidomics and applications in disease biomarker discovery. , 2009, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[127]  T. Hashimoto,et al.  PEROXISOMAL β-OXIDATION AND PEROXISOME PROLIFERATOR–ACTIVATED RECEPTOR α: An Adaptive Metabolic System , 2001 .

[128]  Xianlin Han,et al.  Shotgun lipidomics of neutral lipids as an enabling technology for elucidation of lipid-related diseases. , 2009, American journal of physiology. Endocrinology and metabolism.

[129]  W. Wahli,et al.  Peroxisome proliferator–activated receptor α mediates the adaptive response to fasting , 1999 .

[130]  D. Glower,et al.  Metabolomic Profiling Reveals Distinct Patterns of Myocardial Substrate Use in Humans With Coronary Artery Disease or Left Ventricular Dysfunction During Surgical Ischemia/Reperfusion , 2009, Circulation.

[131]  K. Petersen,et al.  Reversal of nonalcoholic hepatic steatosis, hepatic insulin resistance, and hyperglycemia by moderate weight reduction in patients with type 2 diabetes. , 2005, Diabetes.

[132]  Xianlin Han,et al.  Shotgun lipidomics: electrospray ionization mass spectrometric analysis and quantitation of cellular lipidomes directly from crude extracts of biological samples. , 2005, Mass spectrometry reviews.