MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1/m_MTBLS1_metabolite_profiling_NMR_spectroscopy_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1/a_MTBLS1_metabolite_profiling_NMR_spectroscopy.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1/s_MTBLS1.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1/FILESftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS1primaryOK200120110Jules GriffinReza SalekMetaboLightsPublicNuclear Magnetic Resonance (NMR) - - - BrukerA metabolomic comparison of urinary changes in type 2 diabetes in mouse, rat, and human. 10.1152/physiolgenomics.00194.2006. PMID:17190852Imperial College LondonInternational Agency for Research on CancerHomo sapiensNMR spectroscopyFor the human studies, midstream urine (∼15 ml) samples were collected and frozen from each volunteer. In total, 84 samples were collected from 12 healthy volunteers (7 time points, 8 males and 4 females) and 50 samples from 30 T2DM patients (1–3 time points, 17 males and 13 females) with well-controlled blood glucose maintained at normal concentrations by diet, following the guidelines issued by the American Diabetes Association, rather than medication. The healthy subjects were aged 18–55 yr, had a body mass index (BMI) ≥19 and ≤30 kg/m2 and a body mass ≥50 kg and ≤113 kg, and were free from any major disease or pregnancy. The T2DM patients were aged 30–65 yr (mean 56 ± 9 yr), had a BMI >25 and <40 kg/m2, weighed between 65 and 140 kg (mean 95 ± 19 kg), and were taking at most one oral anti-diabetic drug. T2DM patients agreed to stop treatment with oral anti-diabetic agents during the study. Subjects went through a washout period of 4 wk before sample collection and abstained from alcohol during the study; diet was controlled throughout the study.https://www.ebi.ac.uk/metabolights/MTBLS1Jules Griffin. University of Cambridge. The Department of Biochemistry, The Sanger Building, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.. jlg40@cam.ac.uk. 01223674922.Reza Salek. University of Cambridge. The Department of Biochemistry, The Sanger Building, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.. rms72@cam.ac.uk. 01223674948.Spectra were processed using ACD/1D NMR Manager 8.0 with Intelligent Bucketing Integration (Advanced Chemistry Development, Toronto, ON, Canada). Spectra were integrated 0.20-9.30 ppm excluding water (4.24-5.04 ppm), glucose (3.19-3.99 ppm, 5.21-5.27 ppm), and urea (5.04-6.00 ppm). Intelligent bucketing ensures that bucket edges do not coincide with peak maxima, preventing resonances from being split across separate integral regions; a 0.04 ppm bucket width and a 50% looseness factor were used. All spectra were normalized to total area excluding the water, urea, and glucose regions.GenderMetabolic syndromeFor the human studies, midstream urine (~15 ml) samples were collected and frozen from each volunteer. In total, 84 samples were collected from 12 healthy volunteers (7 time points, 8 males and 4 females) and 50 samples from 30 T2DM patients (1-3 time points, 17 males and 13 females) with well-controlled blood glucose maintained at normal concentrations by diet, following the guidelines issued by the American Diabetes Association, rather than medication. T2DM patients agreed to stop treatment with oral anti-diabetic agents during the study. Subjects went through a washout period of 4 wk before sample collection and abstained from alcohol during the study; diet was controlled throughout the study.Aliquots of 400 µl urine samples were made up to 600 µl with phosphate buffer (0.2 M, pH 7.4) and any precipitate removed by centrifugation. In total, 500 µl of supernatant were transferred to 5-mm NMR tubes with 100 µl of sodium 3-trimethylsilyl-(2,2,3,3-2H4)-1-propionate (TSP)/D2O/sodium azide solution (0.05% wt/vol TSP in D2O and 1% wt/vol sodium azide).A 1D NOESY presaturation pulse sequence was used to analyze the urine samples. For each sample 128 transients were collected into 64k data points using a spectral width of 14.005 kHz (20 ppm) and an acquisition time of 2.34 s per FID.MetabolomicsThe spectra of human urine samples were acquired on a Bruker DRX700 NMR spectrometer using a 5 mm TXI ATMA probe at a proton frequency of 700.1 MHz and ambient temperature of 27 °C.metabolic syndromeHuman Study SubjectUrine global profilingnuclear magnetic resonance spectroscopyuntargeted metabolitesdiabetes mellitusmetabolic syndromeHuman Study SubjectUrine global profilingnuclear magnetic resonance spectroscopyuntargeted metabolitesdiabetes mellitusurineAssignments were confirmed by two dimensional spectroscopy including homonuclear 1H-1H Correlation Spectroscopy (COSY), 1H-13C Heteronuclear Signal Quantum Coherence (HSQC) and 1H-13C Heteronuclear Multiple Bond Correlation (HMBC) Spectroscopy.unknown_d_1.235acetonesarcosinea-Hydroxy-n-butyraten-methylnicotinatemethyl guanidineindoxyl sulphatecitrullinateisoleucinen-methyl-4-pyridone-5-carboxamidehippurateindoxyl sulfateunknown_s_1.27a-hydroxyisobutyrateglutamatea-Hydroxy-n-valeratemethyl N-acetylvalinateleucinen-butyratea-Hydroxyisovalerateb-hydroxy-n-butyraten-methylnicotinamideadenosinelysineN-acetylglutamatemalatecreatineallantoinunknown_s_2.04caproylglyunknown_s_1.35Uridineoxalacetatetyrosinemhppa sulphate?unknown_s_6.68n-caproylglycinea-keto-b-methyl-N valeratedesaminotyrosine4-aminohippurateisocaproateglutaminedimethylglycineethanolN-acetylaspartatemethylmalonateuridineacetatesuccinate3,4-dihydroxymandelateformateornithinemhppa sulphatecreatinineunknown_t_1.205n-methyl-2-pyridone-5-carboxamidetrimethylaminedimethylamineketoglutaratealanineunknown_m_2.97glycolatedihydroxymandelateunknown_s_7.02unassigned pyrimidineunknown_s_2.05_NA-groupcitratephenylalanineureidopropionateb-hydroxybutyrateunknown_d_6.98unknown_m_8.3352-oxoisovalerateUnknown_q_5.03pyruvateb-aminoisobutyrateUnknown_d_5.11acetoacetatenicotinateN-acetyl groups (glycoproteins)valinelactatehistidinefumarateType 2 diabetes mellitus is the result of a combination of impaired insulin secretion with reduced insulin sensitivity of target tissues. There are an estimated 150 million affected individuals worldwide, of whom a large proportion remains undiagnosed because of a lack of specific symptoms early in this disorder and inadequate diagnostics. In this study, NMR-based metabolomic analysis in conjunction with multivariate statistics was applied to examine the urinary metabolic changes in two rodent models of type 2 diabetes mellitus as well as unmedicated human sufferers. The db/db mouse and obese Zucker (fa/fa) rat have autosomal recessive defects in the leptin receptor gene, causing type 2 diabetes. 1H-NMR spectra of urine were used in conjunction with uni- and multivariate statistics to identify disease-related metabolic changes in these two animal models and human sufferers. This study demonstrates metabolic similarities between the three species examined, including metabolic responses associated with general systemic stress, changes in the TCA cycle, and perturbations in nucleotide metabolism and in methylamine metabolism. All three species demonstrated profound changes in nucleotide metabolism, including that of N-methylnicotinamide and N-methyl-2-pyridone-5-carboxamide, which may provide unique biomarkers for following type 2 diabetes mellitus progression.A metabolomic comparison of urinary changes in type 2 diabetes in mouse, rat, and human.Salek R M RM, Maguire M L ML, Bentley E E, Rubtsov D V DV, Hough T T, Cheeseman M M, Nunez D D, Sweatman B C BC, Haselden J N JN, Cox R D RD, Connor S C SC, Griffin J L JLT2DM - Type 2 Diabetes mellitus, Adult-Onset Diabetes Mellitus, Rat, human being, norvegicus, Laboratory, Noninsulin Dependent, mouse, Metabonomic, MODY, Norway Rat, Type 2 Diabetes Mellitus, Metabonomics, Rattus, DIABETES MELLITUS TYPE 02, diabetes mellitus type 2, Norway Rats, Adult-Onset, rat, Mus norvegicus, T2D, Rats, Adult-Onset Diabetes, Mus, Laboratory Rats, Non-Insulin-Dependent, Maturity-Onset, Slow-Onset Diabetes Mellitus, Non-Insulin Dependent Diabetes Mellitus, non-insulin-dependent diabetes mellitus, Type 2 Diabetes Mellitus Non-Insulin Dependent, Norway rat, Ketosis Resistant, Metabolomic, Slow Onset, human., Maturity-Onset Diabetes Mellitus, brown rat, T2DM, Type 2 Diabetes, Rattus norvegicus, mice, Type II Diabetes, Maturity Onset Diabetes Mellitus, Type 2, rats, man, NIDDM, Ketosis-Resistant Diabetes Mellitus, Laboratory Rat, Non-Insulin Dependent Diabetes, Stable, Diabetes Mellitus, Ketosis-Resistant, Mouse, Norway, house mouse, Type II, Non Insulin Dependent, Slow-Onset, Diabetes, Stable Diabetes Mellitus, Maturity Onset, Adult OnsetModb1, Soluble, big, CD295, type 2, other disease, Insulin B Chain, criteria, human being, maturity-onset diabetes, adult onset diabetes, determination, noninsulin-dependent diabetes mellitus, Glucose, Blood, Mbp1, Metabonomic, impaired, insuline humaine, Metabonomics, Insulin B, guidelines, froggy, noninsulin dependent diabetes, Gyltl1a, inadequate, dysfunctional, School-Age, large, LEPRD, reduced, diseases, sensitive, human insulin, Associations, Insulin, Diets, Diabetes mellitus (disorder), disease or disorder, 2, diseases and disorders, non-insulin-dependent diabetes mellitus, DmF2, tiny, Metabolomic, myd, Fs(3)Hor, Leprb, noninsulin-dependent, sensitivity, DM - Diabetes mellitus, lod, Chain, study, School-Age Populations, type II diabetes mellitus, insulin, proportion, DmelCG2684, human disease, non-insulin dependent diabetes, reference sample, proportionality to, MDDGB6, dysfunction, Tissue, hypoplasia, Mbp-1, Statistic, obese-like, Papers., non-insulin dependent diabetes mellitus, type 2 diabetes mellitus, NTef2, Population, Estimated, man, NIDDM, adult-onset diabetes, non-neoplastic, allergic reaction, BPFD#36, Iletin, Secretion, insulin human, type 2 diabetes mellitus non-insulin dependent, Diabetes mellitus, Insulin recombinant, grupos, Clients, great, disorder, type 2 diabetes, Homo sapiens disease, severe, School Age Population, association with, Diabetes, diabetes, Controlled, db, small, Blood glucose, insulinum humanum, diabetes mellitis type II, Controlling, Regular, grupo, PLATEST, DM, gyltl1b-b, disorders, T2DM - type 2 diabetes mellitus, medical condition, late onset, Client, partial functionality, Fs(3)Sz11, group, Regular Insulin, diabetes mellitus (disease), OB-RGRP, insulin (recombinant), Sodium Insulin, MDDGA6, School Age, mKIAA0609, chemical analysis, Insulin A Chain, condition, NOS, LEP-R, Sodium, Novolin, Soluble Insulin, fg, Blood Sugar, LEPROT, Populations, Exubera, gyltl1b, diabetes mellitus, underdeveloped, Dietary, susceptibility to, INS, mdc1d, proportionality, insulina humana, expanded, Rest, rate, lacks function of type, Lds, School Age Populations, insulin resistance, insulin resistance-related, human, low functionality, early, Sugar, Obr, OBR, diabetes mellitis type 2, Diabetes NOS, type II, MDC1D, disease, having decreased function, non-insulin-dependent, enr, enlarged, Patient, Blood Glucose, Horka, type II diabetes, quotient, CG2684, Fs(3)Horka, protection against, digenic, assay, School-Age Population, obl, groupe, Platelets, OB-R, hypertension, GruppeT2DM - Type 2 Diabetes mellitus, Adult-Onset Diabetes Mellitus, Controlling, human being, grupo, Noninsulin Dependent, Metabonomic, MODY, Type 2 Diabetes Mellitus, Metabonomics, DIABETES MELLITUS TYPE 02, diabetes mellitus type 2, Adult-Onset, T2D, Adult-Onset Diabetes, Non-Insulin-Dependent, Maturity-Onset, Slow-Onset Diabetes Mellitus, Non-Insulin Dependent Diabetes Mellitus, group., non-insulin-dependent diabetes mellitus, Type 2 Diabetes Mellitus Non-Insulin Dependent, Ketosis Resistant, Metabolomic, Slow Onset, Maturity-Onset Diabetes Mellitus, T2DM, study, Type 2 Diabetes, reference sample, Type II Diabetes, Rest, Maturity Onset Diabetes Mellitus, Type 2, man, human, NIDDM, Ketosis-Resistant Diabetes Mellitus, Non-Insulin Dependent Diabetes, Stable, grupos, Diabetes Mellitus, Ketosis-Resistant, Type II, Non Insulin Dependent, Slow-Onset, Diabetes, Stable Diabetes Mellitus, groupe, Maturity Onset, Controlled, Gruppe, Adult OnsetSoluble, Adult-Onset Diabetes Mellitus, type 2, Insulin B Chain, Rat, Biological Markers, Viral Marker, Ob Gene Product, adult onset diabetes, Surrogate Endpoints, determination, noninsulin-dependent diabetes mellitus, Laboratory, Mbp1, Metabonomic, Biochemical, Norway Rat, Endpoint, insuline humaine, Type 2 Diabetes Mellitus, Metabonomics, Serum, Norway Rats, diabetes mellitus type 2, methylamine metabolism, Adiposis, inadequate, dysfunctional, Obese, rat, met4PY, Laboratory Markers, unspecified, Tier, diseases, Biological, human insulin, Non-Insulin Dependent Diabetes Mellitus, 2, diseases and disorders, Norway rat, myd, Fs(3)Hor, animal, Slow Onset, type II diabetes mellitus, insulin, DmelCG2684, human disease, non-insulin dependent diabetes, proportionality to, Tissue, hypoplasia, Mbp-1, Statistic, non-insulin dependent diabetes mellitus, type 2 diabetes mellitus, NTef2, Estimated, NIDDM, Laboratory Rat, Ketosis-Resistant Diabetes Mellitus, allergic reaction, Secretion, insulin human, type 2 diabetes mellitus non-insulin dependent, Immune, Markers, urinary aspects, Viral Markers, type 2 diabetes, Homo sapiens disease, house mouse, severe, Non Insulin Dependent, Obese Protein, insulinum humanum, diabetes mellitis type II, Regular, Viral, PLATEST, Surrogate Endpoint, gyltl1b-b, rodents, obese, animalia, Krebs cycle, N-methylnicotinamide monohydrochloride, mouse, T2DM - type 2 diabetes mellitus, Biochemical Markers, Rattus, late onset, Biologic Marker, partial functionality, Fs(3)Sz11, Regular Insulin, Adult-Onset, insulin (recombinant), organism, Mus norvegicus, Adult-Onset Diabetes, Marker, MDDGA6, mKIAA0609, Obesity [Ambiguous], Obese (finding), Diseases, type II., Ketosis Resistant, Soluble Insulin, Maturity-Onset Diabetes Mellitus, fg, gyltl1b, whole organism, End Points, mice, susceptibility to, mdc1d, proportionality, insulina humana, expanded, Type II Diabetes, Adiposity, rate, OBESITY NOS, INSDC_feature:gene, Immunologic, Type 2, lacks function of type, Laboratory Marker, rats, insulin resistance, insulin resistance-related, human, low functionality, early, type II, MDC1D, disease, non-insulin-dependent, enr, Non-Insulin Dependent Diabetes, enlarged, Biochemical Marker, Horka, type II diabetes, Koerper, CG2684, Fs(3)Horka, Mouse, Slow-Onset, Platelets, hypertension, big, T2DM - Type 2 Diabetes mellitus, other disease, human being, multi-cellular organism, maturity-onset diabetes, norvegicus, Clinical Markers, Noninsulin Dependent, Clinical Marker, obesity disease, impaired, Insulin B, rodent, DIABETES MELLITUS TYPE 02, Gene Product, froggy, noninsulin dependent diabetes, Gyltl1a, Surrogate End Points, Obesity, Surrogate Markers, large, Rats, reduced, sensitive, Insulin, Non-Insulin-Dependent, disease or disorder, citric acid cycle, non-insulin-dependent diabetes mellitus, DmF2, tiny, Metabolomic, noninsulin-dependent, sensitivity, lod, brown rat, Chain, T2DM, study, Biomarker, proportion, urinary levels, Rattus norvegicus, Clinical, nucleotide metabolism, MDDGB6, dysfunction, Biological Marker, metazoa, Overweight and obesity, Maturity Onset Diabetes Mellitus, man, adult-onset diabetes, non-neoplastic, BPFD#36, Iletin, Immunologic Markers, Insulin recombinant, Stable, Diabetes Mellitus, great, disorder, methylammonium metabolism, Ketosis-Resistant, species, association with, Immunologic Marker, Diabetes, diabetes, Biologic, Adult Onset, small, methylammonium metabolic process, body, Serum Markers, disorders, MODY, End Point, whole body, medical condition, obesity, Immune Marker, T2D, Sodium Insulin, Mus, Surrogate End Point, Laboratory Rats, chemical analysis, Insulin A Chain, Maturity-Onset, Slow-Onset Diabetes Mellitus, condition, Sodium, Type 2 Diabetes Mellitus Non-Insulin Dependent, Novolin, obesity disorder, Obesity (disorder), Biologic Markers, Serum Marker, Exubera, Type 2 Diabetes, N-METHYL-NICOTINAMIDE, diabetes mellitus, underdeveloped, Surrogate, INS, Obesity NOS, Endpoints, OB, ob, Lds, Surrogate Marker, diabetes mellitis type 2, having decreased function, Obese Gene Product, TCA cycle, quotient, protection against, digenic, Ob, assay, Norway, Type II, Ob Protein, Stable Diabetes Mellitus, Maturity Onset, Immune Markers1112trueA metabolomic study of urinary changes in type 2 diabetes in human compared to the control groupType 2 diabetes mellitus is the result of a combination of impaired insulin secretion with reduced insulin sensitivity of target tissues. There are an estimated 150 million affected individuals worldwide, of whom a large proportion remains undiagnosed because of a lack of specific symptoms early in this disorder and inadequate diagnostics. In this study, NMR-based metabolomic analysis in conjunction with uni- and multivariate statistics was applied to examine the urinary metabolic changes in Human type 2 diabetes mellitus patients compared to the control group. The human population were un medicated diabetic patients who have good daily dietary control over their blood glucose concentrations by following the guidelines on diet issued by the American Diabetes Association. Note: This is part of a larger study, please refer to the original paper below.2012-02-142012-02-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:1148CHEBI:60647CHEBI:24898CHEBI:25017CHEBI:27266CHEBI:60645CHEBI:35932CHEBI:16530CHEBI:16236CHEBI:20067CHEBI:30860CHEBI:78320CHEBI:50129CHEBI:16449CHEBI:64390CHEBI:74911CHEBI:25094CHEBI:18257CHEBI:18211CHEBI:15366CHEBI:17533CHEBI:21547CHEBI:28300CHEBI:30772CHEBI:15347CHEBI:15344CHEBI:74903CHEBI:30744CHEBI:32816CHEBI:18261CHEBI:6650CHEBI:15741CHEBI:30915CHEBI:18237CHEBI:30769CHEBI:17170CHEBI:15611CHEBI:17724CHEBI:16628CHEBI:18139CHEBI:16810CHEBI:16919CHEBI:16737CHEBI:27389CHEBI:17497CHEBI:16704CHEBI:64399CHEBI:15676CHEBI:16335CHEBI:18012CHEBI:27838CHEBI:27410CHEBI:32980CHEBI:104011CHEBI:27637CHEBI:18186CHEBI:27570CHEBI:43355CHEBI:64414CHEBI:28044CHEBI:18089CHEBI:18123CHEBI:15940CHEBI:30751