Effects of Histidine Supplementation on Global Serum and Urine 1 H NMR-based. Metabolomics and Serum Amino Acid Profiles in Obese Women from a
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1 Supporting information for Effects of Histidine Supplementation on Global Serum and Urine 1 H NMR-based Metabolomics and Serum Amino Acid Profiles in Obese Women from a Randomized Controlled Study Shanshan Du 1,, Shuhong Sun 2,, Liyan Liu 1,, Qiao Zhang 1, Fuchuan Guo 3, Chunlong Li 4, Rennan Feng 1,*, and Changhao Sun 1,* 1 Department of Nutrition and Food Hygiene, School of Public Health, Harbin Medical University, Harbin, China 2 Department of Psychiatry, the First Affiliated Hospital of Harbin Medical University, Harbin, China 3 Department of Nutrition and Food Safety, School of Public Health, Fujian Medical University, FuZhou, China. 4 Department of General Surgery, the Second Affiliated Hospital of Harbin Medical University, Harbin, China Table of contents Table S1 The demographic, dietary and clinical data at baseline and 12 weeks in histidine supplementation and placebo groups. Table S2 Metabolites and chemical shifts in serum 1 H NMR spectra. Table S3 Metabolites and chemical shifts in urine 1 H NMR spectra. Figure S1 Score plots derived from serum and urine 1 H NMR metabolomics and serum AA profiles of placebo and histidine supplementation groups. S1
2 Table S1 The demographic, dietary and clinical data at baseline and 12 weeks in histidine supplementation and placebo groups. Variable Placebo (32) Histidine (37) Baseline 12 weeks Baseline 12 weeks Age (years) 48±11 46±10 BMI (kg/m 2 ) 30.56± ±2.92 WC (cm) 96.43± ±6.94 Smoke (%) Alcohol (%) Physical Activity (%) None Moderate Heavy Dietary intakes Energy (kj/d) 11,082±4,113 10,956±4,026 Protein (g/d) 89±39 90±35 Fat (g/d) 102±47 100±38 Carbohydrate (g/d) 370± ±142 BMI (kg/m 2 ) 30.49± ± ± ±2.70 WC (cm) 94.02± ± ± ±4.10 FM (kg) 28.56± ± ± ±3.74 SBP (mmhg) ± ± ± ±14.61 DBP (mmhg) 89.93± ± ± ±8.79 FBG (mmol/l) 5.77± ± ± ±1.40 2h-PG (mmol/l) 7.42± ± ± ±2.58 Insulin (IU/mL) 14.28± ± ± ±5.92 HOMA-IR 3.48± ± ± ±1.06 TG (mmol/l) 1.68± ± ± ±0.70 TC (mmol/l) 5.49± ± ± ±1.20 HDL-c (mmol/l) 1.35± ± ± ±0.37 LDL-c (mmol/l) 3.22± ± ± ±0.70 NFFA (µmol/l) ± ± ± ± * Adiponectin (ng/ml) 6.46± ± ± ±2.85 * TNF-α (pg/ml) 14.22± ± ± ±4.77 * IL-6 (pg/ml) 5.37± ± ± ±3.43 CRP (µg/ml) 5.19± ± ± ±2.05 * MDA (nmol/ml) 2.83± ± ± ±0.94 GSH-Px (nmol/ml) ± ± ± ±28.11 SOD (U/mL) 93.14± ± ± ±16.98 * All values are mean±sd. * P < 0.05, compared with baseline values (paired samples t test); P < 0.05, comparing the two groups at 12 weeks (ANCOVA; the covariates included age, body mass index, physical activities, tobacco use, drink, dietary protein intake and baseline values). S2
3 Table S2 Metabolites and chemical shifts in serum 1 H NMR spectra. Metabolites Assignment Chem shifts Cholesterol/LDL/VLDL CH 3 CH 2 CH 2 C= 0.86(m) Isoleucine δch 3, γch (t),0.9997(d) Leucine δch 3, δch (d),0.9596(d) Valine γ'ch 3, γ'ch (d),1.0315(d) 3-hydroxybutyrate γch 3, αch, αch (d), (dd), 2.419(m) Lipids (triglycerides and fatty acids) CH 3 (CH 2 )n, CH 2 CH 2 CO, CH 2 -C=C, CH 2 -C=O, =C-CH 2 -C= Lactate βch 3,αCH (d), (q) 1.30(m),1.57(m), 2.01(m), 2.23(m), 2.74(m) Lysine γch 2, δch 3, βch 3, (m), (m), (m) Alanine βch 3, αch (d), (q) Acetate βch (s) N-Acetyl glycoproteins CH (s) Glutamine βch 2, γch (m), (m) Methionine S-CH 3, γch (s), (t) Acetone CH (s) Citrate Half CH 2, Half CH (d), (d) Aspartate half β-ch 2, half β-ch (dd), (dd), (dd) Dimethylglycine N-CH (s) Creatine CH (s) Creatinine CH 3, CH (s), (s) Choline N-(CH 3 ) (s) Histidine half β-ch (dd) TMAO CH (s) Scyllo-inositol CHOH (s) Myo-inositol 1,3-CH, 4,6-CH 3.531(dd), (t) Glycine CH (s) Threonine γch 3, αch, β-ch (d), (d), (m) Ethanol CH (dd) β-glucose CH (d) α-glucose CH (d) Unsaturated lipids -CH=CH (m) Urea NH (s) Tyrosine half β-ch 2, αch, CH, CH (dd), (dd), (d), (d) 1-Methylhistidine 4-CH, 2-CH (s), (dd), (d), (d) Formate CH (s) S3
4 Table S3 Metabolites and chemical shifts in urine 1 H NMR spectra. Metabolites Chem shifts Butyrate (t), (t) Valine (d), (d) Isoleucine (d), (d) 3-hydroxyisobutyrate (d), (m) 3-aminoisobutanoic acid (d) 1-methylmalonate (d) 3-hydroxyisovalerate (s), (s) Lactate/Threonine (d) Lactate (d) Threonine (m) α-hydroxyisobutyrate (s) Alanine (d), (dd) Lysine (m), (t) Glutamine (m), (m) Acetone (s) Succinate (s) 2-oxoglutarate (t) Dimethylamine (s) Citrate (d), (d) Methylguanidine (s), (s) Trimethylamine (s) Dimethylglycine (s) Creatinine/Creatine (s) Creatine (s) Creatinine (s) Taurine (s), 3.433(t) Betaine/TMAO (s) 3-hydroxyphenylacetate (s), (d) Glycine (s) a-galactose (dd) Galactitol (m) 3-methylhistidine (s) Erythritol (d) Guanidoacetic acid (s) Adenosine (dd), (dd), (s) Hippurate (d), (m), (tt), (dd) S4
5 Trigonelline Urea Histidine Formate (s) (s) (d), (d) (s) S5
6 Figure S1 Score plots derived from serum and urine 1 H NMR spectra and serum AA profiles of placebo and histidine supplementation groups. A: PCA score plot of serum 1 H NMR spectra at baseline and 12th week; B: PCA score plot of serum 1 H NMR spectra at baseline; C: validation for PLS-DA of serum 1 H NMR spectra at 12th week; D: PCA score plot of urine 1 H NMR spectra at baseline and 12th week; E: PCA score plot of urine 1 H NMR spectra at baseline; F: validation for PLS-DA of urine 1 H NMR spectra at 12th week; G: PCA score plot of serum AA profiles at baseline and 12th week; H: PCA score plot of serum AA profiles at baseline; I: validation for PLS-DA of serum AA profiles at 12th week; red box: histidine supplementation group at 12th week; black diamond: placebo group at 12th week; hollow red box: histidine supplementation group at baseline; hollow black diamond: placebo group at baseline; green triangle: R2Y; blue box: Q2Y. S6
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