Summary of Study ST000483

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000367. The data can be accessed directly via it's Project DOI: 10.21228/M8MC7X This work is supported by NIH grant, U2C- DK119886.

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Study IDST000483
Study TitleAmino Acid Quantifcation of obese patients on a 16 week caloric restriction from Plasma
Study Typetimecourse, quantitative measurements of amino acid
Study SummaryCaloric restriction (CR) improves insulin sensitivity and reduces the incidence of diabetes in obese individuals. The underlying mechanisms whereby CR improves insulin sensitivity are not clear. We evaluated the effect of 16 weeks of CR on whole-body insulin sensitivity by pancreatic clamp before and after CR in 11 obese participants (BMI = 35 kg/m2) compared with 9 matched control subjects (BMI = 34 kg/m2). Compared with the control subjects, CR increased the glucose infusion rate needed to maintain euglycemia during hyperinsulinemia, indicating enhancement of peripheral insulin sensitivity. This improvement in insulin sensitivity was not accompanied by changes in skeletal muscle mitochondrial oxidative capacity or oxidant emissions, nor were there changes in skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels. However, CR lowered insulin-stimulated thioredoxin-interacting protein (TXNIP) levels and enhanced nonoxidative glucose disposal. These results support a role for TXNIP in mediating the improvement in peripheral insulin sensitivity after CR.
Institute
Mayo Clinic
DepartmentEndocrinology
LaboratoryMayo Clinic Metabolomics Resource Core
Last NameNair
First NameSreekumaran
Address200 First Street SW, Rochester, MN 55905
EmailNair.K@mayo.edu
Phone507-285-2415
Submit Date2016-09-23
PublicationsMechanism by Which Caloric Restriction Improves Insulin Sensitivity in Sedentary Obese Adults. DOI: 10.2337/db15-0675
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2016-12-22
Release Version1
Sreekumaran Nair Sreekumaran Nair
https://dx.doi.org/10.21228/M8MC7X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000367
Project DOI:doi: 10.21228/M8MC7X
Project Title:Mechanism by Which Caloric Restriction Improves Insulin Sensitivity in Sedentary Obese Adults
Project Type:skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels
Project Summary:effect of caloric restriction on insulin sensitivity through skeletal muscle ceramide, diacylglycerol, or amino acid metabolite levels
Institute:Mayo Clinic
Department:Endocrinology
Laboratory:Mayo Clinic Metabolomics Resource Core
Last Name:Nair
First Name:Sreekumaran
Address:200 First Street SW, Rochester, MN 55905
Email:Nair.K@mayo.edu
Phone:507-285-2415

Subject:

Subject ID:SU000504
Subject Type:Animal
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

Subject type: Animal; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id time (mins) Category
SA024582ms5361-111 Caloric Restriction
SA024583ms5361-331 Caloric Restriction
SA024584ms5361-311 Caloric Restriction
SA024585ms5361-231 Caloric Restriction
SA024586ms5361-11 Caloric Restriction
SA024587ms5361-191 Caloric Restriction
SA024588ms5361-131 Caloric Restriction
SA024589ms5361-91 Caloric Restriction
SA024590ms5361-51 Caloric Restriction
SA024591ms5361-31 Caloric Restriction
SA024592ms5361-71 Caloric Restriction
SA024593ms5361-391 Control
SA024594ms5361-251 Control
SA024595ms5361-291 Control
SA024596ms5361-371 Control
SA024597ms5361-351 Control
SA024598ms5361-271 Control
SA024599ms5361-211 Control
SA024600ms5361-171 Control
SA024601ms5361-151 Control
SA024602ms5361-122 Caloric Restriction
SA024603ms5361-202 Caloric Restriction
SA024604ms5361-322 Caloric Restriction
SA024605ms5361-102 Caloric Restriction
SA024606ms5361-82 Caloric Restriction
SA024607ms5361-342 Caloric Restriction
SA024608ms5361-142 Caloric Restriction
SA024609ms5361-62 Caloric Restriction
SA024610ms5361-242 Caloric Restriction
SA024611ms5361-22 Caloric Restriction
SA024612ms5361-42 Caloric Restriction
SA024613ms5361-382 Control
SA024614ms5361-362 Control
SA024615ms5361-402 Control
SA024616ms5361-302 Control
SA024617ms5361-182 Control
SA024618ms5361-222 Control
SA024619ms5361-162 Control
SA024620ms5361-262 Control
SA024621ms5361-282 Control
Showing results 1 to 40 of 40

Collection:

Collection ID:CO000498
Collection Summary:Blood samples were collected in a heated hotbox (131°F) through a retrograde intravenous catheter at baseline for glucose and hormone levels, and every 10 min during the clamp to maintain euglycemia. In addition, blood samples were collected every 20 min from 0600 to 0700, 0900 to 1000, and 1200 to 1300 to measure plasma [6,62H2]glucose. At 1330 h, a percutaneous needle muscle biopsy specimen (350–400 mg) was obtained from the vastus lateralis muscle under local anesthesia, immediately frozen in liquid nitrogen, and stored at −80°F for future analysis (27). This biopsy sample was used for analysis of TXNIP mRNA and protein content. The participant remained in the CRU through the remainder of the day and was given a weight-maintenance diet until 2200 h. At 0700 h the following morning, a second muscle biopsy specimen was obtained under local anesthesia, and ∼100 mg was used immediately for mitochondrial function measurements of isolated mitochondria and mtH2O2 emissions (28). The remainder was immediately frozen in liquid nitrogen and stored at −80°F for future analysis, including DAG, ceramide, and amino acid measurements (Fig. 1).
Sample Type:Blood. Plasma was isolated for MS analysis.

Treatment:

Treatment ID:TR000518
Treatment Summary:Before and after 16 weeks of CR or CON, two outpatient visits and one inpatient visit were scheduled. Before the outpatient visits, participants were instructed to fast overnight from 10:00 p.m. the evening before and to avoid strenuous exercise for 24 h preceding the visits. One outpatient visit consisted of an MRI to measure subcutaneous and visceral fat distribution and magnetic resonance spectroscopy to measure skeletal muscle oxidative capacity (25). The second outpatient visit was for measurements of resting energy expenditure (REE) for the calculation of a weight-maintenance diet (Parvo Medics TrueOne 2400 Canopy system), DEXA scan (Lunar DPX-L; Lunar Radiation, Madison, WI), and VO2peak test on a bicycle ergometer (Fig. 1). Participants were admitted to the Clinical Research Unit (CRU) on the evening of the fifth day of the weight-maintaining diet provided by the CRU metabolic kitchen (Supplementary Fig. 1). The weight-maintenance meals (diet composition: 20% protein, 30% fat, 50% carbohydrate) were monitored daily to ensure that the correct calorie level was achieved. Upon admission to the CRU, no calories were consumed after 2100 h to achieve a 10-h fast before the two-stage insulin euglycemic pancreatic clamp the following morning, as previously published (26), with modifications as follows: the following morning at 0400 h, a primed [6,62H2]glucose bolus (6 mg ⋅ kg fat-free mass[FFM]−1) was administered, followed by a 9-h continuous infusion of [6,62H2]glucose (started at 4 mg ⋅ kgFFM−1 ⋅ h−1 then titrated downward over the infusion time period to match anticipated changes in endogenous glucose production [EGP]). At 0600 h, gas exchange was measured by indirect calorimetry for 30 min for REE determination. Then at 0700 h, glucagon (0.001 μg ⋅ kgFFM−1 ⋅ min−1), somatostatin (0.093 μg ⋅ kgFFM−1 ⋅ min−1), and growth hormone (0.0047 μg ⋅ kgFFM−1 ⋅ min−1) were infused for 6 h. Insulin was infused from 0700 to 1000 h at 0.62 mU ⋅ kgFFM−1 ⋅ min−1 and then from 1000 to 1300 h at 2.3 mU ⋅ kgFFM−1 ⋅ min−1. A 40% dextrose with 2% enrichment of [6,62H2]glucose was infused as needed to maintain blood glucose above 4.7 mmol/L from 0700 to 1000 h and then between 4.7 and 5.3 mmol/L from 1000 to 1300 h.

Sample Preparation:

Sampleprep ID:SP000511
Sampleprep Summary:Plasma samples and amino acid calibration standards were prepared with MassTrak Amino Acid Analysis Solution (AAA) kit from Waters according to instructions with slight modifications for detection on a mass spectrometer. A 10 point standard concentration curve was made from the calibration standard solution to calculate amino acid concentrations in plasma samples. A solution containing U-13C4-L-aspartic acid, U-13C3-L-alanine, U-13C4-L-threonine, U-13C5-L-proline, U-13C5-L-valine, U-13C6-leucine, U-13C6-phenylalanine all from Cambridge Isotope Laboratories, 13C6-tyrosine from Isotec, L-arginine (15N2, 2H2) from MassTrace, norvaline from Sigma dissolved in 0.01N HCl was used as the internal standard solution. Frozen plasma samples were thawed, spiked with internal standard then deproteinized with cold MeOH followed by centrifugation at 10,000 g for 5 minutes prior to derivatization according to MassTrak instructions. The amino acid derivatizing reagent used was 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.

Combined analysis:

Analysis ID AN000749
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity
Column C18
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Thermo Quantum Ultra
Ion Mode POSITIVE
Units micromolar

Chromatography:

Chromatography ID:CH000537
Chromatography Summary:High resolution separation was done using an Acquity UPLC system, injecting 1 µl of derviatized solution, with a UPLC BEH C18 1.7 micron 2.1×150 mm column from Waters. Column flow was set to 400 µl/min with a gradient from 99.9%A to 98%B where buffer A is 1% acetonitrile in 0.1% formic acid and buffer B is 100% acetonitrile. A column temp of 43 degrees Celsius and a sample tray temp of 6% Celsius. Mass detection was completed on a TSQ Ultra Quantum from Thermo Finnigan running in ESI positive mode. A scan width of 0.002, scan time of 0.04 seconds per transition mass, collision energy of 25, collision gas pressure of 1.5 mTorr, tube lens value set to 90, monitoring a signature ion of the derivitized amines at m/z 171.04 by selected reaction monitoring.
Instrument Name:Waters Acquity
Column Name:C18
Chromatography Type:Reversed phase

MS:

MS ID:MS000663
Analysis ID:AN000749
Instrument Name:Thermo Quantum Ultra
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:POSITIVE
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