Summary of Study ST001251

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 PR000838. The data can be accessed directly via it's Project DOI: 10.21228/M8C671 This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST001251
Study Title	The effects of a training program encompassing cold exposure, breathing exercises, and meditation on plasma metabomics during experimental human endotoxemia
Study Type	Experimental human endotoxemia study
Study Summary	Study to investigate the effects of a training program encompassing cold exposure, breathing exercises, and meditation on plasma metabolomic during experimental human endotoxemia.
Institute	Radboud University Medical Centre
Department	Intensive Care Medicine
Laboratory	Metabolomic Discoveries (acquired by Metabolon in September 2017)
Last Name	Kox
First Name	Matthijs
Address	Intensive Care Medicine (710), Geert Grooteplein 10
Email	matthijs.kox@radboudumc.nl
Phone	243653881
Submit Date	2019-09-16
Num Groups	2
Total Subjects	24
Num Males	24
Analysis Type Detail	LC-MS
Release Date	2019-10-04
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR000838
Project DOI:	doi: 10.21228/M8C671
Project Title:	The effects of a training program encompassing cold exposure, breathing exercises, and meditation on the inflammatory response
Project Type:	Experimental human endotoxemia project
Project Summary:	Project to investigate the effects of a training program encompassing cold exposure, breathing exercises, and meditation on the inflammatory response during experimental human endotoxemia.
Institute:	Radboud University Medical Centre
Department:	Intensive Care Medicine
Last Name:	Kox
First Name:	Matthijs
Address:	Intensive Care Medicine (710), Geert Grooteplein 10, 6500 HB, Nijmegen, the Netherlands
Email:	matthijs.kox@radboudumc.nl
Phone:	+31243653881
Funding Source:	Dutch Arthritis Society (ReumaNederland, https://reumanederland.nl/) Serendipity Grant, no. 12-1-101
Publications:	https://www.pnas.org/content/111/20/7379.long; https://link.springer.com/article/10.1007%2Fs10067-015-3009-8

Subject:

Subject ID:	SU001319
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	19-27
Weight Or Weight Range:	58-92
Height Or Height Range:	172-190
Gender:	Male
Human Race:	Caucasian
Human Trial Type:	Prospective randomized open label controlled intervention study
Human Medications:	None
Human Prescription Otc:	None
Human Smoking Status:	Non smoker
Human Alcohol Drug Use:	Not in
Human Nutrition:	Fasted for 12 hours before the first sample was obtained.
Human Inclusion Criteria:	Healthy (normal physical examination, electrocardiography, and routine laboratory values).
Human Exclusion Criteria:	Febrile illness during the 2 weeks before the endotoxemia experiment, taking any prescription medication, history of spontaneous vagal collapse, practicing or experience with any kind of meditation, or participation in a previous trial where LPS was administered.

Factors:

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

mb_sample_id	local_sample_id	treatment
SA091016	17_30 _t_240	control
SA091017	17_30 _t_480	control
SA091018	17_30 _t_120	control
SA091019	17_30 _t_60	control
SA091020	17_22 _t_480	control
SA091021	17_30 _t_minus_50	control
SA091022	17_30 _t_0	control
SA091023	17_02 _t_480	control
SA091024	17_02 _t_240	control
SA091025	17_33 _t_120	control
SA091026	17_33 _t_60	control
SA091027	17_33 _t_0	control
SA091028	17_33 _t_240	control
SA091029	17_02 _t_minus_50	control
SA091030	17_02 _t_120	control
SA091031	17_02 _t_60	control
SA091032	17_02 _t_0	control
SA091033	17_22 _t_240	control
SA091034	17_22 _t_120	control
SA091035	17_13 _t_240	control
SA091036	17_29 _t_480	control
SA091037	17_29 _t_240	control
SA091038	17_13 _t_120	control
SA091039	17_13 _t_60	control
SA091040	17_13 _t_minus_50	control
SA091041	17_13 _t_0	control
SA091042	17_29 _t_120	control
SA091043	17_29 _t_60	control
SA091044	17_22 _t_0	control
SA091045	17_41 _t_minus_50	control
SA091046	17_22 _t_60	control
SA091047	17_22 _t_minus_50	control
SA091048	17_13 _t_480	control
SA091049	17_29 _t_0	control
SA091050	17_29 _t_minus_50	control
SA091051	17_33 _t_minus_50	control
SA091052	17_33 _t_480	control
SA091053	17_09_t_0	control
SA091054	17_09_t_120	control
SA091055	17_09_t_240	control
SA091056	17_09_t_480	control
SA091057	17_09_t_minus_50	control
SA091058	17_03_t_480	control
SA091059	17_40 _t_0	control
SA091060	17_03_t_60	control
SA091061	17_03_t_120	control
SA091062	17_03_t_240	control
SA091063	17_42 _t_480	control
SA091064	17_42 _t_240	control
SA091065	17_41 _t_240	control
SA091066	17_41 _t_120	control
SA091067	17_41 _t_60	control
SA091068	17_41 _t_0	control
SA091069	17_41 _t_480	control
SA091070	17_42 _t_minus_50	control
SA091071	17_42 _t_120	control
SA091072	17_42 _t_60	control
SA091073	17_42 _t_0	control
SA091074	17_40 _t_120	control
SA091075	17_40 _t_60	control
SA091076	17_08_t_240	control
SA091077	17_08_t_480	control
SA091078	17_03_t_minus_50	control
SA091079	17_08_t_120	control
SA091080	17_08_t_60	control
SA091081	17_08_t_minus_50	control
SA091082	17_08_t_0	control
SA091083	17_40 _t_240	control
SA091084	17_03_t_0	control
SA091085	17_40 _t_480	control
SA091086	17_15 _t_480	trained
SA091087	17_15 _t_120	trained
SA091088	17_15 _t_240	trained
SA091089	17_20 _t_480	trained
SA091090	17_20 _t_120	trained
SA091091	17_20 _t_240	trained
SA091092	17_20 _t_60	trained
SA091093	17_10 _t_minus_50	trained
SA091094	17_10 _t_480	trained
SA091095	17_15 _t_60	trained
SA091096	17_15 _t_0	trained
SA091097	17_10 _t_240	trained
SA091098	17_10 _t_120	trained
SA091099	17_10 _t_0	trained
SA091100	17_10 _t_60	trained
SA091101	17_15 _t_minus_50	trained
SA091102	17_34 _t_60	trained
SA091103	17_23 _t_minus_50	trained
SA091104	17_23 _t_60	trained
SA091105	17_14 _t_480	trained
SA091106	17_14 _t_240	trained
SA091107	17_14 _t_120	trained
SA091108	17_23 _t_120	trained
SA091109	17_23 _t_240	trained
SA091110	17_27 _t_60	trained
SA091111	17_27 _t_0	trained
SA091112	17_27 _t_minus_50	trained
SA091113	17_23 _t_480	trained
SA091114	17_14 _t_60	trained
SA091115	17_14 _t_0	trained

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Collection:

Collection ID:	CO001313
Collection Summary:	Ethylenediaminetetraacetic acid (EDTA) anti-coagulated blood was obtained one hour before LPS administration (baseline), and at T=0, T=1, T=2, T=4 and T=8 h. Blood was immediately centrifuged at 2000 g for 10 minutes at 4 °C after which plasma was stored at –80 °C until analysis.
Sample Type:	Blood (plasma)
Collection Method:	Arterial canulla
Collection Location:	Radial artery
Collection Frequency:	6
Collection Duration:	1 minute
Storage Conditions:	-80℃
Collection Vials:	Vacutainer blood collection tubes (BD)
Storage Vials:	Micronic vials (polypropylene)
Collection Tube Temp:	Room temperature
Additives:	Ethylenediaminetetraacetic acid (EDTA)

Treatment:

Treatment ID:	TR001334
Treatment Summary:	The subjects were randomly allocated to the trained group (n=12) or the control group (n=12) by the opening of sealed envelopes prepared by unblinded staff not involved in the study. The trained group underwent a 10-day training program provided by Dutch individual Wim Hof, which consisted of three main elements: meditation, exposure to cold, and breathing exercises (see https://www.pnas.org/content/111/20/7379.long for a detailed description). After completion of the training, subjects underwent experimental human endotoxemia at our intensive care research unit, consisting of administration of an intravenous bolus of 2 ng/kg of US Standard Reference Escherichia coli endotoxin (E. coli O:113 [LPS], Clinical Center Reference Endotoxin; National Institutes of Health, Bethesda, MD, USA). The experimental human endotoxemia protocol is provided in detail elsewhere (https://www.ncbi.nlm.nih.gov/pubmed/29936295; https://www.sciencedirect.com/science/article/pii/S0300908418301718?via%3Dihub). The control group did not undergo any training procedures and also underwent experimental human endotoxemia. Subjects in the trained group started practicing the breathing exercises acquired during the training program 30 minutes before LPS administration until two-and-a-half hours afterwards. The control group did not practice any techniques throughout the endotoxemia experiment. As described in detail elsewhere (https://www.pnas.org/content/111/20/7379.long), the breathing techniques consisted of two exercises. In brief, the first exercise comprised cycles of vigorous hyperventilation (approximately 30 breaths) followed by breath holding for several minutes at the discretion of the subject. The second exercise was similar, but at the end of the hyperventilation period, breath was only held for 10 seconds during all body muscles were tightened.
Treatment:	Nonpharmacological training intervention
Treatment Compound:	n/a.
Treatment Route:	n/a.
Treatment Dose:	n/a.
Treatment Dosevolume:	n/a.
Treatment Doseduration:	10 days
Treatment Vehicle:	control group
Human Fasting:	Yes, 12 hours before obtaining the first sample.

Sample Preparation:

Sampleprep ID:	SP001327
Sampleprep Summary:	Sample extraction was performed as described previously (https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0154597), with some modifications. Briefly, 50 µL of plasma was mixed with 450 µL of 90% (v/v) methanol containing internal standards and incubated for 15 min at 37°C with 1150 rpm. Precipitated proteins were separated from the extract by centrifugation for 12 min at 15000 rpm, after which supernatants were stored at -80 °C until further analysis.
Processing Storage Conditions:	-80℃
Extraction Method:	Ethanol precipitation

Combined analysis:

Analysis ID	AN002077
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Agilent 1290
Column	Discovery HS F5-3 (15cm x 2.1 mm,3um) Supelco,Sigma Aldrich)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	Agilent 6540 QTOF
Ion Mode	UNSPECIFIED
Units	relative abundance

Chromatography:

Chromatography ID:	CH001514
Instrument Name:	Agilent 1290
Column Name:	Discovery HS F5-3 (15cm x 2.1 mm,3um) Supelco,Sigma Aldrich)
Flow Gradient:	5% B from 0-0.1min to 35% B at 1.5 min, to 95% B at 2.05 min which was kept constant until 3.2 min, to 5% B at 3.21 min and washing until 4.3 min with 5% B.
Flow Rate:	The flow rate was kept constant at 700 µL/min from 0 min to 2.2 min, and increased up to 900 µL/min from 2.2 min to 2.5 min, after which flow rate was kept constant until 3.2 min.The flow rate was decreased from 900 µL/min to 800 µL/min from 3.2 min to 3.1 min and kept constant until 3.7 min, when flow rate was changed to 700 µL/min.
Injection Temperature:	40 °C
Solvent A:	95% water/5% acetonitrile; 0.1% formic acid; 10 mM ammonium formate
Solvent B:	95% acetonitrile/5% water; 0.1% formic acid; 10 mM ammonium formate
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001928
Analysis ID:	AN002077
Instrument Name:	Agilent 6540 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	The 6540 QTOF/MS Detector (Agilent) operated in both positive and negative ESI mode in a detection range of 50 to 1700 m/z at 2 GHz in extended dynamic range. The DualAJS ESI source was set to the following parameters: Gas temperature 200 °C, drying gas 8 L/min, nebulizer 35 psig, sheath gas temp: 350 °C, sheath gas flow 11 L/min, VCAp 3500 V and nozzle voltage of 0 V. Online calibration of the instrument was performed throughout the data acquisition using Agilent ES-TOF Reference Mass Solution Kit. Chromatograms were generated by the LC-MS instrument in .d format. Raw data were converted into mzXML and chromatogram peaks were extracted using XCMS v1.42.0 (https://pubs.acs.org/doi/10.1021/ac051437y), which was optimized using the IPO R package (https://bmcbioinformatics.biomedcentral.com/articles/10.1186/s12859-015-0562-8) with the following settings: peakwidth=c(10, 70), ppm= 20, snthresh=10, mzdiff=0.0034, prefilter=c(3, 100), noise=100, gapInit=0.8448, gapExtend=2.0544, bw=5, mzwid=0.015, minfrac=0.5, max=50. All further analyses were performed in R programming language, Metaboanalyst 4.0 (https://www.nature.com/articles/nprot.2011.319), and GraphPad Prism version 5.0 (GraphPad Software). IDEOM software (http://mzmatch.sourceforge.net/ideom.php; https://academic.oup.com/bioinformatics/article/28/7/1048/210126) was used to eliminate noise and for putative peak annotation by exact mass within ±10 ppm against the Metabolomic Discoveries in house metabolite library (https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0125044) in negative and positive ESI mode, respectively. Retention time prediction was applied to aid metabolite annotation.
Ion Mode:	UNSPECIFIED