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MB Sample ID: SA091033
| Local Sample ID: | 17_22 _t_240 |
| 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. |
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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:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 17_22 _t_240 | SA091033 | FL013119 | control | treatment |
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 |
