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MB Sample ID: SA094082
| Local Sample ID: | 233_17 |
| Subject ID: | SU001368 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 32-67 |
| Gender: | Male and female |
| Human Race: | Caucasian (C) and African American (AA) |
| Human Ethnicity: | non-hispanic |
| Human Inclusion Criteria: | Sepsis: 1) Suspected or confirmed infection; 2) Any two of four criteria of systemic inflammatory response in ED(18); 3) Age ≥ 18; 4) Lactate ≥ 2.0 mmol/L; 5) Enrollment within 2 hours of initiation of quantitative resuscitation protocol. Controls: admitted to the emergency department and had no medical conditions that required chronic administration of medication expected to affect platelet function (aspirin, PGY12 inhibitors, etc) |
| Human Exclusion Criteria: | Exclusion criteria were:1) any primary diagnosis other than sepsis; 2) established Do Not Resuscitate status; 3) transferred from another hospital with sepsis therapy already initiated; 4) cardiopulmonary resuscitation (chest compression or defibrillation) prior to enrollment; 5) patient or legal representative unable to understand and sign informed consent. |
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Subject:
| Subject ID: | SU001368 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 32-67 |
| Gender: | Male and female |
| Human Race: | Caucasian (C) and African American (AA) |
| Human Ethnicity: | non-hispanic |
| Human Inclusion Criteria: | Sepsis: 1) Suspected or confirmed infection; 2) Any two of four criteria of systemic inflammatory response in ED(18); 3) Age ≥ 18; 4) Lactate ≥ 2.0 mmol/L; 5) Enrollment within 2 hours of initiation of quantitative resuscitation protocol. Controls: admitted to the emergency department and had no medical conditions that required chronic administration of medication expected to affect platelet function (aspirin, PGY12 inhibitors, etc) |
| Human Exclusion Criteria: | Exclusion criteria were:1) any primary diagnosis other than sepsis; 2) established Do Not Resuscitate status; 3) transferred from another hospital with sepsis therapy already initiated; 4) cardiopulmonary resuscitation (chest compression or defibrillation) prior to enrollment; 5) patient or legal representative unable to understand and sign informed consent. |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 233_17 | SA094082 | FL013453 | Control | Condition |
Collection:
| Collection ID: | CO001363 |
| Collection Summary: | WB samples (12 mL each) were collected by direct venipuncture or from an indwelling line into K2 EDTA-containing tubes. Platelets were then isolated. centrifuged (200x gfor 6 min at room temperature).The platelet-rich plasma layer was transferred to another tube and centrifuged (4500x gfor 5 minat 4C) to pellet the platelets.The nearly cell-free plasma layer was transferred to a new tube, leaving approximately 0.25 mL of plasma in the original tube. Remaining plasma was used to resuspend the platelet pellet to produce ultra-rich plasma. |
| Sample Type: | Platelets |
| Collection Method: | direct venipuncture or indwelling line |
| Collection Frequency: | once |
| Volumeoramount Collected: | 12mL |
| Storage Conditions: | Described in summary |
| Collection Vials: | K2 EDTA-containing Vacutainer tubes (BD 134367863; Becton-Dickinson, Franklin Lakes, NJ USA) |
| Additives: | K2 EDTA |
Treatment:
| Treatment ID: | TR001383 |
| Treatment Summary: | No treatment. Sepsis and non-sepsis controls had platelets isolated. Mitochondrial function was measured via mitochondrial oxygen consumption (mOCR) |
Sample Preparation:
| Sampleprep ID: | SP001376 |
| Sampleprep Summary: | Methanol chlorform extraction of platelet pellet after 2 freeze-thaw cycles in the presence of methanol. |
| Sampleprep Protocol ID: | Platelet Pellet MeOHCHCl3 Extraction |
| Sampleprep Protocol Filename: | 233.2.1_-_HAPS_Platelet_Pellet_MeOHCHCl3_Extraction |
| Processing Storage Conditions: | On ice |
| Extraction Method: | Methanol chloroform extraction |
| Extract Enrichment: | Lyophilization |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 500uL 50mM sodium phosphate buffer in D2O |
| Sample Spiking: | 4.99 mM DSS internal standard |
Analysis:
| MB Sample ID: | SA094082 |
| Analysis ID: | AN002155 |
| Laboratory Name: | University of Michigan BioNMR core |
| Analysis Type: | NMR |
| Acquisition Date: | 04/02/2018, 04/03 /2018, 04/04/2018, 04/05/2018 |
| Software Version: | TopSpin 4.0.7 |
| Operator Name: | Larisa Yeomans, Jae Hyun Kim |
| Data Format: | .fid |
| Num Factors: | 2 |
| Num Metabolites: | 19 |
| Units: | µM |
NMR:
| NMR ID: | NM000157 |
| Analysis ID: | AN002155 |
| Instrument Name: | Bruker 18.8 Tesla (800 MHz) NMR spectrometer ascend |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D-1H |
| NMR Comments: | Arrayed for water saturation frequency and 90deg. pulse width for each sample |
| Standard Concentration: | 10% (~0.5mM) |
| Spectrometer Frequency: | 800 MHz |
| NMR Probe: | Triple resonance inverse detection TCI cryoprobe |
| NMR Solvent: | D20 |
| NMR Tube Size: | 5mm |
| Shimming Method: | Auto shim (gradient shimming) |
| Pulse Sequence: | noesygppr1d |
| Water Suppression: | saturation at 80 Hz induced field strength |
| Pulse Width: | 5.5ms |
| Offset Frequency: | around -178Hz |
| Presaturation Power Level: | 80db |
| Chemical Shift Ref Cpd: | DSS-d6 |
| Temperature: | 25 |
| Number Of Scans: | 128 |
