Summary of Study ST001021

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 PR000683. The data can be accessed directly via it's Project DOI: 10.21228/M8D11B This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001021
Study TitleCharacterization of metabolomics profile changes during development of post-traumatic epilepsy in Rat Cerebrospinal Fluid (part-II)
Study SummaryCharacterize the metabolomics profile changes during progression of the transition from traumatic brain injury (TBI) to post-traumatic epilepsy (PTE). To do so, three experiments will be performed. PTE animal model will be developed using ferrous chloride injections. Metabolomics profile changes will be obtained before TBI, after TBI, and after PTE development These temporal changes in metabolomics profile during the course of PTE development will be collected. We will also collect cerebrospinal fluid (CSF) at each time point. In addition, we will collect the brain tissue from the center of injury, around the injury, and from the non-injured area for mass spectrometry. In this study, Rat CSF is analyzed at end of study.
Institute
Mayo Clinic
Last NameSu-youne
First NameChang
Address200 1st Street SW Rochester, MN 55905, USA
EmailChang.SuYoune@mayo.edu
Phone1-507-293-0511
Submit Date2018-07-17
Analysis Type DetailLC-MS
Release Date2020-07-15
Release Version1
Chang Su-youne Chang Su-youne
https://dx.doi.org/10.21228/M8D11B
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000683
Project DOI:doi: 10.21228/M8D11B
Project Title:Mayo Pilot and Feasibility: Characterization of metabolomics profile changes during development of post-traumatic epilepsy
Project Summary:According to the report from the Centers for Disease Control and Prevention in 2014, traumatic brain injury (TBI) accounts for 30% of all injury-related deaths in the U.S. Developing epilepsy after severe head injury is as high as 40%-50% in some settings. Importantly, many TBI victims develop epilepsy months or years following the initial injury. However, it has not been fully identified how to predict who will develop epilepsy and/or the underlying mechanism of post traumatic epilesy (PTE) development. The main goal of this proposal is to identify the metabolomics biomarker of TBI-induced epilepsy and to investigate the underlying mechanism of the transition from TBI to PTE. To do so, we will develop PTE using two TBI animal models: ferrous chloride injection model and cortical undercut model. Once the brain damage is made, electroencephalogram (EEG) and video monitoring will be performed to determine the onset point of epilepsy. Then, metabolomics profile changes will be analyzed and compared before and after PTE development.
Institute:Mayo Clinic
Last Name:Su-youne
First Name:Chang
Address:200 1st Street SW Rochester, MN 55905, USA
Email:Chang.SuYoune@mayo.edu
Phone:1-507-293-0511

Subject:

Subject ID:SU001060
Subject Type:Mammal
Subject Species:Rattus norvegicus
Taxonomy ID:10116

Factors:

Subject type: Mammal; Subject species: Rattus norvegicus (Factor headings shown in green)

mb_sample_id local_sample_id groups
SA064108nC18-22aug17-001C
SA064109pC18-21aug17-009C
SA064110pC18-21aug17-005C
SA064111nhilic-18aug17-005_1C
SA064112philic-17aug17-001C
SA064113philic-17aug17-009C
SA064114philic-17aug17-005C
SA064115pC18-21aug17-001C
SA064116nhilic-18aug17-009_1C
SA064117nhilic-18aug17-001_1C
SA064118nC18-22aug17-009C
SA064119nC18-22aug17-005C
SA064120nhilic-18aug17-004_1F
SA064121nC18-22aug17-007F
SA064122nhilic-18aug17-003_1F
SA064123philic-17aug17-008F
SA064124philic-17aug17-007F
SA064125nhilic-18aug17-007_1F
SA064126philic-17aug17-004F
SA064127nhilic-18aug17-011_1F
SA064128nC18-22aug17-004F
SA064129nhilic-18aug17-008_1F
SA064130nC18-22aug17-008F
SA064131nC18-22aug17-003F
SA064132pC18-21aug17-004F
SA064133pC18-21aug17-003F
SA064134nC18-22aug17-011F
SA064135philic-17aug17-003F
SA064136philic-17aug17-011F
SA064137pC18-21aug17-011F
SA064138pC18-21aug17-007F
SA064139pC18-21aug17-008F
SA064140philic-17aug17-012S
SA064141nC18-22aug17-002S
SA064142philic-17aug17-010S
SA064143philic-17aug17-006S
SA064144nhilic-18aug17-006_1S
SA064145pC18-21aug17-010S
SA064146pC18-21aug17-006S
SA064147nC18-22aug17-010S
SA064148pC18-21aug17-002S
SA064149pC18-21aug17-012S
SA064150nhilic-18aug17-002_1S
SA064151nhilic-18aug17-012_1S
SA064152nhilic-18aug17-010_1S
SA064153nC18-22aug17-006S
SA064154nC18-22aug17-012S
SA064155philic-17aug17-002S
Showing results 1 to 48 of 48

Collection:

Collection ID:CO001054
Collection Summary:Rats will either be controls, injected with saline, or injected with ferrous chloride to influence PTE. Study Groupings: C=control, S=Saline treated, F=ferrous chloride treated Experimental Flow Day0: baseline pre TBI. Blood and CSF collected Day1: Surgery for TBI. Injections of Ferrous Cloride or Saline Day2: CSF collected Weeks1-3: montoring to determine PTE starting point. Blood and CSF collected 1 Month: montoring of PTE. Blood and CSF collected 2 Month: Animal is euthanized and blood, CSF, and tissue harvested
Sample Type:CSF

Treatment:

Treatment ID:TR001074
Treatment Summary:Rats will either be controls, injected with saline, or injected with ferrous chloride to influence PTE. Trauma-Induced Epilepsy Model: Ferrous chloride injection model: Ferrous chloride solution (5 μl of 100 mM with saline) will be injected at a rate of 0.5 μl/min through a Hamilton micro-syringe controlled by a micro-pump (UMP3, WPI, FL). Once the ferrous chloride solution injection is completed, the syringe will remain in position for 5 minutes, and then it will be removed slowly. The burr holes will be closed with light-curing dental cement. The dose of ferrous chloride injection was determined from prior published reports from mouse, rat, and cat. They all used 100 mM ferrous chloride aqueous solution and volumes were various: 1 μl for mouse,14 5 μl for rat (200-300 g),15 and 10 μl for cat (2-4 Kg).15 Video monitoring: The use of 24 x 7 video monitoring and review means that we do not have to rely on the rats having seizures during daily rounding or at some other time when a human happens to be present in the home cage. Normally, the video will be watched in time lapse, fast-forward mode to scan for potential seizures. The reviewer can then stop the video, rewind and watch the behavioral episode in real-time or slow motion to determine whether a seizure actually occurred. Behavioral seizures will be identified by any combination or sequence of the following: loss of postural control (opisthotonus), tonic flexion or extension of limbs or head/neck, and clonic movements of limbs or head/neck. Often, behavioral seizures in rats may be accompanied by drooling, urination and facial twitches, although these may not always be observable on video. In addition, seizures will likely be followed by a postictal phase, which may include a period of running, jumping and general agitation. Video monitoring cannot detect subclinical or electrographic seizures (i.e., seizures without a behavioral component). Video will be reviewed in this way for each rat in order to establish that a cortical injured rat does indeed have epilepsy, to establish the “typical seizure” pattern in each rat, and to help establish a seizure frequency baseline, although it is understood that video monitoring alone might occasionally miss a seizure. EEG monitoring: To prevent imaging distortion and ferromagnetic interference, graphite carbon electrodes will be fabricated and/or purchased. A total of five electrodes will be implanted for EEG monitoring on the skull. EEG will be monitored with the Open EPhys System.18 While EEG recording, EEG electrodes will be connected to wires attached to the ceiling of a cage. In trauma-induced epilepsy rats, spontaneous neural activity will be recorded using a wide bandwidth (0-9 kHz) recording system. Post-analysis will be used to identify epilepsy signals.

Sample Preparation:

Sampleprep ID:SP001067
Sampleprep Summary:large scale profiling of rat cerebral spinal fluid The brain tissue and CSF will be collected for mass spectrometry. To prepare samples, proteins will be removed from collected dialysates by adding cold methanol:water (8:1, v/v) mixture containing 5.0 μg internal standard (IS), myristic-d27 acid, at ambient temperature. Samples will be vortexed for 1 min, incubated on ice for 15 min, and then centrifuged. The supernatant will be completely dried in a SpeedVac, and the lyophilized sample will be subsequently methoxiaminated using 20 μl of a 20 mg/ml solution of methoxyamine hydrochloride in pyridine at 30°C for 90 min and derivatized using 80 μL of N-methyl-N-trimethylsilyltrifluoroacetamide with 1% trimethylchloro-silane (MSTFA + 1% TMCS, Pierce) at 37°C for 30 min.

Combined analysis:

Analysis ID AN001672 AN001673 AN001674 AN001675
Analysis type MS MS MS MS
Chromatography type HILIC HILIC Reversed phase Reversed phase
Chromatography system Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity Agilent 1290 Infinity
Column Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity BEH Amide (150 x 2.1mm,1.7um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um) Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
MS Type ESI ESI ESI ESI
MS instrument type QTOF QTOF QTOF QTOF
MS instrument name Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF Agilent 6550 QTOF
Ion Mode POSITIVE NEGATIVE POSITIVE NEGATIVE
Units intensity intensity intensity intensity

Chromatography:

Chromatography ID:CH001177
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Chromatography Type:HILIC
  
Chromatography ID:CH001178
Instrument Name:Agilent 1290 Infinity
Column Name:Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
Chromatography Type:Reversed phase

MS:

MS ID:MS001547
Analysis ID:AN001672
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001548
Analysis ID:AN001673
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS001549
Analysis ID:AN001674
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:POSITIVE
  
MS ID:MS001550
Analysis ID:AN001675
Instrument Name:Agilent 6550 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
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