Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA064103

Local Sample ID:	philic-17jul17-001
Subject ID:	SU001059
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116

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Subject:

Subject ID:	SU001059
Subject Type:	Mammal
Subject Species:	Rattus norvegicus
Taxonomy ID:	10116

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
philic-17jul17-001	SA064103	FL010940	S	groups
philic-17jul17-001	SA064103	FL010940	Pre	time

Collection:

Collection ID:	CO001053
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 Time: pre=day0, 2W=2 weeks, M1(4W)=1 month 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:	Blood (plasma)

Treatment:

Treatment ID:	TR001073
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.

Treatment ID:

TR001073

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:	SP001066
Sampleprep Summary:	large scale profiling using rat plasma

Combined analysis:

Analysis ID	AN001668	AN001669	AN001670	AN001671
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:	CH001175
Instrument Name:	Agilent 1290 Infinity
Column Name:	Waters Acquity BEH Amide (150 x 2.1mm,1.7um)
Chromatography Type:	HILIC

Chromatography ID:	CH001176
Instrument Name:	Agilent 1290 Infinity
Column Name:	Waters Acquity HSS C18 (150 x 2.1mm,1.8um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001543
Analysis ID:	AN001668
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	POSITIVE

MS ID:	MS001544
Analysis ID:	AN001669
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	NEGATIVE

MS ID:	MS001545
Analysis ID:	AN001670
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	POSITIVE

MS ID:	MS001546
Analysis ID:	AN001671
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
Ion Mode:	NEGATIVE