#METABOLOMICS WORKBENCH METADATA_TEMPLATE.xlsm DATATRACK_ID:243 STUDY_ID:ST000165 ANALYSIS_ID:AN000258
VERSION                          	1
CREATED_ON                       	02-08-2024
#PROJECT
PR:PROJECT_TITLE                 	Sparing of muscle mass and function by passive loading in an experimental
PR:PROJECT_TITLE                 	intensive care unit model
PR:PROJECT_SUMMARY               	The response to mechanical stimuli, i.e. tensegrity, plays an important role in
PR:PROJECT_SUMMARY               	regulating cell physiological and pathophysiological function, and the
PR:PROJECT_SUMMARY               	mechanical silencing observed in intensive care unit (ICU) patients leads to a
PR:PROJECT_SUMMARY               	severe and specific muscle wasting condition. This study aims to unravel the
PR:PROJECT_SUMMARY               	underlying mechanisms and the effects of passive mechanical loading on skeletal
PR:PROJECT_SUMMARY               	muscle mass and function at the gene, protein and cellular levels. A unique
PR:PROJECT_SUMMARY               	experimental rat ICU model has been used allowing long-term (weeks)
PR:PROJECT_SUMMARY               	time-resolved analyses of the effects of standardized unilateral passive
PR:PROJECT_SUMMARY               	mechanical loading on skeletal muscle size and function and underlying
PR:PROJECT_SUMMARY               	mechanisms. Results show that passive mechanical loading alleviated the muscle
PR:PROJECT_SUMMARY               	wasting and the loss of force-generation associated with the ICU intervention,
PR:PROJECT_SUMMARY               	resulting in a doubling of the functional capacity of the loaded versus the
PR:PROJECT_SUMMARY               	unloaded muscles after a 2-week ICU intervention. We demonstrate that the
PR:PROJECT_SUMMARY               	improved maintenance of muscle mass and function is probably a consequence of a
PR:PROJECT_SUMMARY               	reduced oxidative stress revealed by lower levels of carbonylated proteins, and
PR:PROJECT_SUMMARY               	a reduced loss of the molecular motor protein myosin. A complex temporal gene
PR:PROJECT_SUMMARY               	expression pattern, delineated by microarray analysis, was observed with
PR:PROJECT_SUMMARY               	loading-induced changes in transcript levels of sarcomeric proteins, muscle
PR:PROJECT_SUMMARY               	developmental processes, stress response, extracellular matrix/cell adhesion
PR:PROJECT_SUMMARY               	proteins and metabolism. Thus, the results from this study show that passive
PR:PROJECT_SUMMARY               	mechanical loading alleviates the severe negative consequences on muscle size
PR:PROJECT_SUMMARY               	and function associated with the mechanical silencing in ICU patients, strongly
PR:PROJECT_SUMMARY               	supporting early and intense physical therapy in immobilized ICU patients.
PR:INSTITUTE                     	Uppsala University
PR:DEPARTMENT                    	Department of Neuroscience
PR:LAST_NAME                     	Larsson
PR:FIRST_NAME                    	Lars
PR:EMAIL                         	Lars.larsson@neuro.uu.se
PR:DOI                           	http://dx.doi.org/10.21228/M8GP4N
#STUDY
ST:STUDY_TITLE                   	Sparing of muscle mass and function by passive loading in an experimental
ST:STUDY_TITLE                   	intensive care unit model
ST:STUDY_TYPE                    	time course + intervention
ST:STUDY_SUMMARY                 	A unique experimental rat ICU model has been used allowing long-term (weeks)
ST:STUDY_SUMMARY                 	time-resolved analyses of the effects of standardized unilateral passive
ST:STUDY_SUMMARY                 	mechanical loading on skeletal muscle size and function and underlying
ST:STUDY_SUMMARY                 	mechanisms. Results show that passive mechanical loading alleviated the muscle
ST:STUDY_SUMMARY                 	wasting and the loss of force-generation associated with the ICU intervention,
ST:STUDY_SUMMARY                 	resulting in a doubling of the functional capacity of the loaded versus the
ST:STUDY_SUMMARY                 	unloaded muscles after a 2-week ICU intervention.
ST:INSTITUTE                     	Uppsala University
ST:DEPARTMENT                    	Department of Neuroscience
ST:LAST_NAME                     	Larsson
ST:FIRST_NAME                    	Lars
ST:EMAIL                         	Lars.larsson@neuro.uu.se
ST:SUBMIT_DATE                   	2015-05-14
#SUBJECT
SU:SUBJECT_TYPE                  	Animal
SU:SUBJECT_SPECIES               	Rattus norvegicus
SU:TAXONOMY_ID                   	10116
SU:GENOTYPE_STRAIN               	Sprague–Dawley
SU:GENDER                        	female
SU:SPECIES_GROUP                 	Mammal
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data
SUBJECT_SAMPLE_FACTORS           	-	34	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial superficial; ID=MEK080318; tissue fluid(mpe)=22.73; mmp(mpe)=0.0027; FSR (%/hr)=0.0467
SUBJECT_SAMPLE_FACTORS           	-	36	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial deep; ID=MEK080318; tissue fluid(mpe)=25.51; mmp(mpe)=0.0143; FSR (%/hr)=0.2240
SUBJECT_SAMPLE_FACTORS           	-	38	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral superficial; ID=MEK080318; tissue fluid(mpe)=23.89; mmp(mpe)=0.0073; FSR (%/hr)=0.1214
SUBJECT_SAMPLE_FACTORS           	-	40	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral deep; ID=MEK080318; tissue fluid(mpe)=24.76; mmp(mpe)=0.0130; FSR (%/hr)=0.2098
SUBJECT_SAMPLE_FACTORS           	-	42	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial superficial; ID=MEK080507; tissue fluid(mpe)=20.24; mmp(mpe)=0.0067; FSR (%/hr)=0.1330
SUBJECT_SAMPLE_FACTORS           	-	44	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial deep; ID=MEK080507; tissue fluid(mpe)=21.01; mmp(mpe)=0.0150; FSR (%/hr)=0.2855
SUBJECT_SAMPLE_FACTORS           	-	46	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral superficial; ID=MEK080507; tissue fluid(mpe)=19.27; mmp(mpe)=0.0063; FSR (%/hr)=0.1317
SUBJECT_SAMPLE_FACTORS           	-	48	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral deep; ID=MEK080507; tissue fluid(mpe)=22.78; mmp(mpe)=0.0189; FSR (%/hr)=0.3318
SUBJECT_SAMPLE_FACTORS           	-	50	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial superficial; ID=MEK080519; tissue fluid(mpe)=15.00; mmp(mpe)=0.0016; FSR (%/hr)=0.0425
SUBJECT_SAMPLE_FACTORS           	-	52	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial deep; ID=MEK080519; tissue fluid(mpe)=15.96; mmp(mpe)=0.0059; FSR (%/hr)=0.1485
SUBJECT_SAMPLE_FACTORS           	-	54	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral superficial; ID=MEK080519; tissue fluid(mpe)=14.81; mmp(mpe)=0.0027; FSR (%/hr)=0.0736
SUBJECT_SAMPLE_FACTORS           	-	56	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral deep; ID=MEK080519; tissue fluid(mpe)=17.40; mmp(mpe)=0.0114; FSR (%/hr)=0.2615
SUBJECT_SAMPLE_FACTORS           	-	66	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial superficial; ID=UU13; tissue fluid(mpe)=18.99; mmp(mpe)=0.0065; FSR (%/hr)=0.1379
SUBJECT_SAMPLE_FACTORS           	-	68	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. medial deep; ID=UU13; tissue fluid(mpe)=17.47; mmp(mpe)=0.0176; FSR (%/hr)=0.4036
SUBJECT_SAMPLE_FACTORS           	-	70	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral superficial; ID=UU13; tissue fluid(mpe)=17.67; mmp(mpe)=0.0101; FSR (%/hr)=0.2287
SUBJECT_SAMPLE_FACTORS           	-	72	Mechanical loading hours/day:12 | Group:Passive Mechanical Loading	Tissue=Gast=L. lateral deep; ID=UU13; tissue fluid(mpe)=19.57; mmp(mpe)=0.0113; FSR (%/hr)=0.2306
SUBJECT_SAMPLE_FACTORS           	-	33	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial superficial; ID=MEK080318; tissue fluid(mpe)=21.64; mmp(mpe)=0.0029; FSR (%/hr)=0.0536
SUBJECT_SAMPLE_FACTORS           	-	35	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial deep; ID=MEK080318; tissue fluid(mpe)=23.67; mmp(mpe)=0.0088; FSR (%/hr)=0.1482
SUBJECT_SAMPLE_FACTORS           	-	37	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral superficial; ID=MEK080318; tissue fluid(mpe)=26.03; mmp(mpe)=0.0051; FSR (%/hr)=0.0787
SUBJECT_SAMPLE_FACTORS           	-	39	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral deep; ID=MEK080318; tissue fluid(mpe)=26.62; mmp(mpe)=0.0066; FSR (%/hr)=0.0987
SUBJECT_SAMPLE_FACTORS           	-	41	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial superficial; ID=MEK080507; tissue fluid(mpe)=21.61; mmp(mpe)=0.0017; FSR (%/hr)=0.0319
SUBJECT_SAMPLE_FACTORS           	-	43	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial deep; ID=MEK080507; tissue fluid(mpe)=23.74; mmp(mpe)=0.0083; FSR (%/hr)=0.1391
SUBJECT_SAMPLE_FACTORS           	-	45	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral superficial; ID=MEK080507; tissue fluid(mpe)=22.76; mmp(mpe)=0.0091; FSR (%/hr)=0.1596
SUBJECT_SAMPLE_FACTORS           	-	47	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral deep; ID=MEK080507; tissue fluid(mpe)=22.40; mmp(mpe)=0.0086; FSR (%/hr)=0.1530
SUBJECT_SAMPLE_FACTORS           	-	49	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial superficial; ID=MEK080519; tissue fluid(mpe)=19.68; mmp(mpe)=0.0004; FSR (%/hr)=0.0089
SUBJECT_SAMPLE_FACTORS           	-	51	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial deep; ID=MEK080519; tissue fluid(mpe)=18.73; mmp(mpe)=0.0143; FSR (%/hr)=0.3059
SUBJECT_SAMPLE_FACTORS           	-	53	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral superficial; ID=MEK080519; tissue fluid(mpe)=19.30; mmp(mpe)=0.0025; FSR (%/hr)=0.0509
SUBJECT_SAMPLE_FACTORS           	-	55	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral deep; ID=MEK080519; tissue fluid(mpe)=19.13; mmp(mpe)=0.0153; FSR (%/hr)=0.3198
SUBJECT_SAMPLE_FACTORS           	-	65	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial superficial; ID=UU13; tissue fluid(mpe)=20.06; mmp(mpe)=0.0118; FSR (%/hr)=0.2362
SUBJECT_SAMPLE_FACTORS           	-	67	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. medial deep; ID=UU13; tissue fluid(mpe)=19.64; mmp(mpe)=0.0064; FSR (%/hr)=0.1304
SUBJECT_SAMPLE_FACTORS           	-	69	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral superficial; ID=UU13; tissue fluid(mpe)=19.03; mmp(mpe)=0.0118; FSR (%/hr)=0.2485
SUBJECT_SAMPLE_FACTORS           	-	71	Mechanical loading hours/day:None | Group:Passive Mechanical Loading	Tissue=Gast=R. lateral deep; ID=UU13; tissue fluid(mpe)=19.19; mmp(mpe)=0.0107; FSR (%/hr)=0.2231
SUBJECT_SAMPLE_FACTORS           	-	1	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=080617-1; tissue fluid(mpe)=26.02; mmp(mpe)=0.0169; FSR (%/hr)=0.2595
SUBJECT_SAMPLE_FACTORS           	-	10	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=080617-2; tissue fluid(mpe)=20.46; mmp(mpe)=0.0125; FSR (%/hr)=0.2453
SUBJECT_SAMPLE_FACTORS           	-	100	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU17; tissue fluid(mpe)=30.48; mmp(mpe)=0.0187; FSR (%/hr)=0.2457
SUBJECT_SAMPLE_FACTORS           	-	101	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU17; tissue fluid(mpe)=34.48; mmp(mpe)=0.0183; FSR (%/hr)=0.2125
SUBJECT_SAMPLE_FACTORS           	-	102	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU17; tissue fluid(mpe)=31.05; mmp(mpe)=0.0179; FSR (%/hr)=0.2310
SUBJECT_SAMPLE_FACTORS           	-	103	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU17; tissue fluid(mpe)=34.57; mmp(mpe)=0.0240; FSR (%/hr)=0.2782
SUBJECT_SAMPLE_FACTORS           	-	104	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU17; tissue fluid(mpe)=30.37; mmp(mpe)=0.0250; FSR (%/hr)=0.3288
SUBJECT_SAMPLE_FACTORS           	-	11	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=080617-2; tissue fluid(mpe)=26.73; mmp(mpe)=0.0112; FSR (%/hr)=0.1676
SUBJECT_SAMPLE_FACTORS           	-	12	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=080617-2; tissue fluid(mpe)=22.31; mmp(mpe)=0.0178; FSR (%/hr)=0.3199
SUBJECT_SAMPLE_FACTORS           	-	13	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=080617-2; tissue fluid(mpe)=27.76; mmp(mpe)=0.0170; FSR (%/hr)=0.2449
SUBJECT_SAMPLE_FACTORS           	-	14	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=080617-2; tissue fluid(mpe)=23.57; mmp(mpe)=0.0156; FSR (%/hr)=0.2645
SUBJECT_SAMPLE_FACTORS           	-	15	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=080617-2; tissue fluid(mpe)=25.98; mmp(mpe)=0.0229; FSR (%/hr)=0.3525
SUBJECT_SAMPLE_FACTORS           	-	16	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=080617-2; tissue fluid(mpe)=25.42; mmp(mpe)=0.0198; FSR (%/hr)=0.3121
SUBJECT_SAMPLE_FACTORS           	-	17	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU11; tissue fluid(mpe)=15.80; mmp(mpe)=0.0063; FSR (%/hr)=0.1588
SUBJECT_SAMPLE_FACTORS           	-	18	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU11; tissue fluid(mpe)=15.35; mmp(mpe)=0.0045; FSR (%/hr)=0.1173
SUBJECT_SAMPLE_FACTORS           	-	19	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU11; tissue fluid(mpe)=15.72; mmp(mpe)=0.0069; FSR (%/hr)=0.1755
SUBJECT_SAMPLE_FACTORS           	-	2	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=080617-1; tissue fluid(mpe)=29.02; mmp(mpe)=0.0165; FSR (%/hr)=0.2274
SUBJECT_SAMPLE_FACTORS           	-	20	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU11; tissue fluid(mpe)=12.51; mmp(mpe)=0.0042; FSR (%/hr)=0.1338
SUBJECT_SAMPLE_FACTORS           	-	21	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU11; tissue fluid(mpe)=14.78; mmp(mpe)=0.0048; FSR (%/hr)=0.1287
SUBJECT_SAMPLE_FACTORS           	-	22	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU11; tissue fluid(mpe)=13.47; mmp(mpe)=0.0060; FSR (%/hr)=0.1770
SUBJECT_SAMPLE_FACTORS           	-	23	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU11; tissue fluid(mpe)=16.60; mmp(mpe)=0.0092; FSR (%/hr)=0.2215
SUBJECT_SAMPLE_FACTORS           	-	24	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU11; tissue fluid(mpe)=15.47; mmp(mpe)=0.0120; FSR (%/hr)=0.3102
SUBJECT_SAMPLE_FACTORS           	-	25	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU12; tissue fluid(mpe)=24.10; mmp(mpe)=0.0171; FSR (%/hr)=0.2835
SUBJECT_SAMPLE_FACTORS           	-	26	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU12; tissue fluid(mpe)=16.25; mmp(mpe)=0.0082; FSR (%/hr)=0.2020
SUBJECT_SAMPLE_FACTORS           	-	27	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU12; tissue fluid(mpe)=22.97; mmp(mpe)=0.0235; FSR (%/hr)=0.4087
SUBJECT_SAMPLE_FACTORS           	-	28	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU12; tissue fluid(mpe)=19.81; mmp(mpe)=0.0207; FSR (%/hr)=0.4188
SUBJECT_SAMPLE_FACTORS           	-	29	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU12; tissue fluid(mpe)=23.32; mmp(mpe)=0.0115; FSR (%/hr)=0.1966
SUBJECT_SAMPLE_FACTORS           	-	3	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=080617-1; tissue fluid(mpe)=26.14; mmp(mpe)=0.0127; FSR (%/hr)=0.1942
SUBJECT_SAMPLE_FACTORS           	-	30	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU12; tissue fluid(mpe)=15.88; mmp(mpe)=0.0022; FSR (%/hr)=0.0557
SUBJECT_SAMPLE_FACTORS           	-	31	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU12; tissue fluid(mpe)=24.52; mmp(mpe)=0.0162; FSR (%/hr)=0.2647
SUBJECT_SAMPLE_FACTORS           	-	32	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU12; tissue fluid(mpe)=15.23; mmp(mpe)=0.0040; FSR (%/hr)=0.1053
SUBJECT_SAMPLE_FACTORS           	-	4	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=080617-1; tissue fluid(mpe)=28.66; mmp(mpe)=0.0131; FSR (%/hr)=0.1825
SUBJECT_SAMPLE_FACTORS           	-	5	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=080617-1; tissue fluid(mpe)=28.39; mmp(mpe)=0.0178; FSR (%/hr)=0.2511
SUBJECT_SAMPLE_FACTORS           	-	57	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=071220-3; tissue fluid(mpe)=28.82; mmp(mpe)=0.0116; FSR (%/hr)=0.1604
SUBJECT_SAMPLE_FACTORS           	-	58	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=071220-3; tissue fluid(mpe)=25.39; mmp(mpe)=0.0127; FSR (%/hr)=0.1995
SUBJECT_SAMPLE_FACTORS           	-	59	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=071220-3; tissue fluid(mpe)=30.68; mmp(mpe)=0.0158; FSR (%/hr)=0.2055
SUBJECT_SAMPLE_FACTORS           	-	6	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=080617-1; tissue fluid(mpe)=28.02; mmp(mpe)=0.0181; FSR (%/hr)=0.2588
SUBJECT_SAMPLE_FACTORS           	-	60	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=071220-3; tissue fluid(mpe)=28.22; mmp(mpe)=0.0166; FSR (%/hr)=0.2353
SUBJECT_SAMPLE_FACTORS           	-	61	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=071220-3; tissue fluid(mpe)=24.72; mmp(mpe)=0.0070; FSR (%/hr)=0.1133
SUBJECT_SAMPLE_FACTORS           	-	62	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=071220-3; tissue fluid(mpe)=24.03; mmp(mpe)=0.0063; FSR (%/hr)=0.1055
SUBJECT_SAMPLE_FACTORS           	-	63	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=071220-3; tissue fluid(mpe)=30.66; mmp(mpe)=0.0175; FSR (%/hr)=0.2284
SUBJECT_SAMPLE_FACTORS           	-	64	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=071220-3; tissue fluid(mpe)=29.48; mmp(mpe)=0.0202; FSR (%/hr)=0.2735
SUBJECT_SAMPLE_FACTORS           	-	7	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=080617-1; tissue fluid(mpe)=27.05; mmp(mpe)=0.0229; FSR (%/hr)=0.3394
SUBJECT_SAMPLE_FACTORS           	-	73	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU14; tissue fluid(mpe)=21.74; mmp(mpe)=0.0082; FSR (%/hr)=0.1503
SUBJECT_SAMPLE_FACTORS           	-	74	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU14; tissue fluid(mpe)=21.84; mmp(mpe)=0.0167; FSR (%/hr)=0.3064
SUBJECT_SAMPLE_FACTORS           	-	75	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU14; tissue fluid(mpe)=24.08; mmp(mpe)=0.0226; FSR (%/hr)=0.3749
SUBJECT_SAMPLE_FACTORS           	-	76	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU14; tissue fluid(mpe)=22.22; mmp(mpe)=0.0208; FSR (%/hr)=0.3749
SUBJECT_SAMPLE_FACTORS           	-	77	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU14; tissue fluid(mpe)=21.14; mmp(mpe)=0.0092; FSR (%/hr)=0.1748
SUBJECT_SAMPLE_FACTORS           	-	78	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU14; tissue fluid(mpe)=22.41; mmp(mpe)=0.0158; FSR (%/hr)=0.2818
SUBJECT_SAMPLE_FACTORS           	-	79	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU14; tissue fluid(mpe)=24.22; mmp(mpe)=0.0238; FSR (%/hr)=0.3926
SUBJECT_SAMPLE_FACTORS           	-	8	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=080617-1; tissue fluid(mpe)=25.90; mmp(mpe)=0.0266; FSR (%/hr)=0.4110
SUBJECT_SAMPLE_FACTORS           	-	80	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU14; tissue fluid(mpe)=22.65; mmp(mpe)=0.0215; FSR (%/hr)=0.3797
SUBJECT_SAMPLE_FACTORS           	-	81	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU15; tissue fluid(mpe)=16.91; mmp(mpe)=0.0218; FSR (%/hr)=0.5153
SUBJECT_SAMPLE_FACTORS           	-	82	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU15; tissue fluid(mpe)=15.88; mmp(mpe)=0.0169; FSR (%/hr)=0.4253
SUBJECT_SAMPLE_FACTORS           	-	83	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU15; tissue fluid(mpe)=16.23; mmp(mpe)=0.0305; FSR (%/hr)=0.7529
SUBJECT_SAMPLE_FACTORS           	-	84	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU15; tissue fluid(mpe)=16.57; mmp(mpe)=0.0241; FSR (%/hr)=0.5814
SUBJECT_SAMPLE_FACTORS           	-	85	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU15; tissue fluid(mpe)=15.47; mmp(mpe)=0.0205; FSR (%/hr)=0.5306
SUBJECT_SAMPLE_FACTORS           	-	86	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU15; tissue fluid(mpe)=17.58; mmp(mpe)=0.0223; FSR (%/hr)=0.5072
SUBJECT_SAMPLE_FACTORS           	-	87	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU15; tissue fluid(mpe)=16.11; mmp(mpe)=0.0306; FSR (%/hr)=0.7598
SUBJECT_SAMPLE_FACTORS           	-	88	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU15; tissue fluid(mpe)=16.75; mmp(mpe)=0.0201; FSR (%/hr)=0.4805
SUBJECT_SAMPLE_FACTORS           	-	89	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU16; tissue fluid(mpe)=14.92; mmp(mpe)=0.0377; FSR (%/hr)=1.0119
SUBJECT_SAMPLE_FACTORS           	-	9	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=080617-2; tissue fluid(mpe)=25.58; mmp(mpe)=0.0205; FSR (%/hr)=0.3209
SUBJECT_SAMPLE_FACTORS           	-	90	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU16; tissue fluid(mpe)=14.54; mmp(mpe)=0.0108; FSR (%/hr)=0.2960
SUBJECT_SAMPLE_FACTORS           	-	91	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU16; tissue fluid(mpe)=15.31; mmp(mpe)=0.0569; FSR (%/hr)=1.4853
SUBJECT_SAMPLE_FACTORS           	-	92	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial deep; ID=UU16; tissue fluid(mpe)=17.92; mmp(mpe)=0.0229; FSR (%/hr)=0.5116
SUBJECT_SAMPLE_FACTORS           	-	93	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral superficial; ID=UU16; tissue fluid(mpe)=17.25; mmp(mpe)=0.0142; FSR (%/hr)=0.3293
SUBJECT_SAMPLE_FACTORS           	-	94	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral superficial; ID=UU16; tissue fluid(mpe)=14.13; mmp(mpe)=0.0037; FSR (%/hr)=0.1061
SUBJECT_SAMPLE_FACTORS           	-	95	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. lateral deep; ID=UU16; tissue fluid(mpe)=17.81; mmp(mpe)=0.0230; FSR (%/hr)=0.5170
SUBJECT_SAMPLE_FACTORS           	-	96	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. lateral deep; ID=UU16; tissue fluid(mpe)=14.92; mmp(mpe)=0.0025; FSR (%/hr)=0.0682
SUBJECT_SAMPLE_FACTORS           	-	97	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial superficial; ID=UU17; tissue fluid(mpe)=31.62; mmp(mpe)=0.0165; FSR (%/hr)=0.2090
SUBJECT_SAMPLE_FACTORS           	-	98	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=L. medial superficial; ID=UU17; tissue fluid(mpe)=30.41; mmp(mpe)=0.0180; FSR (%/hr)=0.2369
SUBJECT_SAMPLE_FACTORS           	-	99	Mechanical loading hours/day:None | Group:Sham	Tissue=Gast=R. medial deep; ID=UU17; tissue fluid(mpe)=35.19; mmp(mpe)=0.0194; FSR (%/hr)=0.2200
#COLLECTION
CO:COLLECTION_SUMMARY            	The tibialis anterior (TA), extensor digitorum longus (EDL), plantaris,
CO:COLLECTION_SUMMARY            	gastrocnemius and soleus muscles were dissected from the loaded left leg and the
CO:COLLECTION_SUMMARY            	unloaded right leg immediately after death. One half of the soleus and EDL
CO:COLLECTION_SUMMARY            	muscles together with TA and gastrocnemius were quickly frozen in liquid propane
CO:COLLECTION_SUMMARY            	cooled by liquid nitrogen, and stored at ?160°C for further analyses. In the
CO:COLLECTION_SUMMARY            	other halves of the soleus and EDL muscles, bundles of approximately 50 fibres
CO:COLLECTION_SUMMARY            	were dissected from the muscles in relaxing solution at 4°C and tied to glass
CO:COLLECTION_SUMMARY            	capillaries, stretched to about 110% of their resting slack length. The bundles
CO:COLLECTION_SUMMARY            	were chemically skinned for 24 h in relaxing solution containing 50% (v/v)
CO:COLLECTION_SUMMARY            	glycerol for 24 h at 4°C and were subsequently stored at ?20°C (Larsson &
CO:COLLECTION_SUMMARY            	Moss, 1993). All the bundles were cryo-protected within 1 week after skinning by
CO:COLLECTION_SUMMARY            	transferring the bundles every 30 min to relax solution containing increasing
CO:COLLECTION_SUMMARY            	concentrations of sucrose, i.e. 0, 0.5, 1.0, 1.5 and 2.0 m, and subsequently
CO:COLLECTION_SUMMARY            	frozen in liquid propane chilled with liquid nitrogen (Frontera & Larsson,
CO:COLLECTION_SUMMARY            	1997). The frozen bundles were stored at ?160°C pending use. One day before the
CO:COLLECTION_SUMMARY            	experiments, a bundle was transferred to a 2.0 m sucrose solution for 30 min,
CO:COLLECTION_SUMMARY            	subsequently incubated in solutions of decreasing sucrose concentration
CO:COLLECTION_SUMMARY            	(1.5–0.5 m) and finally kept in a skinning solution at ?20°C.
CO:SAMPLE_TYPE                   	Muscle
#TREATMENT
TR:TREATMENT_SUMMARY             	Mechanical stimulation|Sham operation
TR:TREATMENT_PROTOCOL_COMMENTS   	Sham operated controls and 46 anaesthetized and mechanically ventilated female
TR:TREATMENT_PROTOCOL_COMMENTS   	Sprague–Dawley rats treated with ?-cobratoxin for durations varying from 6 h to
TR:TREATMENT_PROTOCOL_COMMENTS   	14 days were included in this study. The experimental model has previously been
TR:TREATMENT_PROTOCOL_COMMENTS   	described in detail (Dworkin & Dworkin, 1990, 2004). For mechanical stimulation
TR:TREATMENT_PROTOCOL_COMMENTS   	animals, left leg was mechanically stimulated and right leg was not. |Sham
TR:TREATMENT_PROTOCOL_COMMENTS   	operated controls and 46 anaesthetized and mechanically ventilated female
TR:TREATMENT_PROTOCOL_COMMENTS   	Sprague–Dawley rats treated with ?-cobratoxin for durations varying from 6 h to
TR:TREATMENT_PROTOCOL_COMMENTS   	14 days were included in this study. The experimental model has previously been
TR:TREATMENT_PROTOCOL_COMMENTS   	described in detail (Dworkin & Dworkin, 1990, 2004). For sham operated controls,
TR:TREATMENT_PROTOCOL_COMMENTS   	no stimulation was performed on either legs
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	An i.v. bolus dose of [ring-13C6]phenylalanine (15 ?g g?1 body weight) was given
SP:SAMPLEPREP_SUMMARY            	15 min prior to the animals being killed. Immediately after death, the
SP:SAMPLEPREP_SUMMARY            	gastrocnemius muscle was removed from the left and right hindlimb and split into
SP:SAMPLEPREP_SUMMARY            	a medial and lateral deep red and superficial white portion and frozen in liquid
SP:SAMPLEPREP_SUMMARY            	propane chilled by liquid nitrogen. Tissue fluid and mixed gastrocnemius muscle
SP:SAMPLEPREP_SUMMARY            	proteins were isolated according to the method of Ljungqvist et al. (1997). The
SP:SAMPLEPREP_SUMMARY            	mixed muscle protein precipitate from the isolation was hydrolysed by heating
SP:SAMPLEPREP_SUMMARY            	with 6 m HCl overnight at 110°C. Both tissue fluid and hydrolysed mixed
SP:SAMPLEPREP_SUMMARY            	gastrocnemius muscle amino acids were purified using a BioRad AG-50 × 8 ion
SP:SAMPLEPREP_SUMMARY            	exchange resin prior to mass spectrometry analysis.
Tissue fluid
The level SP:SAMPLEPREP_SUMMARY of enrichment of [ring-13C6]phenylalanine in tissue fluid was analysed using SP:SAMPLEPREP_SUMMARY ThermoFisher Quantum gas chromatography tandem mass spectrometry (GC/MS/MS) (San SP:SAMPLEPREP_SUMMARY Jose, CA, USA). The heptafluorobutyryl isobutyl ester derivative was prepared as SP:SAMPLEPREP_SUMMARY described by Ford et al. (1985) and the amino acids were measured under negative SP:SAMPLEPREP_SUMMARY ion chemical ionisation conditions using isobutane as reactant gas. The [M-HF]? SP:SAMPLEPREP_SUMMARY fragments reflecting the m0 and m+6 species were monitored (m/z transitions SP:SAMPLEPREP_SUMMARY 397?377 and 403?383, respectively) and the enrichment of the label was measured SP:SAMPLEPREP_SUMMARY against a calibration curve prepared from known amounts of labelled and SP:SAMPLEPREP_SUMMARY unlabelled phenylalanine (range 0–30%). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The level of enrichment of [ring-13C6]phenylalanine derived from hydrolysed CH:CHROMATOGRAPHY_SUMMARY mixed muscle proteins were analysed using a ThermoFisher DeltaPlus Isotope Ratio CH:CHROMATOGRAPHY_SUMMARY mass spectrometer (IR/MS) (Bremen, Germany) fitted with an on-line gas CH:CHROMATOGRAPHY_SUMMARY chromatograph with oxidation and reduction furnaces as previously described CH:CHROMATOGRAPHY_SUMMARY (Balagopal et al. 1996). The amino acids were derivatized to their trimethyl CH:CHROMATOGRAPHY_SUMMARY acetyl, methyl esters according to the method of Metges et al. (1996). Any amino CH:CHROMATOGRAPHY_SUMMARY acid eluting from the gas chromatograph is converted to CO2 and N2 prior to CH:CHROMATOGRAPHY_SUMMARY entry into the IR/MS. The amino acids were derivatized to their trimethyl CH:CHROMATOGRAPHY_SUMMARY acetyl, methyl esters according to the method of Metges et al. (1996). CH:CHROMATOGRAPHY_SUMMARY Enrichment of the tracer was measured by monitoring the ratio of 13C to 12CO2 in CH:CHROMATOGRAPHY_SUMMARY the IR/MS and again referenced to a calibration curve (00.1%). CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:LABORATORY_NAME Mayo Clinic Metabolomics Core AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo DeltaPlus Isotope Ratio MS MS:INSTRUMENT_TYPE IR MS MS:MS_TYPE IR MS MS:MS_COMMENTS the amino acids were measured under negative ion chemical ionisation conditions MS:MS_COMMENTS using isobutane as reactant gas MS:ION_MODE NEGATIVE #END