PMC:1448208 JSON TXT 7 Projects

Annnotations TAB TSV DIC JSON TextAE

Id	Subject	Object	Predicate	Lexical cue
T834	0-10	NN	denotes	Production
T835	0-102	sentence	denotes	Production and characterization of murine models of classic and intermediate maple syrup urine disease
T836	11-14	CC	denotes	and
T837	15-31	NN	denotes	characterization
T838	32-34	IN	denotes	of
T839	35-41	JJ	denotes	murine
T840	42-48	NNS	denotes	models
T841	49-51	IN	denotes	of
T842	52-59	JJ	denotes	classic
T844	60-63	CC	denotes	and
T845	64-76	JJ	denotes	intermediate
T846	77-82	NN	denotes	maple
T847	83-88	NN	denotes	syrup
T848	89-94	NN	denotes	urine
T843	95-102	NN	denotes	disease
T849	102-103	sentence	denotes
T850	113-123	sentence	denotes	Background
T851	113-123	NN	denotes	Background
T852	123-255	sentence	denotes	Maple Syrup Urine Disease (MSUD) is an inborn error of metabolism caused by a deficiency of branched-chain keto acid dehydrogenase.
T853	124-129	NN	denotes	Maple
T854	130-135	NN	denotes	Syrup
T856	136-141	NN	denotes	Urine
T855	142-149	NN	denotes	Disease
T858	150-151	-LRB-	denotes	(
T859	151-155	NN	denotes	MSUD
T860	155-156	-RRB-	denotes	)
T857	157-159	VBZ	denotes	is
T861	160-162	DT	denotes	an
T863	163-169	JJ	denotes	inborn
T862	170-175	NN	denotes	error
T864	176-178	IN	denotes	of
T865	179-189	NN	denotes	metabolism
T866	190-196	VBN	denotes	caused
T867	197-199	IN	denotes	by
T868	200-201	DT	denotes	a
T869	202-212	NN	denotes	deficiency
T870	213-215	IN	denotes	of
T871	216-224	VBN	denotes	branched
T873	224-225	HYPH	denotes	-
T872	225-230	NN	denotes	chain
T875	231-235	NN	denotes	keto
T876	236-240	NN	denotes	acid
T874	241-254	NN	denotes	dehydrogenase
T877	254-255	.	denotes	.
T878	255-338	sentence	denotes	MSUD has several clinical phenotypes depending on the degree of enzyme deficiency.
T879	256-260	NN	denotes	MSUD
T880	261-264	VBZ	denotes	has
T881	265-272	JJ	denotes	several
T883	273-281	JJ	denotes	clinical
T882	282-292	NNS	denotes	phenotypes
T884	293-302	VBG	denotes	depending
T885	303-305	IN	denotes	on
T886	306-309	DT	denotes	the
T887	310-316	NN	denotes	degree
T888	317-319	IN	denotes	of
T889	320-326	NN	denotes	enzyme
T890	327-337	NN	denotes	deficiency
T891	337-338	.	denotes	.
T892	338-429	sentence	denotes	Current treatments are not satisfactory and require new approaches to combat this disease.
T893	339-346	JJ	denotes	Current
T894	347-357	NNS	denotes	treatments
T895	358-361	VBP	denotes	are
T896	362-365	RB	denotes	not
T897	366-378	JJ	denotes	satisfactory
T898	379-382	CC	denotes	and
T899	383-390	VBP	denotes	require
T900	391-394	JJ	denotes	new
T901	395-405	NNS	denotes	approaches
T902	406-408	TO	denotes	to
T903	409-415	VB	denotes	combat
T904	416-420	DT	denotes	this
T905	421-428	NN	denotes	disease
T906	428-429	.	denotes	.
T907	429-519	sentence	denotes	A major hurdle in developing new treatments has been the lack of a suitable animal model.
T908	430-431	DT	denotes	A
T910	432-437	JJ	denotes	major
T909	438-444	NN	denotes	hurdle
T912	445-447	IN	denotes	in
T913	448-458	VBG	denotes	developing
T914	459-462	JJ	denotes	new
T915	463-473	NNS	denotes	treatments
T916	474-477	VBZ	denotes	has
T911	478-482	VBN	denotes	been
T917	483-486	DT	denotes	the
T918	487-491	NN	denotes	lack
T919	492-494	IN	denotes	of
T920	495-496	DT	denotes	a
T922	497-505	JJ	denotes	suitable
T923	506-512	NN	denotes	animal
T921	513-518	NN	denotes	model
T924	518-519	.	denotes	.
T925	519-528	sentence	denotes	Methods
T926	521-528	NNS	denotes	Methods
T927	528-737	sentence	denotes	To create a murine model of classic MSUD, we used gene targeting and embryonic stem cell technologies to create a mouse line that lacked a functional E2 subunit gene of branched-chain keto acid dehydrogenase.
T928	529-531	TO	denotes	To
T929	532-538	VB	denotes	create
T931	539-540	DT	denotes	a
T933	541-547	JJ	denotes	murine
T932	548-553	NN	denotes	model
T934	554-556	IN	denotes	of
T935	557-564	JJ	denotes	classic
T936	565-569	NN	denotes	MSUD
T937	569-571	,	denotes	,
T938	571-573	PRP	denotes	we
T930	574-578	VBD	denotes	used
T939	579-583	NN	denotes	gene
T940	584-593	NN	denotes	targeting
T941	594-597	CC	denotes	and
T942	598-607	JJ	denotes	embryonic
T944	608-612	NN	denotes	stem
T943	613-617	NN	denotes	cell
T945	618-630	NNS	denotes	technologies
T946	631-633	TO	denotes	to
T947	634-640	VB	denotes	create
T948	641-642	DT	denotes	a
T950	643-648	NN	denotes	mouse
T949	649-653	NN	denotes	line
T951	654-658	WDT	denotes	that
T952	659-665	VBD	denotes	lacked
T953	666-667	DT	denotes	a
T955	668-678	JJ	denotes	functional
T956	679-681	NN	denotes	E2
T957	682-689	NN	denotes	subunit
T954	690-694	NN	denotes	gene
T958	695-697	IN	denotes	of
T959	698-706	VBN	denotes	branched
T961	706-707	HYPH	denotes	-
T960	707-712	NN	denotes	chain
T963	713-717	NN	denotes	keto
T964	718-722	NN	denotes	acid
T962	723-736	NN	denotes	dehydrogenase
T965	736-737	.	denotes	.
T966	737-869	sentence	denotes	To create a murine model of intermediate MSUD, we used transgenic technology to express a human E2 cDNA on the knockout background.
T967	738-740	TO	denotes	To
T968	741-747	VB	denotes	create
T970	748-749	DT	denotes	a
T972	750-756	JJ	denotes	murine
T971	757-762	NN	denotes	model
T973	763-765	IN	denotes	of
T974	766-778	JJ	denotes	intermediate
T975	779-783	NN	denotes	MSUD
T976	783-785	,	denotes	,
T977	785-787	PRP	denotes	we
T969	788-792	VBD	denotes	used
T978	793-803	JJ	denotes	transgenic
T979	804-814	NN	denotes	technology
T980	815-817	TO	denotes	to
T981	818-825	VB	denotes	express
T982	826-827	DT	denotes	a
T984	828-833	JJ	denotes	human
T985	834-836	NN	denotes	E2
T983	837-841	NN	denotes	cDNA
T986	842-844	IN	denotes	on
T987	845-848	DT	denotes	the
T989	849-857	NN	denotes	knockout
T988	858-868	NN	denotes	background
T990	868-869	.	denotes	.
T991	869-964	sentence	denotes	Mice of both models were characterized at the molecular, biochemical, and whole animal levels.
T992	870-874	NNS	denotes	Mice
T994	875-877	IN	denotes	of
T995	878-882	DT	denotes	both
T996	883-889	NNS	denotes	models
T997	890-894	VBD	denotes	were
T993	895-908	VBN	denotes	characterized
T998	909-911	IN	denotes	at
T999	912-915	DT	denotes	the
T1001	916-925	JJ	denotes	molecular
T1002	925-927	,	denotes	,
T1003	927-938	JJ	denotes	biochemical
T1004	938-940	,	denotes	,
T1005	940-943	CC	denotes	and
T1006	944-949	JJ	denotes	whole
T1007	950-956	NN	denotes	animal
T1000	957-963	NNS	denotes	levels
T1008	963-964	.	denotes	.
T1009	964-973	sentence	denotes	Results
T1010	966-973	NNS	denotes	Results
T1011	973-1106	sentence	denotes	By disrupting the E2 subunit gene of branched-chain keto acid dehydrogenase, we created a gene knockout mouse model of classic MSUD.
T1012	974-976	IN	denotes	By
T1014	977-987	VBG	denotes	disrupting
T1015	988-991	DT	denotes	the
T1017	992-994	NN	denotes	E2
T1018	995-1002	NN	denotes	subunit
T1016	1003-1007	NN	denotes	gene
T1019	1008-1010	IN	denotes	of
T1020	1011-1019	VBN	denotes	branched
T1022	1019-1020	HYPH	denotes	-
T1021	1020-1025	NN	denotes	chain
T1024	1026-1030	NN	denotes	keto
T1025	1031-1035	NN	denotes	acid
T1023	1036-1049	NN	denotes	dehydrogenase
T1026	1049-1051	,	denotes	,
T1027	1051-1053	PRP	denotes	we
T1013	1054-1061	VBD	denotes	created
T1028	1062-1063	DT	denotes	a
T1030	1064-1068	NN	denotes	gene
T1031	1069-1077	NN	denotes	knockout
T1032	1078-1083	NN	denotes	mouse
T1029	1084-1089	NN	denotes	model
T1033	1090-1092	IN	denotes	of
T1034	1093-1100	JJ	denotes	classic
T1035	1101-1105	NN	denotes	MSUD
T1036	1105-1106	.	denotes	.
T1037	1106-1281	sentence	denotes	The homozygous knockout mice lacked branched-chain keto acid dehydrogenase activity, E2 immunoreactivity, and had a 3-fold increase in circulating branched-chain amino acids.
T1038	1107-1110	DT	denotes	The
T1040	1111-1121	JJ	denotes	homozygous
T1041	1122-1130	NN	denotes	knockout
T1039	1131-1135	NNS	denotes	mice
T1042	1136-1142	VBD	denotes	lacked
T1043	1143-1151	VBN	denotes	branched
T1045	1151-1152	HYPH	denotes	-
T1044	1152-1157	NN	denotes	chain
T1047	1158-1162	NN	denotes	keto
T1048	1163-1167	NN	denotes	acid
T1046	1168-1181	NN	denotes	dehydrogenase
T1049	1182-1190	NN	denotes	activity
T1050	1190-1192	,	denotes	,
T1051	1192-1194	NN	denotes	E2
T1052	1195-1211	NN	denotes	immunoreactivity
T1053	1211-1213	,	denotes	,
T1054	1213-1216	CC	denotes	and
T1055	1217-1220	VBD	denotes	had
T1056	1221-1222	DT	denotes	a
T1058	1223-1224	CD	denotes	3
T1059	1224-1225	HYPH	denotes	-
T1060	1225-1229	JJ	denotes	fold
T1057	1230-1238	NN	denotes	increase
T1061	1239-1241	IN	denotes	in
T1062	1242-1253	VBG	denotes	circulating
T1064	1254-1262	VBN	denotes	branched
T1066	1262-1263	HYPH	denotes	-
T1065	1263-1268	NN	denotes	chain
T1067	1269-1274	NN	denotes	amino
T1063	1275-1280	NNS	denotes	acids
T1068	1280-1281	.	denotes	.
T1069	1281-1342	sentence	denotes	These metabolic derangements resulted in neonatal lethality.
T1070	1282-1287	DT	denotes	These
T1072	1288-1297	JJ	denotes	metabolic
T1071	1298-1310	NNS	denotes	derangements
T1073	1311-1319	VBD	denotes	resulted
T1074	1320-1322	IN	denotes	in
T1075	1323-1331	JJ	denotes	neonatal
T1076	1332-1341	NN	denotes	lethality
T1077	1341-1342	.	denotes	.
T1078	1342-1462	sentence	denotes	Transgenic expression of a human E2 cDNA in the liver of the E2 knockout animals produced a model of intermediate MSUD.
T1079	1343-1353	JJ	denotes	Transgenic
T1080	1354-1364	NN	denotes	expression
T1082	1365-1367	IN	denotes	of
T1083	1368-1369	DT	denotes	a
T1085	1370-1375	JJ	denotes	human
T1086	1376-1378	NN	denotes	E2
T1084	1379-1383	NN	denotes	cDNA
T1087	1384-1386	IN	denotes	in
T1088	1387-1390	DT	denotes	the
T1089	1391-1396	NN	denotes	liver
T1090	1397-1399	IN	denotes	of
T1091	1400-1403	DT	denotes	the
T1093	1404-1406	NN	denotes	E2
T1094	1407-1415	NN	denotes	knockout
T1092	1416-1423	NNS	denotes	animals
T1081	1424-1432	VBD	denotes	produced
T1095	1433-1434	DT	denotes	a
T1096	1435-1440	NN	denotes	model
T1097	1441-1443	IN	denotes	of
T1098	1444-1456	JJ	denotes	intermediate
T1099	1457-1461	NN	denotes	MSUD
T1100	1461-1462	.	denotes	.
T1101	1462-1723	sentence	denotes	Branched-chain keto acid dehydrogenase activity was 5–6% of normal and was sufficient to allow survival, but was insufficient to normalize circulating branched-chain amino acids levels, which were intermediate between wildtype and the classic MSUD mouse model.
T1102	1463-1471	VBN	denotes	Branched
T1104	1471-1472	HYPH	denotes	-
T1103	1472-1477	NN	denotes	chain
T1106	1478-1482	NN	denotes	keto
T1107	1483-1487	NN	denotes	acid
T1105	1488-1501	NN	denotes	dehydrogenase
T1108	1502-1510	NN	denotes	activity
T1109	1511-1514	VBD	denotes	was
T1110	1515-1516	CD	denotes	5
T1112	1516-1517	SYM	denotes	–
T1111	1517-1518	CD	denotes	6
T1113	1518-1519	NN	denotes	%
T1114	1520-1522	IN	denotes	of
T1115	1523-1529	JJ	denotes	normal
T1116	1530-1533	CC	denotes	and
T1117	1534-1537	VBD	denotes	was
T1118	1538-1548	JJ	denotes	sufficient
T1119	1549-1551	TO	denotes	to
T1120	1552-1557	VB	denotes	allow
T1121	1558-1566	NN	denotes	survival
T1122	1566-1568	,	denotes	,
T1123	1568-1571	CC	denotes	but
T1124	1572-1575	VBD	denotes	was
T1125	1576-1588	JJ	denotes	insufficient
T1126	1589-1591	TO	denotes	to
T1128	1592-1601	VB	denotes	normalize
T1127	1602-1613	VBG	denotes	circulating
T1129	1614-1622	VBN	denotes	branched
T1131	1622-1623	HYPH	denotes	-
T1130	1623-1628	NN	denotes	chain
T1133	1629-1634	NN	denotes	amino
T1132	1635-1640	NNS	denotes	acids
T1134	1641-1647	NNS	denotes	levels
T1135	1647-1649	,	denotes	,
T1136	1649-1654	WDT	denotes	which
T1137	1655-1659	VBD	denotes	were
T1138	1660-1672	JJ	denotes	intermediate
T1139	1673-1680	IN	denotes	between
T1140	1681-1689	NN	denotes	wildtype
T1141	1690-1693	CC	denotes	and
T1142	1694-1697	DT	denotes	the
T1144	1698-1705	JJ	denotes	classic
T1145	1706-1710	NN	denotes	MSUD
T1146	1711-1716	NN	denotes	mouse
T1143	1717-1722	NN	denotes	model
T1147	1722-1723	.	denotes	.
T1148	1723-1735	sentence	denotes	Conclusion
T1149	1725-1735	NN	denotes	Conclusion
T1150	1735-1878	sentence	denotes	These mice represent important animal models that closely approximate the phenotype of humans with the classic and intermediate forms of MSUD.
T1151	1736-1741	DT	denotes	These
T1152	1742-1746	NNS	denotes	mice
T1153	1747-1756	VBP	denotes	represent
T1154	1757-1766	JJ	denotes	important
T1156	1767-1773	NN	denotes	animal
T1155	1774-1780	NNS	denotes	models
T1157	1781-1785	WDT	denotes	that
T1159	1786-1793	RB	denotes	closely
T1158	1794-1805	VBP	denotes	approximate
T1160	1806-1809	DT	denotes	the
T1161	1810-1819	NN	denotes	phenotype
T1162	1820-1822	IN	denotes	of
T1163	1823-1829	NNS	denotes	humans
T1164	1830-1834	IN	denotes	with
T1165	1835-1838	DT	denotes	the
T1167	1839-1846	JJ	denotes	classic
T1168	1847-1850	CC	denotes	and
T1169	1851-1863	JJ	denotes	intermediate
T1166	1864-1869	NNS	denotes	forms
T1170	1870-1872	IN	denotes	of
T1171	1873-1877	NN	denotes	MSUD
T1172	1877-1878	.	denotes	.
T1173	1878-2102	sentence	denotes	These animals provide useful models to further characterize the pathogenesis of MSUD, as well as models to test novel therapeutic strategies, such as gene and cellular therapies, to treat this devastating metabolic disease.
T1174	1879-1884	DT	denotes	These
T1175	1885-1892	NNS	denotes	animals
T1176	1893-1900	VBP	denotes	provide
T1177	1901-1907	JJ	denotes	useful
T1178	1908-1914	NNS	denotes	models
T1179	1915-1917	TO	denotes	to
T1181	1918-1925	RB	denotes	further
T1180	1926-1938	VB	denotes	characterize
T1182	1939-1942	DT	denotes	the
T1183	1943-1955	NN	denotes	pathogenesis
T1184	1956-1958	IN	denotes	of
T1185	1959-1963	NN	denotes	MSUD
T1186	1963-1965	,	denotes	,
T1187	1965-1967	RB	denotes	as
T1189	1968-1972	RB	denotes	well
T1188	1973-1975	IN	denotes	as
T1190	1976-1982	NNS	denotes	models
T1191	1983-1985	TO	denotes	to
T1192	1986-1990	VB	denotes	test
T1193	1991-1996	JJ	denotes	novel
T1195	1997-2008	JJ	denotes	therapeutic
T1194	2009-2019	NNS	denotes	strategies
T1196	2019-2021	,	denotes	,
T1197	2021-2025	JJ	denotes	such
T1198	2026-2028	IN	denotes	as
T1199	2029-2033	NN	denotes	gene
T1201	2034-2037	CC	denotes	and
T1202	2038-2046	JJ	denotes	cellular
T1200	2047-2056	NNS	denotes	therapies
T1203	2056-2058	,	denotes	,
T1204	2058-2060	TO	denotes	to
T1205	2061-2066	VB	denotes	treat
T1206	2067-2071	DT	denotes	this
T1208	2072-2083	VBG	denotes	devastating
T1209	2084-2093	JJ	denotes	metabolic
T1207	2094-2101	NN	denotes	disease
T1210	2101-2102	.	denotes	.
T3304	2115-2120	NN	denotes	Maple
T3305	2121-2126	NN	denotes	Syrup
T3307	2127-2132	NN	denotes	Urine
T3306	2133-2140	NN	denotes	Disease
T3309	2141-2142	-LRB-	denotes	(
T3310	2142-2146	NN	denotes	MSUD
T3311	2146-2147	-RRB-	denotes	)
T3308	2148-2150	VBZ	denotes	is
T3312	2151-2152	DT	denotes	a
T3314	2153-2160	JJ	denotes	genetic
T3313	2161-2169	NN	denotes	disorder
T3315	2170-2176	VBN	denotes	caused
T3316	2177-2179	IN	denotes	by
T3317	2180-2181	DT	denotes	a
T3318	2182-2192	NN	denotes	deficiency
T3319	2193-2195	IN	denotes	of
T3320	2196-2204	VBN	denotes	branched
T3322	2204-2205	HYPH	denotes	-
T3321	2205-2210	NN	denotes	chain
T3324	2211-2215	NN	denotes	keto
T3325	2216-2220	NN	denotes	acid
T3323	2221-2234	NN	denotes	dehydrogenase
T3326	2235-2236	-LRB-	denotes	(
T3327	2236-2241	NN	denotes	BCKDH
T3328	2241-2242	-RRB-	denotes	)
T3329	2242-2244	,	denotes	,
T3330	2244-2245	DT	denotes	a
T3332	2246-2259	JJ	denotes	mitochondrial
T3333	2260-2271	NN	denotes	multienzyme
T3331	2272-2279	NN	denotes	complex
T3334	2280-2291	JJ	denotes	responsible
T3335	2292-2295	IN	denotes	for
T3336	2296-2299	DT	denotes	the
T3338	2300-2309	JJ	denotes	oxidative
T3337	2310-2325	NN	denotes	decarboxylation
T3339	2326-2328	IN	denotes	of
T3340	2329-2337	VBN	denotes	branched
T3342	2337-2338	HYPH	denotes	-
T3341	2338-2343	NN	denotes	chain
T3344	2344-2348	NN	denotes	keto
T3343	2349-2354	NNS	denotes	acids
T3345	2355-2362	VBN	denotes	derived
T3346	2363-2367	IN	denotes	from
T3347	2368-2376	VBN	denotes	branched
T3349	2376-2377	HYPH	denotes	-
T3348	2377-2382	NN	denotes	chain
T3351	2383-2388	NN	denotes	amino
T3350	2389-2394	NNS	denotes	acids
T3352	2395-2396	-LRB-	denotes	(
T3353	2396-2400	NN	denotes	BCAA
T3354	2400-2401	-RRB-	denotes	)
T3355	2401-2403	,	denotes	,
T3356	2403-2410	NN	denotes	leucine
T3357	2410-2412	,	denotes	,
T3358	2412-2422	NN	denotes	isoleucine
T3359	2423-2426	CC	denotes	and
T3360	2427-2433	NN	denotes	valine
T3361	2434-2435	-LRB-	denotes	(
T3363	2435-2438	IN	denotes	for
T3364	2439-2445	NN	denotes	review
T3365	2445-2447	,	denotes	,
T3362	2447-2450	VB	denotes	see
T3366	2450-2452	:	denotes	:
T3367	2452-2453	-LRB-	denotes	[
T3368	2453-2454	CD	denotes	1
T3369	2454-2455	-RRB-	denotes	]
T3370	2455-2456	-RRB-	denotes	)
T3371	2456-2457	.	denotes	.
T3372	2457-2601	sentence	denotes	Patients with MSUD, depending on the mutation, show variable degrees of enzyme deficiency leading to several different clinical phenotypes [1].
T3373	2458-2466	NNS	denotes	Patients
T3375	2467-2471	IN	denotes	with
T3376	2472-2476	NN	denotes	MSUD
T3377	2476-2478	,	denotes	,
T3378	2478-2487	VBG	denotes	depending
T3379	2488-2490	IN	denotes	on
T3380	2491-2494	DT	denotes	the
T3381	2495-2503	NN	denotes	mutation
T3382	2503-2505	,	denotes	,
T3374	2505-2509	VBP	denotes	show
T3383	2510-2518	JJ	denotes	variable
T3384	2519-2526	NNS	denotes	degrees
T3385	2527-2529	IN	denotes	of
T3386	2530-2536	NN	denotes	enzyme
T3387	2537-2547	NN	denotes	deficiency
T3388	2548-2555	VBG	denotes	leading
T3389	2556-2558	IN	denotes	to
T3390	2559-2566	JJ	denotes	several
T3392	2567-2576	JJ	denotes	different
T3393	2577-2585	JJ	denotes	clinical
T3391	2586-2596	NNS	denotes	phenotypes
T3394	2597-2598	-LRB-	denotes	[
T3395	2598-2599	CD	denotes	1
T3396	2599-2600	-RRB-	denotes	]
T3397	2600-2601	.	denotes	.
T3398	2601-2729	sentence	denotes	Approximately 75% of MSUD patients have the classic form of the disease with BCKDH activity in the range of 0–2% of normal [1].
T3399	2602-2615	RB	denotes	Approximately
T3400	2616-2618	CD	denotes	75
T3401	2618-2619	NN	denotes	%
T3403	2620-2622	IN	denotes	of
T3404	2623-2627	NN	denotes	MSUD
T3405	2628-2636	NNS	denotes	patients
T3402	2637-2641	VBP	denotes	have
T3406	2642-2645	DT	denotes	the
T3408	2646-2653	JJ	denotes	classic
T3407	2654-2658	NN	denotes	form
T3409	2659-2661	IN	denotes	of
T3410	2662-2665	DT	denotes	the
T3411	2666-2673	NN	denotes	disease
T3412	2674-2678	IN	denotes	with
T3413	2679-2684	NN	denotes	BCKDH
T3414	2685-2693	NN	denotes	activity
T3415	2694-2696	IN	denotes	in
T3416	2697-2700	DT	denotes	the
T3417	2701-2706	NN	denotes	range
T3418	2707-2709	IN	denotes	of
T3419	2710-2711	CD	denotes	0
T3421	2711-2712	SYM	denotes	–
T3420	2712-2713	CD	denotes	2
T3422	2713-2714	NN	denotes	%
T3423	2715-2717	IN	denotes	of
T3424	2718-2724	JJ	denotes	normal
T3425	2725-2726	-LRB-	denotes	[
T3426	2726-2727	CD	denotes	1
T3427	2727-2728	-RRB-	denotes	]
T3428	2728-2729	.	denotes	.
T3429	2729-2818	sentence	denotes	These patients show markedly elevated levels of BCAA in blood and other body fluids [1].
T3430	2730-2735	DT	denotes	These
T3431	2736-2744	NNS	denotes	patients
T3432	2745-2749	VBP	denotes	show
T3433	2750-2758	RB	denotes	markedly
T3434	2759-2767	VBN	denotes	elevated
T3435	2768-2774	NNS	denotes	levels
T3436	2775-2777	IN	denotes	of
T3437	2778-2782	NN	denotes	BCAA
T3438	2783-2785	IN	denotes	in
T3439	2786-2791	NN	denotes	blood
T3440	2792-2795	CC	denotes	and
T3441	2796-2801	JJ	denotes	other
T3443	2802-2806	NN	denotes	body
T3442	2807-2813	NNS	denotes	fluids
T3444	2814-2815	-LRB-	denotes	[
T3445	2815-2816	CD	denotes	1
T3446	2816-2817	-RRB-	denotes	]
T3447	2817-2818	.	denotes	.
T3448	2818-2885	sentence	denotes	Besides the classic form, there are other variants of the disease.
T3449	2819-2826	IN	denotes	Besides
T3451	2827-2830	DT	denotes	the
T3453	2831-2838	JJ	denotes	classic
T3452	2839-2843	NN	denotes	form
T3454	2843-2845	,	denotes	,
T3455	2845-2850	EX	denotes	there
T3450	2851-2854	VBP	denotes	are
T3456	2855-2860	JJ	denotes	other
T3457	2861-2869	NNS	denotes	variants
T3458	2870-2872	IN	denotes	of
T3459	2873-2876	DT	denotes	the
T3460	2877-2884	NN	denotes	disease
T3461	2884-2885	.	denotes	.
T3462	2885-2989	sentence	denotes	Patients with the intermediate form of the disease show BCKDH activity in the range of 3–30% of normal.
T3463	2886-2894	NNS	denotes	Patients
T3465	2895-2899	IN	denotes	with
T3466	2900-2903	DT	denotes	the
T3468	2904-2916	JJ	denotes	intermediate
T3467	2917-2921	NN	denotes	form
T3469	2922-2924	IN	denotes	of
T3470	2925-2928	DT	denotes	the
T3471	2929-2936	NN	denotes	disease
T3464	2937-2941	VBP	denotes	show
T3472	2942-2947	NN	denotes	BCKDH
T3473	2948-2956	NN	denotes	activity
T3474	2957-2959	IN	denotes	in
T3475	2960-2963	DT	denotes	the
T3476	2964-2969	NN	denotes	range
T3477	2970-2972	IN	denotes	of
T3478	2973-2974	CD	denotes	3
T3480	2974-2975	SYM	denotes	–
T3479	2975-2977	CD	denotes	30
T3481	2977-2978	NN	denotes	%
T3482	2979-2981	IN	denotes	of
T3483	2982-2988	JJ	denotes	normal
T3484	2988-2989	.	denotes	.
T3485	2989-3099	sentence	denotes	In such patients the onset of the disease is delayed, but there are persistently elevated levels of BCAA [1].
T3486	2990-2992	IN	denotes	In
T3488	2993-2997	JJ	denotes	such
T3489	2998-3006	NNS	denotes	patients
T3490	3007-3010	DT	denotes	the
T3491	3011-3016	NN	denotes	onset
T3492	3017-3019	IN	denotes	of
T3493	3020-3023	DT	denotes	the
T3494	3024-3031	NN	denotes	disease
T3495	3032-3034	VBZ	denotes	is
T3487	3035-3042	VBN	denotes	delayed
T3496	3042-3044	,	denotes	,
T3497	3044-3047	CC	denotes	but
T3498	3048-3053	EX	denotes	there
T3499	3054-3057	VBP	denotes	are
T3500	3058-3070	RB	denotes	persistently
T3501	3071-3079	VBN	denotes	elevated
T3502	3080-3086	NNS	denotes	levels
T3503	3087-3089	IN	denotes	of
T3504	3090-3094	NN	denotes	BCAA
T3505	3095-3096	-LRB-	denotes	[
T3506	3096-3097	CD	denotes	1
T3507	3097-3098	-RRB-	denotes	]
T3508	3098-3099	.	denotes	.
T3509	3099-3257	sentence	denotes	Patients with the intermittent form of MSUD show BCKDH activity in the range of 5–20% and during the asymptomatic phase the blood BCAA levels are normal [1].
T3510	3100-3108	NNS	denotes	Patients
T3512	3109-3113	IN	denotes	with
T3513	3114-3117	DT	denotes	the
T3515	3118-3130	JJ	denotes	intermittent
T3514	3131-3135	NN	denotes	form
T3516	3136-3138	IN	denotes	of
T3517	3139-3143	NN	denotes	MSUD
T3511	3144-3148	VBP	denotes	show
T3518	3149-3154	NN	denotes	BCKDH
T3519	3155-3163	NN	denotes	activity
T3520	3164-3166	IN	denotes	in
T3521	3167-3170	DT	denotes	the
T3522	3171-3176	NN	denotes	range
T3523	3177-3179	IN	denotes	of
T3524	3180-3181	CD	denotes	5
T3526	3181-3182	SYM	denotes	–
T3525	3182-3184	CD	denotes	20
T3527	3184-3185	NN	denotes	%
T3528	3186-3189	CC	denotes	and
T3529	3190-3196	IN	denotes	during
T3531	3197-3200	DT	denotes	the
T3533	3201-3213	JJ	denotes	asymptomatic
T3532	3214-3219	NN	denotes	phase
T3534	3220-3223	DT	denotes	the
T3536	3224-3229	NN	denotes	blood
T3537	3230-3234	NN	denotes	BCAA
T3535	3235-3241	NNS	denotes	levels
T3530	3242-3245	VBP	denotes	are
T3538	3246-3252	JJ	denotes	normal
T3539	3253-3254	-LRB-	denotes	[
T3540	3254-3255	CD	denotes	1
T3541	3255-3256	-RRB-	denotes	]
T3542	3256-3257	.	denotes	.
T3543	3257-3456	sentence	denotes	The overall incidence of MSUD in the general population is 1:185,000 [1], and in certain population groups, such as Mennonites of Pennsylvania, the incidence is estimated to be as high as 1:176 [2].
T3544	3258-3261	DT	denotes	The
T3546	3262-3269	JJ	denotes	overall
T3545	3270-3279	NN	denotes	incidence
T3548	3280-3282	IN	denotes	of
T3549	3283-3287	NN	denotes	MSUD
T3550	3288-3290	IN	denotes	in
T3551	3291-3294	DT	denotes	the
T3553	3295-3302	JJ	denotes	general
T3552	3303-3313	NN	denotes	population
T3547	3314-3316	VBZ	denotes	is
T3554	3317-3318	CD	denotes	1
T3556	3318-3319	SYM	denotes	:
T3555	3319-3326	CD	denotes	185,000
T3557	3327-3328	-LRB-	denotes	[
T3558	3328-3329	CD	denotes	1
T3559	3329-3330	-RRB-	denotes	]
T3560	3330-3332	,	denotes	,
T3561	3332-3335	CC	denotes	and
T3562	3336-3338	IN	denotes	in
T3564	3339-3346	JJ	denotes	certain
T3566	3347-3357	NN	denotes	population
T3565	3358-3364	NNS	denotes	groups
T3567	3364-3366	,	denotes	,
T3568	3366-3370	JJ	denotes	such
T3569	3371-3373	IN	denotes	as
T3570	3374-3384	NNS	denotes	Mennonites
T3571	3385-3387	IN	denotes	of
T3572	3388-3400	NNP	denotes	Pennsylvania
T3573	3400-3402	,	denotes	,
T3574	3402-3405	DT	denotes	the
T3575	3406-3415	NN	denotes	incidence
T3576	3416-3418	VBZ	denotes	is
T3563	3419-3428	VBN	denotes	estimated
T3577	3429-3431	TO	denotes	to
T3578	3432-3434	VB	denotes	be
T3579	3435-3437	IN	denotes	as
T3580	3438-3442	JJ	denotes	high
T3581	3443-3445	IN	denotes	as
T3582	3446-3447	CD	denotes	1
T3584	3447-3448	SYM	denotes	:
T3583	3448-3451	CD	denotes	176
T3585	3452-3453	-LRB-	denotes	[
T3586	3453-3454	CD	denotes	2
T3587	3454-3455	-RRB-	denotes	]
T3588	3455-3456	.	denotes	.
T3589	3456-3640	sentence	denotes	The BCKDH complex, the deficient enzyme in MSUD, consists of three catalytic proteins, a decarboxylase (E1), a dihydrolipoyl transacylase (E2), and a dihydrolipoyl dehydrogenase (E3).
T3590	3457-3460	DT	denotes	The
T3592	3461-3466	NN	denotes	BCKDH
T3591	3467-3474	NN	denotes	complex
T3594	3474-3476	,	denotes	,
T3595	3476-3479	DT	denotes	the
T3597	3480-3489	JJ	denotes	deficient
T3596	3490-3496	NN	denotes	enzyme
T3598	3497-3499	IN	denotes	in
T3599	3500-3504	NN	denotes	MSUD
T3600	3504-3506	,	denotes	,
T3593	3506-3514	VBZ	denotes	consists
T3601	3515-3517	IN	denotes	of
T3602	3518-3523	CD	denotes	three
T3604	3524-3533	JJ	denotes	catalytic
T3603	3534-3542	NN	denotes	proteins
T3605	3542-3544	,	denotes	,
T3606	3544-3545	DT	denotes	a
T3607	3546-3559	NN	denotes	decarboxylase
T3608	3560-3561	-LRB-	denotes	(
T3609	3561-3563	NN	denotes	E1
T3610	3563-3564	-RRB-	denotes	)
T3611	3564-3566	,	denotes	,
T3612	3566-3567	DT	denotes	a
T3614	3568-3581	NN	denotes	dihydrolipoyl
T3613	3582-3594	NN	denotes	transacylase
T3615	3595-3596	-LRB-	denotes	(
T3616	3596-3598	NN	denotes	E2
T3617	3598-3599	-RRB-	denotes	)
T3618	3599-3601	,	denotes	,
T3619	3601-3604	CC	denotes	and
T3620	3605-3606	DT	denotes	a
T3622	3607-3620	NN	denotes	dihydrolipoyl
T3621	3621-3634	NN	denotes	dehydrogenase
T3623	3635-3636	-LRB-	denotes	(
T3624	3636-3638	NN	denotes	E3
T3625	3638-3639	-RRB-	denotes	)
T3626	3639-3640	.	denotes	.
T3627	3640-3719	sentence	denotes	The E1 component is a heterotetramer composed of two α and two β subunits [3].
T3628	3641-3644	DT	denotes	The
T3630	3645-3647	NN	denotes	E1
T3629	3648-3657	NN	denotes	component
T3631	3658-3660	VBZ	denotes	is
T3632	3661-3662	DT	denotes	a
T3633	3663-3677	NN	denotes	heterotetramer
T3634	3678-3686	VBN	denotes	composed
T3635	3687-3689	IN	denotes	of
T3636	3690-3693	CD	denotes	two
T3637	3694-3695	NN	denotes	α
T3639	3696-3699	CC	denotes	and
T3640	3700-3703	CD	denotes	two
T3641	3704-3705	NN	denotes	β
T3638	3706-3714	NNS	denotes	subunits
T3642	3715-3716	-LRB-	denotes	[
T3643	3716-3717	CD	denotes	3
T3644	3717-3718	-RRB-	denotes	]
T3645	3718-3719	.	denotes	.
T3646	3719-3888	sentence	denotes	The E1 and E2 components are specific to BCKDH, whereas E3 is also used by pyruvate and α-ketoglutarate dehydrogenase complexes [3] and the glycine cleavage system [4].
T3647	3720-3723	DT	denotes	The
T3649	3724-3726	NN	denotes	E1
T3650	3727-3730	CC	denotes	and
T3651	3731-3733	NN	denotes	E2
T3648	3734-3744	NNS	denotes	components
T3652	3745-3748	VBP	denotes	are
T3653	3749-3757	JJ	denotes	specific
T3654	3758-3760	IN	denotes	to
T3655	3761-3766	NN	denotes	BCKDH
T3656	3766-3768	,	denotes	,
T3657	3768-3775	IN	denotes	whereas
T3659	3776-3778	NN	denotes	E3
T3660	3779-3781	VBZ	denotes	is
T3661	3782-3786	RB	denotes	also
T3658	3787-3791	VBN	denotes	used
T3662	3792-3794	IN	denotes	by
T3663	3795-3803	NN	denotes	pyruvate
T3665	3804-3807	CC	denotes	and
T3666	3808-3809	NN	denotes	α
T3668	3809-3810	HYPH	denotes	-
T3667	3810-3823	NN	denotes	ketoglutarate
T3669	3824-3837	NN	denotes	dehydrogenase
T3664	3838-3847	NNS	denotes	complexes
T3670	3848-3849	-LRB-	denotes	[
T3671	3849-3850	CD	denotes	3
T3672	3850-3851	-RRB-	denotes	]
T3673	3852-3855	CC	denotes	and
T3674	3856-3859	DT	denotes	the
T3676	3860-3867	NN	denotes	glycine
T3677	3868-3876	NN	denotes	cleavage
T3675	3877-3883	NN	denotes	system
T3678	3884-3885	-LRB-	denotes	[
T3679	3885-3886	CD	denotes	4
T3680	3886-3887	-RRB-	denotes	]
T3681	3887-3888	.	denotes	.
T3682	3888-4131	sentence	denotes	BCKDH is also associated with two regulatory proteins, a specific kinase and a phosphatase which regulate the activity of this enzyme through a phosphorylation (inactivation) and dephosphorylation (activation) cycle of the E1 α subunit [5,6].
T3683	3889-3894	NN	denotes	BCKDH
T3685	3895-3897	VBZ	denotes	is
T3686	3898-3902	RB	denotes	also
T3684	3903-3913	VBN	denotes	associated
T3687	3914-3918	IN	denotes	with
T3688	3919-3922	CD	denotes	two
T3690	3923-3933	JJ	denotes	regulatory
T3689	3934-3942	NN	denotes	proteins
T3691	3942-3944	,	denotes	,
T3692	3944-3945	DT	denotes	a
T3694	3946-3954	JJ	denotes	specific
T3693	3955-3961	NN	denotes	kinase
T3695	3962-3965	CC	denotes	and
T3696	3966-3967	DT	denotes	a
T3697	3968-3979	NN	denotes	phosphatase
T3698	3980-3985	WDT	denotes	which
T3699	3986-3994	VBP	denotes	regulate
T3700	3995-3998	DT	denotes	the
T3701	3999-4007	NN	denotes	activity
T3702	4008-4010	IN	denotes	of
T3703	4011-4015	DT	denotes	this
T3704	4016-4022	NN	denotes	enzyme
T3705	4023-4030	IN	denotes	through
T3706	4031-4032	DT	denotes	a
T3708	4033-4048	NN	denotes	phosphorylation
T3709	4049-4050	-LRB-	denotes	(
T3710	4050-4062	NN	denotes	inactivation
T3711	4062-4063	-RRB-	denotes	)
T3712	4064-4067	CC	denotes	and
T3713	4068-4085	NN	denotes	dephosphorylation
T3714	4086-4087	-LRB-	denotes	(
T3715	4087-4097	NN	denotes	activation
T3716	4097-4098	-RRB-	denotes	)
T3707	4099-4104	NN	denotes	cycle
T3717	4105-4107	IN	denotes	of
T3718	4108-4111	DT	denotes	the
T3720	4112-4114	NN	denotes	E1
T3721	4115-4116	NN	denotes	α
T3719	4117-4124	NN	denotes	subunit
T3722	4125-4126	-LRB-	denotes	[
T3724	4126-4127	CD	denotes	5
T3725	4127-4128	,	denotes	,
T3723	4128-4129	CD	denotes	6
T3726	4129-4130	-RRB-	denotes	]
T3727	4130-4131	.	denotes	.
T3728	4131-4299	sentence	denotes	Mutations in the genes of the E1 and E2 subunits of BCKDH have been described, however, the majority of MSUD mutations identified thus far are in the E2 subunit [1,7].
T3729	4132-4141	NNS	denotes	Mutations
T3731	4142-4144	IN	denotes	in
T3732	4145-4148	DT	denotes	the
T3733	4149-4154	NNS	denotes	genes
T3734	4155-4157	IN	denotes	of
T3735	4158-4161	DT	denotes	the
T3737	4162-4164	NN	denotes	E1
T3738	4165-4168	CC	denotes	and
T3739	4169-4171	NN	denotes	E2
T3736	4172-4180	NNS	denotes	subunits
T3740	4181-4183	IN	denotes	of
T3741	4184-4189	NN	denotes	BCKDH
T3742	4190-4194	VBP	denotes	have
T3743	4195-4199	VBN	denotes	been
T3730	4200-4209	VBN	denotes	described
T3745	4209-4211	,	denotes	,
T3746	4211-4218	RB	denotes	however
T3747	4218-4220	,	denotes	,
T3748	4220-4223	DT	denotes	the
T3749	4224-4232	NN	denotes	majority
T3750	4233-4235	IN	denotes	of
T3751	4236-4240	NN	denotes	MSUD
T3752	4241-4250	NNS	denotes	mutations
T3753	4251-4261	VBN	denotes	identified
T3754	4262-4266	RB	denotes	thus
T3755	4267-4270	RB	denotes	far
T3744	4271-4274	VBP	denotes	are
T3756	4275-4277	IN	denotes	in
T3757	4278-4281	DT	denotes	the
T3759	4282-4284	NN	denotes	E2
T3758	4285-4292	NN	denotes	subunit
T3760	4293-4294	-LRB-	denotes	[
T3762	4294-4295	CD	denotes	1
T3763	4295-4296	,	denotes	,
T3761	4296-4297	CD	denotes	7
T3764	4297-4298	-RRB-	denotes	]
T3765	4298-4299	.	denotes	.
T3766	4299-4406	sentence	denotes	To date, cases of MSUD have not been associated with defects in the regulatory kinase and phosphatase [1].
T3767	4300-4302	IN	denotes	To
T3769	4303-4307	NN	denotes	date
T3770	4307-4309	,	denotes	,
T3771	4309-4314	NNS	denotes	cases
T3772	4315-4317	IN	denotes	of
T3773	4318-4322	NN	denotes	MSUD
T3774	4323-4327	VBP	denotes	have
T3775	4328-4331	RB	denotes	not
T3776	4332-4336	VBN	denotes	been
T3768	4337-4347	VBN	denotes	associated
T3777	4348-4352	IN	denotes	with
T3778	4353-4360	NNS	denotes	defects
T3779	4361-4363	IN	denotes	in
T3780	4364-4367	DT	denotes	the
T3782	4368-4378	JJ	denotes	regulatory
T3781	4379-4385	NN	denotes	kinase
T3783	4386-4389	CC	denotes	and
T3784	4390-4401	NN	denotes	phosphatase
T3785	4402-4403	-LRB-	denotes	[
T3786	4403-4404	CD	denotes	1
T3787	4404-4405	-RRB-	denotes	]
T3788	4405-4406	.	denotes	.
T3789	4406-4516	sentence	denotes	Current management of MSUD patients relies on a strict lifelong dietary restriction of protein or BCAA [1,8].
T3790	4407-4414	JJ	denotes	Current
T3791	4415-4425	NN	denotes	management
T3793	4426-4428	IN	denotes	of
T3794	4429-4433	NN	denotes	MSUD
T3795	4434-4442	NNS	denotes	patients
T3792	4443-4449	VBZ	denotes	relies
T3796	4450-4452	IN	denotes	on
T3797	4453-4454	DT	denotes	a
T3799	4455-4461	JJ	denotes	strict
T3800	4462-4470	JJ	denotes	lifelong
T3801	4471-4478	JJ	denotes	dietary
T3798	4479-4490	NN	denotes	restriction
T3802	4491-4493	IN	denotes	of
T3803	4494-4501	NN	denotes	protein
T3804	4502-4504	CC	denotes	or
T3805	4505-4509	NN	denotes	BCAA
T3806	4510-4511	-LRB-	denotes	[
T3808	4511-4512	CD	denotes	1
T3809	4512-4513	,	denotes	,
T3807	4513-4514	CD	denotes	8
T3810	4514-4515	-RRB-	denotes	]
T3811	4515-4516	.	denotes	.
T3812	4516-4692	sentence	denotes	Such a dietary management of the disease, however, is not entirely satisfactory especially in times of metabolic decompensation due to infection, injuries and other stressors.
T3813	4517-4521	PDT	denotes	Such
T3815	4522-4523	DT	denotes	a
T3816	4524-4531	JJ	denotes	dietary
T3814	4532-4542	NN	denotes	management
T3818	4543-4545	IN	denotes	of
T3819	4546-4549	DT	denotes	the
T3820	4550-4557	NN	denotes	disease
T3821	4557-4559	,	denotes	,
T3822	4559-4566	RB	denotes	however
T3823	4566-4568	,	denotes	,
T3817	4568-4570	VBZ	denotes	is
T3824	4571-4574	RB	denotes	not
T3825	4575-4583	RB	denotes	entirely
T3826	4584-4596	JJ	denotes	satisfactory
T3827	4597-4607	RB	denotes	especially
T3828	4608-4610	IN	denotes	in
T3829	4611-4616	NNS	denotes	times
T3830	4617-4619	IN	denotes	of
T3831	4620-4629	JJ	denotes	metabolic
T3832	4630-4644	NN	denotes	decompensation
T3833	4645-4648	IN	denotes	due
T3834	4649-4651	IN	denotes	to
T3835	4652-4661	NN	denotes	infection
T3836	4661-4663	,	denotes	,
T3837	4663-4671	NNS	denotes	injuries
T3838	4672-4675	CC	denotes	and
T3839	4676-4681	JJ	denotes	other
T3840	4682-4691	NNS	denotes	stressors
T3841	4691-4692	.	denotes	.
T3842	4692-4848	sentence	denotes	In spite of dietary intervention, there is significant mortality associated with MSUD and there is a high incidence of mental retardation in survivors [9].
T3843	4693-4695	IN	denotes	In
T3845	4696-4701	NN	denotes	spite
T3846	4702-4704	IN	denotes	of
T3847	4705-4712	JJ	denotes	dietary
T3848	4713-4725	NN	denotes	intervention
T3849	4725-4727	,	denotes	,
T3850	4727-4732	EX	denotes	there
T3844	4733-4735	VBZ	denotes	is
T3851	4736-4747	JJ	denotes	significant
T3852	4748-4757	NN	denotes	mortality
T3853	4758-4768	VBN	denotes	associated
T3854	4769-4773	IN	denotes	with
T3855	4774-4778	NN	denotes	MSUD
T3856	4779-4782	CC	denotes	and
T3857	4783-4788	EX	denotes	there
T3858	4789-4791	VBZ	denotes	is
T3859	4792-4793	DT	denotes	a
T3861	4794-4798	JJ	denotes	high
T3860	4799-4808	NN	denotes	incidence
T3862	4809-4811	IN	denotes	of
T3863	4812-4818	JJ	denotes	mental
T3864	4819-4830	NN	denotes	retardation
T3865	4831-4833	IN	denotes	in
T3866	4834-4843	NNS	denotes	survivors
T3867	4844-4845	-LRB-	denotes	[
T3868	4845-4846	CD	denotes	9
T3869	4846-4847	-RRB-	denotes	]
T3870	4847-4848	.	denotes	.
T3871	4848-5062	sentence	denotes	Because of the central role of the liver in amino acid metabolism and moderate/high levels of BCKDH activity in human liver [10-12], a few cases of MSUD have recently been treated by liver transplantation [13-16].
T3872	4849-4856	IN	denotes	Because
T3874	4857-4859	IN	denotes	of
T3875	4860-4863	DT	denotes	the
T3877	4864-4871	JJ	denotes	central
T3876	4872-4876	NN	denotes	role
T3878	4877-4879	IN	denotes	of
T3879	4880-4883	DT	denotes	the
T3880	4884-4889	NN	denotes	liver
T3881	4890-4892	IN	denotes	in
T3882	4893-4898	NN	denotes	amino
T3883	4899-4903	NN	denotes	acid
T3884	4904-4914	NN	denotes	metabolism
T3885	4915-4918	CC	denotes	and
T3886	4919-4927	JJ	denotes	moderate
T3888	4927-4928	SYM	denotes	/
T3887	4928-4932	JJ	denotes	high
T3889	4933-4939	NNS	denotes	levels
T3890	4940-4942	IN	denotes	of
T3891	4943-4948	NN	denotes	BCKDH
T3892	4949-4957	NN	denotes	activity
T3893	4958-4960	IN	denotes	in
T3894	4961-4966	JJ	denotes	human
T3895	4967-4972	NN	denotes	liver
T3896	4973-4974	-LRB-	denotes	[
T3897	4974-4976	CD	denotes	10
T3898	4976-4977	SYM	denotes	-
T3899	4977-4979	CD	denotes	12
T3900	4979-4980	-RRB-	denotes	]
T3901	4980-4982	,	denotes	,
T3902	4982-4983	DT	denotes	a
T3904	4984-4987	JJ	denotes	few
T3903	4988-4993	NNS	denotes	cases
T3905	4994-4996	IN	denotes	of
T3906	4997-5001	NN	denotes	MSUD
T3907	5002-5006	VBP	denotes	have
T3908	5007-5015	RB	denotes	recently
T3909	5016-5020	VBN	denotes	been
T3873	5021-5028	VBN	denotes	treated
T3910	5029-5031	IN	denotes	by
T3911	5032-5037	NN	denotes	liver
T3912	5038-5053	NN	denotes	transplantation
T3913	5054-5055	-LRB-	denotes	[
T3914	5055-5057	CD	denotes	13
T3915	5057-5058	SYM	denotes	-
T3916	5058-5060	CD	denotes	16
T3917	5060-5061	-RRB-	denotes	]
T3918	5061-5062	.	denotes	.
T3919	5062-5182	sentence	denotes	While the short-term outcome of liver transplantation is encouraging, long-term effects of this approach are not known.
T3920	5063-5068	IN	denotes	While
T3922	5069-5072	DT	denotes	the
T3924	5073-5078	JJ	denotes	short
T3926	5078-5079	HYPH	denotes	-
T3925	5079-5083	NN	denotes	term
T3923	5084-5091	NN	denotes	outcome
T3927	5092-5094	IN	denotes	of
T3928	5095-5100	NN	denotes	liver
T3929	5101-5116	NN	denotes	transplantation
T3921	5117-5119	VBZ	denotes	is
T3931	5120-5131	JJ	denotes	encouraging
T3932	5131-5133	,	denotes	,
T3933	5133-5137	JJ	denotes	long
T3935	5137-5138	HYPH	denotes	-
T3934	5138-5142	NN	denotes	term
T3936	5143-5150	NNS	denotes	effects
T3937	5151-5153	IN	denotes	of
T3938	5154-5158	DT	denotes	this
T3939	5159-5167	NN	denotes	approach
T3940	5168-5171	VBP	denotes	are
T3941	5172-5175	RB	denotes	not
T3930	5176-5181	VBN	denotes	known
T3942	5181-5182	.	denotes	.
T3943	5182-5321	sentence	denotes	However, these patients are now required to take immunosuppressant drugs for the rest of their lives, often with undesirable side effects.
T3944	5183-5190	RB	denotes	However
T3946	5190-5192	,	denotes	,
T3947	5192-5197	DT	denotes	these
T3948	5198-5206	NNS	denotes	patients
T3949	5207-5210	VBP	denotes	are
T3950	5211-5214	RB	denotes	now
T3945	5215-5223	VBN	denotes	required
T3951	5224-5226	TO	denotes	to
T3952	5227-5231	VB	denotes	take
T3953	5232-5249	NN	denotes	immunosuppressant
T3954	5250-5255	NNS	denotes	drugs
T3955	5256-5259	IN	denotes	for
T3956	5260-5263	DT	denotes	the
T3957	5264-5268	NN	denotes	rest
T3958	5269-5271	IN	denotes	of
T3959	5272-5277	PRP$	denotes	their
T3960	5278-5283	NNS	denotes	lives
T3961	5283-5285	,	denotes	,
T3962	5285-5290	RB	denotes	often
T3963	5291-5295	IN	denotes	with
T3964	5296-5307	JJ	denotes	undesirable
T3966	5308-5312	NN	denotes	side
T3965	5313-5320	NNS	denotes	effects
T3967	5320-5321	.	denotes	.
T3968	5321-5498	sentence	denotes	Moreover, the cost associated with liver transplantation and the availability of donor livers are additional limiting factors for the practicality of treatment of this disease.
T3969	5322-5330	RB	denotes	Moreover
T3971	5330-5332	,	denotes	,
T3972	5332-5335	DT	denotes	the
T3973	5336-5340	NN	denotes	cost
T3974	5341-5351	VBN	denotes	associated
T3975	5352-5356	IN	denotes	with
T3976	5357-5362	NN	denotes	liver
T3977	5363-5378	NN	denotes	transplantation
T3978	5379-5382	CC	denotes	and
T3979	5383-5386	DT	denotes	the
T3980	5387-5399	NN	denotes	availability
T3981	5400-5402	IN	denotes	of
T3982	5403-5408	NN	denotes	donor
T3983	5409-5415	NNS	denotes	livers
T3970	5416-5419	VBP	denotes	are
T3984	5420-5430	JJ	denotes	additional
T3986	5431-5439	VBG	denotes	limiting
T3985	5440-5447	NNS	denotes	factors
T3987	5448-5451	IN	denotes	for
T3988	5452-5455	DT	denotes	the
T3989	5456-5468	NN	denotes	practicality
T3990	5469-5471	IN	denotes	of
T3991	5472-5481	NN	denotes	treatment
T3992	5482-5484	IN	denotes	of
T3993	5485-5489	DT	denotes	this
T3994	5490-5497	NN	denotes	disease
T3995	5497-5498	.	denotes	.
T3996	5498-5634	sentence	denotes	Because of the current unsatisfactory options for the treatment of MSUD, there is a need for improved therapies to combat this disease.
T3997	5499-5506	IN	denotes	Because
T3999	5507-5509	IN	denotes	of
T4000	5510-5513	DT	denotes	the
T4002	5514-5521	JJ	denotes	current
T4003	5522-5536	JJ	denotes	unsatisfactory
T4001	5537-5544	NNS	denotes	options
T4004	5545-5548	IN	denotes	for
T4005	5549-5552	DT	denotes	the
T4006	5553-5562	NN	denotes	treatment
T4007	5563-5565	IN	denotes	of
T4008	5566-5570	NN	denotes	MSUD
T4009	5570-5572	,	denotes	,
T4010	5572-5577	EX	denotes	there
T3998	5578-5580	VBZ	denotes	is
T4011	5581-5582	DT	denotes	a
T4012	5583-5587	NN	denotes	need
T4013	5588-5591	IN	denotes	for
T4014	5592-5600	VBN	denotes	improved
T4015	5601-5610	NNS	denotes	therapies
T4016	5611-5613	TO	denotes	to
T4017	5614-5620	VB	denotes	combat
T4018	5621-5625	DT	denotes	this
T4019	5626-5633	NN	denotes	disease
T4020	5633-5634	.	denotes	.
T4021	5634-5773	sentence	denotes	An obstacle to developing novel treatments for MSUD has been the lack of a suitable animal model to perform necessary preclinical studies.
T4022	5635-5637	DT	denotes	An
T4023	5638-5646	NN	denotes	obstacle
T4025	5647-5649	IN	denotes	to
T4026	5650-5660	VBG	denotes	developing
T4027	5661-5666	JJ	denotes	novel
T4028	5667-5677	NNS	denotes	treatments
T4029	5678-5681	IN	denotes	for
T4030	5682-5686	NN	denotes	MSUD
T4031	5687-5690	VBZ	denotes	has
T4024	5691-5695	VBN	denotes	been
T4032	5696-5699	DT	denotes	the
T4033	5700-5704	NN	denotes	lack
T4034	5705-5707	IN	denotes	of
T4035	5708-5709	DT	denotes	a
T4037	5710-5718	JJ	denotes	suitable
T4038	5719-5725	NN	denotes	animal
T4036	5726-5731	NN	denotes	model
T4039	5732-5734	TO	denotes	to
T4040	5735-5742	VB	denotes	perform
T4041	5743-5752	JJ	denotes	necessary
T4043	5753-5764	JJ	denotes	preclinical
T4042	5765-5772	NNS	denotes	studies
T4044	5772-5773	.	denotes	.
T4045	5773-5928	sentence	denotes	Although a Hereford calf model with MSUD has been described [17-19], this model is neither readily available nor practical to perform preclinical studies.
T4046	5774-5782	IN	denotes	Although
T4048	5783-5784	DT	denotes	a
T4050	5785-5793	NNP	denotes	Hereford
T4051	5794-5798	NN	denotes	calf
T4049	5799-5804	NN	denotes	model
T4052	5805-5809	IN	denotes	with
T4053	5810-5814	NN	denotes	MSUD
T4054	5815-5818	VBZ	denotes	has
T4055	5819-5823	VBN	denotes	been
T4047	5824-5833	VBN	denotes	described
T4057	5834-5835	-LRB-	denotes	[
T4058	5835-5837	CD	denotes	17
T4059	5837-5838	SYM	denotes	-
T4060	5838-5840	CD	denotes	19
T4061	5840-5841	-RRB-	denotes	]
T4062	5841-5843	,	denotes	,
T4063	5843-5847	DT	denotes	this
T4064	5848-5853	NN	denotes	model
T4056	5854-5856	VBZ	denotes	is
T4065	5857-5864	CC	denotes	neither
T4067	5865-5872	RB	denotes	readily
T4066	5873-5882	JJ	denotes	available
T4068	5883-5886	CC	denotes	nor
T4069	5887-5896	JJ	denotes	practical
T4070	5897-5899	TO	denotes	to
T4071	5900-5907	VB	denotes	perform
T4072	5908-5919	JJ	denotes	preclinical
T4073	5920-5927	NNS	denotes	studies
T4074	5927-5928	.	denotes	.
T4075	5928-6118	sentence	denotes	Furthermore, comparison of this animal model with human MSUD has shown some differences in the pathology of the disease [1], making this animal a less desirable model for the human disease.
T4076	5929-5940	RB	denotes	Furthermore
T4078	5940-5942	,	denotes	,
T4079	5942-5952	NN	denotes	comparison
T4080	5953-5955	IN	denotes	of
T4081	5956-5960	DT	denotes	this
T4083	5961-5967	NN	denotes	animal
T4082	5968-5973	NN	denotes	model
T4084	5974-5978	IN	denotes	with
T4085	5979-5984	JJ	denotes	human
T4086	5985-5989	NN	denotes	MSUD
T4087	5990-5993	VBZ	denotes	has
T4077	5994-5999	VBN	denotes	shown
T4088	6000-6004	DT	denotes	some
T4089	6005-6016	NNS	denotes	differences
T4090	6017-6019	IN	denotes	in
T4091	6020-6023	DT	denotes	the
T4092	6024-6033	NN	denotes	pathology
T4093	6034-6036	IN	denotes	of
T4094	6037-6040	DT	denotes	the
T4095	6041-6048	NN	denotes	disease
T4096	6049-6050	-LRB-	denotes	[
T4097	6050-6051	CD	denotes	1
T4098	6051-6052	-RRB-	denotes	]
T4099	6052-6054	,	denotes	,
T4100	6054-6060	VBG	denotes	making
T4101	6061-6065	DT	denotes	this
T4102	6066-6072	NN	denotes	animal
T4104	6073-6074	DT	denotes	a
T4105	6075-6079	RBR	denotes	less
T4106	6080-6089	JJ	denotes	desirable
T4103	6090-6095	NN	denotes	model
T4107	6096-6099	IN	denotes	for
T4108	6100-6103	DT	denotes	the
T4110	6104-6109	JJ	denotes	human
T4109	6110-6117	NN	denotes	disease
T4111	6117-6118	.	denotes	.
T4112	6118-6245	sentence	denotes	Recently, a N-ethyl-N-nitrosourea (ENU)-induced mutant mouse that phenotypically resembles human MSUD has been described [20].
T4113	6119-6127	RB	denotes	Recently
T4115	6127-6129	,	denotes	,
T4116	6129-6130	DT	denotes	a
T4118	6131-6132	NN	denotes	N
T4120	6132-6133	HYPH	denotes	-
T4119	6133-6138	NN	denotes	ethyl
T4121	6138-6139	HYPH	denotes	-
T4122	6139-6140	NN	denotes	N
T4123	6140-6141	HYPH	denotes	-
T4117	6141-6152	NN	denotes	nitrosourea
T4125	6153-6154	-LRB-	denotes	(
T4126	6154-6157	NN	denotes	ENU
T4127	6157-6158	-RRB-	denotes	)
T4128	6158-6159	HYPH	denotes	-
T4124	6159-6166	VBN	denotes	induced
T4130	6167-6173	NN	denotes	mutant
T4129	6174-6179	NN	denotes	mouse
T4131	6180-6184	WDT	denotes	that
T4133	6185-6199	RB	denotes	phenotypically
T4132	6200-6209	VBZ	denotes	resembles
T4134	6210-6215	JJ	denotes	human
T4135	6216-6220	NN	denotes	MSUD
T4136	6221-6224	VBZ	denotes	has
T4137	6225-6229	VBN	denotes	been
T4114	6230-6239	VBN	denotes	described
T4138	6240-6241	-LRB-	denotes	[
T4139	6241-6243	CD	denotes	20
T4140	6243-6244	-RRB-	denotes	]
T4141	6244-6245	.	denotes	.
T4142	6245-6414	sentence	denotes	However, the mutation in this model disrupts a splice site in the mitochondrial branched-chain aminotransferase (BCAT) gene, not in BCKDH, the deficient enzyme in MSUD.
T4143	6246-6253	RB	denotes	However
T4145	6253-6255	,	denotes	,
T4146	6255-6258	DT	denotes	the
T4147	6259-6267	NN	denotes	mutation
T4148	6268-6270	IN	denotes	in
T4149	6271-6275	DT	denotes	this
T4150	6276-6281	NN	denotes	model
T4144	6282-6290	VBZ	denotes	disrupts
T4151	6291-6292	DT	denotes	a
T4153	6293-6299	NN	denotes	splice
T4152	6300-6304	NN	denotes	site
T4154	6305-6307	IN	denotes	in
T4155	6308-6311	DT	denotes	the
T4157	6312-6325	JJ	denotes	mitochondrial
T4158	6326-6334	VBN	denotes	branched
T4160	6334-6335	HYPH	denotes	-
T4159	6335-6340	NN	denotes	chain
T4156	6341-6357	NN	denotes	aminotransferase
T4162	6358-6359	-LRB-	denotes	(
T4163	6359-6363	NN	denotes	BCAT
T4164	6363-6364	-RRB-	denotes	)
T4161	6365-6369	NN	denotes	gene
T4165	6369-6371	,	denotes	,
T4166	6371-6374	RB	denotes	not
T4167	6375-6377	IN	denotes	in
T4168	6378-6383	NN	denotes	BCKDH
T4169	6383-6385	,	denotes	,
T4170	6385-6388	DT	denotes	the
T4172	6389-6398	JJ	denotes	deficient
T4171	6399-6405	NN	denotes	enzyme
T4173	6406-6408	IN	denotes	in
T4174	6409-6413	NN	denotes	MSUD
T4175	6413-6414	.	denotes	.
T4176	6414-6532	sentence	denotes	Because the mutation is not in BCKDH, the validity of this mutant mouse line for modeling human MSUD is questionable.
T4177	6415-6422	IN	denotes	Because
T4179	6423-6426	DT	denotes	the
T4180	6427-6435	NN	denotes	mutation
T4178	6436-6438	VBZ	denotes	is
T4182	6439-6442	RB	denotes	not
T4183	6443-6445	IN	denotes	in
T4184	6446-6451	NN	denotes	BCKDH
T4185	6451-6453	,	denotes	,
T4186	6453-6456	DT	denotes	the
T4187	6457-6465	NN	denotes	validity
T4188	6466-6468	IN	denotes	of
T4189	6469-6473	DT	denotes	this
T4191	6474-6480	NN	denotes	mutant
T4192	6481-6486	NN	denotes	mouse
T4190	6487-6491	NN	denotes	line
T4193	6492-6495	IN	denotes	for
T4194	6496-6504	VBG	denotes	modeling
T4196	6505-6510	JJ	denotes	human
T4195	6511-6515	NN	denotes	MSUD
T4181	6516-6518	VBZ	denotes	is
T4197	6519-6531	JJ	denotes	questionable
T4198	6531-6532	.	denotes	.
T4199	6532-6719	sentence	denotes	The objective of the present study was to create genetically engineered murine models of MSUD that mimic the pathology of the classic and intermediate variant forms of the human disease.
T4200	6533-6536	DT	denotes	The
T4201	6537-6546	NN	denotes	objective
T4203	6547-6549	IN	denotes	of
T4204	6550-6553	DT	denotes	the
T4206	6554-6561	JJ	denotes	present
T4205	6562-6567	NN	denotes	study
T4202	6568-6571	VBD	denotes	was
T4207	6572-6574	TO	denotes	to
T4208	6575-6581	VB	denotes	create
T4209	6582-6593	RB	denotes	genetically
T4210	6594-6604	VBN	denotes	engineered
T4212	6605-6611	JJ	denotes	murine
T4211	6612-6618	NNS	denotes	models
T4213	6619-6621	IN	denotes	of
T4214	6622-6626	NN	denotes	MSUD
T4215	6627-6631	WDT	denotes	that
T4216	6632-6637	VBP	denotes	mimic
T4217	6638-6641	DT	denotes	the
T4218	6642-6651	NN	denotes	pathology
T4219	6652-6654	IN	denotes	of
T4220	6655-6658	DT	denotes	the
T4222	6659-6666	JJ	denotes	classic
T4223	6667-6670	CC	denotes	and
T4224	6671-6683	JJ	denotes	intermediate
T4225	6684-6691	JJ	denotes	variant
T4221	6692-6697	NNS	denotes	forms
T4226	6698-6700	IN	denotes	of
T4227	6701-6704	DT	denotes	the
T4229	6705-6710	JJ	denotes	human
T4228	6711-6718	NN	denotes	disease
T4230	6718-6719	.	denotes	.
T4231	6719-6859	sentence	denotes	The classic model was created by targeted inactivation of the E2 subunit of BCKDH by homologous recombination in embryonic stem (ES) cells.
T4232	6720-6723	DT	denotes	The
T4234	6724-6731	JJ	denotes	classic
T4233	6732-6737	NN	denotes	model
T4236	6738-6741	VBD	denotes	was
T4235	6742-6749	VBN	denotes	created
T4237	6750-6752	IN	denotes	by
T4238	6753-6761	VBN	denotes	targeted
T4239	6762-6774	NN	denotes	inactivation
T4240	6775-6777	IN	denotes	of
T4241	6778-6781	DT	denotes	the
T4243	6782-6784	NN	denotes	E2
T4242	6785-6792	NN	denotes	subunit
T4244	6793-6795	IN	denotes	of
T4245	6796-6801	NN	denotes	BCKDH
T4246	6802-6804	IN	denotes	by
T4247	6805-6815	JJ	denotes	homologous
T4248	6816-6829	NN	denotes	recombination
T4249	6830-6832	IN	denotes	in
T4250	6833-6842	JJ	denotes	embryonic
T4252	6843-6847	NN	denotes	stem
T4253	6848-6849	-LRB-	denotes	(
T4251	6849-6851	NN	denotes	ES
T4255	6851-6852	-RRB-	denotes	)
T4254	6853-6858	NNS	denotes	cells
T4256	6858-6859	.	denotes	.
T4257	6859-6956	sentence	denotes	The model of intermediate MSUD was created by partial transgenic rescue of the E2 gene knockout.
T4258	6860-6863	DT	denotes	The
T4259	6864-6869	NN	denotes	model
T4261	6870-6872	IN	denotes	of
T4262	6873-6885	JJ	denotes	intermediate
T4263	6886-6890	NN	denotes	MSUD
T4264	6891-6894	VBD	denotes	was
T4260	6895-6902	VBN	denotes	created
T4265	6903-6905	IN	denotes	by
T4266	6906-6913	JJ	denotes	partial
T4268	6914-6924	JJ	denotes	transgenic
T4267	6925-6931	NN	denotes	rescue
T4269	6932-6934	IN	denotes	of
T4270	6935-6938	DT	denotes	the
T4272	6939-6941	NN	denotes	E2
T4273	6942-6946	NN	denotes	gene
T4271	6947-6955	NN	denotes	knockout
T4274	6955-6956	.	denotes	.
T4275	6956-7046	sentence	denotes	This report describes the generation and characterization of these murine models of MSUD.
T4276	6957-6961	DT	denotes	This
T4277	6962-6968	NN	denotes	report
T4278	6969-6978	VBZ	denotes	describes
T4279	6979-6982	DT	denotes	the
T4280	6983-6993	NN	denotes	generation
T4281	6994-6997	CC	denotes	and
T4282	6998-7014	NN	denotes	characterization
T4283	7015-7017	IN	denotes	of
T4284	7018-7023	DT	denotes	these
T4286	7024-7030	JJ	denotes	murine
T4285	7031-7037	NNS	denotes	models
T4287	7038-7040	IN	denotes	of
T4288	7041-7045	NN	denotes	MSUD
T4289	7045-7046	.	denotes	.
T4290	7046-7183	sentence	denotes	These models will allow for the development of novel treatment approaches, such as gene or stem cell therapies, to ultimately cure MSUD.
T4291	7047-7052	DT	denotes	These
T4292	7053-7059	NNS	denotes	models
T4294	7060-7064	MD	denotes	will
T4293	7065-7070	VB	denotes	allow
T4295	7071-7074	IN	denotes	for
T4296	7075-7078	DT	denotes	the
T4297	7079-7090	NN	denotes	development
T4298	7091-7093	IN	denotes	of
T4299	7094-7099	JJ	denotes	novel
T4301	7100-7109	NN	denotes	treatment
T4300	7110-7120	NNS	denotes	approaches
T4302	7120-7122	,	denotes	,
T4303	7122-7126	JJ	denotes	such
T4304	7127-7129	IN	denotes	as
T4305	7130-7134	NN	denotes	gene
T4307	7135-7137	CC	denotes	or
T4308	7138-7142	NN	denotes	stem
T4309	7143-7147	NN	denotes	cell
T4306	7148-7157	NNS	denotes	therapies
T4310	7157-7159	,	denotes	,
T4311	7159-7161	TO	denotes	to
T4313	7162-7172	RB	denotes	ultimately
T4312	7173-7177	VB	denotes	cure
T4314	7178-7182	NN	denotes	MSUD
T4315	7182-7183	.	denotes	.
T4359	7193-7196	DT	denotes	All
T4360	7197-7204	NNS	denotes	studies
T4362	7205-7214	VBG	denotes	involving
T4363	7215-7222	NNS	denotes	animals
T4364	7223-7227	VBD	denotes	were
T4361	7228-7236	VBN	denotes	reviewed
T4365	7237-7240	CC	denotes	and
T4366	7241-7249	VBN	denotes	approved
T4367	7250-7252	IN	denotes	by
T4368	7253-7256	DT	denotes	the
T4369	7257-7267	NNP	denotes	University
T4370	7268-7270	IN	denotes	of
T4371	7271-7281	NNP	denotes	Pittsburgh
T4373	7281-7283	POS	denotes	's
T4374	7284-7297	NNP	denotes	Institutional
T4375	7298-7304	NNP	denotes	Animal
T4376	7305-7309	NNP	denotes	Care
T4377	7310-7313	CC	denotes	and
T4378	7314-7317	NNP	denotes	Use
T4372	7318-7327	NNP	denotes	Committee
T4379	7327-7328	.	denotes	.
T5027	7330-7334	NN	denotes	Gene
T5028	7335-7343	NN	denotes	knockout
T5029	7344-7354	NN	denotes	production
T5030	7354-7580	sentence	denotes	A genomic DNA subclone of Strain 129/SvJ DNA that contained a portion of Exons 4 and 5 of the E2 gene and flanking DNA was obtained from a P1 phage library from Genome Systems, Inc., (St. Louis, MO; '3-Hit Mouse ES Library').
T5031	7355-7356	DT	denotes	A
T5033	7357-7364	JJ	denotes	genomic
T5034	7365-7368	NN	denotes	DNA
T5032	7369-7377	NN	denotes	subclone
T5036	7378-7380	IN	denotes	of
T5037	7381-7387	NN	denotes	Strain
T5039	7388-7391	CD	denotes	129
T5041	7391-7392	HYPH	denotes	/
T5040	7392-7395	NN	denotes	SvJ
T5038	7396-7399	NN	denotes	DNA
T5042	7400-7404	WDT	denotes	that
T5043	7405-7414	VBD	denotes	contained
T5044	7415-7416	DT	denotes	a
T5045	7417-7424	NN	denotes	portion
T5046	7425-7427	IN	denotes	of
T5047	7428-7433	NNS	denotes	Exons
T5048	7434-7435	CD	denotes	4
T5049	7436-7439	CC	denotes	and
T5050	7440-7441	CD	denotes	5
T5051	7442-7444	IN	denotes	of
T5052	7445-7448	DT	denotes	the
T5054	7449-7451	NN	denotes	E2
T5053	7452-7456	NN	denotes	gene
T5055	7457-7460	CC	denotes	and
T5056	7461-7469	NN	denotes	flanking
T5057	7470-7473	NN	denotes	DNA
T5058	7474-7477	VBD	denotes	was
T5035	7478-7486	VBN	denotes	obtained
T5059	7487-7491	IN	denotes	from
T5060	7492-7493	DT	denotes	a
T5062	7494-7496	NN	denotes	P1
T5063	7497-7502	NN	denotes	phage
T5061	7503-7510	NN	denotes	library
T5064	7511-7515	IN	denotes	from
T5065	7516-7522	NNP	denotes	Genome
T5067	7523-7530	NNP	denotes	Systems
T5068	7530-7532	,	denotes	,
T5066	7532-7536	NNP	denotes	Inc.
T5069	7536-7538	,	denotes	,
T5070	7538-7539	-LRB-	denotes	(
T5072	7539-7542	NNP	denotes	St.
T5073	7543-7548	NNP	denotes	Louis
T5074	7548-7550	,	denotes	,
T5075	7550-7552	NNP	denotes	MO
T5076	7552-7553	:	denotes	;
T5077	7554-7555	SYM	denotes	'
T5078	7555-7556	CD	denotes	3
T5080	7556-7557	HYPH	denotes	-
T5079	7557-7560	NN	denotes	Hit
T5081	7561-7566	NNP	denotes	Mouse
T5082	7567-7569	NNP	denotes	ES
T5071	7570-7577	NNP	denotes	Library
T5083	7577-7578	POS	denotes	'
T5084	7578-7579	-RRB-	denotes	)
T5085	7579-7580	.	denotes	.
T5086	7580-7820	sentence	denotes	A positive/negative replacement type gene targeting vector [21] was created by replacing a 1.67 kb EcoRV-Smal fragment that included a portion of Exon 4 and all of Exon 5 with the PGKneo marker gene (See Fig. 1A) from the pPNT vector [22].
T5087	7581-7582	DT	denotes	A
T5089	7583-7591	JJ	denotes	positive
T5091	7591-7592	HYPH	denotes	/
T5090	7592-7600	JJ	denotes	negative
T5092	7601-7612	NN	denotes	replacement
T5093	7613-7617	NN	denotes	type
T5094	7618-7622	NN	denotes	gene
T5095	7623-7632	NN	denotes	targeting
T5088	7633-7639	NN	denotes	vector
T5097	7640-7641	-LRB-	denotes	[
T5098	7641-7643	CD	denotes	21
T5099	7643-7644	-RRB-	denotes	]
T5100	7645-7648	VBD	denotes	was
T5096	7649-7656	VBN	denotes	created
T5101	7657-7659	IN	denotes	by
T5102	7660-7669	VBG	denotes	replacing
T5103	7670-7671	DT	denotes	a
T5105	7672-7676	CD	denotes	1.67
T5106	7677-7679	NN	denotes	kb
T5107	7680-7685	NN	denotes	EcoRV
T5109	7685-7686	HYPH	denotes	-
T5108	7686-7690	NN	denotes	Smal
T5104	7691-7699	NN	denotes	fragment
T5110	7700-7704	WDT	denotes	that
T5111	7705-7713	VBD	denotes	included
T5112	7714-7715	DT	denotes	a
T5113	7716-7723	NN	denotes	portion
T5114	7724-7726	IN	denotes	of
T5115	7727-7731	NN	denotes	Exon
T5116	7732-7733	CD	denotes	4
T5117	7734-7737	CC	denotes	and
T5118	7738-7741	DT	denotes	all
T5119	7742-7744	IN	denotes	of
T5120	7745-7749	NN	denotes	Exon
T5121	7750-7751	CD	denotes	5
T5122	7752-7756	IN	denotes	with
T5123	7757-7760	DT	denotes	the
T5125	7761-7767	NN	denotes	PGKneo
T5126	7768-7774	NN	denotes	marker
T5124	7775-7779	NN	denotes	gene
T5127	7780-7781	-LRB-	denotes	(
T5128	7781-7784	VB	denotes	See
T5129	7785-7789	NN	denotes	Fig.
T5130	7790-7792	NN	denotes	1A
T5131	7792-7793	-RRB-	denotes	)
T5132	7794-7798	IN	denotes	from
T5133	7799-7802	DT	denotes	the
T5135	7803-7807	NN	denotes	pPNT
T5134	7808-7814	NN	denotes	vector
T5136	7815-7816	-LRB-	denotes	[
T5137	7816-7818	CD	denotes	22
T5138	7818-7819	-RRB-	denotes	]
T5139	7819-7820	.	denotes	.
T5140	7820-7954	sentence	denotes	The targeting construct was linearized with NotI and electroporated into R1 ES cells [23] under conditions described previously [24].
T5141	7821-7824	DT	denotes	The
T5143	7825-7834	NN	denotes	targeting
T5142	7835-7844	NN	denotes	construct
T5145	7845-7848	VBD	denotes	was
T5144	7849-7859	VBN	denotes	linearized
T5146	7860-7864	IN	denotes	with
T5147	7865-7869	NN	denotes	NotI
T5148	7870-7873	CC	denotes	and
T5149	7874-7888	VBN	denotes	electroporated
T5150	7889-7893	IN	denotes	into
T5151	7894-7896	NN	denotes	R1
T5153	7897-7899	NN	denotes	ES
T5152	7900-7905	NNS	denotes	cells
T5154	7906-7907	-LRB-	denotes	[
T5155	7907-7909	CD	denotes	23
T5156	7909-7910	-RRB-	denotes	]
T5157	7911-7916	IN	denotes	under
T5158	7917-7927	NNS	denotes	conditions
T5159	7928-7937	VBN	denotes	described
T5160	7938-7948	RB	denotes	previously
T5161	7949-7950	-LRB-	denotes	[
T5162	7950-7952	CD	denotes	24
T5163	7952-7953	-RRB-	denotes	]
T5164	7953-7954	.	denotes	.
T5165	7954-8096	sentence	denotes	ES cells were selected with G418 (300 μg/ml; Life Technologies Inc., Gaithersburg, MD) and gancyclovir (2 μM; gift of Syntex, Palo Alto, CA).
T5166	7955-7957	NN	denotes	ES
T5167	7958-7963	NNS	denotes	cells
T5169	7964-7968	VBD	denotes	were
T5168	7969-7977	VBN	denotes	selected
T5170	7978-7982	IN	denotes	with
T5171	7983-7987	NN	denotes	G418
T5172	7988-7989	-LRB-	denotes	(
T5174	7989-7992	CD	denotes	300
T5175	7993-7995	NN	denotes	μg
T5176	7995-7996	SYM	denotes	/
T5177	7996-7998	NN	denotes	ml
T5178	7998-7999	:	denotes	;
T5179	8000-8004	NNP	denotes	Life
T5180	8005-8017	NNP	denotes	Technologies
T5173	8018-8022	NNP	denotes	Inc.
T5181	8022-8024	,	denotes	,
T5182	8024-8036	NNP	denotes	Gaithersburg
T5183	8036-8038	,	denotes	,
T5184	8038-8040	NNP	denotes	MD
T5185	8040-8041	-RRB-	denotes	)
T5186	8042-8045	CC	denotes	and
T5187	8046-8057	NN	denotes	gancyclovir
T5188	8058-8059	-LRB-	denotes	(
T5189	8059-8060	CD	denotes	2
T5190	8061-8063	NN	denotes	μM
T5191	8063-8064	:	denotes	;
T5192	8065-8069	NN	denotes	gift
T5193	8070-8072	IN	denotes	of
T5194	8073-8079	NNP	denotes	Syntex
T5195	8079-8081	,	denotes	,
T5196	8081-8085	NNP	denotes	Palo
T5197	8086-8090	NNP	denotes	Alto
T5198	8090-8092	,	denotes	,
T5199	8092-8094	NNP	denotes	CA
T5200	8094-8095	-RRB-	denotes	)
T5201	8095-8096	.	denotes	.
T5202	8096-8209	sentence	denotes	Doubly resistant clones were screened for gene targeting by Southern blot analysis of BglI digested genomic DNA.
T5203	8097-8103	RB	denotes	Doubly
T5204	8104-8113	JJ	denotes	resistant
T5205	8114-8120	NNS	denotes	clones
T5207	8121-8125	VBD	denotes	were
T5206	8126-8134	VBN	denotes	screened
T5208	8135-8138	IN	denotes	for
T5209	8139-8143	NN	denotes	gene
T5210	8144-8153	NN	denotes	targeting
T5211	8154-8156	IN	denotes	by
T5212	8157-8165	NNP	denotes	Southern
T5213	8166-8170	NN	denotes	blot
T5214	8171-8179	NN	denotes	analysis
T5215	8180-8182	IN	denotes	of
T5216	8183-8187	NN	denotes	BglI
T5218	8188-8196	VBN	denotes	digested
T5219	8197-8204	JJ	denotes	genomic
T5217	8205-8208	NN	denotes	DNA
T5220	8208-8209	.	denotes	.
T5221	8209-8312	sentence	denotes	Blots were hybridized with an Exon 6 specific probe that was external to the gene targeting construct.
T5222	8210-8215	NNS	denotes	Blots
T5224	8216-8220	VBD	denotes	were
T5223	8221-8231	VBN	denotes	hybridized
T5225	8232-8236	IN	denotes	with
T5226	8237-8239	DT	denotes	an
T5228	8240-8244	NN	denotes	Exon
T5230	8245-8246	CD	denotes	6
T5229	8247-8255	JJ	denotes	specific
T5227	8256-8261	NN	denotes	probe
T5231	8262-8266	WDT	denotes	that
T5232	8267-8270	VBD	denotes	was
T5233	8271-8279	JJ	denotes	external
T5234	8280-8282	IN	denotes	to
T5235	8283-8286	DT	denotes	the
T5237	8287-8291	NN	denotes	gene
T5238	8292-8301	NN	denotes	targeting
T5236	8302-8311	NN	denotes	construct
T5239	8311-8312	.	denotes	.
T5240	8312-8432	sentence	denotes	Correctly targeted ES cells were injected into C57BL/6J blastocysts to produce chimeric mice using standard procedures.
T5241	8313-8322	RB	denotes	Correctly
T5242	8323-8331	VBN	denotes	targeted
T5244	8332-8334	NN	denotes	ES
T5243	8335-8340	NNS	denotes	cells
T5246	8341-8345	VBD	denotes	were
T5245	8346-8354	VBN	denotes	injected
T5247	8355-8359	IN	denotes	into
T5248	8360-8365	NN	denotes	C57BL
T5250	8365-8366	HYPH	denotes	/
T5249	8366-8368	NN	denotes	6J
T5251	8369-8380	NNS	denotes	blastocysts
T5252	8381-8383	TO	denotes	to
T5253	8384-8391	VB	denotes	produce
T5254	8392-8400	JJ	denotes	chimeric
T5255	8401-8405	NNS	denotes	mice
T5256	8406-8411	VBG	denotes	using
T5257	8412-8420	JJ	denotes	standard
T5258	8421-8431	NNS	denotes	procedures
T5259	8431-8432	.	denotes	.
T5260	8432-8589	sentence	denotes	Heterozygous offspring from germline competent chimeras were intercrossed to produce wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice.
T5261	8433-8445	JJ	denotes	Heterozygous
T5262	8446-8455	NN	denotes	offspring
T5264	8456-8460	IN	denotes	from
T5265	8461-8469	NN	denotes	germline
T5266	8470-8479	JJ	denotes	competent
T5267	8480-8488	NNS	denotes	chimeras
T5268	8489-8493	VBD	denotes	were
T5263	8494-8506	VBN	denotes	intercrossed
T5269	8507-8509	TO	denotes	to
T5270	8510-8517	VB	denotes	produce
T5271	8518-8522	JJ	denotes	wild
T5272	8523-8527	NN	denotes	type
T5274	8528-8529	-LRB-	denotes	(
T5276	8529-8530	SYM	denotes	+
T5277	8530-8531	HYPH	denotes	/
T5275	8531-8532	SYM	denotes	+
T5278	8532-8533	-RRB-	denotes	)
T5279	8533-8535	,	denotes	,
T5280	8535-8547	JJ	denotes	heterozygous
T5281	8548-8549	-LRB-	denotes	(
T5283	8549-8550	SYM	denotes	+
T5284	8550-8551	HYPH	denotes	/
T5282	8551-8552	SYM	denotes	-
T5285	8552-8553	-RRB-	denotes	)
T5286	8554-8557	CC	denotes	and
T5287	8558-8568	JJ	denotes	homozygous
T5273	8569-8577	NN	denotes	knockout
T5289	8578-8579	-LRB-	denotes	(
T5291	8579-8580	SYM	denotes	-
T5292	8580-8581	HYPH	denotes	/
T5290	8581-8582	SYM	denotes	-
T5293	8582-8583	-RRB-	denotes	)
T5288	8584-8588	NNS	denotes	mice
T5294	8588-8589	.	denotes	.
T5295	8589-8639	sentence	denotes	At all generations, +/- breeding pairs were used.
T5296	8590-8592	IN	denotes	At
T5298	8593-8596	DT	denotes	all
T5299	8597-8608	NNS	denotes	generations
T5300	8608-8610	,	denotes	,
T5301	8610-8611	SYM	denotes	+
T5303	8611-8612	HYPH	denotes	/
T5302	8612-8613	SYM	denotes	-
T5305	8614-8622	VBG	denotes	breeding
T5304	8623-8628	NNS	denotes	pairs
T5306	8629-8633	VBD	denotes	were
T5297	8634-8638	VBN	denotes	used
T5307	8638-8639	.	denotes	.
T5308	8639-8710	sentence	denotes	Results presented here are from mice derived from the F2+ generations.
T5309	8640-8647	NNS	denotes	Results
T5311	8648-8657	VBN	denotes	presented
T5312	8658-8662	RB	denotes	here
T5310	8663-8666	VBP	denotes	are
T5313	8667-8671	IN	denotes	from
T5314	8672-8676	NNS	denotes	mice
T5315	8677-8684	VBN	denotes	derived
T5316	8685-8689	IN	denotes	from
T5317	8690-8693	DT	denotes	the
T5319	8694-8696	NN	denotes	F2
T5320	8696-8697	SYM	denotes	+
T5318	8698-8709	NNS	denotes	generations
T5321	8709-8710	.	denotes	.
T5322	8710-8786	sentence	denotes	All animals were of a mixed C57BL/6J × Strain 129Sv/SvJ genetic background.
T5323	8711-8714	DT	denotes	All
T5324	8715-8722	NNS	denotes	animals
T5325	8723-8727	VBD	denotes	were
T5326	8728-8730	IN	denotes	of
T5327	8731-8732	DT	denotes	a
T5329	8733-8738	VBN	denotes	mixed
T5330	8739-8744	NN	denotes	C57BL
T5332	8744-8745	HYPH	denotes	/
T5331	8745-8747	NN	denotes	6J
T5333	8748-8749	SYM	denotes	×
T5334	8750-8756	NN	denotes	Strain
T5336	8757-8762	NN	denotes	129Sv
T5337	8762-8763	HYPH	denotes	/
T5335	8763-8766	NN	denotes	SvJ
T5338	8767-8774	JJ	denotes	genetic
T5328	8775-8785	NN	denotes	background
T5339	8785-8786	.	denotes	.
T22055	8797-8799	NN	denotes	E2
T22057	8800-8804	NN	denotes	gene
T22058	8805-8813	NN	denotes	knockout
T22059	8814-8819	NN	denotes	mouse
T22056	8820-8830	NN	denotes	production
T22060	8830-8831	.	denotes	.
T22061	8831-8909	sentence	denotes	A, Gene targeting strategy used for targeting the E2 locus in mouse ES cells.
T22062	8832-8833	LS	denotes	A
T22064	8833-8835	,	denotes	,
T22065	8835-8839	NN	denotes	Gene
T22066	8840-8849	NN	denotes	targeting
T22063	8850-8858	NN	denotes	strategy
T22067	8859-8863	VBN	denotes	used
T22068	8864-8867	IN	denotes	for
T22069	8868-8877	VBG	denotes	targeting
T22070	8878-8881	DT	denotes	the
T22072	8882-8884	NN	denotes	E2
T22071	8885-8890	NN	denotes	locus
T22073	8891-8893	IN	denotes	in
T22074	8894-8899	NN	denotes	mouse
T22076	8900-8902	NN	denotes	ES
T22075	8903-8908	NNS	denotes	cells
T22077	8908-8909	.	denotes	.
T22078	8909-9037	sentence	denotes	The targeting construct was designed to delete 1.67 kb of sequence between an EcoRV site in Exon 4 and a Smal site in intron 5.
T22079	8910-8913	DT	denotes	The
T22081	8914-8923	NN	denotes	targeting
T22080	8924-8933	NN	denotes	construct
T22083	8934-8937	VBD	denotes	was
T22082	8938-8946	VBN	denotes	designed
T22084	8947-8949	TO	denotes	to
T22085	8950-8956	VB	denotes	delete
T22086	8957-8961	CD	denotes	1.67
T22087	8962-8964	NN	denotes	kb
T22088	8965-8967	IN	denotes	of
T22089	8968-8976	NN	denotes	sequence
T22090	8977-8984	IN	denotes	between
T22091	8985-8987	DT	denotes	an
T22093	8988-8993	NN	denotes	EcoRV
T22092	8994-8998	NN	denotes	site
T22094	8999-9001	IN	denotes	in
T22095	9002-9006	NN	denotes	Exon
T22096	9007-9008	CD	denotes	4
T22097	9009-9012	CC	denotes	and
T22098	9013-9014	DT	denotes	a
T22100	9015-9019	NN	denotes	Smal
T22099	9020-9024	NN	denotes	site
T22101	9025-9027	IN	denotes	in
T22102	9028-9034	NN	denotes	intron
T22103	9035-9036	CD	denotes	5
T22104	9036-9037	.	denotes	.
T22105	9037-9150	sentence	denotes	The wild type E2 gene contains an ~16 kb BglI restriction fragment that hybridizes to the Exon 6 specific probe.
T22106	9038-9041	DT	denotes	The
T22108	9042-9046	JJ	denotes	wild
T22109	9047-9051	NN	denotes	type
T22110	9052-9054	NN	denotes	E2
T22107	9055-9059	NN	denotes	gene
T22111	9060-9068	VBZ	denotes	contains
T22112	9069-9071	DT	denotes	an
T22114	9072-9073	SYM	denotes	~
T22115	9073-9075	CD	denotes	16
T22116	9076-9078	NN	denotes	kb
T22117	9079-9083	NN	denotes	BglI
T22118	9084-9095	NN	denotes	restriction
T22113	9096-9104	NN	denotes	fragment
T22119	9105-9109	WDT	denotes	that
T22120	9110-9120	VBZ	denotes	hybridizes
T22121	9121-9123	IN	denotes	to
T22122	9124-9127	DT	denotes	the
T22124	9128-9132	NN	denotes	Exon
T22126	9133-9134	CD	denotes	6
T22125	9135-9143	JJ	denotes	specific
T22123	9144-9149	NN	denotes	probe
T22127	9149-9150	.	denotes	.
T22128	9150-9259	sentence	denotes	A correctly targeted E2 locus harbors an ~11 kb BglI restriction fragment that hybridizes to the same probe.
T22129	9151-9152	DT	denotes	A
T22131	9153-9162	RB	denotes	correctly
T22132	9163-9171	VBN	denotes	targeted
T22133	9172-9174	NN	denotes	E2
T22130	9175-9180	NN	denotes	locus
T22134	9181-9188	VBZ	denotes	harbors
T22135	9189-9191	DT	denotes	an
T22137	9192-9193	SYM	denotes	~
T22138	9193-9195	CD	denotes	11
T22139	9196-9198	NN	denotes	kb
T22140	9199-9203	NN	denotes	BglI
T22141	9204-9215	NN	denotes	restriction
T22136	9216-9224	NN	denotes	fragment
T22142	9225-9229	WDT	denotes	that
T22143	9230-9240	VBZ	denotes	hybridizes
T22144	9241-9243	IN	denotes	to
T22145	9244-9247	DT	denotes	the
T22147	9248-9252	JJ	denotes	same
T22146	9253-9258	NN	denotes	probe
T22148	9258-9259	.	denotes	.
T22149	9259-9386	sentence	denotes	Note that the probe will not detect random integration of the targeting vector because it is external to the targeting vector.
T22150	9260-9264	VB	denotes	Note
T22151	9265-9269	IN	denotes	that
T22153	9270-9273	DT	denotes	the
T22154	9274-9279	NN	denotes	probe
T22155	9280-9284	MD	denotes	will
T22156	9285-9288	RB	denotes	not
T22152	9289-9295	VB	denotes	detect
T22157	9296-9302	JJ	denotes	random
T22158	9303-9314	NN	denotes	integration
T22159	9315-9317	IN	denotes	of
T22160	9318-9321	DT	denotes	the
T22162	9322-9331	NN	denotes	targeting
T22161	9332-9338	NN	denotes	vector
T22163	9339-9346	IN	denotes	because
T22165	9347-9349	PRP	denotes	it
T22164	9350-9352	VBZ	denotes	is
T22166	9353-9361	JJ	denotes	external
T22167	9362-9364	IN	denotes	to
T22168	9365-9368	DT	denotes	the
T22170	9369-9378	NN	denotes	targeting
T22169	9379-9385	NN	denotes	vector
T22171	9385-9386	.	denotes	.
T22172	9386-9621	sentence	denotes	B, Southern blot analysis of Bgll digested genomic DNA derived from the parental wild type ES cell line (R1), a heterozygous targeted ES cell line (362), and from wild type (+/+), heterozygous (+/-) and homozygous knockout (-/-) mice.
T22173	9387-9388	LS	denotes	B
T22175	9388-9390	,	denotes	,
T22176	9390-9398	NNP	denotes	Southern
T22177	9399-9403	NN	denotes	blot
T22174	9404-9412	NN	denotes	analysis
T22178	9413-9415	IN	denotes	of
T22179	9416-9420	NN	denotes	Bgll
T22180	9421-9429	VBN	denotes	digested
T22182	9430-9437	JJ	denotes	genomic
T22181	9438-9441	NN	denotes	DNA
T22183	9442-9449	VBN	denotes	derived
T22184	9450-9454	IN	denotes	from
T22185	9455-9458	DT	denotes	the
T22187	9459-9467	JJ	denotes	parental
T22188	9468-9472	JJ	denotes	wild
T22189	9473-9477	NN	denotes	type
T22190	9478-9480	NN	denotes	ES
T22191	9481-9485	NN	denotes	cell
T22186	9486-9490	NN	denotes	line
T22192	9491-9492	-LRB-	denotes	(
T22193	9492-9494	NN	denotes	R1
T22194	9494-9495	-RRB-	denotes	)
T22195	9495-9497	,	denotes	,
T22196	9497-9498	DT	denotes	a
T22198	9499-9511	JJ	denotes	heterozygous
T22199	9512-9520	VBN	denotes	targeted
T22200	9521-9523	NN	denotes	ES
T22201	9524-9528	NN	denotes	cell
T22197	9529-9533	NN	denotes	line
T22202	9534-9535	-LRB-	denotes	(
T22203	9535-9538	CD	denotes	362
T22204	9538-9539	-RRB-	denotes	)
T22205	9539-9541	,	denotes	,
T22206	9541-9544	CC	denotes	and
T22207	9545-9549	IN	denotes	from
T22208	9550-9554	JJ	denotes	wild
T22209	9555-9559	NN	denotes	type
T22211	9560-9561	-LRB-	denotes	(
T22213	9561-9562	SYM	denotes	+
T22214	9562-9563	HYPH	denotes	/
T22212	9563-9564	SYM	denotes	+
T22215	9564-9565	-RRB-	denotes	)
T22216	9565-9567	,	denotes	,
T22217	9567-9579	JJ	denotes	heterozygous
T22218	9580-9581	-LRB-	denotes	(
T22220	9581-9582	SYM	denotes	+
T22221	9582-9583	HYPH	denotes	/
T22219	9583-9584	SYM	denotes	-
T22222	9584-9585	-RRB-	denotes	)
T22223	9586-9589	CC	denotes	and
T22224	9590-9600	JJ	denotes	homozygous
T22210	9601-9609	NN	denotes	knockout
T22226	9610-9611	-LRB-	denotes	(
T22228	9611-9612	SYM	denotes	-
T22229	9612-9613	HYPH	denotes	/
T22227	9613-9614	SYM	denotes	-
T22230	9614-9615	-RRB-	denotes	)
T22225	9616-9620	NNS	denotes	mice
T22231	9620-9621	.	denotes	.
T22232	9621-9676	sentence	denotes	The blot was hybridized with an Exon 6 specific probe.
T22233	9622-9625	DT	denotes	The
T22234	9626-9630	NN	denotes	blot
T22236	9631-9634	VBD	denotes	was
T22235	9635-9645	VBN	denotes	hybridized
T22237	9646-9650	IN	denotes	with
T22238	9651-9653	DT	denotes	an
T22240	9654-9658	NN	denotes	Exon
T22242	9659-9660	CD	denotes	6
T22241	9661-9669	JJ	denotes	specific
T22239	9670-9675	NN	denotes	probe
T22243	9675-9676	.	denotes	.
T22244	9676-9808	sentence	denotes	C, Immunohistochemical analysis of fresh frozen liver sections from control (+/+) and E2 knockout (-/-) postnatal day 1 mouse pups.
T22245	9677-9678	LS	denotes	C
T22247	9678-9680	,	denotes	,
T22248	9680-9699	JJ	denotes	Immunohistochemical
T22246	9700-9708	NN	denotes	analysis
T22249	9709-9711	IN	denotes	of
T22250	9712-9717	JJ	denotes	fresh
T22251	9718-9724	VBN	denotes	frozen
T22253	9725-9730	NN	denotes	liver
T22252	9731-9739	NNS	denotes	sections
T22254	9740-9744	IN	denotes	from
T22255	9745-9752	NN	denotes	control
T22257	9753-9754	-LRB-	denotes	(
T22259	9754-9755	SYM	denotes	+
T22260	9755-9756	HYPH	denotes	/
T22258	9756-9757	SYM	denotes	+
T22261	9757-9758	-RRB-	denotes	)
T22262	9759-9762	CC	denotes	and
T22263	9763-9765	NN	denotes	E2
T22256	9766-9774	NN	denotes	knockout
T22265	9775-9776	-LRB-	denotes	(
T22267	9776-9777	SYM	denotes	-
T22268	9777-9778	HYPH	denotes	/
T22266	9778-9779	SYM	denotes	-
T22269	9779-9780	-RRB-	denotes	)
T22270	9781-9790	JJ	denotes	postnatal
T22271	9791-9794	NN	denotes	day
T22272	9795-9796	CD	denotes	1
T22273	9797-9802	NN	denotes	mouse
T22264	9803-9807	NNS	denotes	pups
T22274	9807-9808	.	denotes	.
T22275	9808-9903	sentence	denotes	Sections were stained for E2 using an E2 specific antibody (green) and a nuclear stain (blue).
T22276	9809-9817	NNS	denotes	Sections
T22278	9818-9822	VBD	denotes	were
T22277	9823-9830	VBN	denotes	stained
T22279	9831-9834	IN	denotes	for
T22280	9835-9837	NN	denotes	E2
T22281	9838-9843	VBG	denotes	using
T22282	9844-9846	DT	denotes	an
T22284	9847-9849	NN	denotes	E2
T22285	9850-9858	JJ	denotes	specific
T22283	9859-9867	NN	denotes	antibody
T22286	9868-9869	-LRB-	denotes	(
T22287	9869-9874	JJ	denotes	green
T22288	9874-9875	-RRB-	denotes	)
T22289	9876-9879	CC	denotes	and
T22290	9880-9881	DT	denotes	a
T22292	9882-9889	JJ	denotes	nuclear
T22291	9890-9895	NN	denotes	stain
T22293	9896-9897	-LRB-	denotes	(
T22294	9897-9901	JJ	denotes	blue
T22295	9901-9902	-RRB-	denotes	)
T22296	9902-9903	.	denotes	.
T22297	9903-9992	sentence	denotes	Note the complete absence of E2 immunoreactivity in the section from the knockout mouse.
T22298	9904-9908	VB	denotes	Note
T22299	9909-9912	DT	denotes	the
T22301	9913-9921	JJ	denotes	complete
T22300	9922-9929	NN	denotes	absence
T22302	9930-9932	IN	denotes	of
T22303	9933-9935	NN	denotes	E2
T22304	9936-9952	NN	denotes	immunoreactivity
T22305	9953-9955	IN	denotes	in
T22306	9956-9959	DT	denotes	the
T22307	9960-9967	NN	denotes	section
T22308	9968-9972	IN	denotes	from
T22309	9973-9976	DT	denotes	the
T22311	9977-9985	NN	denotes	knockout
T22310	9986-9991	NN	denotes	mouse
T22312	9991-9992	.	denotes	.
T22313	9992-10106	sentence	denotes	D, Similar results were observed upon immunohistochemical analysis of primary mouse embryonic fibroblasts (MEFs).
T22314	9993-9994	LS	denotes	D
T22316	9994-9996	,	denotes	,
T22317	9996-10003	JJ	denotes	Similar
T22318	10004-10011	NNS	denotes	results
T22319	10012-10016	VBD	denotes	were
T22315	10017-10025	VBN	denotes	observed
T22320	10026-10030	IN	denotes	upon
T22321	10031-10050	JJ	denotes	immunohistochemical
T22322	10051-10059	NN	denotes	analysis
T22323	10060-10062	IN	denotes	of
T22324	10063-10070	JJ	denotes	primary
T22326	10071-10076	NN	denotes	mouse
T22327	10077-10086	JJ	denotes	embryonic
T22325	10087-10098	NNS	denotes	fibroblasts
T22328	10099-10100	-LRB-	denotes	(
T22329	10100-10104	NNS	denotes	MEFs
T22330	10104-10105	-RRB-	denotes	)
T22331	10105-10106	.	denotes	.
T22332	10106-10266	sentence	denotes	Note that the readily detectable signal for E2 in the control cells was present in a pattern characteristic of mitochondria, the subcellular location of BCKDH.
T22333	10107-10111	VB	denotes	Note
T22334	10112-10116	IN	denotes	that
T22336	10117-10120	DT	denotes	the
T22338	10121-10128	RB	denotes	readily
T22339	10129-10139	JJ	denotes	detectable
T22337	10140-10146	NN	denotes	signal
T22340	10147-10150	IN	denotes	for
T22341	10151-10153	NN	denotes	E2
T22342	10154-10156	IN	denotes	in
T22343	10157-10160	DT	denotes	the
T22345	10161-10168	NN	denotes	control
T22344	10169-10174	NNS	denotes	cells
T22335	10175-10178	VBD	denotes	was
T22346	10179-10186	JJ	denotes	present
T22347	10187-10189	IN	denotes	in
T22348	10190-10191	DT	denotes	a
T22349	10192-10199	NN	denotes	pattern
T22350	10200-10214	JJ	denotes	characteristic
T22351	10215-10217	IN	denotes	of
T22352	10218-10230	NNS	denotes	mitochondria
T22353	10230-10232	,	denotes	,
T22354	10232-10235	DT	denotes	the
T22356	10236-10247	JJ	denotes	subcellular
T22355	10248-10256	NN	denotes	location
T22357	10257-10259	IN	denotes	of
T22358	10260-10265	NN	denotes	BCKDH
T22359	10265-10266	.	denotes	.
T5872	10268-10278	NN	denotes	Production
T5873	10279-10281	IN	denotes	of
T5874	10282-10292	JJ	denotes	transgenic
T5875	10293-10297	NNS	denotes	mice
T5876	10297-10384	sentence	denotes	Standard molecular techniques were used to assemble the transgenic construct, pTRE-E2.
T5877	10298-10306	JJ	denotes	Standard
T5879	10307-10316	JJ	denotes	molecular
T5878	10317-10327	NNS	denotes	techniques
T5881	10328-10332	VBD	denotes	were
T5880	10333-10337	VBN	denotes	used
T5882	10338-10340	TO	denotes	to
T5883	10341-10349	VB	denotes	assemble
T5884	10350-10353	DT	denotes	the
T5886	10354-10364	JJ	denotes	transgenic
T5885	10365-10374	NN	denotes	construct
T5887	10374-10376	,	denotes	,
T5888	10376-10380	NN	denotes	pTRE
T5890	10380-10381	HYPH	denotes	-
T5889	10381-10383	NN	denotes	E2
T5891	10383-10384	.	denotes	.
T5892	10384-11008	sentence	denotes	This transgene contains the tetracycline responsive hCMV*-1 promoter [consisting of the tetracycline responsive element (TRE) and a minimal hCMV promoter] [25] from pTRE2 (Clontech Inc., Mt. View, CA), a chimeric intron from pCI (Promega) to increase message stability and expression [26,27], a Kozak consensus sequence at the initiation codon to optimize translation [28], the human E2 cDNA which has been modified to contain a 4× alanine linker followed by a c-myc epitope tag at the carboxy terminus to facilitate detection, and an SV40 late polyadenylation sequence from pCI for enhanced mRNA stability and translation.
T5893	10385-10389	DT	denotes	This
T5894	10390-10399	NN	denotes	transgene
T5895	10400-10408	VBZ	denotes	contains
T5896	10409-10412	DT	denotes	the
T5898	10413-10425	NN	denotes	tetracycline
T5899	10426-10436	JJ	denotes	responsive
T5900	10437-10441	NN	denotes	hCMV
T5901	10441-10442	SYM	denotes	*
T5902	10442-10443	HYPH	denotes	-
T5903	10443-10444	CD	denotes	1
T5897	10445-10453	NN	denotes	promoter
T5904	10454-10455	-LRB-	denotes	[
T5905	10455-10465	VBG	denotes	consisting
T5906	10466-10468	IN	denotes	of
T5907	10469-10472	DT	denotes	the
T5909	10473-10485	NN	denotes	tetracycline
T5910	10486-10496	JJ	denotes	responsive
T5908	10497-10504	NN	denotes	element
T5911	10505-10506	-LRB-	denotes	(
T5912	10506-10509	NN	denotes	TRE
T5913	10509-10510	-RRB-	denotes	)
T5914	10511-10514	CC	denotes	and
T5915	10515-10516	DT	denotes	a
T5917	10517-10524	JJ	denotes	minimal
T5918	10525-10529	NN	denotes	hCMV
T5916	10530-10538	NN	denotes	promoter
T5919	10538-10539	-RRB-	denotes	]
T5920	10540-10541	-LRB-	denotes	[
T5921	10541-10543	CD	denotes	25
T5922	10543-10544	-RRB-	denotes	]
T5923	10545-10549	IN	denotes	from
T5924	10550-10555	NN	denotes	pTRE2
T5925	10556-10557	-LRB-	denotes	(
T5927	10557-10565	NNP	denotes	Clontech
T5926	10566-10570	NNP	denotes	Inc.
T5928	10570-10572	,	denotes	,
T5929	10572-10575	NNP	denotes	Mt.
T5930	10576-10580	NNP	denotes	View
T5931	10580-10582	,	denotes	,
T5932	10582-10584	NNP	denotes	CA
T5933	10584-10585	-RRB-	denotes	)
T5934	10585-10587	,	denotes	,
T5935	10587-10588	DT	denotes	a
T5937	10589-10597	JJ	denotes	chimeric
T5936	10598-10604	NN	denotes	intron
T5938	10605-10609	IN	denotes	from
T5939	10610-10613	NN	denotes	pCI
T5940	10614-10615	-LRB-	denotes	(
T5941	10615-10622	NNP	denotes	Promega
T5942	10622-10623	-RRB-	denotes	)
T5943	10624-10626	TO	denotes	to
T5944	10627-10635	VB	denotes	increase
T5945	10636-10643	NN	denotes	message
T5946	10644-10653	NN	denotes	stability
T5947	10654-10657	CC	denotes	and
T5948	10658-10668	NN	denotes	expression
T5949	10669-10670	-LRB-	denotes	[
T5951	10670-10672	CD	denotes	26
T5952	10672-10673	,	denotes	,
T5950	10673-10675	CD	denotes	27
T5953	10675-10676	-RRB-	denotes	]
T5954	10676-10678	,	denotes	,
T5955	10678-10679	DT	denotes	a
T5957	10680-10685	NN	denotes	Kozak
T5958	10686-10695	NN	denotes	consensus
T5956	10696-10704	NN	denotes	sequence
T5959	10705-10707	IN	denotes	at
T5960	10708-10711	DT	denotes	the
T5962	10712-10722	NN	denotes	initiation
T5961	10723-10728	NN	denotes	codon
T5963	10729-10731	TO	denotes	to
T5964	10732-10740	VB	denotes	optimize
T5965	10741-10752	NN	denotes	translation
T5966	10753-10754	-LRB-	denotes	[
T5967	10754-10756	CD	denotes	28
T5968	10756-10757	-RRB-	denotes	]
T5969	10757-10759	,	denotes	,
T5970	10759-10762	DT	denotes	the
T5972	10763-10768	JJ	denotes	human
T5973	10769-10771	NN	denotes	E2
T5971	10772-10776	NN	denotes	cDNA
T5974	10777-10782	WDT	denotes	which
T5976	10783-10786	VBZ	denotes	has
T5977	10787-10791	VBN	denotes	been
T5975	10792-10800	VBN	denotes	modified
T5978	10801-10803	TO	denotes	to
T5979	10804-10811	VB	denotes	contain
T5980	10812-10813	DT	denotes	a
T5982	10814-10815	CD	denotes	4
T5983	10815-10816	SYM	denotes	×
T5984	10817-10824	NN	denotes	alanine
T5981	10825-10831	NN	denotes	linker
T5985	10832-10840	VBN	denotes	followed
T5986	10841-10843	IN	denotes	by
T5987	10844-10845	DT	denotes	a
T5989	10846-10847	NN	denotes	c
T5991	10847-10848	HYPH	denotes	-
T5990	10848-10851	NN	denotes	myc
T5992	10852-10859	NN	denotes	epitope
T5988	10860-10863	NN	denotes	tag
T5993	10864-10866	IN	denotes	at
T5994	10867-10870	DT	denotes	the
T5996	10871-10878	NN	denotes	carboxy
T5995	10879-10887	NN	denotes	terminus
T5997	10888-10890	TO	denotes	to
T5998	10891-10901	VB	denotes	facilitate
T5999	10902-10911	NN	denotes	detection
T6000	10911-10913	,	denotes	,
T6001	10913-10916	CC	denotes	and
T6002	10917-10919	DT	denotes	an
T6004	10920-10924	NN	denotes	SV40
T6005	10925-10929	JJ	denotes	late
T6006	10930-10945	NN	denotes	polyadenylation
T6003	10946-10954	NN	denotes	sequence
T6007	10955-10959	IN	denotes	from
T6008	10960-10963	NN	denotes	pCI
T6009	10964-10967	IN	denotes	for
T6010	10968-10976	VBN	denotes	enhanced
T6012	10977-10981	NN	denotes	mRNA
T6011	10982-10991	NN	denotes	stability
T6013	10992-10995	CC	denotes	and
T6014	10996-11007	NN	denotes	translation
T6015	11007-11008	.	denotes	.
T6016	11008-11296	sentence	denotes	The 2.48 kb TRE-E2 transgene was purified from vector sequences following digestion with XhoI and BamHI and injected into C57BL/6J or C57BL/6J × Strain 129SvEv mouse embryos at the transgenic core facilities of the University of Pittsburgh and the University of Cincinnati, respectively.
T6017	11009-11012	DT	denotes	The
T6019	11013-11017	CD	denotes	2.48
T6020	11018-11020	NN	denotes	kb
T6021	11021-11024	NN	denotes	TRE
T6023	11024-11025	HYPH	denotes	-
T6022	11025-11027	NN	denotes	E2
T6018	11028-11037	NN	denotes	transgene
T6025	11038-11041	VBD	denotes	was
T6024	11042-11050	VBN	denotes	purified
T6026	11051-11055	IN	denotes	from
T6027	11056-11062	NN	denotes	vector
T6028	11063-11072	NNS	denotes	sequences
T6029	11073-11082	VBG	denotes	following
T6030	11083-11092	NN	denotes	digestion
T6031	11093-11097	IN	denotes	with
T6032	11098-11102	NN	denotes	XhoI
T6033	11103-11106	CC	denotes	and
T6034	11107-11112	NN	denotes	BamHI
T6035	11113-11116	CC	denotes	and
T6036	11117-11125	VBN	denotes	injected
T6037	11126-11130	IN	denotes	into
T6038	11131-11136	NN	denotes	C57BL
T6040	11136-11137	HYPH	denotes	/
T6039	11137-11139	NN	denotes	6J
T6042	11140-11142	CC	denotes	or
T6043	11143-11148	NN	denotes	C57BL
T6045	11148-11149	HYPH	denotes	/
T6044	11149-11151	NN	denotes	6J
T6046	11152-11153	SYM	denotes	×
T6041	11154-11160	NN	denotes	Strain
T6048	11161-11168	NN	denotes	129SvEv
T6049	11169-11174	NN	denotes	mouse
T6047	11175-11182	NNS	denotes	embryos
T6050	11183-11185	IN	denotes	at
T6051	11186-11189	DT	denotes	the
T6053	11190-11200	JJ	denotes	transgenic
T6054	11201-11205	NN	denotes	core
T6052	11206-11216	NNS	denotes	facilities
T6055	11217-11219	IN	denotes	of
T6056	11220-11223	DT	denotes	the
T6057	11224-11234	NNP	denotes	University
T6058	11235-11237	IN	denotes	of
T6059	11238-11248	NNP	denotes	Pittsburgh
T6060	11249-11252	CC	denotes	and
T6061	11253-11256	DT	denotes	the
T6062	11257-11267	NNP	denotes	University
T6063	11268-11270	IN	denotes	of
T6064	11271-11281	NNP	denotes	Cincinnati
T6065	11281-11283	,	denotes	,
T6066	11283-11295	RB	denotes	respectively
T6067	11295-11296	.	denotes	.
T6068	11296-11493	sentence	denotes	Genomic DNA from the tail of mice was screened by Southern blot analysis following digestion with EcoRI and hybridization to an ~400 bp probe derived from the SV40 portion of the TRE-E2 transgene.
T6069	11297-11304	JJ	denotes	Genomic
T6070	11305-11308	NN	denotes	DNA
T6072	11309-11313	IN	denotes	from
T6073	11314-11317	DT	denotes	the
T6074	11318-11322	NN	denotes	tail
T6075	11323-11325	IN	denotes	of
T6076	11326-11330	NNS	denotes	mice
T6077	11331-11334	VBD	denotes	was
T6071	11335-11343	VBN	denotes	screened
T6078	11344-11346	IN	denotes	by
T6079	11347-11355	NNP	denotes	Southern
T6080	11356-11360	NN	denotes	blot
T6081	11361-11369	NN	denotes	analysis
T6082	11370-11379	VBG	denotes	following
T6083	11380-11389	NN	denotes	digestion
T6084	11390-11394	IN	denotes	with
T6085	11395-11400	NN	denotes	EcoRI
T6086	11401-11404	CC	denotes	and
T6087	11405-11418	NN	denotes	hybridization
T6088	11419-11421	IN	denotes	to
T6089	11422-11424	DT	denotes	an
T6091	11425-11426	SYM	denotes	~
T6092	11426-11429	CD	denotes	400
T6093	11430-11432	NN	denotes	bp
T6090	11433-11438	NN	denotes	probe
T6094	11439-11446	VBN	denotes	derived
T6095	11447-11451	IN	denotes	from
T6096	11452-11455	DT	denotes	the
T6098	11456-11460	NN	denotes	SV40
T6097	11461-11468	NN	denotes	portion
T6099	11469-11471	IN	denotes	of
T6100	11472-11475	DT	denotes	the
T6102	11476-11479	NN	denotes	TRE
T6104	11479-11480	HYPH	denotes	-
T6103	11480-11482	NN	denotes	E2
T6101	11483-11492	NN	denotes	transgene
T6105	11492-11493	.	denotes	.
T6426	11495-11505	NN	denotes	Production
T6427	11506-11508	IN	denotes	of
T6428	11509-11521	JJ	denotes	intermediate
T6430	11522-11526	NN	denotes	MSUD
T6431	11527-11533	JJ	denotes	murine
T6429	11534-11539	NN	denotes	model
T6432	11539-11796	sentence	denotes	The various TRE-E2 transgenic lines produced were mated independently to mice that were heterozygous for both the E2 knockout and the LAP-tTA transgene [Tg(tTALap)Bjd/J; Stock 3272; The Jackson Laboratory, Bar Harbor, ME; NMRI × FVB × C57BL/6J background].
T6433	11540-11543	DT	denotes	The
T6435	11544-11551	JJ	denotes	various
T6436	11552-11555	NN	denotes	TRE
T6438	11555-11556	HYPH	denotes	-
T6437	11556-11558	NN	denotes	E2
T6439	11559-11569	JJ	denotes	transgenic
T6434	11570-11575	NNS	denotes	lines
T6441	11576-11584	VBN	denotes	produced
T6442	11585-11589	VBD	denotes	were
T6440	11590-11595	VBN	denotes	mated
T6443	11596-11609	RB	denotes	independently
T6444	11610-11612	IN	denotes	to
T6445	11613-11617	NNS	denotes	mice
T6446	11618-11622	WDT	denotes	that
T6447	11623-11627	VBD	denotes	were
T6448	11628-11640	JJ	denotes	heterozygous
T6449	11641-11644	IN	denotes	for
T6450	11645-11649	CC	denotes	both
T6452	11650-11653	DT	denotes	the
T6453	11654-11656	NN	denotes	E2
T6451	11657-11665	NN	denotes	knockout
T6454	11666-11669	CC	denotes	and
T6455	11670-11673	DT	denotes	the
T6457	11674-11677	NN	denotes	LAP
T6459	11677-11678	HYPH	denotes	-
T6458	11678-11681	NN	denotes	tTA
T6456	11682-11691	NN	denotes	transgene
T6460	11692-11693	-LRB-	denotes	[
T6462	11693-11695	NN	denotes	Tg
T6464	11695-11696	-LRB-	denotes	(
T6465	11696-11702	NN	denotes	tTALap
T6466	11702-11703	-RRB-	denotes	)
T6467	11703-11706	NN	denotes	Bjd
T6469	11706-11707	HYPH	denotes	/
T6468	11707-11708	NN	denotes	J
T6470	11708-11709	:	denotes	;
T6463	11710-11715	NN	denotes	Stock
T6471	11716-11720	CD	denotes	3272
T6472	11720-11721	:	denotes	;
T6473	11722-11725	DT	denotes	The
T6475	11726-11733	NNP	denotes	Jackson
T6474	11734-11744	NNP	denotes	Laboratory
T6476	11744-11746	,	denotes	,
T6477	11746-11749	NNP	denotes	Bar
T6478	11750-11756	NNP	denotes	Harbor
T6479	11756-11758	,	denotes	,
T6480	11758-11760	NNP	denotes	ME
T6481	11760-11761	:	denotes	;
T6482	11762-11766	NN	denotes	NMRI
T6483	11767-11768	SYM	denotes	×
T6484	11769-11772	NN	denotes	FVB
T6485	11773-11774	SYM	denotes	×
T6486	11775-11780	NN	denotes	C57BL
T6488	11780-11781	HYPH	denotes	/
T6487	11781-11783	NN	denotes	6J
T6461	11784-11794	NN	denotes	background
T6489	11794-11795	-RRB-	denotes	]
T6490	11795-11796	.	denotes	.
T6491	11796-12097	sentence	denotes	Interbreeding of animals that were heterozygous for both transgenes and the knockout resulted in the production of mice with a variety of genotypes including some animals that were homozygous for the knockout and were positive for both transgenes (we refer to this genotype as the "rescue" genotype).
T6492	11797-11810	NN	denotes	Interbreeding
T6494	11811-11813	IN	denotes	of
T6495	11814-11821	NNS	denotes	animals
T6496	11822-11826	WDT	denotes	that
T6497	11827-11831	VBD	denotes	were
T6498	11832-11844	JJ	denotes	heterozygous
T6499	11845-11848	IN	denotes	for
T6500	11849-11853	DT	denotes	both
T6501	11854-11864	NNS	denotes	transgenes
T6502	11865-11868	CC	denotes	and
T6503	11869-11872	DT	denotes	the
T6504	11873-11881	NN	denotes	knockout
T6493	11882-11890	VBD	denotes	resulted
T6505	11891-11893	IN	denotes	in
T6506	11894-11897	DT	denotes	the
T6507	11898-11908	NN	denotes	production
T6508	11909-11911	IN	denotes	of
T6509	11912-11916	NNS	denotes	mice
T6510	11917-11921	IN	denotes	with
T6511	11922-11923	DT	denotes	a
T6512	11924-11931	NN	denotes	variety
T6513	11932-11934	IN	denotes	of
T6514	11935-11944	NNS	denotes	genotypes
T6515	11945-11954	VBG	denotes	including
T6516	11955-11959	DT	denotes	some
T6517	11960-11967	NNS	denotes	animals
T6518	11968-11972	WDT	denotes	that
T6519	11973-11977	VBD	denotes	were
T6520	11978-11988	JJ	denotes	homozygous
T6521	11989-11992	IN	denotes	for
T6522	11993-11996	DT	denotes	the
T6523	11997-12005	NN	denotes	knockout
T6524	12006-12009	CC	denotes	and
T6525	12010-12014	VBD	denotes	were
T6526	12015-12023	JJ	denotes	positive
T6527	12024-12027	IN	denotes	for
T6528	12028-12032	DT	denotes	both
T6529	12033-12043	NNS	denotes	transgenes
T6530	12044-12045	-LRB-	denotes	(
T6532	12045-12047	PRP	denotes	we
T6531	12048-12053	VBP	denotes	refer
T6533	12054-12056	IN	denotes	to
T6534	12057-12061	DT	denotes	this
T6535	12062-12070	NN	denotes	genotype
T6536	12071-12073	IN	denotes	as
T6537	12074-12077	DT	denotes	the
T6539	12078-12079	``	denotes	"
T6540	12079-12085	NN	denotes	rescue
T6541	12085-12086	''	denotes	"
T6538	12087-12095	NN	denotes	genotype
T6542	12095-12096	-RRB-	denotes	)
T6543	12096-12097	.	denotes	.
T6544	12097-12295	sentence	denotes	If our strategy for rescuing the neonatal lethal phenotype of the knockout were successful, then those homozygous knockout animals that had both transgenes would survive beyond the neonatal period.
T6545	12098-12100	IN	denotes	If
T6547	12101-12104	PRP$	denotes	our
T6548	12105-12113	NN	denotes	strategy
T6549	12114-12117	IN	denotes	for
T6550	12118-12126	VBG	denotes	rescuing
T6551	12127-12130	DT	denotes	the
T6553	12131-12139	JJ	denotes	neonatal
T6554	12140-12146	JJ	denotes	lethal
T6552	12147-12156	NN	denotes	phenotype
T6555	12157-12159	IN	denotes	of
T6556	12160-12163	DT	denotes	the
T6557	12164-12172	NN	denotes	knockout
T6546	12173-12177	VBD	denotes	were
T6559	12178-12188	JJ	denotes	successful
T6560	12188-12190	,	denotes	,
T6561	12190-12194	RB	denotes	then
T6562	12195-12200	DT	denotes	those
T6564	12201-12211	JJ	denotes	homozygous
T6565	12212-12220	NN	denotes	knockout
T6563	12221-12228	NNS	denotes	animals
T6566	12229-12233	WDT	denotes	that
T6567	12234-12237	VBD	denotes	had
T6568	12238-12242	DT	denotes	both
T6569	12243-12253	NNS	denotes	transgenes
T6570	12254-12259	MD	denotes	would
T6558	12260-12267	VB	denotes	survive
T6571	12268-12274	IN	denotes	beyond
T6572	12275-12278	DT	denotes	the
T6574	12279-12287	JJ	denotes	neonatal
T6573	12288-12294	NN	denotes	period
T6575	12294-12295	.	denotes	.
T6670	12297-12307	NN	denotes	Genotyping
T6671	12307-12358	sentence	denotes	All mice were genotyped by Southern blot analysis.
T6672	12308-12311	DT	denotes	All
T6673	12312-12316	NNS	denotes	mice
T6675	12317-12321	VBD	denotes	were
T6674	12322-12331	VBN	denotes	genotyped
T6676	12332-12334	IN	denotes	by
T6677	12335-12343	NNP	denotes	Southern
T6678	12344-12348	NN	denotes	blot
T6679	12349-12357	NN	denotes	analysis
T6680	12357-12358	.	denotes	.
T6681	12358-12556	sentence	denotes	Genomic DNAs prepared from tail snips were digested with an appropriate restriction enzyme, size fractionated by agarose gel electrophoresis, blotted to nylon, and probed using standard procedures.
T6682	12359-12366	JJ	denotes	Genomic
T6683	12367-12371	NNS	denotes	DNAs
T6685	12372-12380	VBN	denotes	prepared
T6686	12381-12385	IN	denotes	from
T6687	12386-12390	NN	denotes	tail
T6688	12391-12396	NNS	denotes	snips
T6689	12397-12401	VBD	denotes	were
T6684	12402-12410	VBN	denotes	digested
T6690	12411-12415	IN	denotes	with
T6691	12416-12418	DT	denotes	an
T6693	12419-12430	JJ	denotes	appropriate
T6694	12431-12442	NN	denotes	restriction
T6692	12443-12449	NN	denotes	enzyme
T6695	12449-12451	,	denotes	,
T6696	12451-12455	NN	denotes	size
T6697	12456-12468	VBN	denotes	fractionated
T6698	12469-12471	IN	denotes	by
T6699	12472-12479	NN	denotes	agarose
T6700	12480-12483	NN	denotes	gel
T6701	12484-12499	NN	denotes	electrophoresis
T6702	12499-12501	,	denotes	,
T6703	12501-12508	VBN	denotes	blotted
T6704	12509-12511	IN	denotes	to
T6705	12512-12517	NN	denotes	nylon
T6706	12517-12519	,	denotes	,
T6707	12519-12522	CC	denotes	and
T6708	12523-12529	VBN	denotes	probed
T6709	12530-12535	VBG	denotes	using
T6710	12536-12544	JJ	denotes	standard
T6711	12545-12555	NNS	denotes	procedures
T6712	12555-12556	.	denotes	.
T7183	12558-12578	NN	denotes	Immunohistochemistry
T7184	12578-12685	sentence	denotes	Primary mouse embryonic fibroblasts were prepared from embryonic day ~16.5–18.5 fetuses as described [29].
T7185	12579-12586	JJ	denotes	Primary
T7187	12587-12592	NN	denotes	mouse
T7188	12593-12602	JJ	denotes	embryonic
T7186	12603-12614	NNS	denotes	fibroblasts
T7190	12615-12619	VBD	denotes	were
T7189	12620-12628	VBN	denotes	prepared
T7191	12629-12633	IN	denotes	from
T7192	12634-12643	JJ	denotes	embryonic
T7194	12644-12647	NN	denotes	day
T7196	12648-12649	SYM	denotes	~
T7195	12649-12653	CD	denotes	16.5
T7197	12653-12654	SYM	denotes	–
T7198	12654-12658	CD	denotes	18.5
T7193	12659-12666	NNS	denotes	fetuses
T7199	12667-12669	IN	denotes	as
T7200	12670-12679	VBN	denotes	described
T7201	12680-12681	-LRB-	denotes	[
T7202	12681-12683	CD	denotes	29
T7203	12683-12684	-RRB-	denotes	]
T7204	12684-12685	.	denotes	.
T7205	12685-12941	sentence	denotes	Fibroblasts were passed onto glass, fixed in 2% paraformaldehyde in PBS for 10 minutes, permeabilized in 2% paraformaldehyde containing 0.1% Triton X100 for 10 minutes and washed three times in PBS containing 0.5% BSA and 0.15% glycine, pH 7.4 (Buffer A).
T7206	12686-12697	NNS	denotes	Fibroblasts
T7208	12698-12702	VBD	denotes	were
T7207	12703-12709	VBN	denotes	passed
T7209	12710-12714	IN	denotes	onto
T7210	12715-12720	NN	denotes	glass
T7211	12720-12722	,	denotes	,
T7212	12722-12727	VBN	denotes	fixed
T7213	12728-12730	IN	denotes	in
T7214	12731-12732	CD	denotes	2
T7215	12732-12733	NN	denotes	%
T7216	12734-12750	NN	denotes	paraformaldehyde
T7217	12751-12753	IN	denotes	in
T7218	12754-12757	NN	denotes	PBS
T7219	12758-12761	IN	denotes	for
T7220	12762-12764	CD	denotes	10
T7221	12765-12772	NNS	denotes	minutes
T7222	12772-12774	,	denotes	,
T7223	12774-12787	VBN	denotes	permeabilized
T7224	12788-12790	IN	denotes	in
T7225	12791-12792	CD	denotes	2
T7226	12792-12793	NN	denotes	%
T7227	12794-12810	NN	denotes	paraformaldehyde
T7228	12811-12821	VBG	denotes	containing
T7229	12822-12825	CD	denotes	0.1
T7230	12825-12826	NN	denotes	%
T7232	12827-12833	NN	denotes	Triton
T7231	12834-12838	NN	denotes	X100
T7233	12839-12842	IN	denotes	for
T7234	12843-12845	CD	denotes	10
T7235	12846-12853	NNS	denotes	minutes
T7236	12854-12857	CC	denotes	and
T7237	12858-12864	VBD	denotes	washed
T7238	12865-12870	CD	denotes	three
T7239	12871-12876	NNS	denotes	times
T7240	12877-12879	IN	denotes	in
T7241	12880-12883	NN	denotes	PBS
T7242	12884-12894	VBG	denotes	containing
T7243	12895-12898	CD	denotes	0.5
T7244	12898-12899	NN	denotes	%
T7245	12900-12903	NN	denotes	BSA
T7246	12904-12907	CC	denotes	and
T7247	12908-12912	CD	denotes	0.15
T7248	12912-12913	NN	denotes	%
T7249	12914-12921	NN	denotes	glycine
T7250	12921-12923	,	denotes	,
T7251	12923-12925	NN	denotes	pH
T7252	12926-12929	CD	denotes	7.4
T7253	12930-12931	-LRB-	denotes	(
T7255	12931-12937	NN	denotes	Buffer
T7254	12938-12939	NN	denotes	A
T7256	12939-12940	-RRB-	denotes	)
T7257	12940-12941	.	denotes	.
T7258	12941-13248	sentence	denotes	Following a 30 min incubation with purified goat IgG (50 (μg/ml) at 25°C and three additional washes with Buffer A, cells were incubated for 60 min with E2-specific antiserum [30] at 1 μg/ml followed by three washes in Buffer A and 60 minute incubation in fluorescently labeled second antibody (1–2 μg/ml).
T7259	12942-12951	VBG	denotes	Following
T7261	12952-12953	DT	denotes	a
T7263	12954-12956	CD	denotes	30
T7264	12957-12960	NN	denotes	min
T7262	12961-12971	NN	denotes	incubation
T7265	12972-12976	IN	denotes	with
T7266	12977-12985	VBN	denotes	purified
T7268	12986-12990	NN	denotes	goat
T7267	12991-12994	NN	denotes	IgG
T7269	12995-12996	-LRB-	denotes	(
T7271	12996-12998	CD	denotes	50
T7272	12999-13000	-LRB-	denotes	(
T7270	13000-13002	NN	denotes	μg
T7273	13002-13003	SYM	denotes	/
T7274	13003-13005	NN	denotes	ml
T7275	13005-13006	-RRB-	denotes	)
T7276	13007-13009	IN	denotes	at
T7277	13010-13012	CD	denotes	25
T7278	13012-13014	NN	denotes	°C
T7279	13015-13018	CC	denotes	and
T7280	13019-13024	CD	denotes	three
T7282	13025-13035	JJ	denotes	additional
T7281	13036-13042	NNS	denotes	washes
T7283	13043-13047	IN	denotes	with
T7284	13048-13054	NN	denotes	Buffer
T7285	13055-13056	NN	denotes	A
T7286	13056-13058	,	denotes	,
T7287	13058-13063	NNS	denotes	cells
T7288	13064-13068	VBD	denotes	were
T7260	13069-13078	VBN	denotes	incubated
T7289	13079-13082	IN	denotes	for
T7290	13083-13085	CD	denotes	60
T7291	13086-13089	NN	denotes	min
T7292	13090-13094	IN	denotes	with
T7293	13095-13097	NN	denotes	E2
T7295	13097-13098	HYPH	denotes	-
T7294	13098-13106	JJ	denotes	specific
T7296	13107-13116	NN	denotes	antiserum
T7297	13117-13118	-LRB-	denotes	[
T7298	13118-13120	CD	denotes	30
T7299	13120-13121	-RRB-	denotes	]
T7300	13122-13124	IN	denotes	at
T7301	13125-13126	CD	denotes	1
T7302	13127-13129	NN	denotes	μg
T7303	13129-13130	SYM	denotes	/
T7304	13130-13132	NN	denotes	ml
T7305	13133-13141	VBN	denotes	followed
T7306	13142-13144	IN	denotes	by
T7307	13145-13150	CD	denotes	three
T7308	13151-13157	NNS	denotes	washes
T7309	13158-13160	IN	denotes	in
T7310	13161-13167	NN	denotes	Buffer
T7311	13168-13169	NN	denotes	A
T7312	13170-13173	CC	denotes	and
T7313	13174-13176	CD	denotes	60
T7314	13177-13183	NN	denotes	minute
T7315	13184-13194	NN	denotes	incubation
T7316	13195-13197	IN	denotes	in
T7317	13198-13211	RB	denotes	fluorescently
T7318	13212-13219	VBN	denotes	labeled
T7320	13220-13226	JJ	denotes	second
T7319	13227-13235	NN	denotes	antibody
T7321	13236-13237	-LRB-	denotes	(
T7323	13237-13238	CD	denotes	1
T7325	13238-13239	SYM	denotes	–
T7324	13239-13240	CD	denotes	2
T7322	13241-13243	NN	denotes	μg
T7326	13243-13244	SYM	denotes	/
T7327	13244-13246	NN	denotes	ml
T7328	13246-13247	-RRB-	denotes	)
T7329	13247-13248	.	denotes	.
T7330	13248-13361	sentence	denotes	The cells were then washed six times (5 min/wash) in Buffer A and then mounted in gelvatol (Monsanto, St Louis).
T7331	13249-13252	DT	denotes	The
T7332	13253-13258	NNS	denotes	cells
T7334	13259-13263	VBD	denotes	were
T7335	13264-13268	RB	denotes	then
T7333	13269-13275	VBN	denotes	washed
T7336	13276-13279	CD	denotes	six
T7337	13280-13285	NNS	denotes	times
T7338	13286-13287	-LRB-	denotes	(
T7340	13287-13288	CD	denotes	5
T7339	13289-13292	NN	denotes	min
T7341	13292-13293	SYM	denotes	/
T7342	13293-13297	NN	denotes	wash
T7343	13297-13298	-RRB-	denotes	)
T7344	13299-13301	IN	denotes	in
T7345	13302-13308	NN	denotes	Buffer
T7346	13309-13310	NN	denotes	A
T7347	13311-13314	CC	denotes	and
T7348	13315-13319	RB	denotes	then
T7349	13320-13327	VBN	denotes	mounted
T7350	13328-13330	IN	denotes	in
T7351	13331-13339	NN	denotes	gelvatol
T7352	13340-13341	-LRB-	denotes	(
T7353	13341-13349	NNP	denotes	Monsanto
T7354	13349-13351	,	denotes	,
T7355	13351-13353	NNP	denotes	St
T7356	13354-13359	NNP	denotes	Louis
T7357	13359-13360	-RRB-	denotes	)
T7358	13360-13361	.	denotes	.
T7359	13361-13566	sentence	denotes	When livers from newborn pups were examined, fixation was by immersion in 2% paraformaldehyde followed by cryoprotection and shock freezing in liquid nitrogen cooled isopentane and sectioning (5 microns).
T7360	13362-13366	WRB	denotes	When
T7362	13367-13373	NNS	denotes	livers
T7363	13374-13378	IN	denotes	from
T7364	13379-13386	JJ	denotes	newborn
T7365	13387-13391	NNS	denotes	pups
T7366	13392-13396	VBD	denotes	were
T7361	13397-13405	VBN	denotes	examined
T7368	13405-13407	,	denotes	,
T7369	13407-13415	NN	denotes	fixation
T7367	13416-13419	VBD	denotes	was
T7370	13420-13422	IN	denotes	by
T7371	13423-13432	NN	denotes	immersion
T7372	13433-13435	IN	denotes	in
T7373	13436-13437	CD	denotes	2
T7374	13437-13438	NN	denotes	%
T7375	13439-13455	NN	denotes	paraformaldehyde
T7376	13456-13464	VBN	denotes	followed
T7377	13465-13467	IN	denotes	by
T7378	13468-13482	NN	denotes	cryoprotection
T7379	13483-13486	CC	denotes	and
T7380	13487-13492	NN	denotes	shock
T7381	13493-13501	NN	denotes	freezing
T7382	13502-13504	IN	denotes	in
T7383	13505-13511	JJ	denotes	liquid
T7384	13512-13520	NN	denotes	nitrogen
T7385	13521-13527	VBN	denotes	cooled
T7386	13528-13538	NN	denotes	isopentane
T7387	13539-13542	CC	denotes	and
T7388	13543-13553	NN	denotes	sectioning
T7389	13554-13555	-LRB-	denotes	(
T7391	13555-13556	CD	denotes	5
T7390	13557-13564	NNS	denotes	microns
T7392	13564-13565	-RRB-	denotes	)
T7393	13565-13566	.	denotes	.
T7394	13566-13664	sentence	denotes	Otherwise processing was as for the cells above (without the fixation and permabilization steps).
T7395	13567-13576	RB	denotes	Otherwise
T7397	13577-13587	NN	denotes	processing
T7396	13588-13591	VBD	denotes	was
T7398	13592-13594	IN	denotes	as
T7399	13595-13598	IN	denotes	for
T7400	13599-13602	DT	denotes	the
T7401	13603-13608	NNS	denotes	cells
T7402	13609-13614	RB	denotes	above
T7403	13615-13616	-LRB-	denotes	(
T7404	13616-13623	IN	denotes	without
T7405	13624-13627	DT	denotes	the
T7407	13628-13636	NN	denotes	fixation
T7408	13637-13640	CC	denotes	and
T7409	13641-13656	NN	denotes	permabilization
T7406	13657-13662	NNS	denotes	steps
T7410	13662-13663	-RRB-	denotes	)
T7411	13663-13664	.	denotes	.
T7545	13666-13671	NN	denotes	Amino
T7546	13672-13676	NN	denotes	acid
T7547	13677-13685	NN	denotes	analysis
T7548	13685-13853	sentence	denotes	Blood was collected from the retroorbital sinus or tail vein of mice and spotted on a filter paper routinely used for blood amino acid analysis for prenatal screening.
T7549	13686-13691	NN	denotes	Blood
T7551	13692-13695	VBD	denotes	was
T7550	13696-13705	VBN	denotes	collected
T7552	13706-13710	IN	denotes	from
T7553	13711-13714	DT	denotes	the
T7555	13715-13727	JJ	denotes	retroorbital
T7556	13728-13733	NN	denotes	sinus
T7557	13734-13736	CC	denotes	or
T7558	13737-13741	NN	denotes	tail
T7554	13742-13746	NN	denotes	vein
T7559	13747-13749	IN	denotes	of
T7560	13750-13754	NNS	denotes	mice
T7561	13755-13758	CC	denotes	and
T7562	13759-13766	VBN	denotes	spotted
T7563	13767-13769	IN	denotes	on
T7564	13770-13771	DT	denotes	a
T7566	13772-13778	NN	denotes	filter
T7565	13779-13784	NN	denotes	paper
T7567	13785-13794	RB	denotes	routinely
T7568	13795-13799	VBN	denotes	used
T7569	13800-13803	IN	denotes	for
T7570	13804-13809	NN	denotes	blood
T7572	13810-13815	NN	denotes	amino
T7573	13816-13820	NN	denotes	acid
T7571	13821-13829	NN	denotes	analysis
T7574	13830-13833	IN	denotes	for
T7575	13834-13842	JJ	denotes	prenatal
T7576	13843-13852	NN	denotes	screening
T7577	13852-13853	.	denotes	.
T7578	13853-13991	sentence	denotes	Concentrations of BCAA and other amino acids in blood were determined by tandem mass spectrometry (Pediatrix Screening, Bridgeville, PA).
T7579	13854-13868	NNS	denotes	Concentrations
T7581	13869-13871	IN	denotes	of
T7582	13872-13876	NN	denotes	BCAA
T7583	13877-13880	CC	denotes	and
T7584	13881-13886	JJ	denotes	other
T7586	13887-13892	NN	denotes	amino
T7585	13893-13898	NNS	denotes	acids
T7587	13899-13901	IN	denotes	in
T7588	13902-13907	NN	denotes	blood
T7589	13908-13912	VBD	denotes	were
T7580	13913-13923	VBN	denotes	determined
T7590	13924-13926	IN	denotes	by
T7591	13927-13933	JJ	denotes	tandem
T7593	13934-13938	NN	denotes	mass
T7592	13939-13951	NN	denotes	spectrometry
T7594	13952-13953	-LRB-	denotes	(
T7596	13953-13962	NNP	denotes	Pediatrix
T7595	13963-13972	NNP	denotes	Screening
T7597	13972-13974	,	denotes	,
T7598	13974-13985	NNP	denotes	Bridgeville
T7599	13985-13987	,	denotes	,
T7600	13987-13989	NNP	denotes	PA
T7601	13989-13990	-RRB-	denotes	)
T7602	13990-13991	.	denotes	.
T8105	13993-13998	NN	denotes	Assay
T8106	13999-14001	IN	denotes	of
T8107	14002-14007	NN	denotes	BCKDH
T8108	14008-14016	NN	denotes	activity
T8109	14016-14085	sentence	denotes	Livers were removed, frozen in liquid nitrogen, and stored at -80°C.
T8110	14017-14023	NNS	denotes	Livers
T8112	14024-14028	VBD	denotes	were
T8111	14029-14036	VBN	denotes	removed
T8113	14036-14038	,	denotes	,
T8114	14038-14044	VBN	denotes	frozen
T8115	14045-14047	IN	denotes	in
T8116	14048-14054	JJ	denotes	liquid
T8117	14055-14063	NN	denotes	nitrogen
T8118	14063-14065	,	denotes	,
T8119	14065-14068	CC	denotes	and
T8120	14069-14075	VBN	denotes	stored
T8121	14076-14078	IN	denotes	at
T8122	14079-14080	HYPH	denotes	-
T8123	14080-14082	CD	denotes	80
T8124	14082-14084	NN	denotes	°C
T8125	14084-14085	.	denotes	.
T8126	14085-14204	sentence	denotes	At the time of enzyme assay, livers were thawed, and homogenized (1:9, w/v) in 0.25 M sucrose, 10 mM Tris-HC1, pH 7.4.
T8127	14086-14088	IN	denotes	At
T8129	14089-14092	DT	denotes	the
T8130	14093-14097	NN	denotes	time
T8131	14098-14100	IN	denotes	of
T8132	14101-14107	NN	denotes	enzyme
T8133	14108-14113	NN	denotes	assay
T8134	14113-14115	,	denotes	,
T8135	14115-14121	NNS	denotes	livers
T8136	14122-14126	VBD	denotes	were
T8128	14127-14133	VBN	denotes	thawed
T8137	14133-14135	,	denotes	,
T8138	14135-14138	CC	denotes	and
T8139	14139-14150	VBN	denotes	homogenized
T8140	14151-14152	-LRB-	denotes	(
T8142	14152-14153	CD	denotes	1
T8144	14153-14154	SYM	denotes	:
T8143	14154-14155	CD	denotes	9
T8145	14155-14157	,	denotes	,
T8146	14157-14158	NN	denotes	w
T8147	14158-14159	HYPH	denotes	/
T8141	14159-14160	NN	denotes	v
T8148	14160-14161	-RRB-	denotes	)
T8149	14162-14164	IN	denotes	in
T8150	14165-14169	CD	denotes	0.25
T8151	14170-14171	NN	denotes	M
T8152	14172-14179	NN	denotes	sucrose
T8153	14179-14181	,	denotes	,
T8154	14181-14183	CD	denotes	10
T8155	14184-14186	NN	denotes	mM
T8157	14187-14191	NN	denotes	Tris
T8158	14191-14192	HYPH	denotes	-
T8156	14192-14195	NN	denotes	HC1
T8159	14195-14197	,	denotes	,
T8160	14197-14199	NN	denotes	pH
T8161	14200-14203	CD	denotes	7.4
T8162	14203-14204	.	denotes	.
T8163	14204-14340	sentence	denotes	Liver homogenates were centrifuged at 600 × g for 10 min at 4°C and the supernatant fraction was saved to determine the BCKDH activity.
T8164	14205-14210	NN	denotes	Liver
T8165	14211-14222	NNS	denotes	homogenates
T8167	14223-14227	VBD	denotes	were
T8166	14228-14239	VBN	denotes	centrifuged
T8168	14240-14242	IN	denotes	at
T8169	14243-14246	CD	denotes	600
T8171	14247-14248	SYM	denotes	×
T8170	14249-14250	NN	denotes	g
T8172	14251-14254	IN	denotes	for
T8173	14255-14257	CD	denotes	10
T8174	14258-14261	NN	denotes	min
T8175	14262-14264	IN	denotes	at
T8176	14265-14266	CD	denotes	4
T8177	14266-14268	NN	denotes	°C
T8178	14269-14272	CC	denotes	and
T8179	14273-14276	DT	denotes	the
T8181	14277-14288	NN	denotes	supernatant
T8180	14289-14297	NN	denotes	fraction
T8183	14298-14301	VBD	denotes	was
T8182	14302-14307	VBN	denotes	saved
T8184	14308-14310	TO	denotes	to
T8185	14311-14320	VB	denotes	determine
T8186	14321-14324	DT	denotes	the
T8188	14325-14330	NN	denotes	BCKDH
T8187	14331-14339	NN	denotes	activity
T8189	14339-14340	.	denotes	.
T8190	14340-14464	sentence	denotes	The use of tissue homogenates was necessitated by the limited availability of liver tissue, particularly from newborn pups.
T8191	14341-14344	DT	denotes	The
T8192	14345-14348	NN	denotes	use
T8194	14349-14351	IN	denotes	of
T8195	14352-14358	NN	denotes	tissue
T8196	14359-14370	NNS	denotes	homogenates
T8197	14371-14374	VBD	denotes	was
T8193	14375-14387	VBN	denotes	necessitated
T8198	14388-14390	IN	denotes	by
T8199	14391-14394	DT	denotes	the
T8201	14395-14402	JJ	denotes	limited
T8200	14403-14415	NN	denotes	availability
T8202	14416-14418	IN	denotes	of
T8203	14419-14424	NN	denotes	liver
T8204	14425-14431	NN	denotes	tissue
T8205	14431-14433	,	denotes	,
T8206	14433-14445	RB	denotes	particularly
T8207	14446-14450	IN	denotes	from
T8208	14451-14458	JJ	denotes	newborn
T8209	14459-14463	NNS	denotes	pups
T8210	14463-14464	.	denotes	.
T8211	14464-14590	sentence	denotes	BCKDH activity was determined by measuring the release of 14CO2 from α-keto [1-14C] isocaproate as described previously [31].
T8212	14465-14470	NN	denotes	BCKDH
T8213	14471-14479	NN	denotes	activity
T8215	14480-14483	VBD	denotes	was
T8214	14484-14494	VBN	denotes	determined
T8216	14495-14497	IN	denotes	by
T8217	14498-14507	VBG	denotes	measuring
T8218	14508-14511	DT	denotes	the
T8219	14512-14519	NN	denotes	release
T8220	14520-14522	IN	denotes	of
T8221	14523-14528	NN	denotes	14CO2
T8222	14529-14533	IN	denotes	from
T8223	14534-14535	NN	denotes	α
T8225	14535-14536	HYPH	denotes	-
T8224	14536-14540	NN	denotes	keto
T8227	14541-14542	-LRB-	denotes	[
T8228	14542-14543	CD	denotes	1
T8229	14543-14544	SYM	denotes	-
T8230	14544-14547	NN	denotes	14C
T8231	14547-14548	-RRB-	denotes	]
T8226	14549-14560	NN	denotes	isocaproate
T8232	14561-14563	IN	denotes	as
T8233	14564-14573	VBN	denotes	described
T8234	14574-14584	RB	denotes	previously
T8235	14585-14586	-LRB-	denotes	[
T8236	14586-14588	CD	denotes	31
T8237	14588-14589	-RRB-	denotes	]
T8238	14589-14590	.	denotes	.
T8239	14590-14915	sentence	denotes	The complete reaction mixture contained (final volume 1 ml) 30 mM potassium phosphate buffer, pH 7.4, 0.20 mM α-ketoisocaproate, 0.40 mM CoASH, 0.40 mM thiamin pyrophosphate, 2 mM NAD+, 2 mM dithiothreitol, 5 mM Mg2+, approximately 250,000 DPM of α-keto [1-14C] isocaproate, and 0.10 ml of liver homogenate (2–3 mg protein).
T8240	14591-14594	DT	denotes	The
T8242	14595-14603	JJ	denotes	complete
T8243	14604-14612	NN	denotes	reaction
T8241	14613-14620	NN	denotes	mixture
T8244	14621-14630	VBD	denotes	contained
T8245	14631-14632	-LRB-	denotes	(
T8247	14632-14637	JJ	denotes	final
T8246	14638-14644	NN	denotes	volume
T8248	14645-14646	CD	denotes	1
T8249	14647-14649	NN	denotes	ml
T8250	14649-14650	-RRB-	denotes	)
T8251	14651-14653	CD	denotes	30
T8252	14654-14656	NN	denotes	mM
T8254	14657-14666	NN	denotes	potassium
T8255	14667-14676	NN	denotes	phosphate
T8253	14677-14683	NN	denotes	buffer
T8256	14683-14685	,	denotes	,
T8257	14685-14687	NN	denotes	pH
T8258	14688-14691	CD	denotes	7.4
T8259	14691-14693	,	denotes	,
T8260	14693-14697	CD	denotes	0.20
T8261	14698-14700	NN	denotes	mM
T8263	14701-14702	NN	denotes	α
T8264	14702-14703	HYPH	denotes	-
T8262	14703-14718	NN	denotes	ketoisocaproate
T8265	14718-14720	,	denotes	,
T8266	14720-14724	CD	denotes	0.40
T8267	14725-14727	NN	denotes	mM
T8268	14728-14733	NN	denotes	CoASH
T8269	14733-14735	,	denotes	,
T8270	14735-14739	CD	denotes	0.40
T8271	14740-14742	NN	denotes	mM
T8273	14743-14750	NN	denotes	thiamin
T8272	14751-14764	NN	denotes	pyrophosphate
T8274	14764-14766	,	denotes	,
T8275	14766-14767	CD	denotes	2
T8276	14768-14770	NN	denotes	mM
T8277	14771-14774	NN	denotes	NAD
T8278	14774-14775	SYM	denotes	+
T8279	14775-14777	,	denotes	,
T8280	14777-14778	CD	denotes	2
T8281	14779-14781	NN	denotes	mM
T8282	14782-14796	NN	denotes	dithiothreitol
T8283	14796-14798	,	denotes	,
T8284	14798-14799	CD	denotes	5
T8285	14800-14802	NN	denotes	mM
T8286	14803-14806	NN	denotes	Mg2
T8287	14806-14807	SYM	denotes	+
T8288	14807-14809	,	denotes	,
T8289	14809-14822	RB	denotes	approximately
T8290	14823-14830	CD	denotes	250,000
T8291	14831-14834	NN	denotes	DPM
T8292	14835-14837	IN	denotes	of
T8293	14838-14839	NN	denotes	α
T8295	14839-14840	HYPH	denotes	-
T8294	14840-14844	NN	denotes	keto
T8297	14845-14846	-LRB-	denotes	[
T8298	14846-14847	CD	denotes	1
T8299	14847-14848	SYM	denotes	-
T8300	14848-14851	NN	denotes	14C
T8301	14851-14852	-RRB-	denotes	]
T8296	14853-14864	NN	denotes	isocaproate
T8302	14864-14866	,	denotes	,
T8303	14866-14869	CC	denotes	and
T8304	14870-14874	CD	denotes	0.10
T8305	14875-14877	NN	denotes	ml
T8306	14878-14880	IN	denotes	of
T8307	14881-14886	NN	denotes	liver
T8308	14887-14897	NN	denotes	homogenate
T8309	14898-14899	-LRB-	denotes	(
T8311	14899-14900	CD	denotes	2
T8313	14900-14901	SYM	denotes	–
T8312	14901-14902	CD	denotes	3
T8314	14903-14905	NN	denotes	mg
T8310	14906-14913	NN	denotes	protein
T8315	14913-14914	-RRB-	denotes	)
T8316	14914-14915	.	denotes	.
T8317	14915-15073	sentence	denotes	Assays were carried out for 15 min at 37°C, 14CO2 was trapped in hydroxide of Hyamine, and radioactivity was determined by liquid scintillation spectrometry.
T8318	14916-14922	NNS	denotes	Assays
T8320	14923-14927	VBD	denotes	were
T8319	14928-14935	VBN	denotes	carried
T8321	14936-14939	RP	denotes	out
T8322	14940-14943	IN	denotes	for
T8323	14944-14946	CD	denotes	15
T8324	14947-14950	NN	denotes	min
T8325	14951-14953	IN	denotes	at
T8326	14954-14956	CD	denotes	37
T8327	14956-14958	NN	denotes	°C
T8328	14958-14960	,	denotes	,
T8329	14960-14965	NN	denotes	14CO2
T8331	14966-14969	VBD	denotes	was
T8330	14970-14977	VBN	denotes	trapped
T8332	14978-14980	IN	denotes	in
T8333	14981-14990	NN	denotes	hydroxide
T8334	14991-14993	IN	denotes	of
T8335	14994-15001	NN	denotes	Hyamine
T8336	15001-15003	,	denotes	,
T8337	15003-15006	CC	denotes	and
T8338	15007-15020	NN	denotes	radioactivity
T8340	15021-15024	VBD	denotes	was
T8339	15025-15035	VBN	denotes	determined
T8341	15036-15038	IN	denotes	by
T8342	15039-15045	JJ	denotes	liquid
T8343	15046-15059	NN	denotes	scintillation
T8344	15060-15072	NN	denotes	spectrometry
T8345	15072-15073	.	denotes	.
T8817	15075-15082	NNP	denotes	Western
T8818	15083-15087	NN	denotes	blot
T8819	15088-15096	NN	denotes	analysis
T8820	15096-15239	sentence	denotes	Protein extracts were isolated from homogenized liver (freshly harvested and flash frozen) of wildtype, Line 525A, and Line A transgenic mice.
T8821	15097-15104	NN	denotes	Protein
T8822	15105-15113	NNS	denotes	extracts
T8824	15114-15118	VBD	denotes	were
T8823	15119-15127	VBN	denotes	isolated
T8825	15128-15132	IN	denotes	from
T8826	15133-15144	VBN	denotes	homogenized
T8827	15145-15150	NN	denotes	liver
T8828	15151-15152	-LRB-	denotes	(
T8829	15152-15159	RB	denotes	freshly
T8830	15160-15169	VBN	denotes	harvested
T8831	15170-15173	CC	denotes	and
T8832	15174-15179	NN	denotes	flash
T8833	15180-15186	VBN	denotes	frozen
T8834	15186-15187	-RRB-	denotes	)
T8835	15188-15190	IN	denotes	of
T8836	15191-15199	NN	denotes	wildtype
T8838	15199-15201	,	denotes	,
T8839	15201-15205	NN	denotes	Line
T8840	15206-15210	NN	denotes	525A
T8841	15210-15212	,	denotes	,
T8842	15212-15215	CC	denotes	and
T8843	15216-15220	NN	denotes	Line
T8844	15221-15222	NN	denotes	A
T8837	15223-15233	JJ	denotes	transgenic
T8845	15234-15238	NNS	denotes	mice
T8846	15238-15239	.	denotes	.
T8847	15239-15426	sentence	denotes	Protein (25 μg per sample) was analyzed by electrophoresis on a 10% SDS-PAGE Ready Gel (Bio-Rad, Hercules, CA) and transferred to PVDF membrane (Sequi-Blot; Bio-Rad) via electroblotting.
T8848	15240-15247	NN	denotes	Protein
T8850	15248-15249	-LRB-	denotes	(
T8852	15249-15251	CD	denotes	25
T8851	15252-15254	NN	denotes	μg
T8853	15255-15258	IN	denotes	per
T8854	15259-15265	NN	denotes	sample
T8855	15265-15266	-RRB-	denotes	)
T8856	15267-15270	VBD	denotes	was
T8849	15271-15279	VBN	denotes	analyzed
T8857	15280-15282	IN	denotes	by
T8858	15283-15298	NN	denotes	electrophoresis
T8859	15299-15301	IN	denotes	on
T8860	15302-15303	DT	denotes	a
T8862	15304-15306	CD	denotes	10
T8863	15306-15307	NN	denotes	%
T8864	15308-15311	NN	denotes	SDS
T8866	15311-15312	HYPH	denotes	-
T8865	15312-15316	NN	denotes	PAGE
T8867	15317-15322	JJ	denotes	Ready
T8861	15323-15326	NN	denotes	Gel
T8868	15327-15328	-LRB-	denotes	(
T8870	15328-15331	NNP	denotes	Bio
T8871	15331-15332	HYPH	denotes	-
T8869	15332-15335	NNP	denotes	Rad
T8872	15335-15337	,	denotes	,
T8873	15337-15345	NNP	denotes	Hercules
T8874	15345-15347	,	denotes	,
T8875	15347-15349	NNP	denotes	CA
T8876	15349-15350	-RRB-	denotes	)
T8877	15351-15354	CC	denotes	and
T8878	15355-15366	VBN	denotes	transferred
T8879	15367-15369	IN	denotes	to
T8880	15370-15374	NN	denotes	PVDF
T8881	15375-15383	NN	denotes	membrane
T8882	15384-15385	-LRB-	denotes	(
T8884	15385-15390	NNP	denotes	Sequi
T8886	15390-15391	HYPH	denotes	-
T8885	15391-15395	NNP	denotes	Blot
T8887	15395-15396	:	denotes	;
T8888	15397-15400	NNP	denotes	Bio
T8889	15400-15401	HYPH	denotes	-
T8883	15401-15404	NNP	denotes	Rad
T8890	15404-15405	-RRB-	denotes	)
T8891	15406-15409	IN	denotes	via
T8892	15410-15425	NN	denotes	electroblotting
T8893	15425-15426	.	denotes	.
T8894	15426-15635	sentence	denotes	All blots were probed for E2 protein using polyclonal rabbit E2 antisera (1:5,000), which detects both mouse (~47 kD) and human (~54 kD) E2 subunits ([30]; gift from Dr. Susan Hutson, Wake Forest University).
T8895	15427-15430	DT	denotes	All
T8896	15431-15436	NNS	denotes	blots
T8898	15437-15441	VBD	denotes	were
T8897	15442-15448	VBN	denotes	probed
T8899	15449-15452	IN	denotes	for
T8900	15453-15455	NN	denotes	E2
T8901	15456-15463	NN	denotes	protein
T8902	15464-15469	VBG	denotes	using
T8903	15470-15480	JJ	denotes	polyclonal
T8905	15481-15487	NN	denotes	rabbit
T8906	15488-15490	NN	denotes	E2
T8904	15491-15499	NNS	denotes	antisera
T8907	15500-15501	-LRB-	denotes	(
T8909	15501-15502	CD	denotes	1
T8910	15502-15503	SYM	denotes	:
T8908	15503-15508	CD	denotes	5,000
T8911	15508-15509	-RRB-	denotes	)
T8912	15509-15511	,	denotes	,
T8913	15511-15516	WDT	denotes	which
T8914	15517-15524	VBZ	denotes	detects
T8915	15525-15529	CC	denotes	both
T8917	15530-15535	NN	denotes	mouse
T8918	15536-15537	-LRB-	denotes	(
T8920	15537-15538	SYM	denotes	~
T8921	15538-15540	CD	denotes	47
T8919	15541-15543	NN	denotes	kD
T8922	15543-15544	-RRB-	denotes	)
T8923	15545-15548	CC	denotes	and
T8924	15549-15554	JJ	denotes	human
T8925	15555-15556	-LRB-	denotes	(
T8927	15556-15557	SYM	denotes	~
T8928	15557-15559	CD	denotes	54
T8926	15560-15562	NN	denotes	kD
T8929	15562-15563	-RRB-	denotes	)
T8930	15564-15566	NN	denotes	E2
T8916	15567-15575	NNS	denotes	subunits
T8931	15576-15577	-LRB-	denotes	(
T8933	15577-15578	-LRB-	denotes	[
T8934	15578-15580	CD	denotes	30
T8935	15580-15581	-RRB-	denotes	]
T8936	15581-15582	:	denotes	;
T8932	15583-15587	NN	denotes	gift
T8937	15588-15592	IN	denotes	from
T8938	15593-15596	NNP	denotes	Dr.
T8940	15597-15602	NNP	denotes	Susan
T8939	15603-15609	NNP	denotes	Hutson
T8941	15609-15611	,	denotes	,
T8942	15611-15615	NNP	denotes	Wake
T8943	15616-15622	NNP	denotes	Forest
T8944	15623-15633	NNP	denotes	University
T8945	15633-15634	-RRB-	denotes	)
T8946	15634-15635	.	denotes	.
T8947	15635-15746	sentence	denotes	Blots were re-probed with a rabbit anti-c-myc tag antibody (1:10,000; abCam, Cambridge, MA; cat.# ab9106-100).
T8948	15636-15641	NNS	denotes	Blots
T8950	15642-15646	VBD	denotes	were
T8949	15647-15656	VBN	denotes	re-probed
T8951	15657-15661	IN	denotes	with
T8952	15662-15663	DT	denotes	a
T8954	15664-15670	NN	denotes	rabbit
T8955	15671-15677	JJ	denotes	anti-c
T8957	15677-15678	HYPH	denotes	-
T8956	15678-15681	NN	denotes	myc
T8958	15682-15685	NN	denotes	tag
T8953	15686-15694	NN	denotes	antibody
T8959	15695-15696	-LRB-	denotes	(
T8961	15696-15697	CD	denotes	1
T8963	15697-15698	SYM	denotes	:
T8962	15698-15704	CD	denotes	10,000
T8964	15704-15705	:	denotes	;
T8965	15706-15711	NNP	denotes	abCam
T8966	15711-15713	,	denotes	,
T8967	15713-15722	NNP	denotes	Cambridge
T8968	15722-15724	,	denotes	,
T8969	15724-15726	NNP	denotes	MA
T8970	15726-15727	:	denotes	;
T8971	15728-15732	NN	denotes	cat.
T8972	15732-15733	SYM	denotes	#
T8960	15734-15740	NN	denotes	ab9106
T8973	15740-15741	HYPH	denotes	-
T8974	15741-15744	CD	denotes	100
T8975	15744-15745	-RRB-	denotes	)
T8976	15745-15746	.	denotes	.
T8977	15746-15877	sentence	denotes	Blots were also re-probed with an antibody for β-actin (43 kD; 1:10,000; abCam; cat.# ab8227-50) to allow for loading comparisons.
T8978	15747-15752	NNS	denotes	Blots
T8980	15753-15757	VBD	denotes	were
T8981	15758-15762	RB	denotes	also
T8979	15763-15772	VBN	denotes	re-probed
T8982	15773-15777	IN	denotes	with
T8983	15778-15780	DT	denotes	an
T8984	15781-15789	NN	denotes	antibody
T8985	15790-15793	IN	denotes	for
T8986	15794-15795	NN	denotes	β
T8988	15795-15796	HYPH	denotes	-
T8987	15796-15801	NN	denotes	actin
T8989	15802-15803	-LRB-	denotes	(
T8991	15803-15805	CD	denotes	43
T8992	15806-15808	NN	denotes	kD
T8993	15808-15809	:	denotes	;
T8994	15810-15811	CD	denotes	1
T8996	15811-15812	SYM	denotes	:
T8995	15812-15818	CD	denotes	10,000
T8997	15818-15819	:	denotes	;
T8998	15820-15825	NN	denotes	abCam
T8999	15825-15826	:	denotes	;
T9000	15827-15831	NN	denotes	cat.
T9001	15831-15832	SYM	denotes	#
T8990	15833-15839	NN	denotes	ab8227
T9002	15839-15840	HYPH	denotes	-
T9003	15840-15842	CD	denotes	50
T9004	15842-15843	-RRB-	denotes	)
T9005	15844-15846	TO	denotes	to
T9006	15847-15852	VB	denotes	allow
T9007	15853-15856	IN	denotes	for
T9008	15857-15864	NN	denotes	loading
T9009	15865-15876	NNS	denotes	comparisons
T9010	15876-15877	.	denotes	.
T9011	15877-16131	sentence	denotes	A goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (1:10,000; Novus, Littleton, CO; cat.# NB730-H) was used for detection using the Western Lightning chemiluminescence reagent (Perkin Elmer, Boston, MA) and exposed to X-ray film.
T9012	15878-15879	DT	denotes	A
T9014	15880-15884	NN	denotes	goat
T9015	15885-15896	JJ	denotes	anti-rabbit
T9016	15897-15906	JJ	denotes	secondary
T9013	15907-15915	NN	denotes	antibody
T9018	15916-15926	VBN	denotes	conjugated
T9019	15927-15929	IN	denotes	to
T9020	15930-15941	NN	denotes	horseradish
T9021	15942-15952	NN	denotes	peroxidase
T9022	15953-15954	-LRB-	denotes	(
T9024	15954-15955	CD	denotes	1
T9026	15955-15956	SYM	denotes	:
T9025	15956-15962	CD	denotes	10,000
T9027	15962-15963	:	denotes	;
T9028	15964-15969	NNP	denotes	Novus
T9029	15969-15971	,	denotes	,
T9030	15971-15980	NNP	denotes	Littleton
T9031	15980-15982	,	denotes	,
T9032	15982-15984	NNP	denotes	CO
T9033	15984-15985	:	denotes	;
T9034	15986-15990	NN	denotes	cat.
T9035	15990-15991	SYM	denotes	#
T9036	15992-15997	NN	denotes	NB730
T9037	15997-15998	HYPH	denotes	-
T9023	15998-15999	NN	denotes	H
T9038	15999-16000	-RRB-	denotes	)
T9039	16001-16004	VBD	denotes	was
T9017	16005-16009	VBN	denotes	used
T9040	16010-16013	IN	denotes	for
T9041	16014-16023	NN	denotes	detection
T9042	16024-16029	VBG	denotes	using
T9043	16030-16033	DT	denotes	the
T9045	16034-16041	NNP	denotes	Western
T9046	16042-16051	NNP	denotes	Lightning
T9047	16052-16069	NN	denotes	chemiluminescence
T9044	16070-16077	NN	denotes	reagent
T9048	16078-16079	-LRB-	denotes	(
T9050	16079-16085	NNP	denotes	Perkin
T9049	16086-16091	NNP	denotes	Elmer
T9051	16091-16093	,	denotes	,
T9052	16093-16099	NNP	denotes	Boston
T9053	16099-16101	,	denotes	,
T9054	16101-16103	NNP	denotes	MA
T9055	16103-16104	-RRB-	denotes	)
T9056	16105-16108	CC	denotes	and
T9057	16109-16116	VBN	denotes	exposed
T9058	16117-16119	IN	denotes	to
T9059	16120-16125	NN	denotes	X-ray
T9060	16126-16130	NN	denotes	film
T9061	16130-16131	.	denotes	.
T9141	16133-16144	JJ	denotes	Statistical
T9142	16145-16153	NNS	denotes	analyses
T9143	16153-16264	sentence	denotes	All BCAA and BCKDH enzyme activity data are presented as the mean +/- the standard error of the mean (S.E.M.).
T9144	16154-16157	DT	denotes	All
T9146	16158-16162	NN	denotes	BCAA
T9147	16163-16166	CC	denotes	and
T9148	16167-16172	NN	denotes	BCKDH
T9149	16173-16179	NN	denotes	enzyme
T9150	16180-16188	NN	denotes	activity
T9145	16189-16193	NNS	denotes	data
T9152	16194-16197	VBP	denotes	are
T9151	16198-16207	VBN	denotes	presented
T9153	16208-16210	IN	denotes	as
T9154	16211-16214	DT	denotes	the
T9155	16215-16219	NN	denotes	mean
T9156	16220-16221	SYM	denotes	+
T9157	16221-16222	HYPH	denotes	/
T9158	16222-16223	SYM	denotes	-
T9159	16224-16227	DT	denotes	the
T9161	16228-16236	JJ	denotes	standard
T9160	16237-16242	NN	denotes	error
T9162	16243-16245	IN	denotes	of
T9163	16246-16249	DT	denotes	the
T9164	16250-16254	NN	denotes	mean
T9165	16255-16256	-LRB-	denotes	(
T9166	16256-16262	NN	denotes	S.E.M.
T9167	16262-16263	-RRB-	denotes	)
T9168	16263-16264	.	denotes	.
T9169	16264-16333	sentence	denotes	Differences between genotypes were compared by Student's t test[32].
T9170	16265-16276	NNS	denotes	Differences
T9172	16277-16284	IN	denotes	between
T9173	16285-16294	NNS	denotes	genotypes
T9174	16295-16299	VBD	denotes	were
T9171	16300-16308	VBN	denotes	compared
T9175	16309-16311	IN	denotes	by
T9176	16312-16319	NNP	denotes	Student
T9178	16319-16321	POS	denotes	's
T9179	16322-16323	NN	denotes	t
T9177	16324-16328	NN	denotes	test
T9180	16328-16329	-LRB-	denotes	[
T9181	16329-16331	CD	denotes	32
T9182	16331-16332	-RRB-	denotes	]
T9183	16332-16333	.	denotes	.
T10367	16344-16354	NN	denotes	Production
T10368	16355-16357	IN	denotes	of
T10369	16358-16360	NN	denotes	E2
T10371	16361-16365	NN	denotes	gene
T10372	16366-16374	NN	denotes	knockout
T10370	16375-16379	NNS	denotes	mice
T10373	16379-16472	sentence	denotes	To create E2 knockout mice, we used gene targeting in mouse ES cells to disrupt the E2 gene.
T10374	16380-16382	TO	denotes	To
T10375	16383-16389	VB	denotes	create
T10377	16390-16392	NN	denotes	E2
T10379	16393-16401	NN	denotes	knockout
T10378	16402-16406	NNS	denotes	mice
T10380	16406-16408	,	denotes	,
T10381	16408-16410	PRP	denotes	we
T10376	16411-16415	VBD	denotes	used
T10382	16416-16420	NN	denotes	gene
T10383	16421-16430	NN	denotes	targeting
T10384	16431-16433	IN	denotes	in
T10385	16434-16439	NN	denotes	mouse
T10387	16440-16442	NN	denotes	ES
T10386	16443-16448	NNS	denotes	cells
T10388	16449-16451	TO	denotes	to
T10389	16452-16459	VB	denotes	disrupt
T10390	16460-16463	DT	denotes	the
T10392	16464-16466	NN	denotes	E2
T10391	16467-16471	NN	denotes	gene
T10393	16471-16472	.	denotes	.
T10394	16472-16547	sentence	denotes	The overall strategy for disrupting the E2 gene is illustrated in Fig. 1A.
T10395	16473-16476	DT	denotes	The
T10397	16477-16484	JJ	denotes	overall
T10396	16485-16493	NN	denotes	strategy
T10399	16494-16497	IN	denotes	for
T10400	16498-16508	VBG	denotes	disrupting
T10401	16509-16512	DT	denotes	the
T10403	16513-16515	NN	denotes	E2
T10402	16516-16520	NN	denotes	gene
T10404	16521-16523	VBZ	denotes	is
T10398	16524-16535	VBN	denotes	illustrated
T10405	16536-16538	IN	denotes	in
T10406	16539-16543	NN	denotes	Fig.
T10407	16544-16546	NN	denotes	1A
T10408	16546-16547	.	denotes	.
T10409	16547-16732	sentence	denotes	The gene targeting construct was designed to replace a 1.67 kb EcoRV/Smal genomic DNA fragment encompassing part of Exon 4 and all of Exon 5 with the PGKneo selectable marker cassette.
T10410	16548-16551	DT	denotes	The
T10412	16552-16556	NN	denotes	gene
T10413	16557-16566	NN	denotes	targeting
T10411	16567-16576	NN	denotes	construct
T10415	16577-16580	VBD	denotes	was
T10414	16581-16589	VBN	denotes	designed
T10416	16590-16592	TO	denotes	to
T10417	16593-16600	VB	denotes	replace
T10418	16601-16602	DT	denotes	a
T10420	16603-16607	CD	denotes	1.67
T10421	16608-16610	NN	denotes	kb
T10422	16611-16616	NN	denotes	EcoRV
T10424	16616-16617	HYPH	denotes	/
T10423	16617-16621	NN	denotes	Smal
T10425	16622-16629	JJ	denotes	genomic
T10426	16630-16633	NN	denotes	DNA
T10419	16634-16642	NN	denotes	fragment
T10427	16643-16655	VBG	denotes	encompassing
T10428	16656-16660	NN	denotes	part
T10429	16661-16663	IN	denotes	of
T10430	16664-16668	NN	denotes	Exon
T10431	16669-16670	CD	denotes	4
T10432	16671-16674	CC	denotes	and
T10433	16675-16678	DT	denotes	all
T10434	16679-16681	IN	denotes	of
T10435	16682-16686	NN	denotes	Exon
T10436	16687-16688	CD	denotes	5
T10437	16689-16693	IN	denotes	with
T10438	16694-16697	DT	denotes	the
T10440	16698-16704	NN	denotes	PGKneo
T10441	16705-16715	JJ	denotes	selectable
T10442	16716-16722	NN	denotes	marker
T10439	16723-16731	NN	denotes	cassette
T10443	16731-16732	.	denotes	.
T10444	16732-16924	sentence	denotes	Of 522 ES cell clones screened for targeting by Southern blot analysis, 29 (5%) displayed the predicted restriction fragment length polymorphisms indicative of gene targeting at the E2 locus.
T10445	16733-16735	IN	denotes	Of
T10447	16736-16739	CD	denotes	522
T10449	16740-16742	NN	denotes	ES
T10450	16743-16747	NN	denotes	cell
T10448	16748-16754	NNS	denotes	clones
T10451	16755-16763	VBN	denotes	screened
T10452	16764-16767	IN	denotes	for
T10453	16768-16777	NN	denotes	targeting
T10454	16778-16780	IN	denotes	by
T10455	16781-16789	NNP	denotes	Southern
T10456	16790-16794	NN	denotes	blot
T10457	16795-16803	NN	denotes	analysis
T10458	16803-16805	,	denotes	,
T10459	16805-16807	CD	denotes	29
T10460	16808-16809	-LRB-	denotes	(
T10462	16809-16810	CD	denotes	5
T10461	16810-16811	NN	denotes	%
T10463	16811-16812	-RRB-	denotes	)
T10446	16813-16822	VBD	denotes	displayed
T10464	16823-16826	DT	denotes	the
T10466	16827-16836	VBN	denotes	predicted
T10467	16837-16848	NN	denotes	restriction
T10469	16849-16857	NN	denotes	fragment
T10468	16858-16864	NN	denotes	length
T10465	16865-16878	NNS	denotes	polymorphisms
T10470	16879-16889	JJ	denotes	indicative
T10471	16890-16892	IN	denotes	of
T10472	16893-16897	NN	denotes	gene
T10473	16898-16907	VBG	denotes	targeting
T10474	16908-16910	IN	denotes	at
T10475	16911-16914	DT	denotes	the
T10477	16915-16917	NN	denotes	E2
T10476	16918-16923	NN	denotes	locus
T10478	16923-16924	.	denotes	.
T10479	16924-17146	sentence	denotes	As illustrated in Fig. 1A and 1B, an E2 Exon 6 specific probe, which is external to the targeting construct, hybridizes to only a ~16 kb BglI restriction fragment from the wild type allele in the parental R1 ES cell line.
T10480	16925-16927	IN	denotes	As
T10481	16928-16939	VBN	denotes	illustrated
T10483	16940-16942	IN	denotes	in
T10484	16943-16947	NN	denotes	Fig.
T10485	16948-16950	NN	denotes	1A
T10486	16951-16954	CC	denotes	and
T10487	16955-16957	NN	denotes	1B
T10488	16957-16959	,	denotes	,
T10489	16959-16961	DT	denotes	an
T10491	16962-16964	NN	denotes	E2
T10492	16965-16969	NN	denotes	Exon
T10493	16970-16971	CD	denotes	6
T10494	16972-16980	JJ	denotes	specific
T10490	16981-16986	NN	denotes	probe
T10495	16986-16988	,	denotes	,
T10496	16988-16993	WDT	denotes	which
T10497	16994-16996	VBZ	denotes	is
T10498	16997-17005	JJ	denotes	external
T10499	17006-17008	IN	denotes	to
T10500	17009-17012	DT	denotes	the
T10502	17013-17022	NN	denotes	targeting
T10501	17023-17032	NN	denotes	construct
T10503	17032-17034	,	denotes	,
T10482	17034-17044	VBZ	denotes	hybridizes
T10504	17045-17047	IN	denotes	to
T10505	17048-17052	RB	denotes	only
T10507	17053-17054	DT	denotes	a
T10508	17055-17056	SYM	denotes	~
T10509	17056-17058	CD	denotes	16
T10510	17059-17061	NN	denotes	kb
T10511	17062-17066	NN	denotes	BglI
T10512	17067-17078	NN	denotes	restriction
T10506	17079-17087	NN	denotes	fragment
T10513	17088-17092	IN	denotes	from
T10514	17093-17096	DT	denotes	the
T10516	17097-17101	JJ	denotes	wild
T10517	17102-17106	NN	denotes	type
T10515	17107-17113	NN	denotes	allele
T10518	17114-17116	IN	denotes	in
T10519	17117-17120	DT	denotes	the
T10521	17121-17129	JJ	denotes	parental
T10522	17130-17132	NN	denotes	R1
T10523	17133-17135	NN	denotes	ES
T10524	17136-17140	NN	denotes	cell
T10520	17141-17145	NN	denotes	line
T10525	17145-17146	.	denotes	.
T10526	17146-17244	sentence	denotes	In correctly targeted ES cells, this probe also hybridizes to a ~11 kb BglI restriction fragment.
T10527	17147-17149	IN	denotes	In
T10529	17150-17159	RB	denotes	correctly
T10530	17160-17168	VBN	denotes	targeted
T10532	17169-17171	NN	denotes	ES
T10531	17172-17177	NNS	denotes	cells
T10533	17177-17179	,	denotes	,
T10534	17179-17183	DT	denotes	this
T10535	17184-17189	NN	denotes	probe
T10536	17190-17194	RB	denotes	also
T10528	17195-17205	VBZ	denotes	hybridizes
T10537	17206-17208	IN	denotes	to
T10538	17209-17210	DT	denotes	a
T10540	17211-17212	SYM	denotes	~
T10541	17212-17214	CD	denotes	11
T10542	17215-17217	NN	denotes	kb
T10543	17218-17222	NN	denotes	BglI
T10544	17223-17234	NN	denotes	restriction
T10539	17235-17243	NN	denotes	fragment
T10545	17243-17244	.	denotes	.
T10546	17244-17341	sentence	denotes	Targeting was confirmed with several additional restriction enzymes and probes (data not shown).
T10547	17245-17254	NN	denotes	Targeting
T10549	17255-17258	VBD	denotes	was
T10548	17259-17268	VBN	denotes	confirmed
T10550	17269-17273	IN	denotes	with
T10551	17274-17281	JJ	denotes	several
T10553	17282-17292	JJ	denotes	additional
T10554	17293-17304	NN	denotes	restriction
T10552	17305-17312	NNS	denotes	enzymes
T10555	17313-17316	CC	denotes	and
T10556	17317-17323	NNS	denotes	probes
T10557	17324-17325	-LRB-	denotes	(
T10559	17325-17329	NNS	denotes	data
T10560	17330-17333	RB	denotes	not
T10558	17334-17339	VBN	denotes	shown
T10561	17339-17340	-RRB-	denotes	)
T10562	17340-17341	.	denotes	.
T10563	17341-17431	sentence	denotes	Correctly targeted ES cells were microinjected into blastocysts to produce chimeric mice.
T10564	17342-17351	RB	denotes	Correctly
T10565	17352-17360	VBN	denotes	targeted
T10567	17361-17363	NN	denotes	ES
T10566	17364-17369	NNS	denotes	cells
T10569	17370-17374	VBD	denotes	were
T10568	17375-17388	VBN	denotes	microinjected
T10570	17389-17393	IN	denotes	into
T10571	17394-17405	NNS	denotes	blastocysts
T10572	17406-17408	TO	denotes	to
T10573	17409-17416	VB	denotes	produce
T10574	17417-17425	JJ	denotes	chimeric
T10575	17426-17430	NNS	denotes	mice
T10576	17430-17431	.	denotes	.
T10577	17431-17469	sentence	denotes	Chimerics were bred to C57BL/6J mice.
T10578	17432-17441	NNS	denotes	Chimerics
T10580	17442-17446	VBD	denotes	were
T10579	17447-17451	VBN	denotes	bred
T10581	17452-17454	IN	denotes	to
T10582	17455-17460	NN	denotes	C57BL
T10584	17460-17461	HYPH	denotes	/
T10583	17461-17463	NN	denotes	6J
T10585	17464-17468	NNS	denotes	mice
T10586	17468-17469	.	denotes	.
T10587	17469-17635	sentence	denotes	Following germline transmission of the targeted allele, heterozygous mice were interbred to produce wild type (+/+), heterozygous (+/-) and homozygous (-/-) animals.
T10588	17470-17479	VBG	denotes	Following
T10590	17480-17488	NN	denotes	germline
T10591	17489-17501	NN	denotes	transmission
T10592	17502-17504	IN	denotes	of
T10593	17505-17508	DT	denotes	the
T10595	17509-17517	VBN	denotes	targeted
T10594	17518-17524	NN	denotes	allele
T10596	17524-17526	,	denotes	,
T10597	17526-17538	JJ	denotes	heterozygous
T10598	17539-17543	NNS	denotes	mice
T10599	17544-17548	VBD	denotes	were
T10589	17549-17558	VBN	denotes	interbred
T10600	17559-17561	TO	denotes	to
T10601	17562-17569	VB	denotes	produce
T10602	17570-17574	JJ	denotes	wild
T10603	17575-17579	NN	denotes	type
T10605	17580-17581	-LRB-	denotes	(
T10607	17581-17582	SYM	denotes	+
T10608	17582-17583	HYPH	denotes	/
T10606	17583-17584	SYM	denotes	+
T10609	17584-17585	-RRB-	denotes	)
T10610	17585-17587	,	denotes	,
T10611	17587-17599	JJ	denotes	heterozygous
T10612	17600-17601	-LRB-	denotes	(
T10614	17601-17602	SYM	denotes	+
T10615	17602-17603	HYPH	denotes	/
T10613	17603-17604	SYM	denotes	-
T10616	17604-17605	-RRB-	denotes	)
T10617	17606-17609	CC	denotes	and
T10618	17610-17620	JJ	denotes	homozygous
T10619	17621-17622	-LRB-	denotes	(
T10621	17622-17623	SYM	denotes	-
T10622	17623-17624	HYPH	denotes	/
T10620	17624-17625	SYM	denotes	-
T10623	17625-17626	-RRB-	denotes	)
T10604	17627-17634	NNS	denotes	animals
T10624	17634-17635	.	denotes	.
T10625	17635-17682	sentence	denotes	Mice were genotyped by Southern blot analysis.
T10626	17636-17640	NNS	denotes	Mice
T10628	17641-17645	VBD	denotes	were
T10627	17646-17655	VBN	denotes	genotyped
T10629	17656-17658	IN	denotes	by
T10630	17659-17667	NNP	denotes	Southern
T10631	17668-17672	NN	denotes	blot
T10632	17673-17681	NN	denotes	analysis
T10633	17681-17682	.	denotes	.
T10634	17682-17865	sentence	denotes	The Exon 6 specific probe hybridized to only a ~16 kb BglI restriction fragment in +/+ mice, a ~11 kb BglI fragment in -/- mice, and to both of these fragments in +/- mice (Fig. 1B).
T10635	17683-17686	DT	denotes	The
T10637	17687-17691	NN	denotes	Exon
T10639	17692-17693	CD	denotes	6
T10638	17694-17702	JJ	denotes	specific
T10636	17703-17708	NN	denotes	probe
T10640	17709-17719	VBN	denotes	hybridized
T10641	17720-17722	IN	denotes	to
T10642	17723-17727	RB	denotes	only
T10644	17728-17729	DT	denotes	a
T10645	17730-17731	SYM	denotes	~
T10646	17731-17733	CD	denotes	16
T10647	17734-17736	NN	denotes	kb
T10648	17737-17741	NN	denotes	BglI
T10649	17742-17753	NN	denotes	restriction
T10643	17754-17762	NN	denotes	fragment
T10650	17763-17765	IN	denotes	in
T10651	17766-17767	SYM	denotes	+
T10653	17767-17768	HYPH	denotes	/
T10652	17768-17769	SYM	denotes	+
T10654	17770-17774	NNS	denotes	mice
T10655	17774-17776	,	denotes	,
T10656	17776-17777	DT	denotes	a
T10658	17778-17779	SYM	denotes	~
T10659	17779-17781	CD	denotes	11
T10660	17782-17784	NN	denotes	kb
T10661	17785-17789	NN	denotes	BglI
T10657	17790-17798	NN	denotes	fragment
T10662	17799-17801	IN	denotes	in
T10663	17802-17803	SYM	denotes	-
T10665	17803-17804	HYPH	denotes	/
T10664	17804-17805	SYM	denotes	-
T10666	17806-17810	NNS	denotes	mice
T10667	17810-17812	,	denotes	,
T10668	17812-17815	CC	denotes	and
T10669	17816-17818	IN	denotes	to
T10670	17819-17823	DT	denotes	both
T10671	17824-17826	IN	denotes	of
T10672	17827-17832	DT	denotes	these
T10673	17833-17842	NNS	denotes	fragments
T10674	17843-17845	IN	denotes	in
T10675	17846-17847	SYM	denotes	+
T10677	17847-17848	HYPH	denotes	/
T10676	17848-17849	SYM	denotes	-
T10678	17850-17854	NNS	denotes	mice
T10679	17855-17856	-LRB-	denotes	(
T10681	17856-17860	NN	denotes	Fig.
T10680	17861-17863	NN	denotes	1B
T10682	17863-17864	-RRB-	denotes	)
T10683	17864-17865	.	denotes	.
T10684	17865-17938	sentence	denotes	Mice homozygous for the E2 mutation were born at the expected frequency.
T10685	17866-17870	NNS	denotes	Mice
T10687	17871-17881	JJ	denotes	homozygous
T10688	17882-17885	IN	denotes	for
T10689	17886-17889	DT	denotes	the
T10691	17890-17892	NN	denotes	E2
T10690	17893-17901	NN	denotes	mutation
T10692	17902-17906	VBD	denotes	were
T10686	17907-17911	VBN	denotes	born
T10693	17912-17914	IN	denotes	at
T10694	17915-17918	DT	denotes	the
T10696	17919-17927	VBN	denotes	expected
T10695	17928-17937	NN	denotes	frequency
T10697	17937-17938	.	denotes	.
T10698	17938-18086	sentence	denotes	Genotype analysis of pups derived from +/- by +/- matings revealed that +/+, +/-, and -/- mice were present at nearly the expected 1:2:1 frequency.
T10699	17939-17947	NN	denotes	Genotype
T10700	17948-17956	NN	denotes	analysis
T10702	17957-17959	IN	denotes	of
T10703	17960-17964	NNS	denotes	pups
T10704	17965-17972	VBN	denotes	derived
T10705	17973-17977	IN	denotes	from
T10706	17978-17979	SYM	denotes	+
T10708	17979-17980	HYPH	denotes	/
T10707	17980-17981	SYM	denotes	-
T10710	17982-17984	IN	denotes	by
T10711	17985-17986	SYM	denotes	+
T10713	17986-17987	HYPH	denotes	/
T10712	17987-17988	SYM	denotes	-
T10709	17989-17996	NNS	denotes	matings
T10701	17997-18005	VBD	denotes	revealed
T10714	18006-18010	IN	denotes	that
T10716	18011-18012	SYM	denotes	+
T10718	18012-18013	HYPH	denotes	/
T10717	18013-18014	SYM	denotes	+
T10720	18014-18016	,	denotes	,
T10721	18016-18017	SYM	denotes	+
T10723	18017-18018	HYPH	denotes	/
T10722	18018-18019	SYM	denotes	-
T10724	18019-18021	,	denotes	,
T10725	18021-18024	CC	denotes	and
T10726	18025-18026	SYM	denotes	-
T10728	18026-18027	HYPH	denotes	/
T10727	18027-18028	SYM	denotes	-
T10719	18029-18033	NNS	denotes	mice
T10715	18034-18038	VBD	denotes	were
T10729	18039-18046	JJ	denotes	present
T10730	18047-18049	IN	denotes	at
T10731	18050-18056	RB	denotes	nearly
T10733	18057-18060	DT	denotes	the
T10734	18061-18069	VBN	denotes	expected
T10735	18070-18071	CD	denotes	1
T10737	18071-18072	SYM	denotes	:
T10738	18072-18073	CD	denotes	2
T10739	18073-18074	SYM	denotes	:
T10736	18074-18075	CD	denotes	1
T10732	18076-18085	NN	denotes	frequency
T10740	18085-18086	.	denotes	.
T10741	18086-18184	sentence	denotes	Of the initial 60 animals genotyped, 19 (32%) were +/+, 27 (45%) were +/-, and 14 (23%) were -/-.
T10742	18087-18089	IN	denotes	Of
T10744	18090-18093	DT	denotes	the
T10746	18094-18101	JJ	denotes	initial
T10747	18102-18104	CD	denotes	60
T10745	18105-18112	NNS	denotes	animals
T10748	18113-18122	VBN	denotes	genotyped
T10749	18122-18124	,	denotes	,
T10750	18124-18126	CD	denotes	19
T10751	18127-18128	-LRB-	denotes	(
T10753	18128-18130	CD	denotes	32
T10752	18130-18131	NN	denotes	%
T10754	18131-18132	-RRB-	denotes	)
T10743	18133-18137	VBD	denotes	were
T10755	18138-18139	SYM	denotes	+
T10757	18139-18140	HYPH	denotes	/
T10756	18140-18141	SYM	denotes	+
T10758	18141-18143	,	denotes	,
T10759	18143-18145	CD	denotes	27
T10761	18146-18147	-LRB-	denotes	(
T10763	18147-18149	CD	denotes	45
T10762	18149-18150	NN	denotes	%
T10764	18150-18151	-RRB-	denotes	)
T10760	18152-18156	VBD	denotes	were
T10765	18157-18158	SYM	denotes	+
T10767	18158-18159	HYPH	denotes	/
T10766	18159-18160	SYM	denotes	-
T10768	18160-18162	,	denotes	,
T10769	18162-18165	CC	denotes	and
T10770	18166-18168	CD	denotes	14
T10772	18169-18170	-LRB-	denotes	(
T10774	18170-18172	CD	denotes	23
T10773	18172-18173	NN	denotes	%
T10775	18173-18174	-RRB-	denotes	)
T10771	18175-18179	VBD	denotes	were
T10776	18180-18181	SYM	denotes	-
T10778	18181-18182	HYPH	denotes	/
T10777	18182-18183	SYM	denotes	-
T10779	18183-18184	.	denotes	.
T10780	18184-18252	sentence	denotes	Thus, the E2 gene was dispensable for normal embryonic development.
T10781	18185-18189	RB	denotes	Thus
T10783	18189-18191	,	denotes	,
T10784	18191-18194	DT	denotes	the
T10786	18195-18197	NN	denotes	E2
T10785	18198-18202	NN	denotes	gene
T10782	18203-18206	VBD	denotes	was
T10787	18207-18218	JJ	denotes	dispensable
T10788	18219-18222	IN	denotes	for
T10789	18223-18229	JJ	denotes	normal
T10791	18230-18239	JJ	denotes	embryonic
T10790	18240-18251	NN	denotes	development
T10792	18251-18252	.	denotes	.
T10793	18252-18331	sentence	denotes	However, as expected, nearly all homozygous mice died in the perinatal period.
T10794	18253-18260	RB	denotes	However
T10796	18260-18262	,	denotes	,
T10797	18262-18264	IN	denotes	as
T10798	18265-18273	VBN	denotes	expected
T10799	18273-18275	,	denotes	,
T10800	18275-18281	RB	denotes	nearly
T10802	18282-18285	DT	denotes	all
T10803	18286-18296	JJ	denotes	homozygous
T10801	18297-18301	NNS	denotes	mice
T10795	18302-18306	VBD	denotes	died
T10804	18307-18309	IN	denotes	in
T10805	18310-18313	DT	denotes	the
T10807	18314-18323	JJ	denotes	perinatal
T10806	18324-18330	NN	denotes	period
T10808	18330-18331	.	denotes	.
T10809	18331-18482	sentence	denotes	Immediately following birth, homozygous pups were indistinguishable from their +/+ and +/- littermates; they were vigorous, active and able to suckle.
T10810	18332-18343	RB	denotes	Immediately
T10811	18344-18353	VBG	denotes	following
T10813	18354-18359	NN	denotes	birth
T10814	18359-18361	,	denotes	,
T10815	18361-18371	JJ	denotes	homozygous
T10816	18372-18376	NNS	denotes	pups
T10812	18377-18381	VBD	denotes	were
T10818	18382-18399	JJ	denotes	indistinguishable
T10819	18400-18404	IN	denotes	from
T10820	18405-18410	PRP$	denotes	their
T10822	18411-18412	SYM	denotes	+
T10824	18412-18413	HYPH	denotes	/
T10823	18413-18414	SYM	denotes	+
T10825	18415-18418	CC	denotes	and
T10826	18419-18420	SYM	denotes	+
T10828	18420-18421	HYPH	denotes	/
T10827	18421-18422	SYM	denotes	-
T10821	18423-18434	NNS	denotes	littermates
T10829	18434-18435	:	denotes	;
T10830	18436-18440	PRP	denotes	they
T10817	18441-18445	VBD	denotes	were
T10831	18446-18454	JJ	denotes	vigorous
T10832	18454-18456	,	denotes	,
T10833	18456-18462	JJ	denotes	active
T10834	18463-18466	CC	denotes	and
T10835	18467-18471	JJ	denotes	able
T10836	18472-18474	TO	denotes	to
T10837	18475-18481	VB	denotes	suckle
T10838	18481-18482	.	denotes	.
T10839	18482-18658	sentence	denotes	By mid to late day on postnatal day one, most -/- pups became moribund and were readily identifiable as they were lethargic, pale, and exhibited gasping respiratory movements.
T10840	18483-18485	IN	denotes	By
T10842	18486-18489	JJ	denotes	mid
T10844	18490-18492	IN	denotes	to
T10845	18493-18497	JJ	denotes	late
T10843	18498-18501	NN	denotes	day
T10846	18502-18504	IN	denotes	on
T10847	18505-18514	JJ	denotes	postnatal
T10848	18515-18518	NN	denotes	day
T10849	18519-18522	CD	denotes	one
T10850	18522-18524	,	denotes	,
T10851	18524-18528	RBS	denotes	most
T10853	18529-18530	SYM	denotes	-
T10855	18530-18531	HYPH	denotes	/
T10854	18531-18532	SYM	denotes	-
T10852	18533-18537	NNS	denotes	pups
T10841	18538-18544	VBD	denotes	became
T10856	18545-18553	JJ	denotes	moribund
T10857	18554-18557	CC	denotes	and
T10858	18558-18562	VBD	denotes	were
T10859	18563-18570	RB	denotes	readily
T10860	18571-18583	JJ	denotes	identifiable
T10861	18584-18586	IN	denotes	as
T10863	18587-18591	PRP	denotes	they
T10862	18592-18596	VBD	denotes	were
T10864	18597-18606	JJ	denotes	lethargic
T10866	18606-18608	,	denotes	,
T10865	18608-18612	JJ	denotes	pale
T10867	18612-18614	,	denotes	,
T10868	18614-18617	CC	denotes	and
T10869	18618-18627	VBD	denotes	exhibited
T10870	18628-18635	VBG	denotes	gasping
T10872	18636-18647	JJ	denotes	respiratory
T10871	18648-18657	NNS	denotes	movements
T10873	18657-18658	.	denotes	.
T10874	18658-18719	sentence	denotes	With few exceptions, -/- pups died within 72 hours of birth.
T10875	18659-18663	IN	denotes	With
T10877	18664-18667	JJ	denotes	few
T10878	18668-18678	NNS	denotes	exceptions
T10879	18678-18680	,	denotes	,
T10880	18680-18681	SYM	denotes	-
T10882	18681-18682	HYPH	denotes	/
T10881	18682-18683	SYM	denotes	-
T10883	18684-18688	NNS	denotes	pups
T10876	18689-18693	VBD	denotes	died
T10884	18694-18700	IN	denotes	within
T10885	18701-18703	CD	denotes	72
T10886	18704-18709	NNS	denotes	hours
T10887	18710-18712	IN	denotes	of
T10888	18713-18718	NN	denotes	birth
T10889	18718-18719	.	denotes	.
T10890	18719-18788	sentence	denotes	We have observed one rare -/- pup that survived to postnatal day 13.
T10891	18720-18722	PRP	denotes	We
T10893	18723-18727	VBP	denotes	have
T10892	18728-18736	VBN	denotes	observed
T10894	18737-18740	CD	denotes	one
T10896	18741-18745	JJ	denotes	rare
T10897	18746-18747	SYM	denotes	-
T10899	18747-18748	HYPH	denotes	/
T10898	18748-18749	SYM	denotes	-
T10895	18750-18753	NN	denotes	pup
T10900	18754-18758	WDT	denotes	that
T10901	18759-18767	VBD	denotes	survived
T10902	18768-18770	IN	denotes	to
T10903	18771-18780	JJ	denotes	postnatal
T10904	18781-18784	NN	denotes	day
T10905	18785-18787	CD	denotes	13
T10906	18787-18788	.	denotes	.
T10907	18788-18850	sentence	denotes	The reason for the prolonged survival of this pup is unknown.
T10908	18789-18792	DT	denotes	The
T10909	18793-18799	NN	denotes	reason
T10911	18800-18803	IN	denotes	for
T10912	18804-18807	DT	denotes	the
T10914	18808-18817	VBN	denotes	prolonged
T10913	18818-18826	NN	denotes	survival
T10915	18827-18829	IN	denotes	of
T10916	18830-18834	DT	denotes	this
T10917	18835-18838	NN	denotes	pup
T10910	18839-18841	VBZ	denotes	is
T10918	18842-18849	JJ	denotes	unknown
T10919	18849-18850	.	denotes	.
T12206	18852-18854	NN	denotes	E2
T12207	18855-18864	JJ	denotes	deficient
T12208	18865-18869	NNS	denotes	mice
T12210	18870-18880	RB	denotes	accurately
T12209	18881-18886	VBP	denotes	model
T12211	18887-18894	JJ	denotes	classic
T12212	18895-18899	NN	denotes	MSUD
T12213	18899-19093	sentence	denotes	To demonstrate that the gene targeting event created a true E2 null allele, we determined BCKDH activity in liver homogenates of postnatal day 1 mouse pups derived from +/- by +/- mating pairs.
T12214	18900-18902	TO	denotes	To
T12215	18903-18914	VB	denotes	demonstrate
T12217	18915-18919	IN	denotes	that
T12219	18920-18923	DT	denotes	the
T12221	18924-18928	NN	denotes	gene
T12222	18929-18938	NN	denotes	targeting
T12220	18939-18944	NN	denotes	event
T12218	18945-18952	VBD	denotes	created
T12223	18953-18954	DT	denotes	a
T12225	18955-18959	JJ	denotes	true
T12226	18960-18962	NN	denotes	E2
T12227	18963-18967	JJ	denotes	null
T12224	18968-18974	NN	denotes	allele
T12228	18974-18976	,	denotes	,
T12229	18976-18978	PRP	denotes	we
T12216	18979-18989	VBD	denotes	determined
T12230	18990-18995	NN	denotes	BCKDH
T12231	18996-19004	NN	denotes	activity
T12232	19005-19007	IN	denotes	in
T12233	19008-19013	NN	denotes	liver
T12234	19014-19025	NNS	denotes	homogenates
T12235	19026-19028	IN	denotes	of
T12236	19029-19038	JJ	denotes	postnatal
T12237	19039-19042	NN	denotes	day
T12239	19043-19044	CD	denotes	1
T12240	19045-19050	NN	denotes	mouse
T12238	19051-19055	NNS	denotes	pups
T12241	19056-19063	VBN	denotes	derived
T12242	19064-19068	IN	denotes	from
T12243	19069-19070	SYM	denotes	+
T12245	19070-19071	HYPH	denotes	/
T12244	19071-19072	SYM	denotes	-
T12247	19073-19075	IN	denotes	by
T12248	19076-19077	SYM	denotes	+
T12250	19077-19078	HYPH	denotes	/
T12249	19078-19079	SYM	denotes	-
T12251	19080-19086	NN	denotes	mating
T12246	19087-19092	NNS	denotes	pairs
T12252	19092-19093	.	denotes	.
T12253	19093-19171	sentence	denotes	As shown in Figure 2A, the BCKDH activity in +/+ mice was readily detectable.
T12254	19094-19096	IN	denotes	As
T12255	19097-19102	VBN	denotes	shown
T12257	19103-19105	IN	denotes	in
T12258	19106-19112	NN	denotes	Figure
T12259	19113-19115	NN	denotes	2A
T12260	19115-19117	,	denotes	,
T12261	19117-19120	DT	denotes	the
T12263	19121-19126	NN	denotes	BCKDH
T12262	19127-19135	NN	denotes	activity
T12264	19136-19138	IN	denotes	in
T12265	19139-19140	SYM	denotes	+
T12267	19140-19141	HYPH	denotes	/
T12266	19141-19142	SYM	denotes	+
T12268	19143-19147	NNS	denotes	mice
T12256	19148-19151	VBD	denotes	was
T12269	19152-19159	RB	denotes	readily
T12270	19160-19170	JJ	denotes	detectable
T12271	19170-19171	.	denotes	.
T12272	19171-19249	sentence	denotes	In marked contrast, BCKDH activity was completely absent in -/- mouse livers.
T12273	19172-19174	IN	denotes	In
T12275	19175-19181	JJ	denotes	marked
T12276	19182-19190	NN	denotes	contrast
T12277	19190-19192	,	denotes	,
T12278	19192-19197	NN	denotes	BCKDH
T12279	19198-19206	NN	denotes	activity
T12274	19207-19210	VBD	denotes	was
T12280	19211-19221	RB	denotes	completely
T12281	19222-19228	JJ	denotes	absent
T12282	19229-19231	IN	denotes	in
T12283	19232-19233	SYM	denotes	-
T12285	19233-19234	HYPH	denotes	/
T12284	19234-19235	SYM	denotes	-
T12287	19236-19241	NN	denotes	mouse
T12286	19242-19248	NNS	denotes	livers
T12288	19248-19249	.	denotes	.
T12289	19249-19362	sentence	denotes	As expected, homogenates from +/- mice had approximately half the activity of their +/+ littermates (Figure 2A).
T12290	19250-19252	IN	denotes	As
T12291	19253-19261	VBN	denotes	expected
T12293	19261-19263	,	denotes	,
T12294	19263-19274	NNS	denotes	homogenates
T12295	19275-19279	IN	denotes	from
T12296	19280-19281	SYM	denotes	+
T12298	19281-19282	HYPH	denotes	/
T12297	19282-19283	SYM	denotes	-
T12299	19284-19288	NNS	denotes	mice
T12292	19289-19292	VBD	denotes	had
T12300	19293-19306	RB	denotes	approximately
T12301	19307-19311	PDT	denotes	half
T12303	19312-19315	DT	denotes	the
T12302	19316-19324	NN	denotes	activity
T12304	19325-19327	IN	denotes	of
T12305	19328-19333	PRP$	denotes	their
T12307	19334-19335	SYM	denotes	+
T12309	19335-19336	HYPH	denotes	/
T12308	19336-19337	SYM	denotes	+
T12306	19338-19349	NNS	denotes	littermates
T12310	19350-19351	-LRB-	denotes	(
T12312	19351-19357	NN	denotes	Figure
T12311	19358-19360	NN	denotes	2A
T12313	19360-19361	-RRB-	denotes	)
T12314	19361-19362	.	denotes	.
T12315	19362-19450	sentence	denotes	The results from -/- mice are similar to that observed in humans with classic MSUD [1].
T12316	19363-19366	DT	denotes	The
T12317	19367-19374	NNS	denotes	results
T12319	19375-19379	IN	denotes	from
T12320	19380-19381	SYM	denotes	-
T12322	19381-19382	HYPH	denotes	/
T12321	19382-19383	SYM	denotes	-
T12323	19384-19388	NNS	denotes	mice
T12318	19389-19392	VBP	denotes	are
T12324	19393-19400	JJ	denotes	similar
T12325	19401-19403	IN	denotes	to
T12326	19404-19408	DT	denotes	that
T12327	19409-19417	VBN	denotes	observed
T12328	19418-19420	IN	denotes	in
T12329	19421-19427	NNS	denotes	humans
T12330	19428-19432	IN	denotes	with
T12331	19433-19440	JJ	denotes	classic
T12332	19441-19445	NN	denotes	MSUD
T12333	19446-19447	-LRB-	denotes	[
T12334	19447-19448	CD	denotes	1
T12335	19448-19449	-RRB-	denotes	]
T12336	19449-19450	.	denotes	.
T12337	19450-19451	sentence	denotes
T22807	19461-19472	JJ	denotes	Biochemical
T22808	19473-19489	NN	denotes	characterization
T22809	19490-19492	IN	denotes	of
T22810	19493-19496	DT	denotes	the
T22812	19497-19504	JJ	denotes	classic
T22813	19505-19509	NN	denotes	MSUD
T22814	19510-19516	JJ	denotes	murine
T22811	19517-19522	NN	denotes	model
T22815	19522-19523	.	denotes	.
T22816	19523-19649	sentence	denotes	A, BCKDH enzyme activity in liver of newborn wild type control (+/+), heterozygous (+/-), and homozygous (-/-) knockout mice.
T22817	19524-19525	LS	denotes	A
T22819	19525-19527	,	denotes	,
T22820	19527-19532	NN	denotes	BCKDH
T22821	19533-19539	NN	denotes	enzyme
T22818	19540-19548	NN	denotes	activity
T22822	19549-19551	IN	denotes	in
T22823	19552-19557	NN	denotes	liver
T22824	19558-19560	IN	denotes	of
T22825	19561-19568	JJ	denotes	newborn
T22827	19569-19573	JJ	denotes	wild
T22828	19574-19578	NN	denotes	type
T22829	19579-19586	NN	denotes	control
T22831	19587-19588	-LRB-	denotes	(
T22833	19588-19589	SYM	denotes	+
T22834	19589-19590	HYPH	denotes	/
T22832	19590-19591	SYM	denotes	+
T22835	19591-19592	-RRB-	denotes	)
T22836	19592-19594	,	denotes	,
T22837	19594-19606	JJ	denotes	heterozygous
T22838	19607-19608	-LRB-	denotes	(
T22840	19608-19609	SYM	denotes	+
T22841	19609-19610	HYPH	denotes	/
T22839	19610-19611	SYM	denotes	-
T22842	19611-19612	-RRB-	denotes	)
T22843	19612-19614	,	denotes	,
T22844	19614-19617	CC	denotes	and
T22845	19618-19628	JJ	denotes	homozygous
T22846	19629-19630	-LRB-	denotes	(
T22848	19630-19631	SYM	denotes	-
T22849	19631-19632	HYPH	denotes	/
T22847	19632-19633	SYM	denotes	-
T22850	19633-19634	-RRB-	denotes	)
T22830	19635-19643	NN	denotes	knockout
T22826	19644-19648	NNS	denotes	mice
T22851	19648-19649	.	denotes	.
T22852	19649-19772	sentence	denotes	Enzyme activity was significantly reduced in +/- liver compared to +/+, and was below the level of detection in -/- liver.
T22853	19650-19656	NN	denotes	Enzyme
T22854	19657-19665	NN	denotes	activity
T22855	19666-19669	VBD	denotes	was
T22856	19670-19683	RB	denotes	significantly
T22857	19684-19691	JJ	denotes	reduced
T22858	19692-19694	IN	denotes	in
T22859	19695-19696	SYM	denotes	+
T22861	19696-19697	HYPH	denotes	/
T22860	19697-19698	SYM	denotes	-
T22862	19699-19704	NN	denotes	liver
T22863	19705-19713	VBN	denotes	compared
T22864	19714-19716	IN	denotes	to
T22865	19717-19718	SYM	denotes	+
T22867	19718-19719	HYPH	denotes	/
T22866	19719-19720	SYM	denotes	+
T22868	19720-19722	,	denotes	,
T22869	19722-19725	CC	denotes	and
T22870	19726-19729	VBD	denotes	was
T22871	19730-19735	IN	denotes	below
T22872	19736-19739	DT	denotes	the
T22873	19740-19745	NN	denotes	level
T22874	19746-19748	IN	denotes	of
T22875	19749-19758	NN	denotes	detection
T22876	19759-19761	IN	denotes	in
T22877	19762-19763	SYM	denotes	-
T22879	19763-19764	HYPH	denotes	/
T22878	19764-19765	SYM	denotes	-
T22880	19766-19771	NN	denotes	liver
T22881	19771-19772	.	denotes	.
T22882	19772-19819	sentence	denotes	B, Total BCAA concentrations in blood of mice.
T22883	19773-19774	LS	denotes	B
T22885	19774-19776	,	denotes	,
T22886	19776-19781	JJ	denotes	Total
T22887	19782-19786	NN	denotes	BCAA
T22884	19787-19801	NNS	denotes	concentrations
T22888	19802-19804	IN	denotes	in
T22889	19805-19810	NN	denotes	blood
T22890	19811-19813	IN	denotes	of
T22891	19814-19818	NNS	denotes	mice
T22892	19818-19819	.	denotes	.
T22893	19819-19884	sentence	denotes	Total BCAA represent the sum of leucine, isoleucine, and valine.
T22894	19820-19825	JJ	denotes	Total
T22895	19826-19830	NN	denotes	BCAA
T22896	19831-19840	VBP	denotes	represent
T22897	19841-19844	DT	denotes	the
T22898	19845-19848	NN	denotes	sum
T22899	19849-19851	IN	denotes	of
T22900	19852-19859	NN	denotes	leucine
T22901	19859-19861	,	denotes	,
T22902	19861-19871	NN	denotes	isoleucine
T22903	19871-19873	,	denotes	,
T22904	19873-19876	CC	denotes	and
T22905	19877-19883	NN	denotes	valine
T22906	19883-19884	.	denotes	.
T22907	19884-19986	sentence	denotes	Total BCAA concentrations in blood from -/- mice were significantly elevated compared to +/+ and +/-.
T22908	19885-19890	JJ	denotes	Total
T22910	19891-19895	NN	denotes	BCAA
T22909	19896-19910	NNS	denotes	concentrations
T22912	19911-19913	IN	denotes	in
T22913	19914-19919	NN	denotes	blood
T22914	19920-19924	IN	denotes	from
T22915	19925-19926	SYM	denotes	-
T22917	19926-19927	HYPH	denotes	/
T22916	19927-19928	SYM	denotes	-
T22918	19929-19933	NNS	denotes	mice
T22911	19934-19938	VBD	denotes	were
T22919	19939-19952	RB	denotes	significantly
T22920	19953-19961	JJ	denotes	elevated
T22921	19962-19970	VBN	denotes	compared
T22922	19971-19973	IN	denotes	to
T22923	19974-19975	SYM	denotes	+
T22925	19975-19976	HYPH	denotes	/
T22924	19976-19977	SYM	denotes	+
T22926	19978-19981	CC	denotes	and
T22927	19982-19983	SYM	denotes	+
T22929	19983-19984	HYPH	denotes	/
T22928	19984-19985	SYM	denotes	-
T22930	19985-19986	.	denotes	.
T22931	19986-20038	sentence	denotes	C, Ratio of total BCAA to alanine in blood of mice.
T22932	19987-19988	LS	denotes	C
T22934	19988-19990	,	denotes	,
T22933	19990-19995	NN	denotes	Ratio
T22935	19996-19998	IN	denotes	of
T22936	19999-20004	JJ	denotes	total
T22937	20005-20009	NN	denotes	BCAA
T22938	20010-20012	IN	denotes	to
T22939	20013-20020	NN	denotes	alanine
T22940	20021-20023	IN	denotes	in
T22941	20024-20029	NN	denotes	blood
T22942	20030-20032	IN	denotes	of
T22943	20033-20037	NNS	denotes	mice
T22944	20037-20038	.	denotes	.
T22945	20038-20118	sentence	denotes	This ratio was significantly elevated in -/- mice compared to +/+ and +/- mice.
T22946	20039-20043	DT	denotes	This
T22947	20044-20049	NN	denotes	ratio
T22948	20050-20053	VBD	denotes	was
T22949	20054-20067	RB	denotes	significantly
T22950	20068-20076	JJ	denotes	elevated
T22951	20077-20079	IN	denotes	in
T22952	20080-20081	SYM	denotes	-
T22954	20081-20082	HYPH	denotes	/
T22953	20082-20083	SYM	denotes	-
T22955	20084-20088	NNS	denotes	mice
T22956	20089-20097	VBN	denotes	compared
T22957	20098-20100	IN	denotes	to
T22958	20101-20102	SYM	denotes	+
T22960	20102-20103	HYPH	denotes	/
T22959	20103-20104	SYM	denotes	+
T22962	20105-20108	CC	denotes	and
T22963	20109-20110	SYM	denotes	+
T22965	20110-20111	HYPH	denotes	/
T22964	20111-20112	SYM	denotes	-
T22961	20113-20117	NNS	denotes	mice
T22966	20117-20118	.	denotes	.
T22967	20118-20181	sentence	denotes	The numbers on the bars indicates the number of mice analyzed.
T22968	20119-20122	DT	denotes	The
T22969	20123-20130	NNS	denotes	numbers
T22971	20131-20133	IN	denotes	on
T22972	20134-20137	DT	denotes	the
T22973	20138-20142	NNS	denotes	bars
T22970	20143-20152	VBZ	denotes	indicates
T22974	20153-20156	DT	denotes	the
T22975	20157-20163	NN	denotes	number
T22976	20164-20166	IN	denotes	of
T22977	20167-20171	NNS	denotes	mice
T22978	20172-20180	VBN	denotes	analyzed
T22979	20180-20181	.	denotes	.
T22980	20181-20288	sentence	denotes	, Significantly different from +/+ (P < 0.001); *, significantly different from +/+ and +/- (P < 0.001).
T22981	20182-20183	SYM	denotes	*
T22983	20183-20185	,	denotes	,
T22984	20185-20198	RB	denotes	Significantly
T22985	20199-20208	JJ	denotes	different
T22986	20209-20213	IN	denotes	from
T22987	20214-20215	SYM	denotes	+
T22989	20215-20216	HYPH	denotes	/
T22988	20216-20217	SYM	denotes	+
T22990	20218-20219	-LRB-	denotes	(
T22992	20219-20220	NN	denotes	P
T22993	20221-20222	SYM	denotes	<
T22991	20223-20228	CD	denotes	0.001
T22994	20228-20229	-RRB-	denotes	)
T22995	20229-20230	:	denotes	;
T22982	20231-20233	SYM	denotes	**
T22996	20233-20235	,	denotes	,
T22997	20235-20248	RB	denotes	significantly
T22998	20249-20258	JJ	denotes	different
T22999	20259-20263	IN	denotes	from
T23000	20264-20265	SYM	denotes	+
T23002	20265-20266	HYPH	denotes	/
T23001	20266-20267	SYM	denotes	+
T23003	20268-20271	CC	denotes	and
T23004	20272-20273	SYM	denotes	+
T23006	20273-20274	HYPH	denotes	/
T23005	20274-20275	SYM	denotes	-
T23007	20276-20277	-LRB-	denotes	(
T23009	20277-20278	NN	denotes	P
T23010	20279-20280	SYM	denotes	<
T23008	20281-20286	CD	denotes	0.001
T23011	20286-20287	-RRB-	denotes	)
T23012	20287-20288	.	denotes	.
T12339	20289-20309	NN	denotes	Immunohistochemistry
T12338	20289-20382	sentence	denotes	Immunohistochemistry with an E2 specific antibody was used to examine E2 protein in the mice.
T12341	20310-20314	IN	denotes	with
T12342	20315-20317	DT	denotes	an
T12344	20318-20320	NN	denotes	E2
T12345	20321-20329	JJ	denotes	specific
T12343	20330-20338	NN	denotes	antibody
T12346	20339-20342	VBD	denotes	was
T12340	20343-20347	VBN	denotes	used
T12347	20348-20350	TO	denotes	to
T12348	20351-20358	VB	denotes	examine
T12349	20359-20361	NN	denotes	E2
T12350	20362-20369	NN	denotes	protein
T12351	20370-20372	IN	denotes	in
T12352	20373-20376	DT	denotes	the
T12353	20377-20381	NNS	denotes	mice
T12354	20381-20382	.	denotes	.
T12355	20382-20498	sentence	denotes	As shown in Figure 1C and 1D, immunoreactive E2 protein was abundant in liver and embryonic fibroblasts of+/+ mice.
T12356	20383-20385	IN	denotes	As
T12357	20386-20391	VBN	denotes	shown
T12359	20392-20394	IN	denotes	in
T12360	20395-20401	NN	denotes	Figure
T12361	20402-20404	NN	denotes	1C
T12362	20405-20408	CC	denotes	and
T12363	20409-20411	NN	denotes	1D
T12364	20411-20413	,	denotes	,
T12365	20413-20427	JJ	denotes	immunoreactive
T12367	20428-20430	NN	denotes	E2
T12366	20431-20438	NN	denotes	protein
T12358	20439-20442	VBD	denotes	was
T12368	20443-20451	JJ	denotes	abundant
T12369	20452-20454	IN	denotes	in
T12370	20455-20460	NN	denotes	liver
T12371	20461-20464	CC	denotes	and
T12372	20465-20474	JJ	denotes	embryonic
T12373	20475-20486	NNS	denotes	fibroblasts
T12374	20487-20489	IN	denotes	of
T12375	20489-20490	SYM	denotes	+
T12377	20490-20491	HYPH	denotes	/
T12376	20491-20492	SYM	denotes	+
T12378	20493-20497	NNS	denotes	mice
T12379	20497-20498	.	denotes	.
T12380	20498-20590	sentence	denotes	In marked contrast, immunoreactive E2 protein was absent in these same tissues of -/- mice.
T12381	20499-20501	IN	denotes	In
T12383	20502-20508	JJ	denotes	marked
T12384	20509-20517	NN	denotes	contrast
T12385	20517-20519	,	denotes	,
T12386	20519-20533	JJ	denotes	immunoreactive
T12388	20534-20536	NN	denotes	E2
T12387	20537-20544	NN	denotes	protein
T12382	20545-20548	VBD	denotes	was
T12389	20549-20555	JJ	denotes	absent
T12390	20556-20558	IN	denotes	in
T12391	20559-20564	DT	denotes	these
T12393	20565-20569	JJ	denotes	same
T12392	20570-20577	NNS	denotes	tissues
T12394	20578-20580	IN	denotes	of
T12395	20581-20582	SYM	denotes	-
T12397	20582-20583	HYPH	denotes	/
T12396	20583-20584	SYM	denotes	-
T12398	20585-20589	NNS	denotes	mice
T12399	20589-20590	.	denotes	.
T12400	20590-20789	sentence	denotes	Homozygous E2 knockout mice had a nearly 3-fold increase in blood (Figure 2B) and urine (data not shown) levels of BCAA (sum of leucine, isoleucine, and valine) as compared to their +/+ littermates.
T12401	20591-20601	JJ	denotes	Homozygous
T12403	20602-20604	NN	denotes	E2
T12402	20605-20613	NN	denotes	knockout
T12404	20614-20618	NNS	denotes	mice
T12405	20619-20622	VBD	denotes	had
T12406	20623-20624	DT	denotes	a
T12408	20625-20631	RB	denotes	nearly
T12409	20632-20633	CD	denotes	3
T12411	20633-20634	HYPH	denotes	-
T12410	20634-20638	JJ	denotes	fold
T12407	20639-20647	NN	denotes	increase
T12412	20648-20650	IN	denotes	in
T12413	20651-20656	NN	denotes	blood
T12415	20657-20658	-LRB-	denotes	(
T12417	20658-20664	NN	denotes	Figure
T12416	20665-20667	NN	denotes	2B
T12418	20667-20668	-RRB-	denotes	)
T12419	20669-20672	CC	denotes	and
T12420	20673-20678	NN	denotes	urine
T12421	20679-20680	-LRB-	denotes	(
T12423	20680-20684	NNS	denotes	data
T12424	20685-20688	RB	denotes	not
T12422	20689-20694	VBN	denotes	shown
T12425	20694-20695	-RRB-	denotes	)
T12414	20696-20702	NNS	denotes	levels
T12426	20703-20705	IN	denotes	of
T12427	20706-20710	NN	denotes	BCAA
T12428	20711-20712	-LRB-	denotes	(
T12429	20712-20715	NN	denotes	sum
T12430	20716-20718	IN	denotes	of
T12431	20719-20726	NN	denotes	leucine
T12432	20726-20728	,	denotes	,
T12433	20728-20738	NN	denotes	isoleucine
T12434	20738-20740	,	denotes	,
T12435	20740-20743	CC	denotes	and
T12436	20744-20750	NN	denotes	valine
T12437	20750-20751	-RRB-	denotes	)
T12438	20752-20754	IN	denotes	as
T12439	20755-20763	VBN	denotes	compared
T12440	20764-20766	IN	denotes	to
T12441	20767-20772	PRP$	denotes	their
T12443	20773-20774	SYM	denotes	+
T12445	20774-20775	HYPH	denotes	/
T12444	20775-20776	SYM	denotes	+
T12442	20777-20788	NNS	denotes	littermates
T12446	20788-20789	.	denotes	.
T12447	20789-20965	sentence	denotes	Because amino acids were analyzed by tandem mass spectrometry, the sum of BCAA shown in Figure 2B also may include alloisoleucine that may have been produced in -/- MSUD mice.
T12448	20790-20797	IN	denotes	Because
T12450	20798-20803	NN	denotes	amino
T12451	20804-20809	NNS	denotes	acids
T12452	20810-20814	VBD	denotes	were
T12449	20815-20823	VBN	denotes	analyzed
T12454	20824-20826	IN	denotes	by
T12455	20827-20833	NN	denotes	tandem
T12457	20834-20838	NN	denotes	mass
T12456	20839-20851	NN	denotes	spectrometry
T12458	20851-20853	,	denotes	,
T12459	20853-20856	DT	denotes	the
T12460	20857-20860	NN	denotes	sum
T12461	20861-20863	IN	denotes	of
T12462	20864-20868	NN	denotes	BCAA
T12463	20869-20874	VBN	denotes	shown
T12464	20875-20877	IN	denotes	in
T12465	20878-20884	NN	denotes	Figure
T12466	20885-20887	NN	denotes	2B
T12467	20888-20892	RB	denotes	also
T12468	20893-20896	MD	denotes	may
T12453	20897-20904	VB	denotes	include
T12469	20905-20919	NN	denotes	alloisoleucine
T12470	20920-20924	WDT	denotes	that
T12472	20925-20928	MD	denotes	may
T12473	20929-20933	VB	denotes	have
T12474	20934-20938	VBN	denotes	been
T12471	20939-20947	VBN	denotes	produced
T12475	20948-20950	IN	denotes	in
T12476	20951-20952	SYM	denotes	-
T12478	20952-20953	HYPH	denotes	/
T12477	20953-20954	SYM	denotes	-
T12480	20955-20959	NN	denotes	MSUD
T12479	20960-20964	NNS	denotes	mice
T12481	20964-20965	.	denotes	.
T12482	20965-21060	sentence	denotes	The metabolism of BCAA is linked with the synthesis of alanine, glutamate, and glutamine [33].
T12483	20966-20969	DT	denotes	The
T12484	20970-20980	NN	denotes	metabolism
T12486	20981-20983	IN	denotes	of
T12487	20984-20988	NN	denotes	BCAA
T12488	20989-20991	VBZ	denotes	is
T12485	20992-20998	VBN	denotes	linked
T12489	20999-21003	IN	denotes	with
T12490	21004-21007	DT	denotes	the
T12491	21008-21017	NN	denotes	synthesis
T12492	21018-21020	IN	denotes	of
T12493	21021-21028	NN	denotes	alanine
T12494	21028-21030	,	denotes	,
T12495	21030-21039	NN	denotes	glutamate
T12496	21039-21041	,	denotes	,
T12497	21041-21044	CC	denotes	and
T12498	21045-21054	NN	denotes	glutamine
T12499	21055-21056	-LRB-	denotes	[
T12500	21056-21058	CD	denotes	33
T12501	21058-21059	-RRB-	denotes	]
T12502	21059-21060	.	denotes	.
T12503	21060-21253	sentence	denotes	Because of impaired metabolism of BCAA in MSUD mice, and to further characterize the abnormal biochemistry in this model, we analyzed the blood levels of the alanine, glutamate, and glutamine.
T12504	21061-21068	IN	denotes	Because
T12506	21069-21071	IN	denotes	of
T12507	21072-21080	JJ	denotes	impaired
T12508	21081-21091	NN	denotes	metabolism
T12509	21092-21094	IN	denotes	of
T12510	21095-21099	NN	denotes	BCAA
T12511	21100-21102	IN	denotes	in
T12512	21103-21107	NN	denotes	MSUD
T12513	21108-21112	NNS	denotes	mice
T12514	21112-21114	,	denotes	,
T12515	21114-21117	CC	denotes	and
T12516	21118-21120	TO	denotes	to
T12518	21121-21128	RB	denotes	further
T12517	21129-21141	VB	denotes	characterize
T12519	21142-21145	DT	denotes	the
T12521	21146-21154	JJ	denotes	abnormal
T12520	21155-21167	NN	denotes	biochemistry
T12522	21168-21170	IN	denotes	in
T12523	21171-21175	DT	denotes	this
T12524	21176-21181	NN	denotes	model
T12525	21181-21183	,	denotes	,
T12526	21183-21185	PRP	denotes	we
T12505	21186-21194	VBD	denotes	analyzed
T12527	21195-21198	DT	denotes	the
T12529	21199-21204	NN	denotes	blood
T12528	21205-21211	NNS	denotes	levels
T12530	21212-21214	IN	denotes	of
T12531	21215-21218	DT	denotes	the
T12532	21219-21226	NN	denotes	alanine
T12533	21226-21228	,	denotes	,
T12534	21228-21237	NN	denotes	glutamate
T12535	21237-21239	,	denotes	,
T12536	21239-21242	CC	denotes	and
T12537	21243-21252	NN	denotes	glutamine
T12538	21252-21253	.	denotes	.
T12539	21253-21385	sentence	denotes	As shown in Table 1, the levels of all three amino acids in homozygous mice were markedly lower than the levels in +/+ or +/- mice.
T12540	21254-21256	IN	denotes	As
T12541	21257-21262	VBN	denotes	shown
T12543	21263-21265	IN	denotes	in
T12544	21266-21271	NN	denotes	Table
T12545	21272-21273	CD	denotes	1
T12546	21273-21275	,	denotes	,
T12547	21275-21278	DT	denotes	the
T12548	21279-21285	NNS	denotes	levels
T12549	21286-21288	IN	denotes	of
T12550	21289-21292	DT	denotes	all
T12552	21293-21298	CD	denotes	three
T12553	21299-21304	NN	denotes	amino
T12551	21305-21310	NNS	denotes	acids
T12554	21311-21313	IN	denotes	in
T12555	21314-21324	JJ	denotes	homozygous
T12556	21325-21329	NNS	denotes	mice
T12542	21330-21334	VBD	denotes	were
T12557	21335-21343	RB	denotes	markedly
T12558	21344-21349	JJR	denotes	lower
T12559	21350-21354	IN	denotes	than
T12560	21355-21358	DT	denotes	the
T12561	21359-21365	NNS	denotes	levels
T12562	21366-21368	IN	denotes	in
T12563	21369-21370	SYM	denotes	+
T12565	21370-21371	HYPH	denotes	/
T12564	21371-21372	SYM	denotes	+
T12567	21373-21375	CC	denotes	or
T12568	21376-21377	SYM	denotes	+
T12570	21377-21378	HYPH	denotes	/
T12569	21378-21379	SYM	denotes	-
T12566	21380-21384	NNS	denotes	mice
T12571	21384-21385	.	denotes	.
T12572	21385-21481	sentence	denotes	The levels of these amino acids in the +/- mice were comparable to those in +/+ mice (Table 1).
T12573	21386-21389	DT	denotes	The
T12574	21390-21396	NNS	denotes	levels
T12576	21397-21399	IN	denotes	of
T12577	21400-21405	DT	denotes	these
T12579	21406-21411	NN	denotes	amino
T12578	21412-21417	NNS	denotes	acids
T12580	21418-21420	IN	denotes	in
T12581	21421-21424	DT	denotes	the
T12583	21425-21426	SYM	denotes	+
T12585	21426-21427	HYPH	denotes	/
T12584	21427-21428	SYM	denotes	-
T12582	21429-21433	NNS	denotes	mice
T12575	21434-21438	VBD	denotes	were
T12586	21439-21449	JJ	denotes	comparable
T12587	21450-21452	IN	denotes	to
T12588	21453-21458	DT	denotes	those
T12589	21459-21461	IN	denotes	in
T12590	21462-21463	SYM	denotes	+
T12592	21463-21464	HYPH	denotes	/
T12591	21464-21465	SYM	denotes	+
T12593	21466-21470	NNS	denotes	mice
T12594	21471-21472	-LRB-	denotes	(
T12595	21472-21477	NN	denotes	Table
T12596	21478-21479	CD	denotes	1
T12597	21479-21480	-RRB-	denotes	)
T12598	21480-21481	.	denotes	.
T12599	21481-21787	sentence	denotes	Because of the abnormal decrease in the blood alanine level and marked rise in blood BCAA levels that are characteristic of MSUD, a recent report on MSUD patients has suggested that the ratio of BCAA/alanine provides a more sensitive measure of the abnormal biochemistry of MSUD than BCAA level alone [8].
T12600	21482-21489	IN	denotes	Because
T12602	21490-21492	IN	denotes	of
T12603	21493-21496	DT	denotes	the
T12605	21497-21505	JJ	denotes	abnormal
T12604	21506-21514	NN	denotes	decrease
T12606	21515-21517	IN	denotes	in
T12607	21518-21521	DT	denotes	the
T12609	21522-21527	NN	denotes	blood
T12610	21528-21535	NN	denotes	alanine
T12608	21536-21541	NN	denotes	level
T12611	21542-21545	CC	denotes	and
T12612	21546-21552	JJ	denotes	marked
T12613	21553-21557	NN	denotes	rise
T12614	21558-21560	IN	denotes	in
T12615	21561-21566	NN	denotes	blood
T12617	21567-21571	NN	denotes	BCAA
T12616	21572-21578	NNS	denotes	levels
T12618	21579-21583	WDT	denotes	that
T12619	21584-21587	VBP	denotes	are
T12620	21588-21602	JJ	denotes	characteristic
T12621	21603-21605	IN	denotes	of
T12622	21606-21610	NN	denotes	MSUD
T12623	21610-21612	,	denotes	,
T12624	21612-21613	DT	denotes	a
T12626	21614-21620	JJ	denotes	recent
T12625	21621-21627	NN	denotes	report
T12627	21628-21630	IN	denotes	on
T12628	21631-21635	NN	denotes	MSUD
T12629	21636-21644	NNS	denotes	patients
T12630	21645-21648	VBZ	denotes	has
T12601	21649-21658	VBN	denotes	suggested
T12631	21659-21663	IN	denotes	that
T12633	21664-21667	DT	denotes	the
T12634	21668-21673	NN	denotes	ratio
T12635	21674-21676	IN	denotes	of
T12636	21677-21681	NN	denotes	BCAA
T12638	21681-21682	HYPH	denotes	/
T12637	21682-21689	NN	denotes	alanine
T12632	21690-21698	VBZ	denotes	provides
T12639	21699-21700	DT	denotes	a
T12641	21701-21705	RBR	denotes	more
T12642	21706-21715	JJ	denotes	sensitive
T12640	21716-21723	NN	denotes	measure
T12643	21724-21726	IN	denotes	of
T12644	21727-21730	DT	denotes	the
T12646	21731-21739	JJ	denotes	abnormal
T12645	21740-21752	NN	denotes	biochemistry
T12647	21753-21755	IN	denotes	of
T12648	21756-21760	NN	denotes	MSUD
T12649	21761-21765	IN	denotes	than
T12650	21766-21770	NN	denotes	BCAA
T12651	21771-21776	NN	denotes	level
T12652	21777-21782	RB	denotes	alone
T12653	21783-21784	-LRB-	denotes	[
T12654	21784-21785	CD	denotes	8
T12655	21785-21786	-RRB-	denotes	]
T12656	21786-21787	.	denotes	.
T12657	21787-21862	sentence	denotes	Therefore, we also expressed the amino acid results as BCAA/alanine ratio.
T12658	21788-21797	RB	denotes	Therefore
T12660	21797-21799	,	denotes	,
T12661	21799-21801	PRP	denotes	we
T12662	21802-21806	RB	denotes	also
T12659	21807-21816	VBD	denotes	expressed
T12663	21817-21820	DT	denotes	the
T12665	21821-21826	NN	denotes	amino
T12666	21827-21831	NN	denotes	acid
T12664	21832-21839	NNS	denotes	results
T12667	21840-21842	IN	denotes	as
T12668	21843-21847	NN	denotes	BCAA
T12670	21847-21848	HYPH	denotes	/
T12669	21848-21855	NN	denotes	alanine
T12671	21856-21861	NN	denotes	ratio
T12672	21861-21862	.	denotes	.
T12673	21862-21965	sentence	denotes	As shown in Figure 2C, this ratio in -/- pups was more than 6-fold higher than +/+ or +/- littermates.
T12674	21863-21865	IN	denotes	As
T12675	21866-21871	VBN	denotes	shown
T12677	21872-21874	IN	denotes	in
T12678	21875-21881	NN	denotes	Figure
T12679	21882-21884	NN	denotes	2C
T12680	21884-21886	,	denotes	,
T12681	21886-21890	DT	denotes	this
T12682	21891-21896	NN	denotes	ratio
T12683	21897-21899	IN	denotes	in
T12684	21900-21901	SYM	denotes	-
T12686	21901-21902	HYPH	denotes	/
T12685	21902-21903	SYM	denotes	-
T12687	21904-21908	NNS	denotes	pups
T12676	21909-21912	VBD	denotes	was
T12688	21913-21917	RBR	denotes	more
T12690	21918-21922	IN	denotes	than
T12689	21923-21924	CD	denotes	6
T12692	21924-21925	HYPH	denotes	-
T12691	21925-21929	RB	denotes	fold
T12693	21930-21936	JJR	denotes	higher
T12694	21937-21941	IN	denotes	than
T12695	21942-21943	SYM	denotes	+
T12697	21943-21944	HYPH	denotes	/
T12696	21944-21945	SYM	denotes	+
T12699	21946-21948	CC	denotes	or
T12700	21949-21950	SYM	denotes	+
T12702	21950-21951	HYPH	denotes	/
T12701	21951-21952	SYM	denotes	-
T12698	21953-21964	NNS	denotes	littermates
T12703	21964-21965	.	denotes	.
T12704	21965-22072	sentence	denotes	These blood amino acid results are consistent with the concentrations seen in patients with MSUD [1,8,15].
T12705	21966-21971	DT	denotes	These
T12707	21972-21977	NN	denotes	blood
T12708	21978-21983	NN	denotes	amino
T12709	21984-21988	NN	denotes	acid
T12706	21989-21996	NNS	denotes	results
T12710	21997-22000	VBP	denotes	are
T12711	22001-22011	JJ	denotes	consistent
T12712	22012-22016	IN	denotes	with
T12713	22017-22020	DT	denotes	the
T12714	22021-22035	NNS	denotes	concentrations
T12715	22036-22040	VBN	denotes	seen
T12716	22041-22043	IN	denotes	in
T12717	22044-22052	NNS	denotes	patients
T12718	22053-22057	IN	denotes	with
T12719	22058-22062	NN	denotes	MSUD
T12720	22063-22064	-LRB-	denotes	[
T12722	22064-22065	CD	denotes	1
T12723	22065-22066	,	denotes	,
T12724	22066-22067	CD	denotes	8
T12725	22067-22068	,	denotes	,
T12721	22068-22070	CD	denotes	15
T12726	22070-22071	-RRB-	denotes	]
T12727	22071-22072	.	denotes	.
T12728	22072-22727	sentence	denotes	Table 1 Summary of additional blood amino acid levels in the classic MSUD model and control littermates as determined by tandem mass spectrometry. All samples were collected on the day of birth. All values are mean +/- SEM. MSUD Genotype Alanine Glutamate Glutamine (μmole/L) +/+ 196.1 ± 23.1 (6) 229.0 ± 12.0 (6) 76.8 ± 3.0 (6) +/- 167.6 ± 9.9 (13) 251.0 ± 12.8 (13) 72.3 ± 1.6 (13) -/- 94.6 ± 11.6* (5) 107.8 ± 4.8* (5) 50.4 ± 2.9* (5) *: Significantly different from +/+ and +/- (P < 0.001). In summary, E2 knockout mice lack BCKDH enzymatic activity, E2 immunoreactivity, and have markedly elevated levels of BCAA in the blood and urine.
T24774	22082-22089	NN	denotes	Summary
T24775	22090-22092	IN	denotes	of
T24776	22093-22103	JJ	denotes	additional
T24778	22104-22109	NN	denotes	blood
T24779	22110-22115	NN	denotes	amino
T24780	22116-22120	NN	denotes	acid
T24777	22121-22127	NNS	denotes	levels
T24781	22128-22130	IN	denotes	in
T24782	22131-22134	DT	denotes	the
T24784	22135-22142	JJ	denotes	classic
T24785	22143-22147	NN	denotes	MSUD
T24783	22148-22153	NN	denotes	model
T24786	22154-22157	CC	denotes	and
T24787	22158-22165	NN	denotes	control
T24788	22166-22177	NNS	denotes	littermates
T24789	22178-22180	IN	denotes	as
T24790	22181-22191	VBN	denotes	determined
T24791	22192-22194	IN	denotes	by
T24792	22195-22201	JJ	denotes	tandem
T24794	22202-22206	NN	denotes	mass
T24793	22207-22219	NN	denotes	spectrometry
T24795	22219-22220	.	denotes	.
T24796	22220-22268	sentence	denotes	All samples were collected on the day of birth.
T24797	22221-22224	DT	denotes	All
T24798	22225-22232	NNS	denotes	samples
T24800	22233-22237	VBD	denotes	were
T24799	22238-22247	VBN	denotes	collected
T24801	22248-22250	IN	denotes	on
T24802	22251-22254	DT	denotes	the
T24803	22255-22258	NN	denotes	day
T24804	22259-22261	IN	denotes	of
T24805	22262-22267	NN	denotes	birth
T24806	22267-22268	.	denotes	.
T24807	22268-22297	sentence	denotes	All values are mean +/- SEM.
T24808	22269-22272	DT	denotes	All
T24809	22273-22279	NNS	denotes	values
T24810	22280-22283	VBP	denotes	are
T24811	22284-22288	NN	denotes	mean
T24812	22289-22290	SYM	denotes	+
T24813	22290-22291	HYPH	denotes	/
T24814	22291-22292	SYM	denotes	-
T24815	22293-22296	NN	denotes	SEM
T24816	22296-22297	.	denotes	.
T12729	22581-22583	IN	denotes	In
T12731	22584-22591	NN	denotes	summary
T12732	22591-22593	,	denotes	,
T12733	22593-22595	NN	denotes	E2
T12735	22596-22604	NN	denotes	knockout
T12734	22605-22609	NNS	denotes	mice
T12730	22610-22614	VBP	denotes	lack
T12736	22615-22620	NN	denotes	BCKDH
T12738	22621-22630	JJ	denotes	enzymatic
T12737	22631-22639	NN	denotes	activity
T12739	22639-22641	,	denotes	,
T12740	22641-22643	NN	denotes	E2
T12741	22644-22660	NN	denotes	immunoreactivity
T12742	22660-22662	,	denotes	,
T12743	22662-22665	CC	denotes	and
T12744	22666-22670	VBP	denotes	have
T12745	22671-22679	RB	denotes	markedly
T12746	22680-22688	VBN	denotes	elevated
T12747	22689-22695	NNS	denotes	levels
T12748	22696-22698	IN	denotes	of
T12749	22699-22703	NN	denotes	BCAA
T12750	22704-22706	IN	denotes	in
T12751	22707-22710	DT	denotes	the
T12752	22711-22716	NN	denotes	blood
T12753	22717-22720	CC	denotes	and
T12754	22721-22726	NN	denotes	urine
T12755	22726-22727	.	denotes	.
T12756	22727-22797	sentence	denotes	These metabolic derangements ultimately result in neonatal lethality.
T12757	22728-22733	DT	denotes	These
T12759	22734-22743	JJ	denotes	metabolic
T12758	22744-22756	NNS	denotes	derangements
T12761	22757-22767	RB	denotes	ultimately
T12760	22768-22774	VBP	denotes	result
T12762	22775-22777	IN	denotes	in
T12763	22778-22786	JJ	denotes	neonatal
T12764	22787-22796	NN	denotes	lethality
T12765	22796-22797	.	denotes	.
T12766	22797-22895	sentence	denotes	These phenotypes are remarkably similar to that observed in humans with the classic form of MSUD.
T12767	22798-22803	DT	denotes	These
T12768	22804-22814	NNS	denotes	phenotypes
T12769	22815-22818	VBP	denotes	are
T12770	22819-22829	RB	denotes	remarkably
T12771	22830-22837	JJ	denotes	similar
T12772	22838-22840	IN	denotes	to
T12773	22841-22845	DT	denotes	that
T12774	22846-22854	VBN	denotes	observed
T12775	22855-22857	IN	denotes	in
T12776	22858-22864	NNS	denotes	humans
T12777	22865-22869	IN	denotes	with
T12778	22870-22873	DT	denotes	the
T12780	22874-22881	JJ	denotes	classic
T12779	22882-22886	NN	denotes	form
T12781	22887-22889	IN	denotes	of
T12782	22890-22894	NN	denotes	MSUD
T12783	22894-22895	.	denotes	.
T12784	22895-22946	sentence	denotes	Thus, E2 knockout mice closely model classic MSUD.
T12785	22896-22900	RB	denotes	Thus
T12787	22900-22902	,	denotes	,
T12788	22902-22904	NN	denotes	E2
T12790	22905-22913	NN	denotes	knockout
T12789	22914-22918	NNS	denotes	mice
T12791	22919-22926	RB	denotes	closely
T12786	22927-22932	VBP	denotes	model
T12792	22933-22940	JJ	denotes	classic
T12793	22941-22945	NN	denotes	MSUD
T12794	22945-22946	.	denotes	.
T15726	22948-22956	NN	denotes	Knockout
T15727	22957-22961	NNS	denotes	mice
T15729	22962-22972	VBG	denotes	expressing
T15730	22973-22974	DT	denotes	a
T15732	22975-22980	JJ	denotes	human
T15733	22981-22983	NN	denotes	E2
T15731	22984-22993	NN	denotes	transgene
T15728	22994-22999	VBP	denotes	model
T15734	23000-23001	DT	denotes	a
T15736	23002-23009	JJ	denotes	variant
T15735	23010-23014	NN	denotes	form
T15737	23015-23017	IN	denotes	of
T15738	23018-23022	NN	denotes	MSUD
T15739	23023-23027	WDT	denotes	that
T15740	23028-23034	VBZ	denotes	mimics
T15741	23035-23047	JJ	denotes	intermediate
T15742	23048-23052	NN	denotes	MSUD
T15743	23052-23256	sentence	denotes	To create a murine model of the intermediate variant form of MSUD, we used a transgenic strategy to rescue the severe elevation of BCAA and neonatal lethality that occurs in the classic MSUD mouse model.
T15744	23053-23055	TO	denotes	To
T15745	23056-23062	VB	denotes	create
T15747	23063-23064	DT	denotes	a
T15749	23065-23071	JJ	denotes	murine
T15748	23072-23077	NN	denotes	model
T15750	23078-23080	IN	denotes	of
T15751	23081-23084	DT	denotes	the
T15753	23085-23097	JJ	denotes	intermediate
T15754	23098-23105	NN	denotes	variant
T15752	23106-23110	NN	denotes	form
T15755	23111-23113	IN	denotes	of
T15756	23114-23118	NN	denotes	MSUD
T15757	23118-23120	,	denotes	,
T15758	23120-23122	PRP	denotes	we
T15746	23123-23127	VBD	denotes	used
T15759	23128-23129	DT	denotes	a
T15761	23130-23140	JJ	denotes	transgenic
T15760	23141-23149	NN	denotes	strategy
T15762	23150-23152	TO	denotes	to
T15763	23153-23159	VB	denotes	rescue
T15764	23160-23163	DT	denotes	the
T15766	23164-23170	JJ	denotes	severe
T15765	23171-23180	NN	denotes	elevation
T15767	23181-23183	IN	denotes	of
T15768	23184-23188	NN	denotes	BCAA
T15769	23189-23192	CC	denotes	and
T15770	23193-23201	JJ	denotes	neonatal
T15771	23202-23211	NN	denotes	lethality
T15772	23212-23216	WDT	denotes	that
T15773	23217-23223	VBZ	denotes	occurs
T15774	23224-23226	IN	denotes	in
T15775	23227-23230	DT	denotes	the
T15777	23231-23238	JJ	denotes	classic
T15778	23239-23243	NN	denotes	MSUD
T15779	23244-23249	NN	denotes	mouse
T15776	23250-23255	NN	denotes	model
T15780	23255-23256	.	denotes	.
T15781	23256-23374	sentence	denotes	Our strategy was composed of a two part transgenic system to express human E2 in liver on the E2 knockout background.
T15782	23257-23260	PRP$	denotes	Our
T15783	23261-23269	NN	denotes	strategy
T15785	23270-23273	VBD	denotes	was
T15784	23274-23282	VBN	denotes	composed
T15786	23283-23285	IN	denotes	of
T15787	23286-23287	DT	denotes	a
T15789	23288-23291	CD	denotes	two
T15790	23292-23296	NN	denotes	part
T15791	23297-23307	JJ	denotes	transgenic
T15788	23308-23314	NN	denotes	system
T15792	23315-23317	TO	denotes	to
T15793	23318-23325	VB	denotes	express
T15794	23326-23331	JJ	denotes	human
T15795	23332-23334	NN	denotes	E2
T15796	23335-23337	IN	denotes	in
T15797	23338-23343	NN	denotes	liver
T15798	23344-23346	IN	denotes	on
T15799	23347-23350	DT	denotes	the
T15801	23351-23353	NN	denotes	E2
T15802	23354-23362	NN	denotes	knockout
T15800	23363-23373	NN	denotes	background
T15803	23373-23374	.	denotes	.
T15804	23374-23467	sentence	denotes	This bi-transgenic system consisted of a LAP-tTA transgene and a TRE-E2 transgene (Fig. 3A).
T15805	23375-23379	DT	denotes	This
T15807	23380-23393	JJ	denotes	bi-transgenic
T15806	23394-23400	NN	denotes	system
T15808	23401-23410	VBD	denotes	consisted
T15809	23411-23413	IN	denotes	of
T15810	23414-23415	DT	denotes	a
T15812	23416-23419	NN	denotes	LAP
T15814	23419-23420	HYPH	denotes	-
T15813	23420-23423	NN	denotes	tTA
T15811	23424-23433	NN	denotes	transgene
T15815	23434-23437	CC	denotes	and
T15816	23438-23439	DT	denotes	a
T15818	23440-23443	NN	denotes	TRE
T15820	23443-23444	HYPH	denotes	-
T15819	23444-23446	NN	denotes	E2
T15817	23447-23456	NN	denotes	transgene
T15821	23457-23458	-LRB-	denotes	(
T15823	23458-23462	NN	denotes	Fig.
T15822	23463-23465	NN	denotes	3A
T15824	23465-23466	-RRB-	denotes	)
T15825	23466-23467	.	denotes	.
T15826	23467-23712	sentence	denotes	The LAP-tTA transgene [34] directs high levels of liver specific expression of the tetracycline-controlled transactivator (tTA), a transcription factor that stimulates expression of promoters that harbor a transactivator response element (TRE).
T15827	23468-23471	DT	denotes	The
T15829	23472-23475	NN	denotes	LAP
T15831	23475-23476	HYPH	denotes	-
T15830	23476-23479	NN	denotes	tTA
T15828	23480-23489	NN	denotes	transgene
T15833	23490-23491	-LRB-	denotes	[
T15834	23491-23493	CD	denotes	34
T15835	23493-23494	-RRB-	denotes	]
T15832	23495-23502	VBZ	denotes	directs
T15836	23503-23507	JJ	denotes	high
T15837	23508-23514	NNS	denotes	levels
T15838	23515-23517	IN	denotes	of
T15839	23518-23523	NN	denotes	liver
T15840	23524-23532	JJ	denotes	specific
T15841	23533-23543	NN	denotes	expression
T15842	23544-23546	IN	denotes	of
T15843	23547-23550	DT	denotes	the
T15845	23551-23563	NN	denotes	tetracycline
T15847	23563-23564	HYPH	denotes	-
T15846	23564-23574	VBN	denotes	controlled
T15844	23575-23589	NN	denotes	transactivator
T15848	23590-23591	-LRB-	denotes	(
T15849	23591-23594	NN	denotes	tTA
T15850	23594-23595	-RRB-	denotes	)
T15851	23595-23597	,	denotes	,
T15852	23597-23598	DT	denotes	a
T15854	23599-23612	NN	denotes	transcription
T15853	23613-23619	NN	denotes	factor
T15855	23620-23624	WDT	denotes	that
T15856	23625-23635	VBZ	denotes	stimulates
T15857	23636-23646	NN	denotes	expression
T15858	23647-23649	IN	denotes	of
T15859	23650-23659	NNS	denotes	promoters
T15860	23660-23664	WDT	denotes	that
T15861	23665-23671	VBP	denotes	harbor
T15862	23672-23673	DT	denotes	a
T15864	23674-23688	NN	denotes	transactivator
T15865	23689-23697	NN	denotes	response
T15863	23698-23705	NN	denotes	element
T15866	23706-23707	-LRB-	denotes	(
T15867	23707-23710	NN	denotes	TRE
T15868	23710-23711	-RRB-	denotes	)
T15869	23711-23712	.	denotes	.
T15870	23712-23837	sentence	denotes	The TRE-E2 transgene was designed to express a human E2 cDNA from a TRE containing minimal promoter upon stimulation by tTA.
T15871	23713-23716	DT	denotes	The
T15873	23717-23720	NN	denotes	TRE
T15875	23720-23721	HYPH	denotes	-
T15874	23721-23723	NN	denotes	E2
T15872	23724-23733	NN	denotes	transgene
T15877	23734-23737	VBD	denotes	was
T15876	23738-23746	VBN	denotes	designed
T15878	23747-23749	TO	denotes	to
T15879	23750-23757	VB	denotes	express
T15880	23758-23759	DT	denotes	a
T15882	23760-23765	JJ	denotes	human
T15883	23766-23768	NN	denotes	E2
T15881	23769-23773	NN	denotes	cDNA
T15884	23774-23778	IN	denotes	from
T15885	23779-23780	DT	denotes	a
T15886	23781-23784	NN	denotes	TRE
T15887	23785-23795	VBG	denotes	containing
T15888	23796-23803	JJ	denotes	minimal
T15889	23804-23812	NN	denotes	promoter
T15890	23813-23817	IN	denotes	upon
T15891	23818-23829	NN	denotes	stimulation
T15892	23830-23832	IN	denotes	by
T15893	23833-23836	NN	denotes	tTA
T15894	23836-23837	.	denotes	.
T15895	23837-23838	sentence	denotes
T23655	23848-23858	JJ	denotes	Transgenic
T23657	23859-23864	NN	denotes	mouse
T23656	23865-23875	NN	denotes	production
T23658	23876-23879	CC	denotes	and
T23659	23880-23896	NN	denotes	characterization
T23660	23896-23897	.	denotes	.
T23661	23897-23964	sentence	denotes	A, Transgenic strategy used to produce mice that express human E2.
T23662	23898-23899	LS	denotes	A
T23664	23899-23901	,	denotes	,
T23665	23901-23911	JJ	denotes	Transgenic
T23663	23912-23920	NN	denotes	strategy
T23666	23921-23925	VBN	denotes	used
T23667	23926-23928	TO	denotes	to
T23668	23929-23936	VB	denotes	produce
T23669	23937-23941	NNS	denotes	mice
T23670	23942-23946	WDT	denotes	that
T23671	23947-23954	VBP	denotes	express
T23672	23955-23960	JJ	denotes	human
T23673	23961-23963	NN	denotes	E2
T23674	23963-23964	.	denotes	.
T23675	23964-24025	sentence	denotes	LAP-tTA transgenic mice have been previously described [34].
T23676	23965-23968	NN	denotes	LAP
T23678	23968-23969	HYPH	denotes	-
T23677	23969-23972	NN	denotes	tTA
T23680	23973-23983	JJ	denotes	transgenic
T23679	23984-23988	NNS	denotes	mice
T23682	23989-23993	VBP	denotes	have
T23683	23994-23998	VBN	denotes	been
T23684	23999-24009	RB	denotes	previously
T23681	24010-24019	VBN	denotes	described
T23685	24020-24021	-LRB-	denotes	[
T23686	24021-24023	CD	denotes	34
T23687	24023-24024	-RRB-	denotes	]
T23688	24024-24025	.	denotes	.
T23689	24025-24131	sentence	denotes	These mice express the tetracycline-controlled transactivator (tTA) from the liver specific LAP promoter.
T23690	24026-24031	DT	denotes	These
T23691	24032-24036	NNS	denotes	mice
T23692	24037-24044	VBP	denotes	express
T23693	24045-24048	DT	denotes	the
T23695	24049-24061	NN	denotes	tetracycline
T23697	24061-24062	HYPH	denotes	-
T23696	24062-24072	VBN	denotes	controlled
T23694	24073-24087	NN	denotes	transactivator
T23698	24088-24089	-LRB-	denotes	(
T23699	24089-24092	NN	denotes	tTA
T23700	24092-24093	-RRB-	denotes	)
T23701	24094-24098	IN	denotes	from
T23702	24099-24102	DT	denotes	the
T23704	24103-24108	NN	denotes	liver
T23705	24109-24117	JJ	denotes	specific
T23706	24118-24121	NN	denotes	LAP
T23703	24122-24130	NN	denotes	promoter
T23707	24130-24131	.	denotes	.
T23708	24131-24360	sentence	denotes	The TRE-E2 transgene contains the tetracycline response element (TRE) as part of the promoter, a synthetic intron (thin line), the human E2 cDNA, an alanine spacer, a c-myc epitope tag, and SV40 derived polyadenylation sequence.
T23709	24132-24135	DT	denotes	The
T23711	24136-24139	NN	denotes	TRE
T23713	24139-24140	HYPH	denotes	-
T23712	24140-24142	NN	denotes	E2
T23710	24143-24152	NN	denotes	transgene
T23714	24153-24161	VBZ	denotes	contains
T23715	24162-24165	DT	denotes	the
T23717	24166-24178	NN	denotes	tetracycline
T23718	24179-24187	NN	denotes	response
T23716	24188-24195	NN	denotes	element
T23719	24196-24197	-LRB-	denotes	(
T23720	24197-24200	NN	denotes	TRE
T23721	24200-24201	-RRB-	denotes	)
T23722	24202-24204	IN	denotes	as
T23723	24205-24209	NN	denotes	part
T23724	24210-24212	IN	denotes	of
T23725	24213-24216	DT	denotes	the
T23726	24217-24225	NN	denotes	promoter
T23727	24225-24227	,	denotes	,
T23728	24227-24228	DT	denotes	a
T23730	24229-24238	JJ	denotes	synthetic
T23729	24239-24245	NN	denotes	intron
T23731	24246-24247	-LRB-	denotes	(
T23733	24247-24251	JJ	denotes	thin
T23732	24252-24256	NN	denotes	line
T23734	24256-24257	-RRB-	denotes	)
T23735	24257-24259	,	denotes	,
T23736	24259-24262	DT	denotes	the
T23738	24263-24268	JJ	denotes	human
T23739	24269-24271	NN	denotes	E2
T23737	24272-24276	NN	denotes	cDNA
T23740	24276-24278	,	denotes	,
T23741	24278-24280	DT	denotes	an
T23743	24281-24288	NN	denotes	alanine
T23742	24289-24295	NN	denotes	spacer
T23744	24295-24297	,	denotes	,
T23745	24297-24298	DT	denotes	a
T23747	24299-24300	NN	denotes	c
T23749	24300-24301	HYPH	denotes	-
T23748	24301-24304	NN	denotes	myc
T23750	24305-24312	NN	denotes	epitope
T23746	24313-24316	NN	denotes	tag
T23751	24316-24318	,	denotes	,
T23752	24318-24321	CC	denotes	and
T23753	24322-24326	NN	denotes	SV40
T23754	24327-24334	VBN	denotes	derived
T23756	24335-24350	NN	denotes	polyadenylation
T23755	24351-24359	NN	denotes	sequence
T23757	24359-24360	.	denotes	.
T23758	24360-24428	sentence	denotes	This construct was used to create several lines of transgenic mice.
T23759	24361-24365	DT	denotes	This
T23760	24366-24375	NN	denotes	construct
T23762	24376-24379	VBD	denotes	was
T23761	24380-24384	VBN	denotes	used
T23763	24385-24387	TO	denotes	to
T23764	24388-24394	VB	denotes	create
T23765	24395-24402	JJ	denotes	several
T23766	24403-24408	NNS	denotes	lines
T23767	24409-24411	IN	denotes	of
T23768	24412-24422	JJ	denotes	transgenic
T23769	24423-24427	NNS	denotes	mice
T23770	24427-24428	.	denotes	.
T23771	24428-24515	sentence	denotes	B, Western blot analysis of E2 protein in liver of control and intermediate MSUD mice.
T23772	24429-24430	LS	denotes	B
T23774	24430-24432	,	denotes	,
T23775	24432-24439	NNP	denotes	Western
T23776	24440-24444	NN	denotes	blot
T23773	24445-24453	NN	denotes	analysis
T23777	24454-24456	IN	denotes	of
T23778	24457-24459	NN	denotes	E2
T23779	24460-24467	NN	denotes	protein
T23780	24468-24470	IN	denotes	in
T23781	24471-24476	NN	denotes	liver
T23782	24477-24479	IN	denotes	of
T23783	24480-24487	NN	denotes	control
T23785	24488-24491	CC	denotes	and
T23786	24492-24504	JJ	denotes	intermediate
T23787	24505-24509	NN	denotes	MSUD
T23784	24510-24514	NNS	denotes	mice
T23788	24514-24515	.	denotes	.
T23789	24515-24730	sentence	denotes	Note that the amount of human E2 protein (predicted MW ~54 Kd) in mice from Lines A and 525 A was variable but in many of the animals the amount was similar to the amount of mouse E2 (MW=~47 Kd) in control animals.
T23790	24516-24520	VB	denotes	Note
T23791	24521-24525	IN	denotes	that
T23793	24526-24529	DT	denotes	the
T23794	24530-24536	NN	denotes	amount
T23795	24537-24539	IN	denotes	of
T23796	24540-24545	JJ	denotes	human
T23798	24546-24548	NN	denotes	E2
T23797	24549-24556	NN	denotes	protein
T23799	24557-24558	-LRB-	denotes	(
T23801	24558-24567	VBN	denotes	predicted
T23800	24568-24570	NN	denotes	MW
T23802	24571-24572	SYM	denotes	~
T23803	24572-24574	CD	denotes	54
T23804	24575-24577	NN	denotes	Kd
T23805	24577-24578	-RRB-	denotes	)
T23806	24579-24581	IN	denotes	in
T23807	24582-24586	NNS	denotes	mice
T23808	24587-24591	IN	denotes	from
T23809	24592-24597	NNS	denotes	Lines
T23810	24598-24599	NN	denotes	A
T23811	24600-24603	CC	denotes	and
T23812	24604-24607	CD	denotes	525
T23813	24608-24609	NN	denotes	A
T23792	24610-24613	VBD	denotes	was
T23814	24614-24622	JJ	denotes	variable
T23815	24623-24626	CC	denotes	but
T23816	24627-24629	IN	denotes	in
T23818	24630-24634	JJ	denotes	many
T23819	24635-24637	IN	denotes	of
T23820	24638-24641	DT	denotes	the
T23821	24642-24649	NNS	denotes	animals
T23822	24650-24653	DT	denotes	the
T23823	24654-24660	NN	denotes	amount
T23817	24661-24664	VBD	denotes	was
T23824	24665-24672	JJ	denotes	similar
T23825	24673-24675	IN	denotes	to
T23826	24676-24679	DT	denotes	the
T23827	24680-24686	NN	denotes	amount
T23828	24687-24689	IN	denotes	of
T23829	24690-24695	NN	denotes	mouse
T23830	24696-24698	NN	denotes	E2
T23831	24699-24700	-LRB-	denotes	(
T23833	24700-24702	NN	denotes	MW
T23834	24702-24703	SYM	denotes	=
T23835	24703-24704	SYM	denotes	~
T23836	24704-24706	CD	denotes	47
T23832	24707-24709	NN	denotes	Kd
T23837	24709-24710	-RRB-	denotes	)
T23838	24711-24713	IN	denotes	in
T23839	24714-24721	NN	denotes	control
T23840	24722-24729	NNS	denotes	animals
T23841	24729-24730	.	denotes	.
T23842	24730-24879	sentence	denotes	Re-probing with a c-myc tag antibody confirmed the presence of the transgene derived, c-myc tagged, human E2 in transgenic mice but not in controls.
T23843	24731-24733	NN	denotes	Re
T23845	24733-24734	HYPH	denotes	-
T23844	24734-24741	NN	denotes	probing
T23847	24742-24746	IN	denotes	with
T23848	24747-24748	DT	denotes	a
T23850	24749-24750	NN	denotes	c
T23852	24750-24751	HYPH	denotes	-
T23851	24751-24754	NN	denotes	myc
T23853	24755-24758	NN	denotes	tag
T23849	24759-24767	NN	denotes	antibody
T23846	24768-24777	VBD	denotes	confirmed
T23854	24778-24781	DT	denotes	the
T23855	24782-24790	NN	denotes	presence
T23856	24791-24793	IN	denotes	of
T23857	24794-24797	DT	denotes	the
T23859	24798-24807	NN	denotes	transgene
T23860	24808-24815	VBN	denotes	derived
T23861	24815-24817	,	denotes	,
T23862	24817-24818	NN	denotes	c
T23864	24818-24819	HYPH	denotes	-
T23863	24819-24822	NN	denotes	myc
T23865	24823-24829	VBN	denotes	tagged
T23866	24829-24831	,	denotes	,
T23867	24831-24836	JJ	denotes	human
T23858	24837-24839	NN	denotes	E2
T23868	24840-24842	IN	denotes	in
T23869	24843-24853	JJ	denotes	transgenic
T23870	24854-24858	NNS	denotes	mice
T23871	24859-24862	CC	denotes	but
T23872	24863-24866	RB	denotes	not
T23873	24867-24869	IN	denotes	in
T23874	24870-24878	NNS	denotes	controls
T23875	24878-24879	.	denotes	.
T23876	24879-25012	sentence	denotes	Western blot analysis of brain (C), kidney (D), and muscle (E) revealed negligible amounts of transgene derived E2 in those tissues.
T23877	24880-24887	NNP	denotes	Western
T23878	24888-24892	NN	denotes	blot
T23879	24893-24901	NN	denotes	analysis
T23881	24902-24904	IN	denotes	of
T23882	24905-24910	NN	denotes	brain
T23883	24911-24912	-LRB-	denotes	(
T23884	24912-24913	NN	denotes	C
T23885	24913-24914	-RRB-	denotes	)
T23886	24914-24916	,	denotes	,
T23887	24916-24922	NN	denotes	kidney
T23888	24923-24924	-LRB-	denotes	(
T23889	24924-24925	NN	denotes	D
T23890	24925-24926	-RRB-	denotes	)
T23891	24926-24928	,	denotes	,
T23892	24928-24931	CC	denotes	and
T23893	24932-24938	NN	denotes	muscle
T23894	24939-24940	-LRB-	denotes	(
T23895	24940-24941	NN	denotes	E
T23896	24941-24942	-RRB-	denotes	)
T23880	24943-24951	VBD	denotes	revealed
T23897	24952-24962	JJ	denotes	negligible
T23898	24963-24970	NNS	denotes	amounts
T23899	24971-24973	IN	denotes	of
T23900	24974-24983	NN	denotes	transgene
T23901	24984-24991	VBN	denotes	derived
T23902	24992-24994	NN	denotes	E2
T23903	24995-24997	IN	denotes	in
T23904	24998-25003	DT	denotes	those
T23905	25004-25011	NNS	denotes	tissues
T23906	25011-25012	.	denotes	.
T23907	25012-25123	sentence	denotes	All blots were re-probed with an actin antibody to allow amount of protein loaded in each lane to be compared.
T23908	25013-25016	DT	denotes	All
T23909	25017-25022	NNS	denotes	blots
T23911	25023-25027	VBD	denotes	were
T23910	25028-25037	VBN	denotes	re-probed
T23912	25038-25042	IN	denotes	with
T23913	25043-25045	DT	denotes	an
T23915	25046-25051	NN	denotes	actin
T23914	25052-25060	NN	denotes	antibody
T23916	25061-25063	TO	denotes	to
T23917	25064-25069	VB	denotes	allow
T23918	25070-25076	NN	denotes	amount
T23920	25077-25079	IN	denotes	of
T23921	25080-25087	NN	denotes	protein
T23922	25088-25094	VBN	denotes	loaded
T23923	25095-25097	IN	denotes	in
T23924	25098-25102	DT	denotes	each
T23925	25103-25107	NN	denotes	lane
T23926	25108-25110	TO	denotes	to
T23927	25111-25113	VB	denotes	be
T23919	25114-25122	VBN	denotes	compared
T23928	25122-25123	.	denotes	.
T15897	25124-25127	NN	denotes	LAP
T15896	25124-25137	sentence	denotes	LAP-tTA mice
T15899	25127-25128	HYPH	denotes	-
T15898	25128-25131	NN	denotes	tTA
T15900	25132-25136	NNS	denotes	mice
T15903	25137-25138	VBD	denotes	w
T15901	25137-25142	sentence	denotes	were
T15902	25138-25139	VBN	denotes	e
T15904	25139-25141	VB	denotes	re
T15906	25142-25145	NN	denotes	pre
T15905	25142-25147	sentence	denotes	previ
T15907	25145-25146	NN	denotes	v
T15908	25146-25147	JJ	denotes	i
T15910	25147-25148	VB	denotes	o
T15909	25147-25153	sentence	denotes	ously
T15911	25148-25149	NN	denotes	u
T15912	25149-25150	NNS	denotes	s
T15913	25150-25151	JJ	denotes	l
T15914	25151-25152	JJ	denotes	y
T15916	25153-25156	NN	denotes	pro
T15915	25153-25160	sentence	denotes	produce
T15917	25156-25157	NN	denotes	d
T15918	25157-25158	JJ	denotes	u
T15919	25158-25159	JJ	denotes	c
T15920	25159-25160	NNS	denotes	e
T15922	25160-25161	NN	denotes	d
T15921	25160-25168	sentence	denotes	d and ch
T15923	25162-25165	CC	denotes	and
T15924	25166-25167	NN	denotes	c
T15925	25167-25168	DT	denotes	h
T15927	25168-25169	DT	denotes	a
T15926	25168-25175	sentence	denotes	aracter
T15929	25169-25170	NN	denotes	r
T15930	25170-25171	VBN	denotes	a
T15931	25171-25172	DT	denotes	c
T15932	25172-25173	TO	denotes	t
T15933	25173-25174	VB	denotes	e
T15928	25174-25175	VBN	denotes	r
T15935	25175-25176	NNS	denotes	i
T15934	25175-25210	sentence	denotes	ized by the Bujard laboratory [34].
T15936	25176-25179	VBN	denotes	zed
T15937	25180-25182	IN	denotes	by
T15938	25183-25186	DT	denotes	the
T15940	25187-25193	NNP	denotes	Bujard
T15939	25194-25204	NN	denotes	laboratory
T15941	25205-25206	-LRB-	denotes	[
T15942	25206-25208	CD	denotes	34
T15943	25208-25209	-RRB-	denotes	]
T15944	25209-25210	.	denotes	.
T15945	25210-25308	sentence	denotes	Pronuclear microinjection was used to produce transgenic mice that harbored the TRE-E2 transgene.
T15946	25211-25221	JJ	denotes	Pronuclear
T15947	25222-25236	NN	denotes	microinjection
T15949	25237-25240	VBD	denotes	was
T15948	25241-25245	VBN	denotes	used
T15950	25246-25248	TO	denotes	to
T15951	25249-25256	VB	denotes	produce
T15952	25257-25267	JJ	denotes	transgenic
T15953	25268-25272	NNS	denotes	mice
T15954	25273-25277	WDT	denotes	that
T15955	25278-25286	VBD	denotes	harbored
T15956	25287-25290	DT	denotes	the
T15958	25291-25294	NN	denotes	TRE
T15960	25294-25295	HYPH	denotes	-
T15959	25295-25297	NN	denotes	E2
T15957	25298-25307	NN	denotes	transgene
T15961	25307-25308	.	denotes	.
T15962	25308-25481	sentence	denotes	From injections at the University of Pittsburgh, 2 transgene positive founders were identified that led to the generation of 3 different transgenic lines (Lines A, B, & D).
T15963	25309-25313	IN	denotes	From
T15965	25314-25324	NNS	denotes	injections
T15966	25325-25327	IN	denotes	at
T15967	25328-25331	DT	denotes	the
T15968	25332-25342	NNP	denotes	University
T15969	25343-25345	IN	denotes	of
T15970	25346-25356	NNP	denotes	Pittsburgh
T15971	25356-25358	,	denotes	,
T15972	25358-25359	CD	denotes	2
T15974	25360-25369	NN	denotes	transgene
T15975	25370-25378	JJ	denotes	positive
T15973	25379-25387	NNS	denotes	founders
T15976	25388-25392	VBD	denotes	were
T15964	25393-25403	VBN	denotes	identified
T15977	25404-25408	WDT	denotes	that
T15978	25409-25412	VBD	denotes	led
T15979	25413-25415	IN	denotes	to
T15980	25416-25419	DT	denotes	the
T15981	25420-25430	NN	denotes	generation
T15982	25431-25433	IN	denotes	of
T15983	25434-25435	CD	denotes	3
T15985	25436-25445	JJ	denotes	different
T15986	25446-25456	JJ	denotes	transgenic
T15984	25457-25462	NNS	denotes	lines
T15987	25463-25464	-LRB-	denotes	(
T15989	25464-25469	NNS	denotes	Lines
T15988	25470-25471	NN	denotes	A
T15990	25471-25473	,	denotes	,
T15991	25473-25474	NN	denotes	B
T15992	25474-25476	,	denotes	,
T15993	25476-25477	CC	denotes	&
T15994	25478-25479	NN	denotes	D
T15995	25479-25480	-RRB-	denotes	)
T15996	25480-25481	.	denotes	.
T15997	25481-25577	sentence	denotes	From the injections at the University of Cincinnati, we obtained 8 transgene positive founders.
T15998	25482-25486	IN	denotes	From
T16000	25487-25490	DT	denotes	the
T16001	25491-25501	NNS	denotes	injections
T16002	25502-25504	IN	denotes	at
T16003	25505-25508	DT	denotes	the
T16004	25509-25519	NNP	denotes	University
T16005	25520-25522	IN	denotes	of
T16006	25523-25533	NNP	denotes	Cincinnati
T16007	25533-25535	,	denotes	,
T16008	25535-25537	PRP	denotes	we
T15999	25538-25546	VBD	denotes	obtained
T16009	25547-25548	CD	denotes	8
T16011	25549-25558	NN	denotes	transgene
T16012	25559-25567	JJ	denotes	positive
T16010	25568-25576	NNS	denotes	founders
T16013	25576-25577	.	denotes	.
T16014	25577-25705	sentence	denotes	Subsequent breeding of these founders revealed that many of the animals had multiple transgene insertion sites that segregated.
T16015	25578-25588	JJ	denotes	Subsequent
T16016	25589-25597	NN	denotes	breeding
T16018	25598-25600	IN	denotes	of
T16019	25601-25606	DT	denotes	these
T16020	25607-25615	NNS	denotes	founders
T16017	25616-25624	VBD	denotes	revealed
T16021	25625-25629	IN	denotes	that
T16023	25630-25634	JJ	denotes	many
T16024	25635-25637	IN	denotes	of
T16025	25638-25641	DT	denotes	the
T16026	25642-25649	NNS	denotes	animals
T16022	25650-25653	VBD	denotes	had
T16027	25654-25662	JJ	denotes	multiple
T16029	25663-25672	NN	denotes	transgene
T16030	25673-25682	NN	denotes	insertion
T16028	25683-25688	NNS	denotes	sites
T16031	25689-25693	WDT	denotes	that
T16032	25694-25704	VBD	denotes	segregated
T16033	25704-25705	.	denotes	.
T16034	25705-25782	sentence	denotes	We established a total of 15 different transgenic lines from these founders.
T16035	25706-25708	PRP	denotes	We
T16036	25709-25720	VBD	denotes	established
T16037	25721-25722	DT	denotes	a
T16038	25723-25728	NN	denotes	total
T16039	25729-25731	IN	denotes	of
T16040	25732-25734	CD	denotes	15
T16042	25735-25744	JJ	denotes	different
T16043	25745-25755	JJ	denotes	transgenic
T16041	25756-25761	NNS	denotes	lines
T16044	25762-25766	IN	denotes	from
T16045	25767-25772	DT	denotes	these
T16046	25773-25781	NNS	denotes	founders
T16047	25781-25782	.	denotes	.
T16048	25782-25857	sentence	denotes	Limited resources allowed us to only focus on a total of 9 of these lines.
T16049	25783-25790	VBN	denotes	Limited
T16050	25791-25800	NNS	denotes	resources
T16051	25801-25808	VBD	denotes	allowed
T16052	25809-25811	PRP	denotes	us
T16054	25812-25814	TO	denotes	to
T16055	25815-25819	RB	denotes	only
T16053	25820-25825	VB	denotes	focus
T16056	25826-25828	IN	denotes	on
T16057	25829-25830	DT	denotes	a
T16058	25831-25836	NN	denotes	total
T16059	25837-25839	IN	denotes	of
T16060	25840-25841	CD	denotes	9
T16061	25842-25844	IN	denotes	of
T16062	25845-25850	DT	denotes	these
T16063	25851-25856	NNS	denotes	lines
T16064	25856-25857	.	denotes	.
T16065	25857-25985	sentence	denotes	These lines differed substantially in transgene copy number as compared by Southern blot analysis of tail DNA (data not shown).
T16066	25858-25863	DT	denotes	These
T16067	25864-25869	NNS	denotes	lines
T16068	25870-25878	VBD	denotes	differed
T16069	25879-25892	RB	denotes	substantially
T16070	25893-25895	IN	denotes	in
T16071	25896-25905	NN	denotes	transgene
T16073	25906-25910	NN	denotes	copy
T16072	25911-25917	NN	denotes	number
T16074	25918-25920	IN	denotes	as
T16075	25921-25929	VBN	denotes	compared
T16076	25930-25932	IN	denotes	by
T16077	25933-25941	NNP	denotes	Southern
T16078	25942-25946	NN	denotes	blot
T16079	25947-25955	NN	denotes	analysis
T16080	25956-25958	IN	denotes	of
T16081	25959-25963	NN	denotes	tail
T16082	25964-25967	NN	denotes	DNA
T16083	25968-25969	-LRB-	denotes	(
T16085	25969-25973	NNS	denotes	data
T16086	25974-25977	RB	denotes	not
T16084	25978-25983	VBN	denotes	shown
T16087	25983-25984	-RRB-	denotes	)
T16088	25984-25985	.	denotes	.
T16089	25985-26166	sentence	denotes	Transgenic TRE-E2 mice from each line (either founders or F1 offspring) were crossed to mice that were positive for the LAP-tTA transgene and were heterozygous for the E2 knockout.
T16090	25986-25996	JJ	denotes	Transgenic
T16092	25997-26000	NN	denotes	TRE
T16094	26000-26001	HYPH	denotes	-
T16093	26001-26003	NN	denotes	E2
T16091	26004-26008	NNS	denotes	mice
T16096	26009-26013	IN	denotes	from
T16097	26014-26018	DT	denotes	each
T16098	26019-26023	NN	denotes	line
T16099	26024-26025	-LRB-	denotes	(
T16101	26025-26031	CC	denotes	either
T16102	26032-26040	NNS	denotes	founders
T16103	26041-26043	CC	denotes	or
T16104	26044-26046	NN	denotes	F1
T16100	26047-26056	NN	denotes	offspring
T16105	26056-26057	-RRB-	denotes	)
T16106	26058-26062	VBD	denotes	were
T16095	26063-26070	VBN	denotes	crossed
T16107	26071-26073	IN	denotes	to
T16108	26074-26078	NNS	denotes	mice
T16109	26079-26083	WDT	denotes	that
T16110	26084-26088	VBD	denotes	were
T16111	26089-26097	JJ	denotes	positive
T16112	26098-26101	IN	denotes	for
T16113	26102-26105	DT	denotes	the
T16115	26106-26109	NN	denotes	LAP
T16117	26109-26110	HYPH	denotes	-
T16116	26110-26113	NN	denotes	tTA
T16114	26114-26123	NN	denotes	transgene
T16118	26124-26127	CC	denotes	and
T16119	26128-26132	VBD	denotes	were
T16120	26133-26145	JJ	denotes	heterozygous
T16121	26146-26149	IN	denotes	for
T16122	26150-26153	DT	denotes	the
T16124	26154-26156	NN	denotes	E2
T16123	26157-26165	NN	denotes	knockout
T16125	26165-26166	.	denotes	.
T16126	26166-26309	sentence	denotes	Ultimately, breeding pairs were used in which the mice were heterozygous for the LAP-tTA transgene, the TRE-E2 transgene, and the E2 knockout.
T16127	26167-26177	RB	denotes	Ultimately
T16129	26177-26179	,	denotes	,
T16130	26179-26187	VBG	denotes	breeding
T16131	26188-26193	NNS	denotes	pairs
T16132	26194-26198	VBD	denotes	were
T16128	26199-26203	VBN	denotes	used
T16133	26204-26206	IN	denotes	in
T16135	26207-26212	WDT	denotes	which
T16136	26213-26216	DT	denotes	the
T16137	26217-26221	NNS	denotes	mice
T16134	26222-26226	VBD	denotes	were
T16138	26227-26239	JJ	denotes	heterozygous
T16139	26240-26243	IN	denotes	for
T16140	26244-26247	DT	denotes	the
T16142	26248-26251	NN	denotes	LAP
T16144	26251-26252	HYPH	denotes	-
T16143	26252-26255	NN	denotes	tTA
T16141	26256-26265	NN	denotes	transgene
T16145	26265-26267	,	denotes	,
T16146	26267-26270	DT	denotes	the
T16148	26271-26274	NN	denotes	TRE
T16150	26274-26275	HYPH	denotes	-
T16149	26275-26277	NN	denotes	E2
T16147	26278-26287	NN	denotes	transgene
T16151	26287-26289	,	denotes	,
T16152	26289-26292	CC	denotes	and
T16153	26293-26296	DT	denotes	the
T16155	26297-26299	NN	denotes	E2
T16154	26300-26308	NN	denotes	knockout
T16156	26308-26309	.	denotes	.
T16157	26309-26446	sentence	denotes	To efficiently screen for the ability of the transgenes to rescue the knockout from neonatal lethality, we genotyped litters at weaning.
T16158	26310-26312	TO	denotes	To
T16160	26313-26324	RB	denotes	efficiently
T16159	26325-26331	VB	denotes	screen
T16162	26332-26335	IN	denotes	for
T16163	26336-26339	DT	denotes	the
T16164	26340-26347	NN	denotes	ability
T16165	26348-26350	IN	denotes	of
T16166	26351-26354	DT	denotes	the
T16167	26355-26365	NNS	denotes	transgenes
T16168	26366-26368	TO	denotes	to
T16169	26369-26375	VB	denotes	rescue
T16170	26376-26379	DT	denotes	the
T16171	26380-26388	NN	denotes	knockout
T16172	26389-26393	IN	denotes	from
T16173	26394-26402	JJ	denotes	neonatal
T16174	26403-26412	NN	denotes	lethality
T16175	26412-26414	,	denotes	,
T16176	26414-26416	PRP	denotes	we
T16161	26417-26426	VBD	denotes	genotyped
T16177	26427-26434	NNS	denotes	litters
T16178	26435-26437	IN	denotes	at
T16179	26438-26445	NN	denotes	weaning
T16180	26445-26446	.	denotes	.
T16181	26446-26624	sentence	denotes	This breeding strategy is expected to result in a theoretical maximum of animals with the rescue genotype (i.e., homozygous E2 knockout and positive for both transgenes) of 14%.
T16182	26447-26451	DT	denotes	This
T16184	26452-26460	VBG	denotes	breeding
T16183	26461-26469	NN	denotes	strategy
T16186	26470-26472	VBZ	denotes	is
T16185	26473-26481	VBN	denotes	expected
T16187	26482-26484	TO	denotes	to
T16188	26485-26491	VB	denotes	result
T16189	26492-26494	IN	denotes	in
T16190	26495-26496	DT	denotes	a
T16192	26497-26508	JJ	denotes	theoretical
T16191	26509-26516	NN	denotes	maximum
T16193	26517-26519	IN	denotes	of
T16194	26520-26527	NNS	denotes	animals
T16195	26528-26532	IN	denotes	with
T16196	26533-26536	DT	denotes	the
T16198	26537-26543	NN	denotes	rescue
T16197	26544-26552	NN	denotes	genotype
T16199	26553-26554	-LRB-	denotes	(
T16201	26554-26558	FW	denotes	i.e.
T16202	26558-26560	,	denotes	,
T16203	26560-26570	JJ	denotes	homozygous
T16204	26571-26573	NN	denotes	E2
T16200	26574-26582	NN	denotes	knockout
T16205	26583-26586	CC	denotes	and
T16206	26587-26595	JJ	denotes	positive
T16207	26596-26599	IN	denotes	for
T16208	26600-26604	DT	denotes	both
T16209	26605-26615	NNS	denotes	transgenes
T16210	26615-26616	-RRB-	denotes	)
T16211	26617-26619	IN	denotes	of
T16212	26620-26622	CD	denotes	14
T16213	26622-26623	NN	denotes	%
T16214	26623-26624	.	denotes	.
T16215	26624-26737	sentence	denotes	Fig. 4A shows the percentage of mice alive at weaning with the rescue genotype from each transgenic line tested.
T16216	26625-26629	NN	denotes	Fig.
T16217	26630-26632	NN	denotes	4A
T16218	26633-26638	VBZ	denotes	shows
T16219	26639-26642	DT	denotes	the
T16220	26643-26653	NN	denotes	percentage
T16221	26654-26656	IN	denotes	of
T16222	26657-26661	NNS	denotes	mice
T16223	26662-26667	JJ	denotes	alive
T16224	26668-26670	IN	denotes	at
T16225	26671-26678	NN	denotes	weaning
T16226	26679-26683	IN	denotes	with
T16227	26684-26687	DT	denotes	the
T16229	26688-26694	NN	denotes	rescue
T16228	26695-26703	NN	denotes	genotype
T16230	26704-26708	IN	denotes	from
T16231	26709-26713	DT	denotes	each
T16233	26714-26724	JJ	denotes	transgenic
T16232	26725-26729	NN	denotes	line
T16234	26730-26736	VBN	denotes	tested
T16235	26736-26737	.	denotes	.
T16236	26737-26790	sentence	denotes	Considerable variability was observed between lines.
T16237	26738-26750	JJ	denotes	Considerable
T16238	26751-26762	NN	denotes	variability
T16240	26763-26766	VBD	denotes	was
T16239	26767-26775	VBN	denotes	observed
T16241	26776-26783	IN	denotes	between
T16242	26784-26789	NNS	denotes	lines
T16243	26789-26790	.	denotes	.
T16244	26790-26869	sentence	denotes	The line with the highest percentage of rescue mice at weaning was line 525 A.
T16245	26791-26794	DT	denotes	The
T16246	26795-26799	NN	denotes	line
T16248	26800-26804	IN	denotes	with
T16249	26805-26808	DT	denotes	the
T16251	26809-26816	JJS	denotes	highest
T16250	26817-26827	NN	denotes	percentage
T16252	26828-26830	IN	denotes	of
T16253	26831-26837	NN	denotes	rescue
T16254	26838-26842	NNS	denotes	mice
T16255	26843-26845	IN	denotes	at
T16256	26846-26853	NN	denotes	weaning
T16247	26854-26857	VBD	denotes	was
T16257	26858-26862	NN	denotes	line
T16259	26863-26866	CD	denotes	525
T16258	26867-26868	NN	denotes	A
T16260	26868-26869	.	denotes	.
T16261	26869-26953	sentence	denotes	Approximately 10% of weaned, surviving pups from this line had the rescue genotype.
T16262	26870-26883	RB	denotes	Approximately
T16263	26884-26886	CD	denotes	10
T16264	26886-26887	NN	denotes	%
T16266	26888-26890	IN	denotes	of
T16267	26891-26897	VBN	denotes	weaned
T16269	26897-26899	,	denotes	,
T16270	26899-26908	VBG	denotes	surviving
T16268	26909-26913	NNS	denotes	pups
T16271	26914-26918	IN	denotes	from
T16272	26919-26923	DT	denotes	this
T16273	26924-26928	NN	denotes	line
T16265	26929-26932	VBD	denotes	had
T16274	26933-26936	DT	denotes	the
T16276	26937-26943	NN	denotes	rescue
T16275	26944-26952	NN	denotes	genotype
T16277	26952-26953	.	denotes	.
T16278	26953-27071	sentence	denotes	Thus, the 525A transgene appears to be highly effective at rescuing the neonatal lethal phenotype of the E2 knockout.
T16279	26954-26958	RB	denotes	Thus
T16281	26958-26960	,	denotes	,
T16282	26960-26963	DT	denotes	the
T16284	26964-26968	NN	denotes	525A
T16283	26969-26978	NN	denotes	transgene
T16280	26979-26986	VBZ	denotes	appears
T16285	26987-26989	TO	denotes	to
T16286	26990-26992	VB	denotes	be
T16287	26993-26999	RB	denotes	highly
T16288	27000-27009	JJ	denotes	effective
T16289	27010-27012	IN	denotes	at
T16290	27013-27021	VBG	denotes	rescuing
T16291	27022-27025	DT	denotes	the
T16293	27026-27034	JJ	denotes	neonatal
T16294	27035-27041	JJ	denotes	lethal
T16292	27042-27051	NN	denotes	phenotype
T16295	27052-27054	IN	denotes	of
T16296	27055-27058	DT	denotes	the
T16298	27059-27061	NN	denotes	E2
T16297	27062-27070	NN	denotes	knockout
T16299	27070-27071	.	denotes	.
T16300	27071-27153	sentence	denotes	In contrast, line 520B completely failed to rescue the neonatal lethal phenotype.
T16301	27072-27074	IN	denotes	In
T16303	27075-27083	NN	denotes	contrast
T16304	27083-27085	,	denotes	,
T16305	27085-27089	NN	denotes	line
T16306	27090-27094	NN	denotes	520B
T16307	27095-27105	RB	denotes	completely
T16302	27106-27112	VBD	denotes	failed
T16308	27113-27115	TO	denotes	to
T16309	27116-27122	VB	denotes	rescue
T16310	27123-27126	DT	denotes	the
T16312	27127-27135	JJ	denotes	neonatal
T16313	27136-27142	JJ	denotes	lethal
T16311	27143-27152	NN	denotes	phenotype
T16314	27152-27153	.	denotes	.
T16315	27153-27258	sentence	denotes	All other lines tested produced surviving rescue animals at a frequency between 1 and 5% of weaned pups.
T16316	27154-27157	DT	denotes	All
T16318	27158-27163	JJ	denotes	other
T16317	27164-27169	NNS	denotes	lines
T16320	27170-27176	VBN	denotes	tested
T16319	27177-27185	VBD	denotes	produced
T16321	27186-27195	VBG	denotes	surviving
T16323	27196-27202	NN	denotes	rescue
T16322	27203-27210	NNS	denotes	animals
T16324	27211-27213	IN	denotes	at
T16325	27214-27215	DT	denotes	a
T16326	27216-27225	NN	denotes	frequency
T16327	27226-27233	IN	denotes	between
T16328	27234-27235	CD	denotes	1
T16330	27236-27239	CC	denotes	and
T16331	27240-27241	CD	denotes	5
T16329	27241-27242	NN	denotes	%
T16332	27243-27245	IN	denotes	of
T16333	27246-27252	VBN	denotes	weaned
T16334	27253-27257	NNS	denotes	pups
T16335	27257-27258	.	denotes	.
T16336	27258-27259	sentence	denotes
T24445	27269-27285	NN	denotes	Characterization
T24446	27286-27288	IN	denotes	of
T24447	27289-27292	DT	denotes	the
T24449	27293-27305	JJ	denotes	intermediate
T24450	27306-27310	NN	denotes	MSUD
T24451	27311-27317	JJ	denotes	murine
T24448	27318-27323	NN	denotes	model
T24452	27323-27324	.	denotes	.
T24453	27324-27373	sentence	denotes	A, Survival analysis of transgenic rescue lines.
T24454	27325-27326	LS	denotes	A
T24456	27326-27328	,	denotes	,
T24457	27328-27336	NN	denotes	Survival
T24455	27337-27345	NN	denotes	analysis
T24458	27346-27348	IN	denotes	of
T24459	27349-27359	JJ	denotes	transgenic
T24461	27360-27366	NN	denotes	rescue
T24460	27367-27372	NNS	denotes	lines
T24462	27372-27373	.	denotes	.
T24463	27373-27592	sentence	denotes	Presented are percentages of mice alive at weaning from each transgenic line tested that had the rescue genotype (i.e., homozygous for knockout of endogenous E2 and positive for both the LAP-tTA and TRE-E2 transgenes).
T24464	27374-27383	VBN	denotes	Presented
T24465	27384-27387	VBP	denotes	are
T24466	27388-27399	NNS	denotes	percentages
T24467	27400-27402	IN	denotes	of
T24468	27403-27407	NNS	denotes	mice
T24469	27408-27413	JJ	denotes	alive
T24470	27414-27416	IN	denotes	at
T24471	27417-27424	VBG	denotes	weaning
T24472	27425-27429	IN	denotes	from
T24473	27430-27434	DT	denotes	each
T24475	27435-27445	JJ	denotes	transgenic
T24474	27446-27450	NN	denotes	line
T24476	27451-27457	VBN	denotes	tested
T24477	27458-27462	WDT	denotes	that
T24478	27463-27466	VBD	denotes	had
T24479	27467-27470	DT	denotes	the
T24481	27471-27477	NN	denotes	rescue
T24480	27478-27486	NN	denotes	genotype
T24482	27487-27488	-LRB-	denotes	(
T24484	27488-27492	FW	denotes	i.e.
T24485	27492-27494	,	denotes	,
T24483	27494-27504	JJ	denotes	homozygous
T24486	27505-27508	IN	denotes	for
T24487	27509-27517	NN	denotes	knockout
T24488	27518-27520	IN	denotes	of
T24489	27521-27531	JJ	denotes	endogenous
T24490	27532-27534	NN	denotes	E2
T24491	27535-27538	CC	denotes	and
T24492	27539-27547	JJ	denotes	positive
T24493	27548-27551	IN	denotes	for
T24494	27552-27556	CC	denotes	both
T24496	27557-27560	DT	denotes	the
T24497	27561-27564	NN	denotes	LAP
T24499	27564-27565	HYPH	denotes	-
T24498	27565-27568	NN	denotes	tTA
T24500	27569-27572	CC	denotes	and
T24501	27573-27576	NN	denotes	TRE
T24503	27576-27577	HYPH	denotes	-
T24502	27577-27579	NN	denotes	E2
T24495	27580-27590	NNS	denotes	transgenes
T24504	27590-27591	-RRB-	denotes	)
T24505	27591-27592	.	denotes	.
T24506	27592-27708	sentence	denotes	Also plotted is the theoretical maximum frequency at which this genotype is expected in this population of animals.
T24507	27593-27597	RB	denotes	Also
T24509	27598-27605	VBN	denotes	plotted
T24508	27606-27608	VBZ	denotes	is
T24510	27609-27612	DT	denotes	the
T24512	27613-27624	JJ	denotes	theoretical
T24513	27625-27632	JJ	denotes	maximum
T24511	27633-27642	NN	denotes	frequency
T24514	27643-27645	IN	denotes	at
T24516	27646-27651	WDT	denotes	which
T24517	27652-27656	DT	denotes	this
T24518	27657-27665	NN	denotes	genotype
T24519	27666-27668	VBZ	denotes	is
T24515	27669-27677	VBN	denotes	expected
T24520	27678-27680	IN	denotes	in
T24521	27681-27685	DT	denotes	this
T24522	27686-27696	NN	denotes	population
T24523	27697-27699	IN	denotes	of
T24524	27700-27707	NNS	denotes	animals
T24525	27707-27708	.	denotes	.
T24526	27708-27785	sentence	denotes	The numbers on the bars indicates the numbers of observations for each line.
T24527	27709-27712	DT	denotes	The
T24528	27713-27720	NNS	denotes	numbers
T24530	27721-27723	IN	denotes	on
T24531	27724-27727	DT	denotes	the
T24532	27728-27732	NNS	denotes	bars
T24529	27733-27742	VBZ	denotes	indicates
T24533	27743-27746	DT	denotes	the
T24534	27747-27754	NNS	denotes	numbers
T24535	27755-27757	IN	denotes	of
T24536	27758-27770	NNS	denotes	observations
T24537	27771-27774	IN	denotes	for
T24538	27775-27779	DT	denotes	each
T24539	27780-27784	NN	denotes	line
T24540	27784-27785	.	denotes	.
T24541	27785-27872	sentence	denotes	B, Ratio of total BCAA to alanine in blood of mice from controls and Lines A and 525A.
T24542	27786-27787	LS	denotes	B
T24544	27787-27789	,	denotes	,
T24543	27789-27794	NN	denotes	Ratio
T24545	27795-27797	IN	denotes	of
T24546	27798-27803	JJ	denotes	total
T24547	27804-27808	NN	denotes	BCAA
T24548	27809-27811	IN	denotes	to
T24549	27812-27819	NN	denotes	alanine
T24550	27820-27822	IN	denotes	in
T24551	27823-27828	NN	denotes	blood
T24552	27829-27831	IN	denotes	of
T24553	27832-27836	NNS	denotes	mice
T24554	27837-27841	IN	denotes	from
T24555	27842-27850	NNS	denotes	controls
T24556	27851-27854	CC	denotes	and
T24557	27855-27860	NNS	denotes	Lines
T24558	27861-27862	NN	denotes	A
T24559	27863-27866	CC	denotes	and
T24560	27867-27871	NN	denotes	525A
T24561	27871-27872	.	denotes	.
T24562	27872-27937	sentence	denotes	Total BCAA represent the sum of leucine, isoleucine, and valine.
T24563	27873-27878	JJ	denotes	Total
T24564	27879-27883	NN	denotes	BCAA
T24565	27884-27893	VBP	denotes	represent
T24566	27894-27897	DT	denotes	the
T24567	27898-27901	NN	denotes	sum
T24568	27902-27904	IN	denotes	of
T24569	27905-27912	NN	denotes	leucine
T24570	27912-27914	,	denotes	,
T24571	27914-27924	NN	denotes	isoleucine
T24572	27924-27926	,	denotes	,
T24573	27926-27929	CC	denotes	and
T24574	27930-27936	NN	denotes	valine
T24575	27936-27937	.	denotes	.
T24576	27937-28046	sentence	denotes	BCAA/alanine values were significantly greater for Lines A and 525A compared to controls at all ages tested.
T24577	27938-27942	NN	denotes	BCAA
T24579	27942-27943	HYPH	denotes	/
T24578	27943-27950	NN	denotes	alanine
T24580	27951-27957	NNS	denotes	values
T24581	27958-27962	VBD	denotes	were
T24582	27963-27976	RB	denotes	significantly
T24583	27977-27984	JJR	denotes	greater
T24584	27985-27988	IN	denotes	for
T24585	27989-27994	NNS	denotes	Lines
T24586	27995-27996	NN	denotes	A
T24587	27997-28000	CC	denotes	and
T24588	28001-28005	NN	denotes	525A
T24589	28006-28014	VBN	denotes	compared
T24590	28015-28017	IN	denotes	to
T24591	28018-28026	NNS	denotes	controls
T24592	28027-28029	IN	denotes	at
T24593	28030-28033	DT	denotes	all
T24594	28034-28038	NNS	denotes	ages
T24595	28039-28045	VBN	denotes	tested
T24596	28045-28046	.	denotes	.
T24597	28046-28113	sentence	denotes	The numbers on the bars indicates the numbers of samples analyzed.
T24598	28047-28050	DT	denotes	The
T24599	28051-28058	NNS	denotes	numbers
T24601	28059-28061	IN	denotes	on
T24602	28062-28065	DT	denotes	the
T24603	28066-28070	NNS	denotes	bars
T24600	28071-28080	VBZ	denotes	indicates
T24604	28081-28084	DT	denotes	the
T24605	28085-28092	NNS	denotes	numbers
T24606	28093-28095	IN	denotes	of
T24607	28096-28103	NNS	denotes	samples
T24608	28104-28112	VBN	denotes	analyzed
T24609	28112-28113	.	denotes	.
T24610	28113-28141	sentence	denotes	, P ≤ 0.01; *, P < 0.005.
T24611	28114-28115	SYM	denotes	*
T24613	28115-28117	,	denotes	,
T24614	28117-28118	NN	denotes	P
T24615	28119-28120	SYM	denotes	≤
T24612	28121-28125	CD	denotes	0.01
T24617	28125-28126	:	denotes	;
T24618	28127-28129	SYM	denotes	**
T24619	28129-28131	,	denotes	,
T24620	28131-28132	NN	denotes	P
T24621	28133-28134	SYM	denotes	<
T24616	28135-28140	CD	denotes	0.005
T24622	28140-28141	.	denotes	.
T24623	28141-28213	sentence	denotes	C, BCKDH enzyme activity in liver of control and Lines A and 525A mice.
T24624	28142-28143	LS	denotes	C
T24626	28143-28145	,	denotes	,
T24627	28145-28150	NN	denotes	BCKDH
T24628	28151-28157	NN	denotes	enzyme
T24625	28158-28166	NN	denotes	activity
T24629	28167-28169	IN	denotes	in
T24630	28170-28175	NN	denotes	liver
T24631	28176-28178	IN	denotes	of
T24632	28179-28186	NN	denotes	control
T24634	28187-28190	CC	denotes	and
T24635	28191-28196	NNS	denotes	Lines
T24636	28197-28198	NN	denotes	A
T24637	28199-28202	CC	denotes	and
T24638	28203-28207	NN	denotes	525A
T24633	28208-28212	NNS	denotes	mice
T24639	28212-28213	.	denotes	.
T24640	28213-28243	sentence	denotes	, P ≤ 0.001; *, P ≤ 0.0001.
T24641	28214-28215	SYM	denotes	*
T24643	28215-28217	,	denotes	,
T24644	28217-28218	NN	denotes	P
T24645	28219-28220	SYM	denotes	≤
T24642	28221-28226	CD	denotes	0.001
T24647	28226-28227	:	denotes	;
T24648	28228-28230	SYM	denotes	**
T24649	28230-28232	,	denotes	,
T24650	28232-28233	NN	denotes	P
T24651	28234-28235	SYM	denotes	≤
T24646	28236-28242	CD	denotes	0.0001
T24652	28242-28243	.	denotes	.
T24653	28243-28294	sentence	denotes	D, Survival curves for mice from Lines A and 525A.
T24654	28244-28245	LS	denotes	D
T24656	28245-28247	,	denotes	,
T24657	28247-28255	NN	denotes	Survival
T24655	28256-28262	NNS	denotes	curves
T24658	28263-28266	IN	denotes	for
T24659	28267-28271	NNS	denotes	mice
T24660	28272-28276	IN	denotes	from
T24661	28277-28282	NNS	denotes	Lines
T24662	28283-28284	NN	denotes	A
T24663	28285-28288	CC	denotes	and
T24664	28289-28293	NN	denotes	525A
T24665	28293-28294	.	denotes	.
T24666	28294-28426	sentence	denotes	Rescue mice alive at weaning were monitored until they were moribund and subsequently sacrificed or were found dead in their cages.
T24667	28295-28301	NN	denotes	Rescue
T24668	28302-28306	NNS	denotes	mice
T24670	28307-28312	JJ	denotes	alive
T24671	28313-28315	IN	denotes	at
T24672	28316-28323	NN	denotes	weaning
T24673	28324-28328	VBD	denotes	were
T24669	28329-28338	VBN	denotes	monitored
T24674	28339-28344	IN	denotes	until
T24676	28345-28349	PRP	denotes	they
T24675	28350-28354	VBD	denotes	were
T24677	28355-28363	JJ	denotes	moribund
T24678	28364-28367	CC	denotes	and
T24679	28368-28380	RB	denotes	subsequently
T24680	28381-28391	VBN	denotes	sacrificed
T24681	28392-28394	CC	denotes	or
T24682	28395-28399	VBD	denotes	were
T24683	28400-28405	VBN	denotes	found
T24684	28406-28410	JJ	denotes	dead
T24685	28411-28413	IN	denotes	in
T24686	28414-28419	PRP$	denotes	their
T24687	28420-28425	NNS	denotes	cages
T24688	28425-28426	.	denotes	.
T16337	28427-28428	sentence	denotes	L
T16338	28427-28428	NN	denotes	L
T16340	28428-28429	VBZ	denotes	i
T16339	28428-28433	sentence	denotes	ines
T16341	28429-28430	NN	denotes	n
T16342	28430-28431	NN	denotes	e
T16343	28431-28432	VBZ	denotes	s
T16345	28433-28434	NN	denotes	A
T16344	28433-28443	sentence	denotes	A and 525A
T16346	28435-28437	NN	denotes	an
T16347	28437-28438	NN	denotes	d
T16348	28439-28443	NN	denotes	525A
T16349	28443-28444	sentence	denotes
T16351	28444-28448	VBD	denotes	were
T16350	28444-28453	sentence	denotes	were sele
T16352	28449-28450	RB	denotes	s
T16353	28450-28451	JJR	denotes	e
T16354	28451-28452	NNS	denotes	l
T16355	28452-28453	NNS	denotes	e
T16357	28453-28456	VBZ	denotes	cte
T16356	28453-28457	sentence	denotes	cted
T16358	28456-28457	VBN	denotes	d
T16359	28457-28458	sentence	denotes
T16361	28458-28459	IN	denotes	f
T16360	28458-28464	sentence	denotes	for de
T16362	28459-28461	NN	denotes	or
T16363	28462-28463	CC	denotes	d
T16364	28463-28464	NNS	denotes	e
T16366	28464-28465	IN	denotes	t
T16365	28464-28474	sentence	denotes	tailed cha
T16367	28465-28467	NN	denotes	ai
T16368	28467-28468	IN	denotes	l
T16369	28468-28469	PRP	denotes	e
T16370	28469-28470	JJ	denotes	d
T16371	28471-28472	VBN	denotes	c
T16372	28472-28473	PRP$	denotes	h
T16373	28473-28474	NNS	denotes	a
T16375	28474-28475	VBD	denotes	r
T16374	28474-28488	sentence	denotes	racterization.
T16376	28475-28476	VBN	denotes	a
T16377	28476-28487	NN	denotes	cterization
T16378	28487-28488	.	denotes	.
T16379	28488-28590	sentence	denotes	Fig. 4B shows the results of blood amino acid analysis presented as a ratio of total BCAA to alanine.
T16380	28489-28493	NN	denotes	Fig.
T16381	28494-28496	NN	denotes	4B
T16382	28497-28502	VBZ	denotes	shows
T16383	28503-28506	DT	denotes	the
T16384	28507-28514	NNS	denotes	results
T16385	28515-28517	IN	denotes	of
T16386	28518-28523	NN	denotes	blood
T16388	28524-28529	NN	denotes	amino
T16389	28530-28534	NN	denotes	acid
T16387	28535-28543	NN	denotes	analysis
T16390	28544-28553	VBN	denotes	presented
T16391	28554-28556	IN	denotes	as
T16392	28557-28558	DT	denotes	a
T16393	28559-28564	NN	denotes	ratio
T16394	28565-28567	IN	denotes	of
T16395	28568-28573	JJ	denotes	total
T16396	28574-28578	NN	denotes	BCAA
T16397	28579-28581	IN	denotes	to
T16398	28582-28589	NN	denotes	alanine
T16399	28589-28590	.	denotes	.
T16400	28590-28725	sentence	denotes	The BCAA/alanine values for mice from both lines were significantly elevated compared to controls for all of the time points analyzed.
T16401	28591-28594	DT	denotes	The
T16403	28595-28599	NN	denotes	BCAA
T16405	28599-28600	HYPH	denotes	/
T16404	28600-28607	NN	denotes	alanine
T16402	28608-28614	NNS	denotes	values
T16407	28615-28618	IN	denotes	for
T16408	28619-28623	NNS	denotes	mice
T16409	28624-28628	IN	denotes	from
T16410	28629-28633	DT	denotes	both
T16411	28634-28639	NNS	denotes	lines
T16406	28640-28644	VBD	denotes	were
T16412	28645-28658	RB	denotes	significantly
T16413	28659-28667	JJ	denotes	elevated
T16414	28668-28676	VBN	denotes	compared
T16415	28677-28679	IN	denotes	to
T16416	28680-28688	NNS	denotes	controls
T16417	28689-28692	IN	denotes	for
T16418	28693-28696	DT	denotes	all
T16419	28697-28699	IN	denotes	of
T16420	28700-28703	DT	denotes	the
T16422	28704-28708	NN	denotes	time
T16421	28709-28715	NNS	denotes	points
T16423	28716-28724	VBN	denotes	analyzed
T16424	28724-28725	.	denotes	.
T16425	28725-28848	sentence	denotes	Note that the BCAA/alanine values for the transgenic mice are intermediate between controls and knockouts (see Figure 2C).
T16426	28726-28730	VB	denotes	Note
T16427	28731-28735	IN	denotes	that
T16429	28736-28739	DT	denotes	the
T16431	28740-28744	NN	denotes	BCAA
T16433	28744-28745	HYPH	denotes	/
T16432	28745-28752	NN	denotes	alanine
T16430	28753-28759	NNS	denotes	values
T16434	28760-28763	IN	denotes	for
T16435	28764-28767	DT	denotes	the
T16437	28768-28778	JJ	denotes	transgenic
T16436	28779-28783	NNS	denotes	mice
T16428	28784-28787	VBP	denotes	are
T16438	28788-28800	JJ	denotes	intermediate
T16439	28801-28808	IN	denotes	between
T16440	28809-28817	NNS	denotes	controls
T16441	28818-28821	CC	denotes	and
T16442	28822-28831	NNS	denotes	knockouts
T16443	28832-28833	-LRB-	denotes	(
T16444	28833-28836	VB	denotes	see
T16445	28837-28843	NN	denotes	Figure
T16446	28844-28846	NN	denotes	2C
T16447	28846-28847	-RRB-	denotes	)
T16448	28847-28848	.	denotes	.
T16449	28848-28995	sentence	denotes	Blood levels of alanine, glutamate, and glutamine were reduced in Line A mice, as was glutamate in Line 525A mice, compared to controls (Table 2).
T16450	28849-28854	NN	denotes	Blood
T16451	28855-28861	NNS	denotes	levels
T16453	28862-28864	IN	denotes	of
T16454	28865-28872	NN	denotes	alanine
T16455	28872-28874	,	denotes	,
T16456	28874-28883	NN	denotes	glutamate
T16457	28883-28885	,	denotes	,
T16458	28885-28888	CC	denotes	and
T16459	28889-28898	NN	denotes	glutamine
T16460	28899-28903	VBD	denotes	were
T16452	28904-28911	VBN	denotes	reduced
T16461	28912-28914	IN	denotes	in
T16462	28915-28919	NN	denotes	Line
T16463	28920-28921	NN	denotes	A
T16464	28922-28926	NNS	denotes	mice
T16465	28926-28928	,	denotes	,
T16466	28928-28930	IN	denotes	as
T16467	28931-28934	VBD	denotes	was
T16468	28935-28944	NN	denotes	glutamate
T16469	28945-28947	IN	denotes	in
T16470	28948-28952	NN	denotes	Line
T16471	28953-28957	NN	denotes	525A
T16472	28958-28962	NNS	denotes	mice
T16473	28962-28964	,	denotes	,
T16474	28964-28972	VBN	denotes	compared
T16475	28973-28975	IN	denotes	to
T16476	28976-28984	NNS	denotes	controls
T16477	28985-28986	-LRB-	denotes	(
T16478	28986-28991	NN	denotes	Table
T16479	28992-28993	CD	denotes	2
T16480	28993-28994	-RRB-	denotes	)
T16481	28994-28995	.	denotes	.
T16482	28995-28996	sentence	denotes
T24914	29005-29012	NN	denotes	Summary
T24915	29013-29015	IN	denotes	of
T24916	29016-29026	JJ	denotes	additional
T24918	29027-29032	NN	denotes	blood
T24919	29033-29038	NN	denotes	amino
T24920	29039-29043	NN	denotes	acid
T24917	29044-29050	NNS	denotes	levels
T24921	29051-29053	IN	denotes	in
T24922	29054-29057	DT	denotes	the
T24924	29058-29070	JJ	denotes	intermediate
T24925	29071-29075	NN	denotes	MSUD
T24923	29076-29081	NN	denotes	model
T24926	29082-29085	CC	denotes	and
T24927	29086-29096	NN	denotes	littermate
T24928	29097-29105	NNS	denotes	controls
T24929	29106-29108	IN	denotes	as
T24930	29109-29119	VBN	denotes	determined
T24931	29120-29122	IN	denotes	by
T24932	29123-29129	JJ	denotes	tandem
T24934	29130-29134	NN	denotes	mass
T24933	29135-29147	NN	denotes	spectrometry
T24935	29147-29148	.	denotes	.
T24936	29148-29209	sentence	denotes	Samples were collected from mice that were 4–6 weeks of age.
T24937	29149-29156	NNS	denotes	Samples
T24939	29157-29161	VBD	denotes	were
T24938	29162-29171	VBN	denotes	collected
T24940	29172-29176	IN	denotes	from
T24941	29177-29181	NNS	denotes	mice
T24942	29182-29186	WDT	denotes	that
T24943	29187-29191	VBD	denotes	were
T24944	29192-29193	CD	denotes	4
T24946	29193-29194	SYM	denotes	–
T24945	29194-29195	CD	denotes	6
T24947	29196-29201	NNS	denotes	weeks
T24948	29202-29204	IN	denotes	of
T24949	29205-29208	NN	denotes	age
T24950	29208-29209	.	denotes	.
T24951	29209-29238	sentence	denotes	All values are mean +/- SEM.
T24952	29210-29213	DT	denotes	All
T24953	29214-29220	NNS	denotes	values
T24954	29221-29224	VBP	denotes	are
T24955	29225-29229	NN	denotes	mean
T24956	29230-29231	SYM	denotes	+
T24957	29231-29232	HYPH	denotes	/
T24958	29232-29233	SYM	denotes	-
T24959	29234-29237	NN	denotes	SEM
T24960	29237-29238	.	denotes	.
T16484	29642-29649	IN	denotes	Because
T16483	29642-29890	sentence	denotes	Because the LAP-tTA mice that were used to drive expression of transgenic human E2 have been demonstrated to produce liver specific expression [34], BCKDH enzyme activity and production of human E2 protein in liver of Lines A and 525A was examined.
T16486	29650-29653	DT	denotes	the
T16488	29654-29657	NN	denotes	LAP
T16490	29657-29658	HYPH	denotes	-
T16489	29658-29661	NN	denotes	tTA
T16487	29662-29666	NNS	denotes	mice
T16491	29667-29671	WDT	denotes	that
T16493	29672-29676	VBD	denotes	were
T16492	29677-29681	VBN	denotes	used
T16494	29682-29684	TO	denotes	to
T16495	29685-29690	VB	denotes	drive
T16496	29691-29701	NN	denotes	expression
T16497	29702-29704	IN	denotes	of
T16498	29705-29715	JJ	denotes	transgenic
T16500	29716-29721	JJ	denotes	human
T16499	29722-29724	NN	denotes	E2
T16501	29725-29729	VBP	denotes	have
T16502	29730-29734	VBN	denotes	been
T16485	29735-29747	VBN	denotes	demonstrated
T16504	29748-29750	TO	denotes	to
T16505	29751-29758	VB	denotes	produce
T16506	29759-29764	NN	denotes	liver
T16507	29765-29773	JJ	denotes	specific
T16508	29774-29784	NN	denotes	expression
T16509	29785-29786	-LRB-	denotes	[
T16510	29786-29788	CD	denotes	34
T16511	29788-29789	-RRB-	denotes	]
T16512	29789-29791	,	denotes	,
T16513	29791-29796	NN	denotes	BCKDH
T16515	29797-29803	NN	denotes	enzyme
T16514	29804-29812	NN	denotes	activity
T16516	29813-29816	CC	denotes	and
T16517	29817-29827	NN	denotes	production
T16518	29828-29830	IN	denotes	of
T16519	29831-29836	JJ	denotes	human
T16521	29837-29839	NN	denotes	E2
T16520	29840-29847	NN	denotes	protein
T16522	29848-29850	IN	denotes	in
T16523	29851-29856	NN	denotes	liver
T16524	29857-29859	IN	denotes	of
T16525	29860-29865	NNS	denotes	Lines
T16526	29866-29867	NN	denotes	A
T16527	29868-29871	CC	denotes	and
T16528	29872-29876	NN	denotes	525A
T16529	29877-29880	VBD	denotes	was
T16503	29881-29889	VBN	denotes	examined
T16530	29889-29890	.	denotes	.
T16531	29890-30037	sentence	denotes	BCKDH enzymatic activity in liver from Lines A and 525A was ~6 and 5%, respectively, of the enzymatic activity present in control liver (Fig. 4C).
T16532	29891-29896	NN	denotes	BCKDH
T16534	29897-29906	JJ	denotes	enzymatic
T16533	29907-29915	NN	denotes	activity
T16536	29916-29918	IN	denotes	in
T16537	29919-29924	NN	denotes	liver
T16538	29925-29929	IN	denotes	from
T16539	29930-29935	NNS	denotes	Lines
T16540	29936-29937	NN	denotes	A
T16541	29938-29941	CC	denotes	and
T16542	29942-29946	NN	denotes	525A
T16535	29947-29950	VBD	denotes	was
T16543	29951-29952	SYM	denotes	~
T16544	29952-29953	CD	denotes	6
T16546	29954-29957	CC	denotes	and
T16547	29958-29959	CD	denotes	5
T16545	29959-29960	NN	denotes	%
T16548	29960-29962	,	denotes	,
T16549	29962-29974	RB	denotes	respectively
T16550	29974-29976	,	denotes	,
T16551	29976-29978	IN	denotes	of
T16552	29979-29982	DT	denotes	the
T16554	29983-29992	JJ	denotes	enzymatic
T16553	29993-30001	NN	denotes	activity
T16555	30002-30009	JJ	denotes	present
T16556	30010-30012	IN	denotes	in
T16557	30013-30020	NN	denotes	control
T16558	30021-30026	NN	denotes	liver
T16559	30027-30028	-LRB-	denotes	(
T16561	30028-30032	NN	denotes	Fig.
T16560	30033-30035	NN	denotes	4C
T16562	30035-30036	-RRB-	denotes	)
T16563	30036-30037	.	denotes	.
T16564	30037-30160	sentence	denotes	As shown in Fig. 3B, the amount of human E2 (predicted MW~54 Kd) in these transgenic mice was quite variable between mice.
T16565	30038-30040	IN	denotes	As
T16566	30041-30046	VBN	denotes	shown
T16568	30047-30049	IN	denotes	in
T16569	30050-30054	NNP	denotes	Fig.
T16570	30055-30057	NN	denotes	3B
T16571	30057-30059	,	denotes	,
T16572	30059-30062	DT	denotes	the
T16573	30063-30069	NN	denotes	amount
T16574	30070-30072	IN	denotes	of
T16575	30073-30078	JJ	denotes	human
T16576	30079-30081	NN	denotes	E2
T16577	30082-30083	-LRB-	denotes	(
T16579	30083-30092	VBN	denotes	predicted
T16580	30093-30095	NN	denotes	MW
T16581	30095-30096	SYM	denotes	~
T16582	30096-30098	NN	denotes	54
T16578	30099-30101	NN	denotes	Kd
T16583	30101-30102	-RRB-	denotes	)
T16584	30103-30105	IN	denotes	in
T16585	30106-30111	DT	denotes	these
T16587	30112-30122	JJ	denotes	transgenic
T16586	30123-30127	NNS	denotes	mice
T16567	30128-30131	VBD	denotes	was
T16588	30132-30137	RB	denotes	quite
T16589	30138-30146	JJ	denotes	variable
T16590	30147-30154	IN	denotes	between
T16591	30155-30159	NNS	denotes	mice
T16592	30159-30160	.	denotes	.
T16593	30160-30324	sentence	denotes	In some of these transgenic mice, the level of human E2 was approximately equal to the amount of mouse E2 (~47 Kd) produced in liver of nontransgenic control mice.
T16594	30161-30163	IN	denotes	In
T16596	30164-30168	DT	denotes	some
T16597	30169-30171	IN	denotes	of
T16598	30172-30177	DT	denotes	these
T16600	30178-30188	JJ	denotes	transgenic
T16599	30189-30193	NNS	denotes	mice
T16601	30193-30195	,	denotes	,
T16602	30195-30198	DT	denotes	the
T16603	30199-30204	NN	denotes	level
T16604	30205-30207	IN	denotes	of
T16605	30208-30213	JJ	denotes	human
T16606	30214-30216	NN	denotes	E2
T16595	30217-30220	VBD	denotes	was
T16607	30221-30234	RB	denotes	approximately
T16608	30235-30240	JJ	denotes	equal
T16609	30241-30243	IN	denotes	to
T16610	30244-30247	DT	denotes	the
T16611	30248-30254	NN	denotes	amount
T16612	30255-30257	IN	denotes	of
T16613	30258-30263	NN	denotes	mouse
T16614	30264-30266	NN	denotes	E2
T16615	30267-30268	-LRB-	denotes	(
T16617	30268-30269	SYM	denotes	~
T16618	30269-30271	CD	denotes	47
T16616	30272-30274	NN	denotes	Kd
T16619	30274-30275	-RRB-	denotes	)
T16620	30276-30284	VBN	denotes	produced
T16621	30285-30287	IN	denotes	in
T16622	30288-30293	NN	denotes	liver
T16623	30294-30296	IN	denotes	of
T16624	30297-30310	JJ	denotes	nontransgenic
T16626	30311-30318	NN	denotes	control
T16625	30319-30323	NNS	denotes	mice
T16627	30323-30324	.	denotes	.
T16628	30324-30511	sentence	denotes	The observations that these near normal amounts of E2 protein result in only ~5–6% of normal BCKDH enzyme activity suggest that transgene derived E2 is functioning at a suboptimal level.
T16629	30325-30328	DT	denotes	The
T16630	30329-30341	NNS	denotes	observations
T16632	30342-30346	IN	denotes	that
T16634	30347-30352	DT	denotes	these
T16636	30353-30357	RB	denotes	near
T16637	30358-30364	JJ	denotes	normal
T16635	30365-30372	NNS	denotes	amounts
T16638	30373-30375	IN	denotes	of
T16639	30376-30378	NN	denotes	E2
T16640	30379-30386	NN	denotes	protein
T16633	30387-30393	VBP	denotes	result
T16641	30394-30396	IN	denotes	in
T16642	30397-30401	RB	denotes	only
T16644	30402-30403	SYM	denotes	~
T16645	30403-30404	CD	denotes	5
T16646	30404-30405	SYM	denotes	–
T16643	30405-30406	CD	denotes	6
T16647	30406-30407	NN	denotes	%
T16648	30408-30410	IN	denotes	of
T16649	30411-30417	JJ	denotes	normal
T16651	30418-30423	NN	denotes	BCKDH
T16652	30424-30430	NN	denotes	enzyme
T16650	30431-30439	NN	denotes	activity
T16631	30440-30447	VBP	denotes	suggest
T16653	30448-30452	IN	denotes	that
T16655	30453-30462	NN	denotes	transgene
T16657	30463-30470	VBN	denotes	derived
T16656	30471-30473	NN	denotes	E2
T16658	30474-30476	VBZ	denotes	is
T16654	30477-30488	VBG	denotes	functioning
T16659	30489-30491	IN	denotes	at
T16660	30492-30493	DT	denotes	a
T16662	30494-30504	JJ	denotes	suboptimal
T16661	30505-30510	NN	denotes	level
T16663	30510-30511	.	denotes	.
T16664	30511-30639	sentence	denotes	It is probable that the c-myc tag at the carboxy terminus of the transgene derived human E2 interfered with enzymatic activity.
T16665	30512-30514	PRP	denotes	It
T16666	30515-30517	VBZ	denotes	is
T16667	30518-30526	JJ	denotes	probable
T16668	30527-30531	IN	denotes	that
T16670	30532-30535	DT	denotes	the
T16672	30536-30537	NN	denotes	c
T16674	30537-30538	HYPH	denotes	-
T16673	30538-30541	NN	denotes	myc
T16671	30542-30545	NN	denotes	tag
T16675	30546-30548	IN	denotes	at
T16676	30549-30552	DT	denotes	the
T16678	30553-30560	NN	denotes	carboxy
T16677	30561-30569	NN	denotes	terminus
T16679	30570-30572	IN	denotes	of
T16680	30573-30576	DT	denotes	the
T16682	30577-30586	NN	denotes	transgene
T16683	30587-30594	VBN	denotes	derived
T16684	30595-30600	JJ	denotes	human
T16681	30601-30603	NN	denotes	E2
T16669	30604-30614	VBD	denotes	interfered
T16685	30615-30619	IN	denotes	with
T16686	30620-30629	JJ	denotes	enzymatic
T16687	30630-30638	NN	denotes	activity
T16688	30638-30639	.	denotes	.
T16689	30639-30977	sentence	denotes	This interpretation is consistent with previous studies which have revealed that the carboxy terminus of an analogous subunit of the pyruvate dehydrogenase complex is essential for subunit interactions [35] and insertion of a Hisx6 tag on the carboxy terminus of an analogous E. coli subunit interfered with normal subunit assembly [36].
T16690	30640-30644	DT	denotes	This
T16691	30645-30659	NN	denotes	interpretation
T16692	30660-30662	VBZ	denotes	is
T16693	30663-30673	JJ	denotes	consistent
T16694	30674-30678	IN	denotes	with
T16695	30679-30687	JJ	denotes	previous
T16696	30688-30695	NNS	denotes	studies
T16697	30696-30701	WDT	denotes	which
T16699	30702-30706	VBP	denotes	have
T16698	30707-30715	VBN	denotes	revealed
T16700	30716-30720	IN	denotes	that
T16702	30721-30724	DT	denotes	the
T16704	30725-30732	NN	denotes	carboxy
T16703	30733-30741	NN	denotes	terminus
T16705	30742-30744	IN	denotes	of
T16706	30745-30747	DT	denotes	an
T16708	30748-30757	JJ	denotes	analogous
T16707	30758-30765	NN	denotes	subunit
T16709	30766-30768	IN	denotes	of
T16710	30769-30772	DT	denotes	the
T16712	30773-30781	NN	denotes	pyruvate
T16713	30782-30795	NN	denotes	dehydrogenase
T16711	30796-30803	NN	denotes	complex
T16701	30804-30806	VBZ	denotes	is
T16714	30807-30816	JJ	denotes	essential
T16715	30817-30820	IN	denotes	for
T16716	30821-30828	NN	denotes	subunit
T16717	30829-30841	NNS	denotes	interactions
T16718	30842-30843	-LRB-	denotes	[
T16719	30843-30845	CD	denotes	35
T16720	30845-30846	-RRB-	denotes	]
T16721	30847-30850	CC	denotes	and
T16722	30851-30860	NN	denotes	insertion
T16724	30861-30863	IN	denotes	of
T16725	30864-30865	DT	denotes	a
T16727	30866-30871	NN	denotes	Hisx6
T16726	30872-30875	NN	denotes	tag
T16728	30876-30878	IN	denotes	on
T16729	30879-30882	DT	denotes	the
T16731	30883-30890	NN	denotes	carboxy
T16730	30891-30899	NN	denotes	terminus
T16732	30900-30902	IN	denotes	of
T16733	30903-30905	DT	denotes	an
T16735	30906-30915	JJ	denotes	analogous
T16736	30916-30918	FW	denotes	E.
T16737	30919-30923	FW	denotes	coli
T16734	30924-30931	NN	denotes	subunit
T16723	30932-30942	VBD	denotes	interfered
T16738	30943-30947	IN	denotes	with
T16739	30948-30954	JJ	denotes	normal
T16741	30955-30962	NN	denotes	subunit
T16740	30963-30971	NN	denotes	assembly
T16742	30972-30973	-LRB-	denotes	[
T16743	30973-30975	CD	denotes	36
T16744	30975-30976	-RRB-	denotes	]
T16745	30976-30977	.	denotes	.
T16746	30977-31082	sentence	denotes	It is also possible that human E2 was not fully functional when complexed with mouse E1 and E3 subunits.
T16747	30978-30980	PRP	denotes	It
T16748	30981-30983	VBZ	denotes	is
T16749	30984-30988	RB	denotes	also
T16750	30989-30997	JJ	denotes	possible
T16751	30998-31002	IN	denotes	that
T16753	31003-31008	JJ	denotes	human
T16754	31009-31011	NN	denotes	E2
T16752	31012-31015	VBD	denotes	was
T16755	31016-31019	RB	denotes	not
T16756	31020-31025	RB	denotes	fully
T16757	31026-31036	JJ	denotes	functional
T16758	31037-31041	WRB	denotes	when
T16759	31042-31051	VBN	denotes	complexed
T16760	31052-31056	IN	denotes	with
T16761	31057-31062	NN	denotes	mouse
T16762	31063-31065	NN	denotes	E1
T16763	31066-31069	CC	denotes	and
T16764	31070-31072	NN	denotes	E3
T16765	31073-31081	NNS	denotes	subunits
T16766	31081-31082	.	denotes	.
T16767	31082-31149	sentence	denotes	Human E2 shares ~88% identity to mouse E2 at the amino acid level.
T16768	31083-31088	JJ	denotes	Human
T16769	31089-31091	NN	denotes	E2
T16770	31092-31098	VBZ	denotes	shares
T16771	31099-31100	SYM	denotes	~
T16772	31100-31102	CD	denotes	88
T16773	31102-31103	NN	denotes	%
T16774	31104-31112	NN	denotes	identity
T16775	31113-31115	IN	denotes	to
T16776	31116-31121	NN	denotes	mouse
T16777	31122-31124	NN	denotes	E2
T16778	31125-31127	IN	denotes	at
T16779	31128-31131	DT	denotes	the
T16781	31132-31137	NN	denotes	amino
T16782	31138-31142	NN	denotes	acid
T16780	31143-31148	NN	denotes	level
T16783	31148-31149	.	denotes	.
T16784	31149-31255	sentence	denotes	We also used western blot analysis to analyze expression of the E2 transgene in brain, kidney and muscle.
T16785	31150-31152	PRP	denotes	We
T16787	31153-31157	RB	denotes	also
T16786	31158-31162	VBD	denotes	used
T16788	31163-31170	NNP	denotes	western
T16789	31171-31175	NN	denotes	blot
T16790	31176-31184	NN	denotes	analysis
T16791	31185-31187	TO	denotes	to
T16792	31188-31195	VB	denotes	analyze
T16793	31196-31206	NN	denotes	expression
T16794	31207-31209	IN	denotes	of
T16795	31210-31213	DT	denotes	the
T16797	31214-31216	NN	denotes	E2
T16796	31217-31226	NN	denotes	transgene
T16798	31227-31229	IN	denotes	in
T16799	31230-31235	NN	denotes	brain
T16800	31235-31237	,	denotes	,
T16801	31237-31243	NN	denotes	kidney
T16802	31244-31247	CC	denotes	and
T16803	31248-31254	NN	denotes	muscle
T16804	31254-31255	.	denotes	.
T16805	31255-31350	sentence	denotes	As shown in Figure 3C, D and 3E respectively, expression of E2 is negligible in those tissues.
T16806	31256-31258	IN	denotes	As
T16807	31259-31264	VBN	denotes	shown
T16809	31265-31267	IN	denotes	in
T16810	31268-31274	NN	denotes	Figure
T16811	31275-31277	NN	denotes	3C
T16812	31277-31279	,	denotes	,
T16813	31279-31280	NN	denotes	D
T16814	31281-31284	CC	denotes	and
T16815	31285-31287	NN	denotes	3E
T16816	31288-31300	RB	denotes	respectively
T16817	31300-31302	,	denotes	,
T16818	31302-31312	NN	denotes	expression
T16819	31313-31315	IN	denotes	of
T16820	31316-31318	NN	denotes	E2
T16808	31319-31321	VBZ	denotes	is
T16821	31322-31332	JJ	denotes	negligible
T16822	31333-31335	IN	denotes	in
T16823	31336-31341	DT	denotes	those
T16824	31342-31349	NNS	denotes	tissues
T16825	31349-31350	.	denotes	.
T16826	31350-31465	sentence	denotes	Long-term survival of the mice from these two transgenic lines that survived beyond weaning is plotted in Fig. 4D.
T16827	31351-31355	JJ	denotes	Long
T16829	31355-31356	HYPH	denotes	-
T16828	31356-31360	NN	denotes	term
T16830	31361-31369	NN	denotes	survival
T16832	31370-31372	IN	denotes	of
T16833	31373-31376	DT	denotes	the
T16834	31377-31381	NNS	denotes	mice
T16835	31382-31386	IN	denotes	from
T16836	31387-31392	DT	denotes	these
T16838	31393-31396	CD	denotes	two
T16839	31397-31407	JJ	denotes	transgenic
T16837	31408-31413	NNS	denotes	lines
T16840	31414-31418	WDT	denotes	that
T16841	31419-31427	VBD	denotes	survived
T16842	31428-31434	IN	denotes	beyond
T16843	31435-31442	NN	denotes	weaning
T16844	31443-31445	VBZ	denotes	is
T16831	31446-31453	VBN	denotes	plotted
T16845	31454-31456	IN	denotes	in
T16846	31457-31461	NN	denotes	Fig.
T16847	31462-31464	NN	denotes	4D
T16848	31464-31465	.	denotes	.
T16849	31465-31600	sentence	denotes	Although survival data for control animals was not collected, it is readily apparent that survival of the rescue mice was compromised.
T16850	31466-31474	IN	denotes	Although
T16852	31475-31483	NN	denotes	survival
T16853	31484-31488	NNS	denotes	data
T16854	31489-31492	IN	denotes	for
T16855	31493-31500	NN	denotes	control
T16856	31501-31508	NNS	denotes	animals
T16857	31509-31512	VBD	denotes	was
T16858	31513-31516	RB	denotes	not
T16851	31517-31526	VBN	denotes	collected
T16860	31526-31528	,	denotes	,
T16861	31528-31530	PRP	denotes	it
T16859	31531-31533	VBZ	denotes	is
T16862	31534-31541	RB	denotes	readily
T16863	31542-31550	JJ	denotes	apparent
T16864	31551-31555	IN	denotes	that
T16866	31556-31564	NN	denotes	survival
T16867	31565-31567	IN	denotes	of
T16868	31568-31571	DT	denotes	the
T16870	31572-31578	NN	denotes	rescue
T16869	31579-31583	NNS	denotes	mice
T16871	31584-31587	VBD	denotes	was
T16865	31588-31599	VBN	denotes	compromised
T16872	31599-31600	.	denotes	.
T16873	31600-31705	sentence	denotes	By 16 weeks, all mice of Line A were moribund and humanely sacrificed, or were found dead in their cage.
T16874	31601-31603	IN	denotes	By
T16876	31604-31606	CD	denotes	16
T16877	31607-31612	NNS	denotes	weeks
T16878	31612-31614	,	denotes	,
T16879	31614-31617	DT	denotes	all
T16880	31618-31622	NNS	denotes	mice
T16881	31623-31625	IN	denotes	of
T16882	31626-31630	NN	denotes	Line
T16883	31631-31632	NN	denotes	A
T16875	31633-31637	VBD	denotes	were
T16884	31638-31646	JJ	denotes	moribund
T16885	31647-31650	CC	denotes	and
T16886	31651-31659	RB	denotes	humanely
T16887	31660-31670	VBN	denotes	sacrificed
T16888	31670-31672	,	denotes	,
T16889	31672-31674	CC	denotes	or
T16890	31675-31679	VBD	denotes	were
T16891	31680-31685	VBN	denotes	found
T16892	31686-31690	JJ	denotes	dead
T16893	31691-31693	IN	denotes	in
T16894	31694-31699	PRP$	denotes	their
T16895	31700-31704	NN	denotes	cage
T16896	31704-31705	.	denotes	.
T16897	31705-31759	sentence	denotes	Survival of Line 525A appeared to be somewhat better.
T16898	31706-31714	NN	denotes	Survival
T16900	31715-31717	IN	denotes	of
T16901	31718-31722	NN	denotes	Line
T16902	31723-31727	NN	denotes	525A
T16899	31728-31736	VBD	denotes	appeared
T16903	31737-31739	TO	denotes	to
T16904	31740-31742	VB	denotes	be
T16905	31743-31751	RB	denotes	somewhat
T16906	31752-31758	JJR	denotes	better
T16907	31758-31759	.	denotes	.
T16908	31759-31820	sentence	denotes	At 20 weeks of age, ~12% of mice (2 of 16) were still alive.
T16909	31760-31762	IN	denotes	At
T16911	31763-31765	CD	denotes	20
T16912	31766-31771	NNS	denotes	weeks
T16913	31772-31774	IN	denotes	of
T16914	31775-31778	NN	denotes	age
T16915	31778-31780	,	denotes	,
T16916	31780-31781	SYM	denotes	~
T16917	31781-31783	CD	denotes	12
T16918	31783-31784	NN	denotes	%
T16919	31785-31787	IN	denotes	of
T16920	31788-31792	NNS	denotes	mice
T16921	31793-31794	-LRB-	denotes	(
T16922	31794-31795	CD	denotes	2
T16923	31796-31798	IN	denotes	of
T16924	31799-31801	CD	denotes	16
T16925	31801-31802	-RRB-	denotes	)
T16910	31803-31807	VBD	denotes	were
T16926	31808-31813	RB	denotes	still
T16927	31814-31819	JJ	denotes	alive
T16928	31819-31820	.	denotes	.
T16929	31820-31877	sentence	denotes	These two rare survivors died at 40 and 60 weeks of age.
T16930	31821-31826	DT	denotes	These
T16932	31827-31830	CD	denotes	two
T16933	31831-31835	JJ	denotes	rare
T16931	31836-31845	NNS	denotes	survivors
T16934	31846-31850	VBD	denotes	died
T16935	31851-31853	IN	denotes	at
T16936	31854-31856	CD	denotes	40
T16938	31857-31860	CC	denotes	and
T16939	31861-31863	CD	denotes	60
T16937	31864-31869	NNS	denotes	weeks
T16940	31870-31872	IN	denotes	of
T16941	31873-31876	NN	denotes	age
T16942	31876-31877	.	denotes	.
T16943	31877-32059	sentence	denotes	In summary, these surviving mice had BCAA/alanine ratios that were intermediate between controls and knockouts and they expressed ~5–6% of normal BCKDH enzyme activity in the liver.
T16944	31878-31880	IN	denotes	In
T16946	31881-31888	NN	denotes	summary
T16947	31888-31890	,	denotes	,
T16948	31890-31895	DT	denotes	these
T16950	31896-31905	VBG	denotes	surviving
T16949	31906-31910	NNS	denotes	mice
T16945	31911-31914	VBD	denotes	had
T16951	31915-31919	NN	denotes	BCAA
T16953	31919-31920	HYPH	denotes	/
T16952	31920-31927	NN	denotes	alanine
T16954	31928-31934	NNS	denotes	ratios
T16955	31935-31939	WDT	denotes	that
T16956	31940-31944	VBD	denotes	were
T16957	31945-31957	JJ	denotes	intermediate
T16958	31958-31965	IN	denotes	between
T16959	31966-31974	NNS	denotes	controls
T16960	31975-31978	CC	denotes	and
T16961	31979-31988	NNS	denotes	knockouts
T16962	31989-31992	CC	denotes	and
T16963	31993-31997	PRP	denotes	they
T16964	31998-32007	VBD	denotes	expressed
T16965	32008-32009	SYM	denotes	~
T16967	32009-32010	CD	denotes	5
T16968	32010-32011	SYM	denotes	–
T16966	32011-32012	CD	denotes	6
T16969	32012-32013	NN	denotes	%
T16970	32014-32016	IN	denotes	of
T16971	32017-32023	JJ	denotes	normal
T16973	32024-32029	NN	denotes	BCKDH
T16974	32030-32036	NN	denotes	enzyme
T16972	32037-32045	NN	denotes	activity
T16975	32046-32048	IN	denotes	in
T16976	32049-32052	DT	denotes	the
T16977	32053-32058	NN	denotes	liver
T16978	32058-32059	.	denotes	.
T16979	32059-32211	sentence	denotes	These phenotypic observations are remarkably similar to the clinical phenotype observed in MSUD patients with the intermediate form of the disease [1].
T16980	32060-32065	DT	denotes	These
T16982	32066-32076	JJ	denotes	phenotypic
T16981	32077-32089	NNS	denotes	observations
T16983	32090-32093	VBP	denotes	are
T16984	32094-32104	RB	denotes	remarkably
T16985	32105-32112	JJ	denotes	similar
T16986	32113-32115	IN	denotes	to
T16987	32116-32119	DT	denotes	the
T16989	32120-32128	JJ	denotes	clinical
T16988	32129-32138	NN	denotes	phenotype
T16990	32139-32147	VBN	denotes	observed
T16991	32148-32150	IN	denotes	in
T16992	32151-32155	NN	denotes	MSUD
T16993	32156-32164	NNS	denotes	patients
T16994	32165-32169	IN	denotes	with
T16995	32170-32173	DT	denotes	the
T16997	32174-32186	JJ	denotes	intermediate
T16996	32187-32191	NN	denotes	form
T16998	32192-32194	IN	denotes	of
T16999	32195-32198	DT	denotes	the
T17000	32199-32206	NN	denotes	disease
T17001	32207-32208	-LRB-	denotes	[
T17002	32208-32209	CD	denotes	1
T17003	32209-32210	-RRB-	denotes	]
T17004	32210-32211	.	denotes	.
T17005	32211-32301	sentence	denotes	Thus, these rescue mice represent a very useful model of the intermediate MSUD phenotype.
T17006	32212-32216	RB	denotes	Thus
T17008	32216-32218	,	denotes	,
T17009	32218-32223	DT	denotes	these
T17011	32224-32230	NN	denotes	rescue
T17010	32231-32235	NNS	denotes	mice
T17007	32236-32245	VBP	denotes	represent
T17012	32246-32247	DT	denotes	a
T17014	32248-32252	RB	denotes	very
T17015	32253-32259	JJ	denotes	useful
T17013	32260-32265	NN	denotes	model
T17016	32266-32268	IN	denotes	of
T17017	32269-32272	DT	denotes	the
T17019	32273-32285	JJ	denotes	intermediate
T17020	32286-32290	NN	denotes	MSUD
T17018	32291-32300	NN	denotes	phenotype
T17021	32300-32301	.	denotes	.
T19826	32314-32315	DT	denotes	A
T19828	32316-32321	JJ	denotes	major
T19827	32322-32328	NN	denotes	hurdle
T19830	32329-32331	IN	denotes	in
T19831	32332-32342	VBG	denotes	developing
T19832	32343-32346	JJ	denotes	new
T19833	32347-32357	NNS	denotes	treatments
T19834	32358-32361	IN	denotes	for
T19835	32362-32366	NN	denotes	MSUD
T19836	32367-32370	VBZ	denotes	has
T19829	32371-32375	VBN	denotes	been
T19837	32376-32379	DT	denotes	the
T19838	32380-32384	NN	denotes	lack
T19839	32385-32387	IN	denotes	of
T19840	32388-32389	DT	denotes	a
T19842	32390-32399	JJ	denotes	practical
T19843	32399-32401	,	denotes	,
T19844	32401-32409	JJ	denotes	accurate
T19845	32410-32416	NN	denotes	animal
T19841	32417-32422	NN	denotes	model
T19846	32423-32425	IN	denotes	of
T19847	32426-32429	DT	denotes	the
T19848	32430-32437	NN	denotes	disease
T19849	32437-32438	.	denotes	.
T19850	32438-32473	sentence	denotes	This hurdle has now been overcome.
T19851	32439-32443	DT	denotes	This
T19852	32444-32450	NN	denotes	hurdle
T19854	32451-32454	VBZ	denotes	has
T19855	32455-32458	RB	denotes	now
T19856	32459-32463	VBN	denotes	been
T19853	32464-32472	VBN	denotes	overcome
T19857	32472-32473	.	denotes	.
T19858	32473-32691	sentence	denotes	In this report, we describe the development and characterization of two genetically engineered mouse models that are phenotypically very similar to MSUD patients with the classic and intermediate forms of the disease.
T19859	32474-32476	IN	denotes	In
T19861	32477-32481	DT	denotes	this
T19862	32482-32488	NN	denotes	report
T19863	32488-32490	,	denotes	,
T19864	32490-32492	PRP	denotes	we
T19860	32493-32501	VBP	denotes	describe
T19865	32502-32505	DT	denotes	the
T19866	32506-32517	NN	denotes	development
T19867	32518-32521	CC	denotes	and
T19868	32522-32538	NN	denotes	characterization
T19869	32539-32541	IN	denotes	of
T19870	32542-32545	CD	denotes	two
T19872	32546-32557	RB	denotes	genetically
T19873	32558-32568	VBN	denotes	engineered
T19874	32569-32574	NN	denotes	mouse
T19871	32575-32581	NNS	denotes	models
T19875	32582-32586	WDT	denotes	that
T19876	32587-32590	VBP	denotes	are
T19877	32591-32605	RB	denotes	phenotypically
T19878	32606-32610	RB	denotes	very
T19879	32611-32618	JJ	denotes	similar
T19880	32619-32621	IN	denotes	to
T19881	32622-32626	NN	denotes	MSUD
T19882	32627-32635	NNS	denotes	patients
T19883	32636-32640	IN	denotes	with
T19884	32641-32644	DT	denotes	the
T19886	32645-32652	JJ	denotes	classic
T19887	32653-32656	CC	denotes	and
T19888	32657-32669	JJ	denotes	intermediate
T19885	32670-32675	NNS	denotes	forms
T19889	32676-32678	IN	denotes	of
T19890	32679-32682	DT	denotes	the
T19891	32683-32690	NN	denotes	disease
T19892	32690-32691	.	denotes	.
T19893	32691-32774	sentence	denotes	Our model of classic MSUD was created by gene knockout of the E2 subunit of BCKDH.
T19894	32692-32695	PRP$	denotes	Our
T19895	32696-32701	NN	denotes	model
T19897	32702-32704	IN	denotes	of
T19898	32705-32712	JJ	denotes	classic
T19899	32713-32717	NN	denotes	MSUD
T19900	32718-32721	VBD	denotes	was
T19896	32722-32729	VBN	denotes	created
T19901	32730-32732	IN	denotes	by
T19902	32733-32737	NN	denotes	gene
T19903	32738-32746	NN	denotes	knockout
T19904	32747-32749	IN	denotes	of
T19905	32750-32753	DT	denotes	the
T19907	32754-32756	NN	denotes	E2
T19906	32757-32764	NN	denotes	subunit
T19908	32765-32767	IN	denotes	of
T19909	32768-32773	NN	denotes	BCKDH
T19910	32773-32774	.	denotes	.
T19911	32774-32865	sentence	denotes	The phenotype of these knockout animals is strikingly similar to humans with classic MSUD.
T19912	32775-32778	DT	denotes	The
T19913	32779-32788	NN	denotes	phenotype
T19915	32789-32791	IN	denotes	of
T19916	32792-32797	DT	denotes	these
T19918	32798-32806	NN	denotes	knockout
T19917	32807-32814	NNS	denotes	animals
T19914	32815-32817	VBZ	denotes	is
T19919	32818-32828	RB	denotes	strikingly
T19920	32829-32836	JJ	denotes	similar
T19921	32837-32839	IN	denotes	to
T19922	32840-32846	NNS	denotes	humans
T19923	32847-32851	IN	denotes	with
T19924	32852-32859	JJ	denotes	classic
T19925	32860-32864	NN	denotes	MSUD
T19926	32864-32865	.	denotes	.
T19927	32865-32945	sentence	denotes	Knockout mice were born at the expected frequency and appeared normal at birth.
T19928	32866-32874	NN	denotes	Knockout
T19929	32875-32879	NNS	denotes	mice
T19931	32880-32884	VBD	denotes	were
T19930	32885-32889	VBN	denotes	born
T19932	32890-32892	IN	denotes	at
T19933	32893-32896	DT	denotes	the
T19935	32897-32905	VBN	denotes	expected
T19934	32906-32915	NN	denotes	frequency
T19936	32916-32919	CC	denotes	and
T19937	32920-32928	VBD	denotes	appeared
T19938	32929-32935	JJ	denotes	normal
T19939	32936-32938	IN	denotes	at
T19940	32939-32944	NN	denotes	birth
T19941	32944-32945	.	denotes	.
T19942	32945-33036	sentence	denotes	Within a day of birth and following suckling, blood levels of BCAA were markedly elevated.
T19943	32946-32952	IN	denotes	Within
T19945	32953-32954	DT	denotes	a
T19946	32955-32958	NN	denotes	day
T19947	32959-32961	IN	denotes	of
T19948	32962-32967	NN	denotes	birth
T19949	32968-32971	CC	denotes	and
T19950	32972-32981	VBG	denotes	following
T19951	32982-32990	NN	denotes	suckling
T19952	32990-32992	,	denotes	,
T19953	32992-32997	NN	denotes	blood
T19954	32998-33004	NNS	denotes	levels
T19955	33005-33007	IN	denotes	of
T19956	33008-33012	NN	denotes	BCAA
T19944	33013-33017	VBD	denotes	were
T19957	33018-33026	RB	denotes	markedly
T19958	33027-33035	JJ	denotes	elevated
T19959	33035-33036	.	denotes	.
T19960	33036-33196	sentence	denotes	Concomitantly, levels of the amino acids alanine, glutamate, and glutamine, whose synthesis is linked with normal metabolism of BCAA, were decreased (Table 1).
T19961	33037-33050	RB	denotes	Concomitantly
T19963	33050-33052	,	denotes	,
T19964	33052-33058	NNS	denotes	levels
T19965	33059-33061	IN	denotes	of
T19966	33062-33065	DT	denotes	the
T19968	33066-33071	NN	denotes	amino
T19967	33072-33077	NNS	denotes	acids
T19969	33078-33085	NN	denotes	alanine
T19970	33085-33087	,	denotes	,
T19971	33087-33096	NN	denotes	glutamate
T19972	33096-33098	,	denotes	,
T19973	33098-33101	CC	denotes	and
T19974	33102-33111	NN	denotes	glutamine
T19975	33111-33113	,	denotes	,
T19976	33113-33118	WP$	denotes	whose
T19977	33119-33128	NN	denotes	synthesis
T19979	33129-33131	VBZ	denotes	is
T19978	33132-33138	VBN	denotes	linked
T19980	33139-33143	IN	denotes	with
T19981	33144-33150	JJ	denotes	normal
T19982	33151-33161	NN	denotes	metabolism
T19983	33162-33164	IN	denotes	of
T19984	33165-33169	NN	denotes	BCAA
T19985	33169-33171	,	denotes	,
T19986	33171-33175	VBD	denotes	were
T19962	33176-33185	VBN	denotes	decreased
T19987	33186-33187	-LRB-	denotes	(
T19988	33187-33192	NN	denotes	Table
T19989	33193-33194	CD	denotes	1
T19990	33194-33195	-RRB-	denotes	)
T19991	33195-33196	.	denotes	.
T19992	33196-33335	sentence	denotes	The activity of BCKDH in livers of homozygous knockout mouse pups was undetectable, accounting for the accumulation of unmetabolized BCAA.
T19993	33197-33200	DT	denotes	The
T19994	33201-33209	NN	denotes	activity
T19996	33210-33212	IN	denotes	of
T19997	33213-33218	NN	denotes	BCKDH
T19998	33219-33221	IN	denotes	in
T19999	33222-33228	NNS	denotes	livers
T20000	33229-33231	IN	denotes	of
T20001	33232-33242	JJ	denotes	homozygous
T20002	33243-33251	NN	denotes	knockout
T20004	33252-33257	NN	denotes	mouse
T20003	33258-33262	NNS	denotes	pups
T19995	33263-33266	VBD	denotes	was
T20005	33267-33279	JJ	denotes	undetectable
T20006	33279-33281	,	denotes	,
T20007	33281-33291	VBG	denotes	accounting
T20008	33292-33295	IN	denotes	for
T20009	33296-33299	DT	denotes	the
T20010	33300-33312	NN	denotes	accumulation
T20011	33313-33315	IN	denotes	of
T20012	33316-33329	JJ	denotes	unmetabolized
T20013	33330-33334	NN	denotes	BCAA
T20014	33334-33335	.	denotes	.
T20015	33335-33417	sentence	denotes	Immunoreactive E2 protein was absent in liver and fibroblasts of homozygous pups.
T20016	33336-33350	JJ	denotes	Immunoreactive
T20018	33351-33353	NN	denotes	E2
T20017	33354-33361	NN	denotes	protein
T20019	33362-33365	VBD	denotes	was
T20020	33366-33372	JJ	denotes	absent
T20021	33373-33375	IN	denotes	in
T20022	33376-33381	NN	denotes	liver
T20023	33382-33385	CC	denotes	and
T20024	33386-33397	NNS	denotes	fibroblasts
T20025	33398-33400	IN	denotes	of
T20026	33401-33411	JJ	denotes	homozygous
T20027	33412-33416	NNS	denotes	pups
T20028	33416-33417	.	denotes	.
T20029	33417-33524	sentence	denotes	The phenotypic behavior of homozygous mouse pups resembles symptoms seen in newborn classic MSUD patients.
T20030	33418-33421	DT	denotes	The
T20032	33422-33432	JJ	denotes	phenotypic
T20031	33433-33441	NN	denotes	behavior
T20034	33442-33444	IN	denotes	of
T20035	33445-33455	JJ	denotes	homozygous
T20037	33456-33461	NN	denotes	mouse
T20036	33462-33466	NNS	denotes	pups
T20033	33467-33476	VBZ	denotes	resembles
T20038	33477-33485	NNS	denotes	symptoms
T20039	33486-33490	VBN	denotes	seen
T20040	33491-33493	IN	denotes	in
T20041	33494-33501	JJ	denotes	newborn
T20043	33502-33509	JJ	denotes	classic
T20044	33510-33514	NN	denotes	MSUD
T20042	33515-33523	NNS	denotes	patients
T20045	33523-33524	.	denotes	.
T20046	33524-33664	sentence	denotes	These include signs of neurologic dysfunction such as seizures, stupor, lethargy, loss of motor activity, and respiratory difficulties [1].
T20047	33525-33530	DT	denotes	These
T20048	33531-33538	VBP	denotes	include
T20049	33539-33544	NNS	denotes	signs
T20050	33545-33547	IN	denotes	of
T20051	33548-33558	JJ	denotes	neurologic
T20052	33559-33570	NN	denotes	dysfunction
T20053	33571-33575	JJ	denotes	such
T20054	33576-33578	IN	denotes	as
T20055	33579-33587	NNS	denotes	seizures
T20056	33587-33589	,	denotes	,
T20057	33589-33595	NN	denotes	stupor
T20058	33595-33597	,	denotes	,
T20059	33597-33605	NN	denotes	lethargy
T20060	33605-33607	,	denotes	,
T20061	33607-33611	NN	denotes	loss
T20062	33612-33614	IN	denotes	of
T20063	33615-33620	NN	denotes	motor
T20064	33621-33629	NN	denotes	activity
T20065	33629-33631	,	denotes	,
T20066	33631-33634	CC	denotes	and
T20067	33635-33646	JJ	denotes	respiratory
T20068	33647-33659	NNS	denotes	difficulties
T20069	33660-33661	-LRB-	denotes	[
T20070	33661-33662	CD	denotes	1
T20071	33662-33663	-RRB-	denotes	]
T20072	33663-33664	.	denotes	.
T20073	33664-33877	sentence	denotes	These neurologic symptoms may result from reduced levels of glutamate, glutamine, alanine, and other similar neuroactive amino acids, which are considered the culprit for MSUD encephalopathies in patients [1,37].
T20074	33665-33670	DT	denotes	These
T20076	33671-33681	JJ	denotes	neurologic
T20075	33682-33690	NNS	denotes	symptoms
T20078	33691-33694	MD	denotes	may
T20077	33695-33701	VB	denotes	result
T20079	33702-33706	IN	denotes	from
T20080	33707-33714	VBN	denotes	reduced
T20081	33715-33721	NNS	denotes	levels
T20082	33722-33724	IN	denotes	of
T20083	33725-33734	NN	denotes	glutamate
T20084	33734-33736	,	denotes	,
T20085	33736-33745	NN	denotes	glutamine
T20086	33745-33747	,	denotes	,
T20087	33747-33754	NN	denotes	alanine
T20088	33754-33756	,	denotes	,
T20089	33756-33759	CC	denotes	and
T20090	33760-33765	JJ	denotes	other
T20092	33766-33773	JJ	denotes	similar
T20093	33774-33785	JJ	denotes	neuroactive
T20094	33786-33791	NN	denotes	amino
T20091	33792-33797	NNS	denotes	acids
T20095	33797-33799	,	denotes	,
T20096	33799-33804	WDT	denotes	which
T20098	33805-33808	VBP	denotes	are
T20097	33809-33819	VBN	denotes	considered
T20099	33820-33823	DT	denotes	the
T20100	33824-33831	NN	denotes	culprit
T20101	33832-33835	IN	denotes	for
T20102	33836-33840	NN	denotes	MSUD
T20103	33841-33857	NNS	denotes	encephalopathies
T20104	33858-33860	IN	denotes	in
T20105	33861-33869	NNS	denotes	patients
T20106	33870-33871	-LRB-	denotes	[
T20108	33871-33872	CD	denotes	1
T20109	33872-33873	,	denotes	,
T20107	33873-33875	CD	denotes	37
T20110	33875-33876	-RRB-	denotes	]
T20111	33876-33877	.	denotes	.
T20112	33877-33951	sentence	denotes	Finally, nearly all of the homozygous pups died within 72 hours of birth.
T20113	33878-33885	RB	denotes	Finally
T20115	33885-33887	,	denotes	,
T20116	33887-33893	RB	denotes	nearly
T20117	33894-33897	DT	denotes	all
T20118	33898-33900	IN	denotes	of
T20119	33901-33904	DT	denotes	the
T20121	33905-33915	JJ	denotes	homozygous
T20120	33916-33920	NNS	denotes	pups
T20114	33921-33925	VBD	denotes	died
T20122	33926-33932	IN	denotes	within
T20123	33933-33935	CD	denotes	72
T20124	33936-33941	NNS	denotes	hours
T20125	33942-33944	IN	denotes	of
T20126	33945-33950	NN	denotes	birth
T20127	33950-33951	.	denotes	.
T20128	33951-34164	sentence	denotes	This neonatal lethality is likely due to the accumulation of BCAA to neurotoxic levels, ketoacidosis, brain edema, dehydration, and malnutrition as observed in the MSUD calf [17] and in classic MSUD patients [1].
T20129	33952-33956	DT	denotes	This
T20131	33957-33965	JJ	denotes	neonatal
T20130	33966-33975	NN	denotes	lethality
T20132	33976-33978	VBZ	denotes	is
T20133	33979-33985	RB	denotes	likely
T20134	33986-33989	IN	denotes	due
T20135	33990-33992	IN	denotes	to
T20136	33993-33996	DT	denotes	the
T20137	33997-34009	NN	denotes	accumulation
T20138	34010-34012	IN	denotes	of
T20139	34013-34017	NN	denotes	BCAA
T20140	34018-34020	IN	denotes	to
T20141	34021-34031	JJ	denotes	neurotoxic
T20142	34032-34038	NNS	denotes	levels
T20143	34038-34040	,	denotes	,
T20144	34040-34052	NN	denotes	ketoacidosis
T20145	34052-34054	,	denotes	,
T20146	34054-34059	NN	denotes	brain
T20147	34060-34065	NN	denotes	edema
T20148	34065-34067	,	denotes	,
T20149	34067-34078	NN	denotes	dehydration
T20150	34078-34080	,	denotes	,
T20151	34080-34083	CC	denotes	and
T20152	34084-34096	NN	denotes	malnutrition
T20153	34097-34099	IN	denotes	as
T20154	34100-34108	VBN	denotes	observed
T20155	34109-34111	IN	denotes	in
T20156	34112-34115	DT	denotes	the
T20158	34116-34120	NN	denotes	MSUD
T20157	34121-34125	NN	denotes	calf
T20159	34126-34127	-LRB-	denotes	[
T20160	34127-34129	CD	denotes	17
T20161	34129-34130	-RRB-	denotes	]
T20162	34131-34134	CC	denotes	and
T20163	34135-34137	IN	denotes	in
T20164	34138-34145	JJ	denotes	classic
T20166	34146-34150	NN	denotes	MSUD
T20165	34151-34159	NNS	denotes	patients
T20167	34160-34161	-LRB-	denotes	[
T20168	34161-34162	CD	denotes	1
T20169	34162-34163	-RRB-	denotes	]
T20170	34163-34164	.	denotes	.
T20171	34164-34292	sentence	denotes	Heterozygous knockouts were normal and had normal levels of BCAA despite having approximately half of BCKDH enzymatic activity.
T20172	34165-34177	JJ	denotes	Heterozygous
T20173	34178-34187	NNS	denotes	knockouts
T20174	34188-34192	VBD	denotes	were
T20175	34193-34199	JJ	denotes	normal
T20176	34200-34203	CC	denotes	and
T20177	34204-34207	VBD	denotes	had
T20178	34208-34214	JJ	denotes	normal
T20179	34215-34221	NNS	denotes	levels
T20180	34222-34224	IN	denotes	of
T20181	34225-34229	NN	denotes	BCAA
T20182	34230-34237	IN	denotes	despite
T20183	34238-34244	VBG	denotes	having
T20184	34245-34258	RB	denotes	approximately
T20185	34259-34263	NN	denotes	half
T20186	34264-34266	IN	denotes	of
T20187	34267-34272	NN	denotes	BCKDH
T20189	34273-34282	JJ	denotes	enzymatic
T20188	34283-34291	NN	denotes	activity
T20190	34291-34292	.	denotes	.
T20191	34292-34524	sentence	denotes	From the characterization studies completed thus far, the null mutation mouse accurately represents a model of classic MSUD and appears to be a faithful model of the human disease with respect to several biochemical phenotypes [1].
T20192	34293-34297	IN	denotes	From
T20194	34298-34301	DT	denotes	the
T20196	34302-34318	NN	denotes	characterization
T20195	34319-34326	NNS	denotes	studies
T20197	34327-34336	VBN	denotes	completed
T20198	34337-34341	RB	denotes	thus
T20199	34342-34345	RB	denotes	far
T20200	34345-34347	,	denotes	,
T20201	34347-34350	DT	denotes	the
T20203	34351-34355	JJ	denotes	null
T20204	34356-34364	NN	denotes	mutation
T20202	34365-34370	NN	denotes	mouse
T20205	34371-34381	RB	denotes	accurately
T20193	34382-34392	VBZ	denotes	represents
T20206	34393-34394	DT	denotes	a
T20207	34395-34400	NN	denotes	model
T20208	34401-34403	IN	denotes	of
T20209	34404-34411	JJ	denotes	classic
T20210	34412-34416	NN	denotes	MSUD
T20211	34417-34420	CC	denotes	and
T20212	34421-34428	VBZ	denotes	appears
T20213	34429-34431	TO	denotes	to
T20214	34432-34434	VB	denotes	be
T20215	34435-34436	DT	denotes	a
T20217	34437-34445	JJ	denotes	faithful
T20216	34446-34451	NN	denotes	model
T20218	34452-34454	IN	denotes	of
T20219	34455-34458	DT	denotes	the
T20221	34459-34464	JJ	denotes	human
T20220	34465-34472	NN	denotes	disease
T20222	34473-34477	IN	denotes	with
T20223	34478-34485	NN	denotes	respect
T20224	34486-34488	IN	denotes	to
T20225	34489-34496	JJ	denotes	several
T20227	34497-34508	JJ	denotes	biochemical
T20226	34509-34519	NNS	denotes	phenotypes
T20228	34520-34521	-LRB-	denotes	[
T20229	34521-34522	CD	denotes	1
T20230	34522-34523	-RRB-	denotes	]
T20231	34523-34524	.	denotes	.
T20232	34524-34648	sentence	denotes	To create a model of intermediate MSUD, we used a transgenic strategy to express human E2 in the liver of E2 knockout mice.
T20233	34525-34527	TO	denotes	To
T20234	34528-34534	VB	denotes	create
T20236	34535-34536	DT	denotes	a
T20237	34537-34542	NN	denotes	model
T20238	34543-34545	IN	denotes	of
T20239	34546-34558	JJ	denotes	intermediate
T20240	34559-34563	NN	denotes	MSUD
T20241	34563-34565	,	denotes	,
T20242	34565-34567	PRP	denotes	we
T20235	34568-34572	VBD	denotes	used
T20243	34573-34574	DT	denotes	a
T20245	34575-34585	JJ	denotes	transgenic
T20244	34586-34594	NN	denotes	strategy
T20246	34595-34597	TO	denotes	to
T20247	34598-34605	VB	denotes	express
T20248	34606-34611	JJ	denotes	human
T20249	34612-34614	NN	denotes	E2
T20250	34615-34617	IN	denotes	in
T20251	34618-34621	DT	denotes	the
T20252	34622-34627	NN	denotes	liver
T20253	34628-34630	IN	denotes	of
T20254	34631-34633	NN	denotes	E2
T20256	34634-34642	NN	denotes	knockout
T20255	34643-34647	NNS	denotes	mice
T20257	34647-34648	.	denotes	.
T20258	34648-34748	sentence	denotes	As mentioned above, E2 knockouts without transgene derived E2 die during the early neonatal period.
T20259	34649-34651	IN	denotes	As
T20260	34652-34661	VBN	denotes	mentioned
T20262	34662-34667	RB	denotes	above
T20263	34667-34669	,	denotes	,
T20264	34669-34671	NN	denotes	E2
T20265	34672-34681	NNS	denotes	knockouts
T20266	34682-34689	IN	denotes	without
T20267	34690-34699	NN	denotes	transgene
T20268	34700-34707	VBN	denotes	derived
T20269	34708-34710	NN	denotes	E2
T20261	34711-34714	VBP	denotes	die
T20270	34715-34721	IN	denotes	during
T20271	34722-34725	DT	denotes	the
T20273	34726-34731	JJ	denotes	early
T20274	34732-34740	JJ	denotes	neonatal
T20272	34741-34747	NN	denotes	period
T20275	34747-34748	.	denotes	.
T20276	34748-34874	sentence	denotes	We show that expression of a human E2 transgene in the liver of these knockout mice is able to rescue the neonatal lethality.
T20277	34749-34751	PRP	denotes	We
T20278	34752-34756	VBP	denotes	show
T20279	34757-34761	IN	denotes	that
T20281	34762-34772	NN	denotes	expression
T20282	34773-34775	IN	denotes	of
T20283	34776-34777	DT	denotes	a
T20285	34778-34783	JJ	denotes	human
T20286	34784-34786	NN	denotes	E2
T20284	34787-34796	NN	denotes	transgene
T20287	34797-34799	IN	denotes	in
T20288	34800-34803	DT	denotes	the
T20289	34804-34809	NN	denotes	liver
T20290	34810-34812	IN	denotes	of
T20291	34813-34818	DT	denotes	these
T20293	34819-34827	NN	denotes	knockout
T20292	34828-34832	NNS	denotes	mice
T20280	34833-34835	VBZ	denotes	is
T20294	34836-34840	JJ	denotes	able
T20295	34841-34843	TO	denotes	to
T20296	34844-34850	VB	denotes	rescue
T20297	34851-34854	DT	denotes	the
T20299	34855-34863	JJ	denotes	neonatal
T20298	34864-34873	NN	denotes	lethality
T20300	34873-34874	.	denotes	.
T20301	34874-34921	sentence	denotes	Many of the rescue mice survived to adulthood.
T20302	34875-34879	JJ	denotes	Many
T20304	34880-34882	IN	denotes	of
T20305	34883-34886	DT	denotes	the
T20307	34887-34893	NN	denotes	rescue
T20306	34894-34898	NNS	denotes	mice
T20303	34899-34907	VBD	denotes	survived
T20308	34908-34910	IN	denotes	to
T20309	34911-34920	NN	denotes	adulthood
T20310	34920-34921	.	denotes	.
T20311	34921-35056	sentence	denotes	It is interesting to note that in these mouse lines only ~5–6% of normal BCKDH activity in the liver was sufficient to allow survival.
T20312	34922-34924	PRP	denotes	It
T20313	34925-34927	VBZ	denotes	is
T20314	34928-34939	JJ	denotes	interesting
T20315	34940-34942	TO	denotes	to
T20316	34943-34947	VB	denotes	note
T20317	34948-34952	IN	denotes	that
T20319	34953-34955	IN	denotes	in
T20320	34956-34961	DT	denotes	these
T20322	34962-34967	NN	denotes	mouse
T20321	34968-34973	NNS	denotes	lines
T20323	34974-34978	RB	denotes	only
T20325	34979-34980	SYM	denotes	~
T20326	34980-34981	CD	denotes	5
T20327	34981-34982	SYM	denotes	–
T20324	34982-34983	CD	denotes	6
T20328	34983-34984	NN	denotes	%
T20329	34985-34987	IN	denotes	of
T20330	34988-34994	JJ	denotes	normal
T20332	34995-35000	NN	denotes	BCKDH
T20331	35001-35009	NN	denotes	activity
T20333	35010-35012	IN	denotes	in
T20334	35013-35016	DT	denotes	the
T20335	35017-35022	NN	denotes	liver
T20318	35023-35026	VBD	denotes	was
T20336	35027-35037	JJ	denotes	sufficient
T20337	35038-35040	TO	denotes	to
T20338	35041-35046	VB	denotes	allow
T20339	35047-35055	NN	denotes	survival
T20340	35055-35056	.	denotes	.
T20341	35056-35174	sentence	denotes	We also demonstrated that BCAA levels in blood of these rescue mice were intermediate between controls and knockouts.
T20342	35057-35059	PRP	denotes	We
T20344	35060-35064	RB	denotes	also
T20343	35065-35077	VBD	denotes	demonstrated
T20345	35078-35082	IN	denotes	that
T20347	35083-35087	NN	denotes	BCAA
T20348	35088-35094	NNS	denotes	levels
T20349	35095-35097	IN	denotes	in
T20350	35098-35103	NN	denotes	blood
T20351	35104-35106	IN	denotes	of
T20352	35107-35112	DT	denotes	these
T20354	35113-35119	NN	denotes	rescue
T20353	35120-35124	NNS	denotes	mice
T20346	35125-35129	VBD	denotes	were
T20355	35130-35142	JJ	denotes	intermediate
T20356	35143-35150	IN	denotes	between
T20357	35151-35159	NNS	denotes	controls
T20358	35160-35163	CC	denotes	and
T20359	35164-35173	NNS	denotes	knockouts
T20360	35173-35174	.	denotes	.
T20361	35174-35322	sentence	denotes	The hallmarks of intermediate MSUD human patients are persistently increased levels of BCAA and BCKDH activity in the range of 3–30% of normal [1].
T20362	35175-35178	DT	denotes	The
T20363	35179-35188	NNS	denotes	hallmarks
T20365	35189-35191	IN	denotes	of
T20366	35192-35204	JJ	denotes	intermediate
T20368	35205-35209	NN	denotes	MSUD
T20369	35210-35215	JJ	denotes	human
T20367	35216-35224	NNS	denotes	patients
T20364	35225-35228	VBP	denotes	are
T20370	35229-35241	RB	denotes	persistently
T20371	35242-35251	VBN	denotes	increased
T20372	35252-35258	NNS	denotes	levels
T20373	35259-35261	IN	denotes	of
T20374	35262-35266	NN	denotes	BCAA
T20376	35267-35270	CC	denotes	and
T20377	35271-35276	NN	denotes	BCKDH
T20375	35277-35285	NN	denotes	activity
T20378	35286-35288	IN	denotes	in
T20379	35289-35292	DT	denotes	the
T20380	35293-35298	NN	denotes	range
T20381	35299-35301	IN	denotes	of
T20382	35302-35303	CD	denotes	3
T20384	35303-35304	SYM	denotes	–
T20383	35304-35306	CD	denotes	30
T20385	35306-35307	NN	denotes	%
T20386	35308-35310	IN	denotes	of
T20387	35311-35317	JJ	denotes	normal
T20388	35318-35319	-LRB-	denotes	[
T20389	35319-35320	CD	denotes	1
T20390	35320-35321	-RRB-	denotes	]
T20391	35321-35322	.	denotes	.
T20392	35322-35509	sentence	denotes	Because of the phenotypic similarities of the rescue mice to the human patients, these genetically engineered mice represent a useful small animal model of the intermediate form of MSUD.
T20393	35323-35330	IN	denotes	Because
T20395	35331-35333	IN	denotes	of
T20396	35334-35337	DT	denotes	the
T20398	35338-35348	JJ	denotes	phenotypic
T20397	35349-35361	NNS	denotes	similarities
T20399	35362-35364	IN	denotes	of
T20400	35365-35368	DT	denotes	the
T20402	35369-35375	NN	denotes	rescue
T20401	35376-35380	NNS	denotes	mice
T20403	35381-35383	IN	denotes	to
T20404	35384-35387	DT	denotes	the
T20406	35388-35393	JJ	denotes	human
T20405	35394-35402	NNS	denotes	patients
T20407	35402-35404	,	denotes	,
T20408	35404-35409	DT	denotes	these
T20410	35410-35421	RB	denotes	genetically
T20411	35422-35432	VBN	denotes	engineered
T20409	35433-35437	NNS	denotes	mice
T20394	35438-35447	VBP	denotes	represent
T20412	35448-35449	DT	denotes	a
T20414	35450-35456	JJ	denotes	useful
T20415	35457-35462	JJ	denotes	small
T20416	35463-35469	NN	denotes	animal
T20413	35470-35475	NN	denotes	model
T20417	35476-35478	IN	denotes	of
T20418	35479-35482	DT	denotes	the
T20420	35483-35495	JJ	denotes	intermediate
T20419	35496-35500	NN	denotes	form
T20421	35501-35503	IN	denotes	of
T20422	35504-35508	NN	denotes	MSUD
T20423	35508-35509	.	denotes	.
T20424	35509-35718	sentence	denotes	The transgenic approach that was used to create the intermediate MSUD model was based on the tetracycline regulated gene switch system that has been used with great success in other studies, for example [34].
T20425	35510-35513	DT	denotes	The
T20427	35514-35524	JJ	denotes	transgenic
T20426	35525-35533	NN	denotes	approach
T20429	35534-35538	WDT	denotes	that
T20431	35539-35542	VBD	denotes	was
T20430	35543-35547	VBN	denotes	used
T20432	35548-35550	TO	denotes	to
T20433	35551-35557	VB	denotes	create
T20434	35558-35561	DT	denotes	the
T20436	35562-35574	JJ	denotes	intermediate
T20437	35575-35579	NN	denotes	MSUD
T20435	35580-35585	NN	denotes	model
T20438	35586-35589	VBD	denotes	was
T20428	35590-35595	VBN	denotes	based
T20439	35596-35598	IN	denotes	on
T20440	35599-35602	DT	denotes	the
T20442	35603-35615	NN	denotes	tetracycline
T20443	35616-35625	VBN	denotes	regulated
T20444	35626-35630	NN	denotes	gene
T20445	35631-35637	NN	denotes	switch
T20441	35638-35644	NN	denotes	system
T20446	35645-35649	WDT	denotes	that
T20448	35650-35653	VBZ	denotes	has
T20449	35654-35658	VBN	denotes	been
T20447	35659-35663	VBN	denotes	used
T20450	35664-35668	IN	denotes	with
T20451	35669-35674	JJ	denotes	great
T20452	35675-35682	NN	denotes	success
T20453	35683-35685	IN	denotes	in
T20454	35686-35691	JJ	denotes	other
T20455	35692-35699	NNS	denotes	studies
T20456	35699-35701	,	denotes	,
T20457	35701-35704	IN	denotes	for
T20458	35705-35712	NN	denotes	example
T20459	35713-35714	-LRB-	denotes	[
T20460	35714-35716	CD	denotes	34
T20461	35716-35717	-RRB-	denotes	]
T20462	35717-35718	.	denotes	.
T20463	35718-35927	sentence	denotes	The strategy behind our approach was to create mice with an intermediate MSUD phenotype that could be converted to the classic phenotype at the investigator's discretion by turning off the rescuing transgene.
T20464	35719-35722	DT	denotes	The
T20465	35723-35731	NN	denotes	strategy
T20467	35732-35738	IN	denotes	behind
T20468	35739-35742	PRP$	denotes	our
T20469	35743-35751	NN	denotes	approach
T20466	35752-35755	VBD	denotes	was
T20470	35756-35758	TO	denotes	to
T20471	35759-35765	VB	denotes	create
T20472	35766-35770	NNS	denotes	mice
T20473	35771-35775	IN	denotes	with
T20474	35776-35778	DT	denotes	an
T20476	35779-35791	JJ	denotes	intermediate
T20477	35792-35796	NN	denotes	MSUD
T20475	35797-35806	NN	denotes	phenotype
T20478	35807-35811	WDT	denotes	that
T20480	35812-35817	MD	denotes	could
T20481	35818-35820	VB	denotes	be
T20479	35821-35830	VBN	denotes	converted
T20482	35831-35833	IN	denotes	to
T20483	35834-35837	DT	denotes	the
T20485	35838-35845	JJ	denotes	classic
T20484	35846-35855	NN	denotes	phenotype
T20486	35856-35858	IN	denotes	at
T20487	35859-35862	DT	denotes	the
T20488	35863-35875	NN	denotes	investigator
T20490	35875-35877	POS	denotes	's
T20489	35878-35888	NN	denotes	discretion
T20491	35889-35891	IN	denotes	by
T20492	35892-35899	VBG	denotes	turning
T20493	35900-35903	RP	denotes	off
T20494	35904-35907	DT	denotes	the
T20496	35908-35916	VBG	denotes	rescuing
T20495	35917-35926	NN	denotes	transgene
T20497	35926-35927	.	denotes	.
T20498	35927-36121	sentence	denotes	However, we were unable to consistently turn off the human E2 expression cassette by the potent tetracycline analogue, doxycycline in either of the two transgenic lines tested (data not shown).
T20499	35928-35935	RB	denotes	However
T20501	35935-35937	,	denotes	,
T20502	35937-35939	PRP	denotes	we
T20500	35940-35944	VBD	denotes	were
T20503	35945-35951	JJ	denotes	unable
T20504	35952-35954	TO	denotes	to
T20506	35955-35967	RB	denotes	consistently
T20505	35968-35972	VB	denotes	turn
T20507	35973-35976	RP	denotes	off
T20508	35977-35980	DT	denotes	the
T20510	35981-35986	JJ	denotes	human
T20511	35987-35989	NN	denotes	E2
T20512	35990-36000	NN	denotes	expression
T20509	36001-36009	NN	denotes	cassette
T20513	36010-36012	IN	denotes	by
T20514	36013-36016	DT	denotes	the
T20516	36017-36023	JJ	denotes	potent
T20517	36024-36036	NN	denotes	tetracycline
T20515	36037-36045	NN	denotes	analogue
T20518	36045-36047	,	denotes	,
T20519	36047-36058	NN	denotes	doxycycline
T20520	36059-36061	IN	denotes	in
T20521	36062-36068	DT	denotes	either
T20522	36069-36071	IN	denotes	of
T20523	36072-36075	DT	denotes	the
T20525	36076-36079	CD	denotes	two
T20526	36080-36090	JJ	denotes	transgenic
T20524	36091-36096	NNS	denotes	lines
T20527	36097-36103	VBN	denotes	tested
T20528	36104-36105	-LRB-	denotes	(
T20530	36105-36109	NNS	denotes	data
T20531	36110-36113	RB	denotes	not
T20529	36114-36119	VBN	denotes	shown
T20532	36119-36120	-RRB-	denotes	)
T20533	36120-36121	.	denotes	.
T20534	36121-36193	sentence	denotes	The reason these two lines were unresponsive to doxycycline is unknown.
T20535	36122-36125	DT	denotes	The
T20536	36126-36132	NN	denotes	reason
T20538	36133-36138	DT	denotes	these
T20540	36139-36142	CD	denotes	two
T20539	36143-36148	NNS	denotes	lines
T20541	36149-36153	VBD	denotes	were
T20542	36154-36166	JJ	denotes	unresponsive
T20543	36167-36169	IN	denotes	to
T20544	36170-36181	NN	denotes	doxycycline
T20537	36182-36184	VBZ	denotes	is
T20545	36185-36192	JJ	denotes	unknown
T20546	36192-36193	.	denotes	.
T20547	36193-36300	sentence	denotes	It is conceivable that the integration site of the transgenes was not permissive for regulated expression.
T20548	36194-36196	PRP	denotes	It
T20549	36197-36199	VBZ	denotes	is
T20550	36200-36211	JJ	denotes	conceivable
T20551	36212-36216	IN	denotes	that
T20553	36217-36220	DT	denotes	the
T20555	36221-36232	NN	denotes	integration
T20554	36233-36237	NN	denotes	site
T20556	36238-36240	IN	denotes	of
T20557	36241-36244	DT	denotes	the
T20558	36245-36255	NNS	denotes	transgenes
T20552	36256-36259	VBD	denotes	was
T20559	36260-36263	RB	denotes	not
T20560	36264-36274	JJ	denotes	permissive
T20561	36275-36278	IN	denotes	for
T20562	36279-36288	VBN	denotes	regulated
T20563	36289-36299	NN	denotes	expression
T20564	36299-36300	.	denotes	.
T20565	36300-36409	sentence	denotes	Screening of additional lines may be needed to find lines that allow for survival and can also be regulated.
T20566	36301-36310	NN	denotes	Screening
T20568	36311-36313	IN	denotes	of
T20569	36314-36324	JJ	denotes	additional
T20570	36325-36330	NNS	denotes	lines
T20571	36331-36334	MD	denotes	may
T20572	36335-36337	VB	denotes	be
T20567	36338-36344	VBN	denotes	needed
T20573	36345-36347	TO	denotes	to
T20574	36348-36352	VB	denotes	find
T20575	36353-36358	NNS	denotes	lines
T20576	36359-36363	WDT	denotes	that
T20577	36364-36369	VBP	denotes	allow
T20578	36370-36373	IN	denotes	for
T20579	36374-36382	NN	denotes	survival
T20580	36383-36386	CC	denotes	and
T20581	36387-36390	MD	denotes	can
T20583	36391-36395	RB	denotes	also
T20584	36396-36398	VB	denotes	be
T20582	36399-36408	VBN	denotes	regulated
T20585	36408-36409	.	denotes	.
T20586	36409-36513	sentence	denotes	The models described in this communication are important advances over the previously described models.
T20587	36410-36413	DT	denotes	The
T20588	36414-36420	NNS	denotes	models
T20590	36421-36430	VBN	denotes	described
T20591	36431-36433	IN	denotes	in
T20592	36434-36438	DT	denotes	this
T20593	36439-36452	NN	denotes	communication
T20589	36453-36456	VBP	denotes	are
T20594	36457-36466	JJ	denotes	important
T20595	36467-36475	NNS	denotes	advances
T20596	36476-36480	IN	denotes	over
T20597	36481-36484	DT	denotes	the
T20599	36485-36495	RB	denotes	previously
T20600	36496-36505	VBN	denotes	described
T20598	36506-36512	NNS	denotes	models
T20601	36512-36513	.	denotes	.
T20602	36513-36619	sentence	denotes	An ENU mouse resembling human MSUD was previously created by disrupting the mitochondrial BCAT gene [20].
T20603	36514-36516	DT	denotes	An
T20605	36517-36520	NN	denotes	ENU
T20604	36521-36526	NN	denotes	mouse
T20607	36527-36537	VBG	denotes	resembling
T20608	36538-36543	JJ	denotes	human
T20609	36544-36548	NN	denotes	MSUD
T20610	36549-36552	VBD	denotes	was
T20611	36553-36563	RB	denotes	previously
T20606	36564-36571	VBN	denotes	created
T20612	36572-36574	IN	denotes	by
T20613	36575-36585	VBG	denotes	disrupting
T20614	36586-36589	DT	denotes	the
T20616	36590-36603	JJ	denotes	mitochondrial
T20617	36604-36608	NN	denotes	BCAT
T20615	36609-36613	NN	denotes	gene
T20618	36614-36615	-LRB-	denotes	[
T20619	36615-36617	CD	denotes	20
T20620	36617-36618	-RRB-	denotes	]
T20621	36618-36619	.	denotes	.
T20622	36619-36745	sentence	denotes	The BCKDH activity in the liver and muscle of this mouse was normal even though the blood BCAA levels were markedly elevated.
T20623	36620-36623	DT	denotes	The
T20625	36624-36629	NN	denotes	BCKDH
T20624	36630-36638	NN	denotes	activity
T20627	36639-36641	IN	denotes	in
T20628	36642-36645	DT	denotes	the
T20629	36646-36651	NN	denotes	liver
T20630	36652-36655	CC	denotes	and
T20631	36656-36662	NN	denotes	muscle
T20632	36663-36665	IN	denotes	of
T20633	36666-36670	DT	denotes	this
T20634	36671-36676	NN	denotes	mouse
T20626	36677-36680	VBD	denotes	was
T20635	36681-36687	JJ	denotes	normal
T20636	36688-36692	RB	denotes	even
T20638	36693-36699	IN	denotes	though
T20639	36700-36703	DT	denotes	the
T20641	36704-36709	NN	denotes	blood
T20642	36710-36714	NN	denotes	BCAA
T20640	36715-36721	NNS	denotes	levels
T20637	36722-36726	VBD	denotes	were
T20643	36727-36735	RB	denotes	markedly
T20644	36736-36744	JJ	denotes	elevated
T20645	36744-36745	.	denotes	.
T20646	36745-36885	sentence	denotes	While this is an interesting model, it is not a true model of MSUD because the mutation is not in BCKDH and the levels of BCKDH are normal.
T20647	36746-36751	IN	denotes	While
T20649	36752-36756	DT	denotes	this
T20648	36757-36759	VBZ	denotes	is
T20651	36760-36762	DT	denotes	an
T20653	36763-36774	JJ	denotes	interesting
T20652	36775-36780	NN	denotes	model
T20654	36780-36782	,	denotes	,
T20655	36782-36784	PRP	denotes	it
T20650	36785-36787	VBZ	denotes	is
T20656	36788-36791	RB	denotes	not
T20657	36792-36793	DT	denotes	a
T20659	36794-36798	JJ	denotes	true
T20658	36799-36804	NN	denotes	model
T20660	36805-36807	IN	denotes	of
T20661	36808-36812	NN	denotes	MSUD
T20662	36813-36820	IN	denotes	because
T20664	36821-36824	DT	denotes	the
T20665	36825-36833	NN	denotes	mutation
T20663	36834-36836	VBZ	denotes	is
T20666	36837-36840	RB	denotes	not
T20667	36841-36843	IN	denotes	in
T20668	36844-36849	NN	denotes	BCKDH
T20669	36850-36853	CC	denotes	and
T20670	36854-36857	DT	denotes	the
T20671	36858-36864	NNS	denotes	levels
T20673	36865-36867	IN	denotes	of
T20674	36868-36873	NN	denotes	BCKDH
T20672	36874-36877	VBP	denotes	are
T20675	36878-36884	JJ	denotes	normal
T20676	36884-36885	.	denotes	.
T20677	36885-36978	sentence	denotes	Furthermore, no case of MSUD has been described attributing this disease to BCAT deficiency.
T20678	36886-36897	RB	denotes	Furthermore
T20680	36897-36899	,	denotes	,
T20681	36899-36901	DT	denotes	no
T20682	36902-36906	NN	denotes	case
T20683	36907-36909	IN	denotes	of
T20684	36910-36914	NN	denotes	MSUD
T20685	36915-36918	VBZ	denotes	has
T20686	36919-36923	VBN	denotes	been
T20679	36924-36933	VBN	denotes	described
T20687	36934-36945	VBG	denotes	attributing
T20688	36946-36950	DT	denotes	this
T20689	36951-36958	NN	denotes	disease
T20690	36959-36961	IN	denotes	to
T20691	36962-36966	NN	denotes	BCAT
T20692	36967-36977	NN	denotes	deficiency
T20693	36977-36978	.	denotes	.
T20694	36978-37075	sentence	denotes	In addition to this mouse model, a cow model of MSUD has also been previously described [17-19].
T20695	36979-36981	IN	denotes	In
T20697	36982-36990	NN	denotes	addition
T20698	36991-36993	IN	denotes	to
T20699	36994-36998	DT	denotes	this
T20701	36999-37004	NN	denotes	mouse
T20700	37005-37010	NN	denotes	model
T20702	37010-37012	,	denotes	,
T20703	37012-37013	DT	denotes	a
T20705	37014-37017	NN	denotes	cow
T20704	37018-37023	NN	denotes	model
T20706	37024-37026	IN	denotes	of
T20707	37027-37031	NN	denotes	MSUD
T20708	37032-37035	VBZ	denotes	has
T20709	37036-37040	RB	denotes	also
T20710	37041-37045	VBN	denotes	been
T20711	37046-37056	RB	denotes	previously
T20696	37057-37066	VBN	denotes	described
T20712	37067-37068	-LRB-	denotes	[
T20713	37068-37070	CD	denotes	17
T20714	37070-37071	SYM	denotes	-
T20715	37071-37073	CD	denotes	19
T20716	37073-37074	-RRB-	denotes	]
T20717	37074-37075	.	denotes	.
T20718	37075-37279	sentence	denotes	However, due to practical constraints imposed by such a large animal model and due to the observation of differences between the MSUD cow model and MSUD humans [1], this model is also of limited utility.
T20719	37076-37083	RB	denotes	However
T20721	37083-37085	,	denotes	,
T20722	37085-37088	IN	denotes	due
T20723	37089-37091	IN	denotes	to
T20724	37092-37101	JJ	denotes	practical
T20725	37102-37113	NNS	denotes	constraints
T20726	37114-37121	VBN	denotes	imposed
T20727	37122-37124	IN	denotes	by
T20728	37125-37129	JJ	denotes	such
T20730	37130-37131	DT	denotes	a
T20731	37132-37137	JJ	denotes	large
T20732	37138-37144	NN	denotes	animal
T20729	37145-37150	NN	denotes	model
T20733	37151-37154	CC	denotes	and
T20734	37155-37158	IN	denotes	due
T20735	37159-37161	IN	denotes	to
T20736	37162-37165	DT	denotes	the
T20737	37166-37177	NN	denotes	observation
T20738	37178-37180	IN	denotes	of
T20739	37181-37192	NNS	denotes	differences
T20740	37193-37200	IN	denotes	between
T20741	37201-37204	DT	denotes	the
T20743	37205-37209	NN	denotes	MSUD
T20744	37210-37213	NN	denotes	cow
T20742	37214-37219	NN	denotes	model
T20745	37220-37223	CC	denotes	and
T20746	37224-37228	NN	denotes	MSUD
T20747	37229-37235	NNS	denotes	humans
T20748	37236-37237	-LRB-	denotes	[
T20749	37237-37238	CD	denotes	1
T20750	37238-37239	-RRB-	denotes	]
T20751	37239-37241	,	denotes	,
T20752	37241-37245	DT	denotes	this
T20753	37246-37251	NN	denotes	model
T20720	37252-37254	VBZ	denotes	is
T20754	37255-37259	RB	denotes	also
T20755	37260-37262	IN	denotes	of
T20756	37263-37270	JJ	denotes	limited
T20757	37271-37278	NN	denotes	utility
T20758	37278-37279	.	denotes	.
T20759	37279-37391	sentence	denotes	In contrast, the murine models described in this report are more valid and appropriate for modeling human MSUD.
T20760	37280-37282	IN	denotes	In
T20762	37283-37291	NN	denotes	contrast
T20763	37291-37293	,	denotes	,
T20764	37293-37296	DT	denotes	the
T20766	37297-37303	JJ	denotes	murine
T20765	37304-37310	NNS	denotes	models
T20767	37311-37320	VBN	denotes	described
T20768	37321-37323	IN	denotes	in
T20769	37324-37328	DT	denotes	this
T20770	37329-37335	NN	denotes	report
T20761	37336-37339	VBP	denotes	are
T20771	37340-37344	RBR	denotes	more
T20772	37345-37350	JJ	denotes	valid
T20773	37351-37354	CC	denotes	and
T20774	37355-37366	JJ	denotes	appropriate
T20775	37367-37370	IN	denotes	for
T20776	37371-37379	VBG	denotes	modeling
T20778	37380-37385	JJ	denotes	human
T20777	37386-37390	NN	denotes	MSUD
T20779	37390-37391	.	denotes	.
T20780	37391-37591	sentence	denotes	The most significant opportunity presented by the MSUD mouse models is to test novel treatments such as gene [38-41] and cell based therapies (e.g., hepatocytes [42,43] or embryonic stem cells [44]).
T20781	37392-37395	DT	denotes	The
T20783	37396-37400	RBS	denotes	most
T20784	37401-37412	JJ	denotes	significant
T20782	37413-37424	NN	denotes	opportunity
T20786	37425-37434	VBN	denotes	presented
T20787	37435-37437	IN	denotes	by
T20788	37438-37441	DT	denotes	the
T20790	37442-37446	NN	denotes	MSUD
T20791	37447-37452	NN	denotes	mouse
T20789	37453-37459	NNS	denotes	models
T20785	37460-37462	VBZ	denotes	is
T20792	37463-37465	TO	denotes	to
T20793	37466-37470	VB	denotes	test
T20794	37471-37476	JJ	denotes	novel
T20795	37477-37487	NNS	denotes	treatments
T20796	37488-37492	JJ	denotes	such
T20797	37493-37495	IN	denotes	as
T20798	37496-37500	NN	denotes	gene
T20800	37501-37502	-LRB-	denotes	[
T20801	37502-37504	CD	denotes	38
T20802	37504-37505	SYM	denotes	-
T20803	37505-37507	CD	denotes	41
T20804	37507-37508	-RRB-	denotes	]
T20805	37509-37512	CC	denotes	and
T20806	37513-37517	NN	denotes	cell
T20799	37518-37523	VBN	denotes	based
T20807	37524-37533	NNS	denotes	therapies
T20808	37534-37535	-LRB-	denotes	(
T20810	37535-37539	FW	denotes	e.g.
T20811	37539-37541	,	denotes	,
T20809	37541-37552	NNS	denotes	hepatocytes
T20812	37553-37554	-LRB-	denotes	[
T20814	37554-37556	CD	denotes	42
T20815	37556-37557	,	denotes	,
T20813	37557-37559	CD	denotes	43
T20816	37559-37560	-RRB-	denotes	]
T20817	37561-37563	CC	denotes	or
T20818	37564-37573	JJ	denotes	embryonic
T20820	37574-37578	NN	denotes	stem
T20819	37579-37584	NNS	denotes	cells
T20821	37585-37586	-LRB-	denotes	[
T20822	37586-37588	CD	denotes	44
T20823	37588-37589	-RRB-	denotes	]
T20824	37589-37590	-RRB-	denotes	)
T20825	37590-37591	.	denotes	.
T20826	37591-37766	sentence	denotes	Relevant to gene therapy, recent problems with gene therapy in humans highlight the importance and critical need of animal models of genetic diseases for preclinical studies.
T20827	37592-37600	JJ	denotes	Relevant
T20829	37601-37603	IN	denotes	to
T20830	37604-37608	NN	denotes	gene
T20831	37609-37616	NN	denotes	therapy
T20832	37616-37618	,	denotes	,
T20833	37618-37624	JJ	denotes	recent
T20834	37625-37633	NNS	denotes	problems
T20835	37634-37638	IN	denotes	with
T20836	37639-37643	NN	denotes	gene
T20837	37644-37651	NN	denotes	therapy
T20838	37652-37654	IN	denotes	in
T20839	37655-37661	NNS	denotes	humans
T20828	37662-37671	VBP	denotes	highlight
T20840	37672-37675	DT	denotes	the
T20841	37676-37686	NN	denotes	importance
T20842	37687-37690	CC	denotes	and
T20843	37691-37699	JJ	denotes	critical
T20844	37700-37704	NN	denotes	need
T20845	37705-37707	IN	denotes	of
T20846	37708-37714	NN	denotes	animal
T20847	37715-37721	NNS	denotes	models
T20848	37722-37724	IN	denotes	of
T20849	37725-37732	JJ	denotes	genetic
T20850	37733-37741	NNS	denotes	diseases
T20851	37742-37745	IN	denotes	for
T20852	37746-37757	JJ	denotes	preclinical
T20853	37758-37765	NNS	denotes	studies
T20854	37765-37766	.	denotes	.
T20855	37766-37954	sentence	denotes	The testing of gene and cell based therapies on appropriate animal model systems provides a preliminary method of establishing not only the efficacy, but also short- and long-term safety.
T20856	37767-37770	DT	denotes	The
T20857	37771-37778	NN	denotes	testing
T20859	37779-37781	IN	denotes	of
T20860	37782-37786	NN	denotes	gene
T20862	37787-37790	CC	denotes	and
T20863	37791-37795	NN	denotes	cell
T20861	37796-37801	VBN	denotes	based
T20864	37802-37811	NNS	denotes	therapies
T20865	37812-37814	IN	denotes	on
T20866	37815-37826	JJ	denotes	appropriate
T20868	37827-37833	NN	denotes	animal
T20869	37834-37839	NN	denotes	model
T20867	37840-37847	NNS	denotes	systems
T20858	37848-37856	VBZ	denotes	provides
T20870	37857-37858	DT	denotes	a
T20872	37859-37870	JJ	denotes	preliminary
T20871	37871-37877	NN	denotes	method
T20873	37878-37880	IN	denotes	of
T20874	37881-37893	VBG	denotes	establishing
T20875	37894-37897	RB	denotes	not
T20877	37898-37902	RB	denotes	only
T20878	37903-37906	DT	denotes	the
T20876	37907-37915	NN	denotes	efficacy
T20879	37915-37917	,	denotes	,
T20880	37917-37920	CC	denotes	but
T20881	37921-37925	RB	denotes	also
T20882	37926-37931	JJ	denotes	short
T20884	37931-37932	HYPH	denotes	-
T20885	37933-37936	CC	denotes	and
T20886	37937-37941	JJ	denotes	long
T20887	37941-37942	HYPH	denotes	-
T20883	37942-37946	NN	denotes	term
T20888	37947-37953	NN	denotes	safety
T20889	37953-37954	.	denotes	.
T20890	37954-38079	sentence	denotes	Success with animal studies is expected to advance such therapeutic approaches and could pave the way for studies in humans.
T20891	37955-37962	NN	denotes	Success
T20893	37963-37967	IN	denotes	with
T20894	37968-37974	NN	denotes	animal
T20895	37975-37982	NNS	denotes	studies
T20896	37983-37985	VBZ	denotes	is
T20892	37986-37994	VBN	denotes	expected
T20897	37995-37997	TO	denotes	to
T20898	37998-38005	VB	denotes	advance
T20899	38006-38010	JJ	denotes	such
T20901	38011-38022	JJ	denotes	therapeutic
T20900	38023-38033	NNS	denotes	approaches
T20902	38034-38037	CC	denotes	and
T20903	38038-38043	MD	denotes	could
T20904	38044-38048	VB	denotes	pave
T20905	38049-38052	DT	denotes	the
T20906	38053-38056	NN	denotes	way
T20907	38057-38060	IN	denotes	for
T20908	38061-38068	NNS	denotes	studies
T20909	38069-38071	IN	denotes	in
T20910	38072-38078	NNS	denotes	humans
T20911	38078-38079	.	denotes	.
T20912	38079-38241	sentence	denotes	In addition to testing various treatments, these models should also be very useful for investigating the underlying pathophysiologic consequences of the disease.
T20913	38080-38082	IN	denotes	In
T20915	38083-38091	NN	denotes	addition
T20916	38092-38094	IN	denotes	to
T20917	38095-38102	VBG	denotes	testing
T20918	38103-38110	JJ	denotes	various
T20919	38111-38121	NNS	denotes	treatments
T20920	38121-38123	,	denotes	,
T20921	38123-38128	DT	denotes	these
T20922	38129-38135	NNS	denotes	models
T20923	38136-38142	MD	denotes	should
T20924	38143-38147	RB	denotes	also
T20914	38148-38150	VB	denotes	be
T20925	38151-38155	RB	denotes	very
T20926	38156-38162	JJ	denotes	useful
T20927	38163-38166	IN	denotes	for
T20928	38167-38180	VBG	denotes	investigating
T20929	38181-38184	DT	denotes	the
T20931	38185-38195	VBG	denotes	underlying
T20932	38196-38212	JJ	denotes	pathophysiologic
T20930	38213-38225	NNS	denotes	consequences
T20933	38226-38228	IN	denotes	of
T20934	38229-38232	DT	denotes	the
T20935	38233-38240	NN	denotes	disease
T20936	38240-38241	.	denotes	.
T20937	38241-38376	sentence	denotes	Such studies are very difficult/impossible to do in human patients for obvious ethical reasons, and often must rely on autopsy tissue.
T20938	38242-38246	JJ	denotes	Such
T20939	38247-38254	NNS	denotes	studies
T20940	38255-38258	VBP	denotes	are
T20941	38259-38263	RB	denotes	very
T20942	38264-38273	JJ	denotes	difficult
T20944	38273-38274	HYPH	denotes	/
T20943	38274-38284	JJ	denotes	impossible
T20945	38285-38287	TO	denotes	to
T20946	38288-38290	VB	denotes	do
T20947	38291-38293	IN	denotes	in
T20948	38294-38299	JJ	denotes	human
T20949	38300-38308	NNS	denotes	patients
T20950	38309-38312	IN	denotes	for
T20951	38313-38320	JJ	denotes	obvious
T20953	38321-38328	JJ	denotes	ethical
T20952	38329-38336	NNS	denotes	reasons
T20954	38336-38338	,	denotes	,
T20955	38338-38341	CC	denotes	and
T20956	38342-38347	RB	denotes	often
T20958	38348-38352	MD	denotes	must
T20957	38353-38357	VB	denotes	rely
T20959	38358-38360	IN	denotes	on
T20960	38361-38368	NN	denotes	autopsy
T20961	38369-38375	NN	denotes	tissue
T20962	38375-38376	.	denotes	.
T20963	38376-38480	sentence	denotes	Additionally, the intermediate MSUD model may also be useful for studies to test the effect of thiamin.
T20964	38377-38389	RB	denotes	Additionally
T20966	38389-38391	,	denotes	,
T20967	38391-38394	DT	denotes	the
T20969	38395-38407	JJ	denotes	intermediate
T20970	38408-38412	NN	denotes	MSUD
T20968	38413-38418	NN	denotes	model
T20971	38419-38422	MD	denotes	may
T20972	38423-38427	RB	denotes	also
T20965	38428-38430	VB	denotes	be
T20973	38431-38437	JJ	denotes	useful
T20974	38438-38441	IN	denotes	for
T20975	38442-38449	NNS	denotes	studies
T20976	38450-38452	TO	denotes	to
T20977	38453-38457	VB	denotes	test
T20978	38458-38461	DT	denotes	the
T20979	38462-38468	NN	denotes	effect
T20980	38469-38471	IN	denotes	of
T20981	38472-38479	NN	denotes	thiamin
T20982	38479-38480	.	denotes	.
T20983	38480-38538	sentence	denotes	A thiamin-responsive form of MSUD has been described [1].
T20984	38481-38482	DT	denotes	A
T20986	38483-38490	NN	denotes	thiamin
T20988	38490-38491	HYPH	denotes	-
T20987	38491-38501	JJ	denotes	responsive
T20985	38502-38506	NN	denotes	form
T20990	38507-38509	IN	denotes	of
T20991	38510-38514	NN	denotes	MSUD
T20992	38515-38518	VBZ	denotes	has
T20993	38519-38523	VBN	denotes	been
T20989	38524-38533	VBN	denotes	described
T20994	38534-38535	-LRB-	denotes	[
T20995	38535-38536	CD	denotes	1
T20996	38536-38537	-RRB-	denotes	]
T20997	38537-38538	.	denotes	.
T20998	38538-38670	sentence	denotes	The phenotype of these patients is heterogeneous and treatment with a wide range of thiamin doses has produced limited success [1].
T20999	38539-38542	DT	denotes	The
T21000	38543-38552	NN	denotes	phenotype
T21002	38553-38555	IN	denotes	of
T21003	38556-38561	DT	denotes	these
T21004	38562-38570	NNS	denotes	patients
T21001	38571-38573	VBZ	denotes	is
T21005	38574-38587	JJ	denotes	heterogeneous
T21006	38588-38591	CC	denotes	and
T21007	38592-38601	NN	denotes	treatment
T21009	38602-38606	IN	denotes	with
T21010	38607-38608	DT	denotes	a
T21012	38609-38613	JJ	denotes	wide
T21011	38614-38619	NN	denotes	range
T21013	38620-38622	IN	denotes	of
T21014	38623-38630	NN	denotes	thiamin
T21015	38631-38636	NNS	denotes	doses
T21016	38637-38640	VBZ	denotes	has
T21008	38641-38649	VBN	denotes	produced
T21017	38650-38657	JJ	denotes	limited
T21018	38658-38665	NN	denotes	success
T21019	38666-38667	-LRB-	denotes	[
T21020	38667-38668	CD	denotes	1
T21021	38668-38669	-RRB-	denotes	]
T21022	38669-38670	.	denotes	.
T21023	38670-38844	sentence	denotes	Recent cell culture studies with MSUD cells have suggested that the thiamin-responsive phenotype is dependent upon the presence of at least one E2 expressing allele [45,46].
T21024	38671-38677	JJ	denotes	Recent
T21026	38678-38682	NN	denotes	cell
T21027	38683-38690	NN	denotes	culture
T21025	38691-38698	NNS	denotes	studies
T21029	38699-38703	IN	denotes	with
T21030	38704-38708	NN	denotes	MSUD
T21031	38709-38714	NNS	denotes	cells
T21032	38715-38719	VBP	denotes	have
T21028	38720-38729	VBN	denotes	suggested
T21033	38730-38734	IN	denotes	that
T21035	38735-38738	DT	denotes	the
T21037	38739-38746	NN	denotes	thiamin
T21039	38746-38747	HYPH	denotes	-
T21038	38747-38757	JJ	denotes	responsive
T21036	38758-38767	NN	denotes	phenotype
T21034	38768-38770	VBZ	denotes	is
T21040	38771-38780	JJ	denotes	dependent
T21041	38781-38785	IN	denotes	upon
T21042	38786-38789	DT	denotes	the
T21043	38790-38798	NN	denotes	presence
T21044	38799-38801	IN	denotes	of
T21045	38802-38804	RB	denotes	at
T21047	38805-38810	RBS	denotes	least
T21046	38811-38814	CD	denotes	one
T21049	38815-38817	NN	denotes	E2
T21050	38818-38828	VBG	denotes	expressing
T21048	38829-38835	NN	denotes	allele
T21051	38836-38837	-LRB-	denotes	[
T21053	38837-38839	CD	denotes	45
T21054	38839-38840	,	denotes	,
T21052	38840-38842	CD	denotes	46
T21055	38842-38843	-RRB-	denotes	]
T21056	38843-38844	.	denotes	.
T21057	38844-39046	sentence	denotes	In light of these newer findings, our intermediate MSUD mouse provides a model to test the effect of thiamin with respect to BCKDH activity and blood amino acid levels at the level of the whole animal.
T21058	38845-38847	IN	denotes	In
T21060	38848-38853	NN	denotes	light
T21061	38854-38856	IN	denotes	of
T21062	38857-38862	DT	denotes	these
T21064	38863-38868	JJR	denotes	newer
T21063	38869-38877	NNS	denotes	findings
T21065	38877-38879	,	denotes	,
T21066	38879-38882	PRP$	denotes	our
T21068	38883-38895	JJ	denotes	intermediate
T21069	38896-38900	NN	denotes	MSUD
T21067	38901-38906	NN	denotes	mouse
T21059	38907-38915	VBZ	denotes	provides
T21070	38916-38917	DT	denotes	a
T21071	38918-38923	NN	denotes	model
T21072	38924-38926	TO	denotes	to
T21073	38927-38931	VB	denotes	test
T21074	38932-38935	DT	denotes	the
T21075	38936-38942	NN	denotes	effect
T21076	38943-38945	IN	denotes	of
T21077	38946-38953	NN	denotes	thiamin
T21078	38954-38958	IN	denotes	with
T21079	38959-38966	NN	denotes	respect
T21080	38967-38969	IN	denotes	to
T21081	38970-38975	NN	denotes	BCKDH
T21082	38976-38984	NN	denotes	activity
T21083	38985-38988	CC	denotes	and
T21084	38989-38994	NN	denotes	blood
T21086	38995-39000	NN	denotes	amino
T21087	39001-39005	NN	denotes	acid
T21085	39006-39012	NNS	denotes	levels
T21088	39013-39015	IN	denotes	at
T21089	39016-39019	DT	denotes	the
T21090	39020-39025	NN	denotes	level
T21091	39026-39028	IN	denotes	of
T21092	39029-39032	DT	denotes	the
T21094	39033-39038	JJ	denotes	whole
T21093	39039-39045	NN	denotes	animal
T21095	39045-39046	.	denotes	.
T21096	39046-39231	sentence	denotes	Because of recent interest in structural analysis of multienzyme complexes [35,36,45,46], the intermediate MSUD model may provide a valuable resource for studies of structural biology.
T21097	39047-39054	IN	denotes	Because
T21099	39055-39057	IN	denotes	of
T21100	39058-39064	JJ	denotes	recent
T21101	39065-39073	NN	denotes	interest
T21102	39074-39076	IN	denotes	in
T21103	39077-39087	JJ	denotes	structural
T21104	39088-39096	NN	denotes	analysis
T21105	39097-39099	IN	denotes	of
T21106	39100-39111	NN	denotes	multienzyme
T21107	39112-39121	NNS	denotes	complexes
T21108	39122-39123	-LRB-	denotes	[
T21110	39123-39125	CD	denotes	35
T21111	39125-39126	,	denotes	,
T21112	39126-39128	CD	denotes	36
T21113	39128-39129	,	denotes	,
T21114	39129-39131	CD	denotes	45
T21115	39131-39132	,	denotes	,
T21109	39132-39134	CD	denotes	46
T21116	39134-39135	-RRB-	denotes	]
T21117	39135-39137	,	denotes	,
T21118	39137-39140	DT	denotes	the
T21120	39141-39153	JJ	denotes	intermediate
T21121	39154-39158	NN	denotes	MSUD
T21119	39159-39164	NN	denotes	model
T21122	39165-39168	MD	denotes	may
T21098	39169-39176	VB	denotes	provide
T21123	39177-39178	DT	denotes	a
T21125	39179-39187	JJ	denotes	valuable
T21124	39188-39196	NN	denotes	resource
T21126	39197-39200	IN	denotes	for
T21127	39201-39208	NNS	denotes	studies
T21128	39209-39211	IN	denotes	of
T21129	39212-39222	JJ	denotes	structural
T21130	39223-39230	NN	denotes	biology
T21131	39230-39231	.	denotes	.
T21132	39231-39439	sentence	denotes	Lastly, the information and knowledge gained from studies with the MSUD models described here will also be applicable and transferable to other mitochondrial disorders due to defects in multisubunit enzymes.
T21133	39232-39238	RB	denotes	Lastly
T21135	39238-39240	,	denotes	,
T21136	39240-39243	DT	denotes	the
T21137	39244-39255	NN	denotes	information
T21138	39256-39259	CC	denotes	and
T21139	39260-39269	NN	denotes	knowledge
T21140	39270-39276	VBN	denotes	gained
T21141	39277-39281	IN	denotes	from
T21142	39282-39289	NNS	denotes	studies
T21143	39290-39294	IN	denotes	with
T21144	39295-39298	DT	denotes	the
T21146	39299-39303	NN	denotes	MSUD
T21145	39304-39310	NNS	denotes	models
T21147	39311-39320	VBN	denotes	described
T21148	39321-39325	RB	denotes	here
T21149	39326-39330	MD	denotes	will
T21150	39331-39335	RB	denotes	also
T21134	39336-39338	VB	denotes	be
T21151	39339-39349	JJ	denotes	applicable
T21152	39350-39353	CC	denotes	and
T21153	39354-39366	JJ	denotes	transferable
T21154	39367-39369	IN	denotes	to
T21155	39370-39375	JJ	denotes	other
T21157	39376-39389	JJ	denotes	mitochondrial
T21156	39390-39399	NNS	denotes	disorders
T21158	39400-39403	IN	denotes	due
T21159	39404-39406	IN	denotes	to
T21160	39407-39414	NNS	denotes	defects
T21161	39415-39417	IN	denotes	in
T21162	39418-39430	JJ	denotes	multisubunit
T21163	39431-39438	NNS	denotes	enzymes
T21164	39438-39439	.	denotes	.
T21291	39452-39454	IN	denotes	In
T21293	39455-39462	NN	denotes	summary
T21294	39462-39464	,	denotes	,
T21295	39464-39468	DT	denotes	this
T21296	39469-39475	NN	denotes	report
T21292	39476-39485	VBZ	denotes	describes
T21297	39486-39489	DT	denotes	the
T21298	39490-39501	NN	denotes	development
T21299	39502-39505	CC	denotes	and
T21300	39506-39522	NN	denotes	characterization
T21301	39523-39525	IN	denotes	of
T21302	39526-39537	RB	denotes	genetically
T21303	39538-39548	VBN	denotes	engineered
T21305	39549-39554	NN	denotes	mouse
T21304	39555-39561	NNS	denotes	models
T21306	39562-39564	IN	denotes	of
T21307	39565-39572	JJ	denotes	classic
T21309	39573-39575	RB	denotes	as
T21311	39576-39580	RB	denotes	well
T21310	39581-39583	IN	denotes	as
T21312	39584-39596	JJ	denotes	intermediate
T21308	39597-39601	NN	denotes	MSUD
T21313	39601-39602	.	denotes	.
T21314	39602-39826	sentence	denotes	These animals provide useful models to further characterize the pathogenesis of MSUD, as well as models to test novel therapeutic strategies, such as gene and cellular therapies, to treat this devastating metabolic disease.
T21315	39603-39608	DT	denotes	These
T21316	39609-39616	NNS	denotes	animals
T21317	39617-39624	VBP	denotes	provide
T21318	39625-39631	JJ	denotes	useful
T21319	39632-39638	NNS	denotes	models
T21320	39639-39641	TO	denotes	to
T21322	39642-39649	RB	denotes	further
T21321	39650-39662	VB	denotes	characterize
T21323	39663-39666	DT	denotes	the
T21324	39667-39679	NN	denotes	pathogenesis
T21325	39680-39682	IN	denotes	of
T21326	39683-39687	NN	denotes	MSUD
T21327	39687-39689	,	denotes	,
T21328	39689-39691	RB	denotes	as
T21330	39692-39696	RB	denotes	well
T21329	39697-39699	IN	denotes	as
T21331	39700-39706	NNS	denotes	models
T21332	39707-39709	TO	denotes	to
T21333	39710-39714	VB	denotes	test
T21334	39715-39720	JJ	denotes	novel
T21336	39721-39732	JJ	denotes	therapeutic
T21335	39733-39743	NNS	denotes	strategies
T21337	39743-39745	,	denotes	,
T21338	39745-39749	JJ	denotes	such
T21339	39750-39752	IN	denotes	as
T21340	39753-39757	NN	denotes	gene
T21342	39758-39761	CC	denotes	and
T21343	39762-39770	JJ	denotes	cellular
T21341	39771-39780	NNS	denotes	therapies
T21344	39780-39782	,	denotes	,
T21345	39782-39784	TO	denotes	to
T21346	39785-39790	VB	denotes	treat
T21347	39791-39795	DT	denotes	this
T21349	39796-39807	VBG	denotes	devastating
T21350	39808-39817	JJ	denotes	metabolic
T21348	39818-39825	NN	denotes	disease
T21351	39825-39826	.	denotes	.
T21373	40814-40817	DT	denotes	The
T21375	40818-40833	JJ	denotes	pre-publication
T21374	40834-40841	NN	denotes	history
T21377	40842-40845	IN	denotes	for
T21378	40846-40850	DT	denotes	this
T21379	40851-40856	NN	denotes	paper
T21380	40857-40860	MD	denotes	can
T21381	40861-40863	VB	denotes	be
T21376	40864-40872	VBN	denotes	accessed
T21382	40873-40877	RB	denotes	here
T21383	40877-40878	:	denotes	:
R1	T836	T834	cc	and,Production
R10	T845	T842	conj	intermediate,classic
R100	T951	T952	dep	that,lacked
R1000	T4006	T4004	pobj	treatment,for
R1001	T4007	T4006	prep	of,treatment
R1002	T3838	T3837	cc	and,injuries
R1003	T4008	T4007	pobj	MSUD,of
R1004	T4009	T3998	punct	", ",is
R1005	T4010	T3998	expl	there,is
R1006	T3839	T3840	amod	other,stressors
R1007	T4011	T4012	det	a,need
R1008	T4012	T3998	attr	need,is
R1009	T4013	T4012	prep	for,need
R101	T952	T949	relcl	lacked,line
R1010	T4014	T4015	amod	improved,therapies
R1011	T3840	T3837	conj	stressors,injuries
R1012	T4015	T4013	pobj	therapies,for
R1013	T4016	T4017	aux	to,combat
R1014	T3841	T3817	punct	.,is
R1015	T4017	T4015	advcl	combat,therapies
R1016	T4018	T4019	det	this,disease
R1017	T4019	T4017	dobj	disease,combat
R1018	T4020	T3998	punct	.,is
R1019	T3843	T3844	prep	In,is
R102	T953	T954	det	a,gene
R1020	T4022	T4023	det	An,obstacle
R1021	T4023	T4024	nsubj	obstacle,been
R1022	T4025	T4023	prep	to,obstacle
R1023	T4057	T4058	punct	[,17
R1024	T4026	T4025	pcomp	developing,to
R1025	T4027	T4028	amod	novel,treatments
R1026	T4028	T4026	dobj	treatments,developing
R1027	T4029	T4028	prep	for,treatments
R1028	T4030	T4029	pobj	MSUD,for
R1029	T4031	T4024	aux	has,been
R103	T954	T952	dobj	gene,lacked
R1030	T4058	T4047	parataxis	17,described
R1031	T4032	T4033	det	the,lack
R1032	T4033	T4024	attr	lack,been
R1033	T4034	T4033	prep	of,lack
R1034	T4059	T4060	punct	-,19
R1035	T4035	T4036	det	a,model
R1036	T4036	T4034	pobj	model,of
R1037	T4037	T4036	amod	suitable,model
R1038	T4038	T4036	compound	animal,model
R1039	T4060	T4058	prep	19,17
R104	T955	T954	amod	functional,gene
R1040	T4039	T4040	aux	to,perform
R1041	T4040	T4036	advcl	perform,model
R1042	T4041	T4042	amod	necessary,studies
R1043	T4061	T4058	punct	],17
R1044	T4042	T4040	dobj	studies,perform
R1045	T4043	T4042	amod	preclinical,studies
R1046	T4044	T4024	punct	.,been
R1047	T4062	T4056	punct	", ",is
R1048	T4046	T4047	mark	Although,described
R1049	T4047	T4056	advcl	described,is
R105	T956	T954	compound	E2,gene
R1050	T4063	T4064	det	this,model
R1051	T4048	T4049	det	a,model
R1052	T4049	T4047	nsubjpass	model,described
R1053	T4050	T4051	compound	Hereford,calf
R1054	T4064	T4056	nsubj	model,is
R1055	T4051	T4049	compound	calf,model
R1056	T4052	T4049	prep	with,model
R1057	T4053	T4052	pobj	MSUD,with
R1058	T4065	T4066	preconj	neither,available
R1059	T4054	T4047	aux	has,described
R106	T957	T954	compound	subunit,gene
R1060	T4055	T4047	auxpass	been,described
R1061	T4066	T4056	acomp	available,is
R1062	T4067	T4066	advmod	readily,available
R1063	T4068	T4066	cc	nor,available
R1064	T4069	T4066	conj	practical,available
R1065	T4162	T4156	punct	(,aminotransferase
R1066	T4070	T4071	aux	to,perform
R1067	T4163	T4156	appos	BCAT,aminotransferase
R1068	T4164	T4161	punct	),gene
R1069	T4165	T4154	punct	", ",in
R107	T958	T954	prep	of,gene
R1070	T4166	T4154	neg	not,in
R1071	T4167	T4154	prep	in,in
R1072	T4071	T4069	xcomp	perform,practical
R1073	T4168	T4167	pobj	BCKDH,in
R1074	T4169	T4168	punct	", ",BCKDH
R1075	T4072	T4073	amod	preclinical,studies
R1076	T4170	T4171	det	the,enzyme
R1077	T4171	T4168	appos	enzyme,BCKDH
R1078	T4172	T4171	amod	deficient,enzyme
R1079	T4173	T4171	prep	in,enzyme
R108	T959	T960	amod	branched,chain
R1080	T4073	T4071	dobj	studies,perform
R1081	T4174	T4173	pobj	MSUD,in
R1082	T4175	T4144	punct	.,disrupts
R1083	T4074	T4056	punct	.,is
R1084	T4177	T4178	mark	Because,is
R1085	T4178	T4181	advcl	is,is
R1086	T4076	T4077	advmod	Furthermore,shown
R1087	T4179	T4180	det	the,mutation
R1088	T4180	T4178	nsubj	mutation,is
R1089	T4182	T4178	neg	not,is
R109	T960	T962	compound	chain,dehydrogenase
R1090	T4078	T4077	punct	", ",shown
R1091	T4183	T4178	prep	in,is
R1092	T4184	T4183	pobj	BCKDH,in
R1093	T4185	T4181	punct	", ",is
R1094	T4186	T4187	det	the,validity
R1095	T4187	T4181	nsubj	validity,is
R1096	T4079	T4077	nsubj	comparison,shown
R1097	T4188	T4187	prep	of,validity
R1098	T4189	T4190	det	this,line
R1099	T4190	T4188	pobj	line,of
R11	T846	T847	compound	maple,syrup
R110	T961	T960	punct	-,chain
R1100	T4191	T4190	compound	mutant,line
R1101	T4080	T4079	prep	of,comparison
R1102	T4192	T4190	compound	mouse,line
R1103	T4193	T4187	prep	for,validity
R1104	T4194	T4195	amod	modeling,MSUD
R1105	T4081	T4082	det	this,model
R1106	T4195	T4193	pobj	MSUD,for
R1107	T4196	T4195	amod	human,MSUD
R1108	T4197	T4181	acomp	questionable,is
R1109	T4082	T4080	pobj	model,of
R111	T962	T958	pobj	dehydrogenase,of
R1110	T4198	T4181	punct	.,is
R1111	T4200	T4201	det	The,objective
R1112	T4083	T4082	compound	animal,model
R1113	T4201	T4202	nsubj	objective,was
R1114	T4203	T4201	prep	of,objective
R1115	T4084	T4082	prep	with,model
R1116	T4204	T4205	det	the,study
R1117	T4205	T4203	pobj	study,of
R1118	T4206	T4205	amod	present,study
R1119	T4085	T4086	amod	human,MSUD
R112	T963	T964	compound	keto,acid
R1120	T4207	T4208	aux	to,create
R1121	T4208	T4202	xcomp	create,was
R1122	T4209	T4210	advmod	genetically,engineered
R1123	T4210	T4211	amod	engineered,models
R1124	T4211	T4208	dobj	models,create
R1125	T4212	T4211	amod	murine,models
R1126	T4086	T4084	pobj	MSUD,with
R1127	T4213	T4211	prep	of,models
R1128	T4214	T4213	pobj	MSUD,of
R1129	T4087	T4077	aux	has,shown
R113	T964	T962	compound	acid,dehydrogenase
R1130	T4215	T4216	dep	that,mimic
R1131	T4216	T4211	relcl	mimic,models
R1132	T4217	T4218	det	the,pathology
R1133	T4088	T4089	det	some,differences
R1134	T4218	T4216	dobj	pathology,mimic
R1135	T4219	T4218	prep	of,pathology
R1136	T4220	T4221	det	the,forms
R1137	T4221	T4219	pobj	forms,of
R1138	T4089	T4077	dobj	differences,shown
R1139	T4222	T4221	amod	classic,forms
R114	T965	T930	punct	.,used
R1140	T4223	T4222	cc	and,classic
R1141	T4224	T4222	conj	intermediate,classic
R1142	T4090	T4089	prep	in,differences
R1143	T4225	T4221	amod	variant,forms
R1144	T4226	T4221	prep	of,forms
R1145	T4091	T4092	det	the,pathology
R1146	T4227	T4228	det	the,disease
R1147	T4228	T4226	pobj	disease,of
R1148	T4229	T4228	amod	human,disease
R1149	T4092	T4090	pobj	pathology,in
R115	T967	T968	aux	To,create
R1150	T4230	T4202	punct	.,was
R1151	T4232	T4233	det	The,model
R1152	T4093	T4092	prep	of,pathology
R1153	T4233	T4235	nsubjpass	model,created
R1154	T4234	T4233	amod	classic,model
R1155	T4094	T4095	det	the,disease
R1156	T4236	T4235	auxpass	was,created
R1157	T4237	T4235	prep	by,created
R1158	T4095	T4093	pobj	disease,of
R1159	T4238	T4239	amod	targeted,inactivation
R116	T968	T969	advcl	create,used
R1160	T4239	T4237	pobj	inactivation,by
R1161	T4240	T4239	prep	of,inactivation
R1162	T4096	T4097	punct	[,1
R1163	T4097	T4077	parataxis	1,shown
R1164	T4241	T4242	det	the,subunit
R1165	T4098	T4097	punct	],1
R1166	T4242	T4240	pobj	subunit,of
R1167	T4243	T4242	compound	E2,subunit
R1168	T4099	T4077	punct	", ",shown
R1169	T4244	T4242	prep	of,subunit
R117	T970	T971	det	a,model
R1170	T4245	T4244	pobj	BCKDH,of
R1171	T4246	T4239	prep	by,inactivation
R1172	T4100	T4077	advcl	making,shown
R1173	T4247	T4248	amod	homologous,recombination
R1174	T4248	T4246	pobj	recombination,by
R1175	T4249	T4239	prep	in,inactivation
R1176	T4250	T4251	nmod	embryonic,ES
R1177	T4251	T4254	nmod	ES,cells
R1178	T4101	T4102	det	this,animal
R1179	T4252	T4251	nmod	stem,ES
R118	T971	T968	dobj	model,create
R1180	T4253	T4251	punct	(,ES
R1181	T4102	T4103	nsubj	animal,model
R1182	T4254	T4249	pobj	cells,in
R1183	T4255	T4254	punct	),cells
R1184	T4256	T4235	punct	.,created
R1185	T4103	T4100	ccomp	model,making
R1186	T4258	T4259	det	The,model
R1187	T4259	T4260	nsubjpass	model,created
R1188	T4104	T4103	det	a,model
R1189	T4261	T4259	prep	of,model
R119	T972	T971	amod	murine,model
R1190	T4262	T4263	amod	intermediate,MSUD
R1191	T4105	T4106	advmod	less,desirable
R1192	T4263	T4261	pobj	MSUD,of
R1193	T4264	T4260	auxpass	was,created
R1194	T4265	T4260	prep	by,created
R1195	T4106	T4103	amod	desirable,model
R1196	T4266	T4267	amod	partial,rescue
R1197	T4267	T4265	pobj	rescue,by
R1198	T4107	T4103	prep	for,model
R1199	T4108	T4109	det	the,disease
R12	T848	T843	compound	urine,disease
R120	T973	T971	prep	of,model
R1200	T4268	T4267	amod	transgenic,rescue
R1201	T4269	T4267	prep	of,rescue
R1202	T4109	T4107	pobj	disease,for
R1203	T4270	T4271	det	the,knockout
R1204	T4271	T4269	pobj	knockout,of
R1205	T4110	T4109	amod	human,disease
R1206	T4272	T4271	compound	E2,knockout
R1207	T4273	T4271	compound	gene,knockout
R1208	T4274	T4260	punct	.,created
R1209	T4111	T4077	punct	.,shown
R121	T974	T975	amod	intermediate,MSUD
R1210	T4276	T4277	det	This,report
R1211	T4277	T4278	nsubj	report,describes
R1212	T4113	T4114	advmod	Recently,described
R1213	T4279	T4280	det	the,generation
R1214	T4280	T4278	dobj	generation,describes
R1215	T4281	T4280	cc	and,generation
R1216	T4282	T4280	conj	characterization,generation
R1217	T4115	T4114	punct	", ",described
R1218	T4283	T4280	prep	of,generation
R1219	T4284	T4285	det	these,models
R122	T975	T973	pobj	MSUD,of
R1220	T4285	T4283	pobj	models,of
R1221	T4286	T4285	amod	murine,models
R1222	T4116	T4117	det	a,nitrosourea
R1223	T4287	T4285	prep	of,models
R1224	T4288	T4287	pobj	MSUD,of
R1225	T4289	T4278	punct	.,describes
R1226	T4291	T4292	det	These,models
R1227	T4117	T4124	npadvmod	nitrosourea,induced
R1228	T4292	T4293	nsubj	models,allow
R1229	T4294	T4293	aux	will,allow
R123	T976	T969	punct	", ",used
R1230	T4295	T4293	prep	for,allow
R1231	T4118	T4119	compound	N,ethyl
R1232	T4296	T4297	det	the,development
R1233	T4297	T4295	pobj	development,for
R1234	T4298	T4297	prep	of,development
R1235	T4119	T4117	compound	ethyl,nitrosourea
R1236	T4299	T4300	amod	novel,approaches
R1237	T4300	T4298	pobj	approaches,of
R1238	T4301	T4300	compound	treatment,approaches
R1239	T4120	T4119	punct	-,ethyl
R124	T977	T969	nsubj	we,used
R1240	T4302	T4300	punct	", ",approaches
R1241	T4303	T4304	amod	such,as
R1242	T4304	T4300	prep	as,approaches
R1243	T4121	T4117	punct	-,nitrosourea
R1244	T4305	T4306	nmod	gene,therapies
R1245	T4306	T4304	pobj	therapies,as
R1246	T4307	T4305	cc	or,gene
R1247	T4122	T4117	compound	N,nitrosourea
R1248	T4308	T4309	compound	stem,cell
R1249	T4309	T4305	conj	cell,gene
R125	T978	T979	amod	transgenic,technology
R1250	T4310	T4293	punct	", ",allow
R1251	T4311	T4312	aux	to,cure
R1252	T4123	T4117	punct	-,nitrosourea
R1253	T4312	T4293	advcl	cure,allow
R1254	T4313	T4312	advmod	ultimately,cure
R1255	T4314	T4312	dobj	MSUD,cure
R1256	T4124	T4129	amod	induced,mouse
R1257	T4315	T4293	punct	.,allow
R1258	T4125	T4117	punct	(,nitrosourea
R1259	T4126	T4117	appos	ENU,nitrosourea
R126	T979	T969	dobj	technology,used
R1260	T4127	T4124	punct	),induced
R1261	T4128	T4124	punct	-,induced
R1262	T4129	T4114	nsubjpass	mouse,described
R1263	T4130	T4129	compound	mutant,mouse
R1264	T4131	T4132	dep	that,resembles
R1265	T4132	T4129	relcl	resembles,mouse
R1266	T4133	T4132	advmod	phenotypically,resembles
R1267	T4134	T4135	amod	human,MSUD
R1268	T4135	T4132	dobj	MSUD,resembles
R1269	T4136	T4114	aux	has,described
R127	T980	T981	aux	to,express
R1270	T4137	T4114	auxpass	been,described
R1271	T4138	T4139	punct	[,20
R1272	T4139	T4114	parataxis	20,described
R1273	T4140	T4139	punct	],20
R1274	T4141	T4114	punct	.,described
R1275	T4143	T4144	advmod	However,disrupts
R1276	T4145	T4144	punct	", ",disrupts
R1277	T4146	T4147	det	the,mutation
R1278	T4147	T4144	nsubj	mutation,disrupts
R1279	T4148	T4147	prep	in,mutation
R128	T981	T969	advcl	express,used
R1280	T4149	T4150	det	this,model
R1281	T4150	T4148	pobj	model,in
R1282	T4151	T4152	det	a,site
R1283	T4152	T4144	dobj	site,disrupts
R1284	T4153	T4152	compound	splice,site
R1285	T4154	T4152	prep	in,site
R1286	T4155	T4156	det	the,aminotransferase
R1287	T4156	T4161	nmod	aminotransferase,gene
R1288	T4157	T4156	amod	mitochondrial,aminotransferase
R1289	T4158	T4159	amod	branched,chain
R129	T982	T983	det	a,cDNA
R1290	T4159	T4156	nmod	chain,aminotransferase
R1291	T4160	T4159	punct	-,chain
R1292	T4161	T4154	pobj	gene,in
R1293	T4359	T4360	det	All,studies
R1294	T4360	T4361	nsubjpass	studies,reviewed
R1295	T4362	T4360	acl	involving,studies
R1296	T4363	T4362	dobj	animals,involving
R1297	T4364	T4361	auxpass	were,reviewed
R1298	T4365	T4361	cc	and,reviewed
R1299	T4366	T4361	conj	approved,reviewed
R13	T853	T854	compound	Maple,Syrup
R130	T983	T981	dobj	cDNA,express
R1300	T4367	T4366	agent	by,approved
R1301	T4368	T4369	det	the,University
R1302	T4369	T4367	pobj	University,by
R1303	T4370	T4369	prep	of,University
R1304	T4371	T4372	poss	Pittsburgh,Committee
R1305	T4372	T4370	pobj	Committee,of
R1306	T4373	T4371	case	's,Pittsburgh
R1307	T4374	T4375	nmod	Institutional,Animal
R1308	T4375	T4376	nmod	Animal,Care
R1309	T4376	T4372	nmod	Care,Committee
R131	T984	T983	amod	human,cDNA
R1310	T4377	T4376	cc	and,Care
R1311	T4378	T4376	conj	Use,Care
R1312	T4379	T4361	punct	.,reviewed
R1313	T5027	T5028	compound	Gene,knockout
R1314	T5028	T5029	compound	knockout,production
R1315	T5031	T5032	det	A,subclone
R1316	T5032	T5035	nsubjpass	subclone,obtained
R1317	T5033	T5032	amod	genomic,subclone
R1318	T5034	T5032	compound	DNA,subclone
R1319	T5036	T5032	prep	of,subclone
R132	T985	T983	compound	E2,cDNA
R1320	T5037	T5038	nmod	Strain,DNA
R1321	T5038	T5036	pobj	DNA,of
R1322	T5039	T5040	nummod	129,SvJ
R1323	T5040	T5038	compound	SvJ,DNA
R1324	T5041	T5040	punct	/,SvJ
R1325	T5042	T5043	dep	that,contained
R1326	T5043	T5032	relcl	contained,subclone
R1327	T5044	T5045	det	a,portion
R1328	T5045	T5043	dobj	portion,contained
R1329	T5046	T5045	prep	of,portion
R133	T986	T983	prep	on,cDNA
R1330	T5047	T5048	nmod	Exons,4
R1331	T5048	T5046	pobj	4,of
R1332	T5049	T5048	cc	and,4
R1333	T5050	T5048	conj	5,4
R1334	T5051	T5048	prep	of,4
R1335	T5052	T5053	det	the,gene
R1336	T5053	T5051	pobj	gene,of
R1337	T5054	T5053	compound	E2,gene
R1338	T5055	T5053	cc	and,gene
R1339	T5056	T5057	compound	flanking,DNA
R134	T987	T988	det	the,background
R1340	T5057	T5053	conj	DNA,gene
R1341	T5058	T5035	auxpass	was,obtained
R1342	T5059	T5035	prep	from,obtained
R1343	T5060	T5061	det	a,library
R1344	T5061	T5059	pobj	library,from
R1345	T5062	T5063	compound	P1,phage
R1346	T5063	T5061	compound	phage,library
R1347	T5064	T5061	prep	from,library
R1348	T5065	T5066	nmod	Genome,Inc.
R1349	T5066	T5064	pobj	Inc.,from
R135	T988	T986	pobj	background,on
R1350	T5067	T5066	nmod	Systems,Inc.
R1351	T5068	T5066	punct	", ",Inc.
R1352	T5069	T5066	punct	", ",Inc.
R1353	T5070	T5071	punct	(,Library
R1354	T5071	T5066	poss	Library,Inc.
R1355	T5072	T5073	compound	St.,Louis
R1356	T5073	T5071	dep	Louis,Library
R1357	T5074	T5073	punct	", ",Louis
R1358	T5075	T5073	npadvmod	MO,Louis
R1359	T5076	T5071	punct	;,Library
R136	T989	T988	compound	knockout,background
R1360	T5077	T5071	punct	',Library
R1361	T5078	T5079	nummod	3,Hit
R1362	T5079	T5071	compound	Hit,Library
R1363	T5080	T5079	punct	-,Hit
R1364	T5081	T5082	compound	Mouse,ES
R1365	T5082	T5071	compound	ES,Library
R1366	T5083	T5071	dep	',Library
R1367	T5084	T5071	punct	),Library
R1368	T5085	T5035	punct	.,obtained
R1369	T5087	T5088	det	A,vector
R137	T990	T969	punct	.,used
R1370	T5088	T5096	nsubjpass	vector,created
R1371	T5089	T5090	amod	positive,negative
R1372	T5090	T5088	amod	negative,vector
R1373	T5091	T5090	punct	/,negative
R1374	T5092	T5093	compound	replacement,type
R1375	T5093	T5088	compound	type,vector
R1376	T5094	T5095	compound	gene,targeting
R1377	T5095	T5088	compound	targeting,vector
R1378	T5097	T5098	punct	[,21
R1379	T5098	T5088	parataxis	21,vector
R138	T992	T993	nsubjpass	Mice,characterized
R1380	T5099	T5098	punct	],21
R1381	T5100	T5096	auxpass	was,created
R1382	T5101	T5096	prep	by,created
R1383	T5102	T5101	pcomp	replacing,by
R1384	T5103	T5104	det	a,fragment
R1385	T5104	T5102	dobj	fragment,replacing
R1386	T5105	T5106	nummod	1.67,kb
R1387	T5106	T5104	compound	kb,fragment
R1388	T5107	T5108	compound	EcoRV,Smal
R1389	T5108	T5104	compound	Smal,fragment
R139	T994	T992	prep	of,Mice
R1390	T5109	T5108	punct	-,Smal
R1391	T5110	T5111	dep	that,included
R1392	T5111	T5104	relcl	included,fragment
R1393	T5112	T5113	det	a,portion
R1394	T5113	T5111	dobj	portion,included
R1395	T5114	T5113	prep	of,portion
R1396	T5115	T5114	pobj	Exon,of
R1397	T5116	T5115	nummod	4,Exon
R1398	T5117	T5113	cc	and,portion
R1399	T5118	T5113	conj	all,portion
R14	T854	T855	compound	Syrup,Disease
R140	T995	T996	det	both,models
R1400	T5119	T5118	prep	of,all
R1401	T5120	T5119	pobj	Exon,of
R1402	T5121	T5120	nummod	5,Exon
R1403	T5122	T5113	prep	with,portion
R1404	T5123	T5124	det	the,gene
R1405	T5124	T5122	pobj	gene,with
R1406	T5125	T5124	compound	PGKneo,gene
R1407	T5126	T5124	compound	marker,gene
R1408	T5127	T5128	punct	(,See
R1409	T5128	T5124	parataxis	See,gene
R141	T996	T994	pobj	models,of
R1410	T5129	T5130	compound	Fig.,1A
R1411	T5130	T5128	dobj	1A,See
R1412	T5131	T5128	punct	),See
R1413	T5132	T5124	prep	from,gene
R1414	T5133	T5134	det	the,vector
R1415	T5134	T5132	pobj	vector,from
R1416	T5135	T5134	compound	pPNT,vector
R1417	T5136	T5137	punct	[,22
R1418	T5137	T5111	parataxis	22,included
R1419	T5138	T5137	punct	],22
R142	T997	T993	auxpass	were,characterized
R1420	T5139	T5096	punct	.,created
R1421	T5141	T5142	det	The,construct
R1422	T5142	T5144	nsubjpass	construct,linearized
R1423	T5143	T5142	compound	targeting,construct
R1424	T5145	T5144	auxpass	was,linearized
R1425	T5146	T5144	prep	with,linearized
R1426	T5147	T5146	pobj	NotI,with
R1427	T5148	T5144	cc	and,linearized
R1428	T5149	T5144	conj	electroporated,linearized
R1429	T5150	T5149	prep	into,electroporated
R143	T998	T993	prep	at,characterized
R1430	T5151	T5152	compound	R1,cells
R1431	T5152	T5150	pobj	cells,into
R1432	T5153	T5152	compound	ES,cells
R1433	T5154	T5155	punct	[,23
R1434	T5155	T5152	parataxis	23,cells
R1435	T5156	T5155	punct	],23
R1436	T5157	T5149	prep	under,electroporated
R1437	T5158	T5157	pobj	conditions,under
R1438	T5159	T5158	acl	described,conditions
R1439	T5160	T5159	advmod	previously,described
R144	T999	T1000	det	the,levels
R1440	T5161	T5162	punct	[,24
R1441	T5162	T5159	parataxis	24,described
R1442	T5163	T5162	punct	],24
R1443	T5164	T5144	punct	.,linearized
R1444	T5166	T5167	compound	ES,cells
R1445	T5167	T5168	nsubjpass	cells,selected
R1446	T5169	T5168	auxpass	were,selected
R1447	T5170	T5168	prep	with,selected
R1448	T5171	T5170	pobj	G418,with
R1449	T5172	T5173	punct	(,Inc.
R145	T1000	T998	pobj	levels,at
R1450	T5173	T5171	parataxis	Inc.,G418
R1451	T5174	T5175	nummod	300,μg
R1452	T5175	T5173	dep	μg,Inc.
R1453	T5176	T5177	punct	/,ml
R1454	T5177	T5175	prep	ml,μg
R1455	T5178	T5173	punct	;,Inc.
R1456	T5179	T5173	compound	Life,Inc.
R1457	T5180	T5173	compound	Technologies,Inc.
R1458	T5181	T5173	punct	", ",Inc.
R1459	T5182	T5173	npadvmod	Gaithersburg,Inc.
R146	T1001	T1000	amod	molecular,levels
R1460	T5183	T5173	punct	", ",Inc.
R1461	T5184	T5173	npadvmod	MD,Inc.
R1462	T5185	T5173	punct	),Inc.
R1463	T5186	T5171	cc	and,G418
R1464	T5187	T5171	conj	gancyclovir,G418
R1465	T5188	T5187	punct	(,gancyclovir
R1466	T5189	T5190	nummod	2,μM
R1467	T5190	T5187	nmod	μM,gancyclovir
R1468	T5191	T5187	punct	;,gancyclovir
R1469	T5192	T5187	nmod	gift,gancyclovir
R147	T1002	T1001	punct	", ",molecular
R1470	T5193	T5192	prep	of,gift
R1471	T5194	T5193	pobj	Syntex,of
R1472	T5195	T5194	punct	", ",Syntex
R1473	T5196	T5197	compound	Palo,Alto
R1474	T5197	T5194	npadvmod	Alto,Syntex
R1475	T5198	T5194	punct	", ",Syntex
R1476	T5199	T5194	npadvmod	CA,Syntex
R1477	T5200	T5187	punct	),gancyclovir
R1478	T5201	T5168	punct	.,selected
R1479	T5203	T5204	advmod	Doubly,resistant
R148	T1003	T1001	conj	biochemical,molecular
R1480	T5204	T5205	amod	resistant,clones
R1481	T5205	T5206	nsubjpass	clones,screened
R1482	T5207	T5206	auxpass	were,screened
R1483	T5208	T5206	prep	for,screened
R1484	T5209	T5210	compound	gene,targeting
R1485	T5210	T5208	pobj	targeting,for
R1486	T5211	T5206	prep	by,screened
R1487	T5212	T5213	compound	Southern,blot
R1488	T5213	T5214	compound	blot,analysis
R1489	T5214	T5211	pobj	analysis,by
R149	T1004	T1003	punct	", ",biochemical
R1490	T5215	T5214	prep	of,analysis
R1491	T5216	T5217	nmod	BglI,DNA
R1492	T5217	T5215	pobj	DNA,of
R1493	T5218	T5217	amod	digested,DNA
R1494	T5219	T5217	amod	genomic,DNA
R1495	T5220	T5206	punct	.,screened
R1496	T5222	T5223	nsubjpass	Blots,hybridized
R1497	T5224	T5223	auxpass	were,hybridized
R1498	T5225	T5223	prep	with,hybridized
R1499	T5226	T5227	det	an,probe
R15	T855	T857	nsubj	Disease,is
R150	T1005	T1003	cc	and,biochemical
R1500	T5227	T5225	pobj	probe,with
R1501	T5228	T5229	npadvmod	Exon,specific
R1502	T5229	T5227	amod	specific,probe
R1503	T5230	T5228	nummod	6,Exon
R1504	T5231	T5232	dep	that,was
R1505	T5232	T5227	relcl	was,probe
R1506	T5233	T5232	acomp	external,was
R1507	T5234	T5233	prep	to,external
R1508	T5235	T5236	det	the,construct
R1509	T5236	T5234	pobj	construct,to
R151	T1006	T1007	amod	whole,animal
R1510	T5237	T5238	compound	gene,targeting
R1511	T5238	T5236	compound	targeting,construct
R1512	T5239	T5223	punct	.,hybridized
R1513	T5241	T5242	advmod	Correctly,targeted
R1514	T5242	T5243	amod	targeted,cells
R1515	T5243	T5245	nsubjpass	cells,injected
R1516	T5244	T5243	compound	ES,cells
R1517	T5246	T5245	auxpass	were,injected
R1518	T5247	T5245	prep	into,injected
R1519	T5248	T5249	compound	C57BL,6J
R152	T1007	T1003	conj	animal,biochemical
R1520	T5249	T5251	compound	6J,blastocysts
R1521	T5250	T5249	punct	/,6J
R1522	T5251	T5247	pobj	blastocysts,into
R1523	T5252	T5253	aux	to,produce
R1524	T5253	T5245	advcl	produce,injected
R1525	T5254	T5255	amod	chimeric,mice
R1526	T5255	T5253	dobj	mice,produce
R1527	T5256	T5253	advcl	using,produce
R1528	T5257	T5258	amod	standard,procedures
R1529	T5258	T5256	dobj	procedures,using
R153	T1008	T993	punct	.,characterized
R1530	T5259	T5245	punct	.,injected
R1531	T5261	T5262	amod	Heterozygous,offspring
R1532	T5262	T5263	nsubjpass	offspring,intercrossed
R1533	T5264	T5262	prep	from,offspring
R1534	T5265	T5266	npadvmod	germline,competent
R1535	T5266	T5267	amod	competent,chimeras
R1536	T5267	T5264	pobj	chimeras,from
R1537	T5268	T5263	auxpass	were,intercrossed
R1538	T5269	T5270	aux	to,produce
R1539	T5270	T5263	advcl	produce,intercrossed
R154	T1012	T1013	prep	By,created
R1540	T5271	T5272	amod	wild,type
R1541	T5272	T5273	nmod	type,knockout
R1542	T5273	T5288	nmod	knockout,mice
R1543	T5274	T5275	punct	(,+
R1544	T5275	T5272	punct	+,type
R1545	T5276	T5275	punct	+,+
R1546	T5277	T5275	punct	/,+
R1547	T5278	T5275	punct	),+
R1548	T5279	T5272	punct	", ",type
R1549	T5280	T5272	conj	heterozygous,type
R155	T1014	T1012	pcomp	disrupting,By
R1550	T5281	T5282	punct	(,-
R1551	T5282	T5280	punct	-,heterozygous
R1552	T5283	T5282	punct	+,-
R1553	T5284	T5282	punct	/,-
R1554	T5285	T5282	punct	),-
R1555	T5286	T5280	cc	and,heterozygous
R1556	T5287	T5280	conj	homozygous,heterozygous
R1557	T5288	T5270	dobj	mice,produce
R1558	T5289	T5290	punct	(,-
R1559	T5290	T5273	punct	-,knockout
R156	T1015	T1016	det	the,gene
R1560	T5291	T5290	punct	-,-
R1561	T5292	T5290	punct	/,-
R1562	T5293	T5290	punct	),-
R1563	T5294	T5263	punct	.,intercrossed
R1564	T5296	T5297	prep	At,used
R1565	T5298	T5299	det	all,generations
R1566	T5299	T5296	pobj	generations,At
R1567	T5300	T5297	punct	", ",used
R1568	T5301	T5302	punct	+,-
R1569	T5302	T5304	punct	-,pairs
R157	T1016	T1014	dobj	gene,disrupting
R1570	T5303	T5302	punct	/,-
R1571	T5304	T5297	nsubjpass	pairs,used
R1572	T5305	T5304	amod	breeding,pairs
R1573	T5306	T5297	auxpass	were,used
R1574	T5307	T5297	punct	.,used
R1575	T5309	T5310	nsubj	Results,are
R1576	T5311	T5309	acl	presented,Results
R1577	T5312	T5311	advmod	here,presented
R1578	T5313	T5310	prep	from,are
R1579	T5314	T5313	pobj	mice,from
R158	T1017	T1016	compound	E2,gene
R1580	T5315	T5314	acl	derived,mice
R1581	T5316	T5315	prep	from,derived
R1582	T5317	T5318	det	the,generations
R1583	T5318	T5316	pobj	generations,from
R1584	T5319	T5318	nmod	F2,generations
R1585	T5320	T5319	punct	+,F2
R1586	T5321	T5310	punct	.,are
R1587	T5323	T5324	det	All,animals
R1588	T5324	T5325	nsubj	animals,were
R1589	T5326	T5325	prep	of,were
R159	T1018	T1016	compound	subunit,gene
R1590	T5327	T5328	det	a,background
R1591	T5328	T5326	pobj	background,of
R1592	T5329	T5328	amod	mixed,background
R1593	T5330	T5331	nmod	C57BL,6J
R1594	T5331	T5328	nmod	6J,background
R1595	T5332	T5331	punct	/,6J
R1596	T5333	T5331	punct	×,6J
R1597	T5334	T5335	compound	Strain,SvJ
R1598	T5335	T5331	appos	SvJ,6J
R1599	T5336	T5335	compound	129Sv,SvJ
R16	T856	T855	compound	Urine,Disease
R160	T1019	T1016	prep	of,gene
R1600	T5337	T5335	punct	/,SvJ
R1601	T5338	T5328	amod	genetic,background
R1602	T5339	T5325	punct	.,were
R1603	T5873	T5872	prep	of,Production
R1604	T5874	T5875	amod	transgenic,mice
R1605	T5875	T5873	pobj	mice,of
R1606	T5877	T5878	amod	Standard,techniques
R1607	T5878	T5880	nsubjpass	techniques,used
R1608	T5879	T5878	amod	molecular,techniques
R1609	T5881	T5880	auxpass	were,used
R161	T1020	T1021	amod	branched,chain
R1610	T5882	T5883	aux	to,assemble
R1611	T5883	T5880	advcl	assemble,used
R1612	T5884	T5885	det	the,construct
R1613	T5885	T5883	dobj	construct,assemble
R1614	T5886	T5885	amod	transgenic,construct
R1615	T5887	T5885	punct	", ",construct
R1616	T5888	T5889	compound	pTRE,E2
R1617	T5889	T5885	appos	E2,construct
R1618	T5890	T5889	punct	-,E2
R1619	T5891	T5880	punct	.,used
R162	T1021	T1023	compound	chain,dehydrogenase
R1620	T5893	T5894	det	This,transgene
R1621	T5894	T5895	nsubj	transgene,contains
R1622	T5896	T5897	det	the,promoter
R1623	T5897	T5895	dobj	promoter,contains
R1624	T5898	T5897	nmod	tetracycline,promoter
R1625	T5899	T5897	amod	responsive,promoter
R1626	T5900	T5897	nmod	hCMV,promoter
R1627	T5901	T5900	punct	*,hCMV
R1628	T5902	T5900	punct	-,hCMV
R1629	T5903	T5900	nummod	1,hCMV
R163	T1022	T1021	punct	-,chain
R1630	T5904	T5897	punct	[,promoter
R1631	T5905	T5897	acl	consisting,promoter
R1632	T5906	T5905	prep	of,consisting
R1633	T5907	T5908	det	the,element
R1634	T5908	T5906	pobj	element,of
R1635	T5909	T5908	nmod	tetracycline,element
R1636	T5910	T5908	amod	responsive,element
R1637	T5911	T5908	punct	(,element
R1638	T5912	T5908	appos	TRE,element
R1639	T5913	T5908	punct	),element
R164	T1023	T1019	pobj	dehydrogenase,of
R1640	T5914	T5908	cc	and,element
R1641	T5915	T5916	det	a,promoter
R1642	T5916	T5908	conj	promoter,element
R1643	T5917	T5916	amod	minimal,promoter
R1644	T5918	T5916	compound	hCMV,promoter
R1645	T5919	T5916	punct	],promoter
R1646	T5920	T5921	punct	[,25
R1647	T5921	T5916	parataxis	25,promoter
R1648	T5922	T5921	punct	],25
R1649	T5923	T5916	prep	from,promoter
R165	T1024	T1025	compound	keto,acid
R1650	T5924	T5923	pobj	pTRE2,from
R1651	T5925	T5926	punct	(,Inc.
R1652	T5926	T5916	parataxis	Inc.,promoter
R1653	T5927	T5926	compound	Clontech,Inc.
R1654	T5928	T5926	punct	", ",Inc.
R1655	T5929	T5930	compound	Mt.,View
R1656	T5930	T5926	npadvmod	View,Inc.
R1657	T5931	T5926	punct	", ",Inc.
R1658	T5932	T5926	npadvmod	CA,Inc.
R1659	T5933	T5926	punct	),Inc.
R166	T1025	T1023	compound	acid,dehydrogenase
R1660	T5934	T5897	punct	", ",promoter
R1661	T5935	T5936	det	a,intron
R1662	T5936	T5897	conj	intron,promoter
R1663	T5937	T5936	amod	chimeric,intron
R1664	T5938	T5936	prep	from,intron
R1665	T5939	T5938	pobj	pCI,from
R1666	T5940	T5941	punct	(,Promega
R1667	T5941	T5936	parataxis	Promega,intron
R1668	T5942	T5941	punct	),Promega
R1669	T5943	T5944	aux	to,increase
R167	T1026	T1013	punct	", ",created
R1670	T5944	T5936	advcl	increase,intron
R1671	T5945	T5946	compound	message,stability
R1672	T5946	T5944	dobj	stability,increase
R1673	T5947	T5946	cc	and,stability
R1674	T5948	T5946	conj	expression,stability
R1675	T5949	T5950	punct	[,27
R1676	T5950	T5944	parataxis	27,increase
R1677	T5951	T5950	nummod	26,27
R1678	T5952	T5950	punct	",",27
R1679	T5953	T5950	punct	],27
R168	T1027	T1013	nsubj	we,created
R1680	T5954	T5936	punct	", ",intron
R1681	T5955	T5956	det	a,sequence
R1682	T5956	T5936	conj	sequence,intron
R1683	T5957	T5956	compound	Kozak,sequence
R1684	T5958	T5956	compound	consensus,sequence
R1685	T5959	T5956	prep	at,sequence
R1686	T5960	T5961	det	the,codon
R1687	T5961	T5959	pobj	codon,at
R1688	T5962	T5961	compound	initiation,codon
R1689	T5963	T5964	aux	to,optimize
R169	T1028	T1029	det	a,model
R1690	T5964	T5956	advcl	optimize,sequence
R1691	T5965	T5964	dobj	translation,optimize
R1692	T5966	T5967	punct	[,28
R1693	T5967	T5964	parataxis	28,optimize
R1694	T5968	T5967	punct	],28
R1695	T5969	T5956	punct	", ",sequence
R1696	T5970	T5971	det	the,cDNA
R1697	T5971	T5956	conj	cDNA,sequence
R1698	T5972	T5971	amod	human,cDNA
R1699	T5973	T5971	compound	E2,cDNA
R17	T858	T855	punct	(,Disease
R170	T1029	T1013	dobj	model,created
R1700	T5974	T5975	dep	which,modified
R1701	T5975	T5971	relcl	modified,cDNA
R1702	T5976	T5975	aux	has,modified
R1703	T5977	T5975	auxpass	been,modified
R1704	T5978	T5979	aux	to,contain
R1705	T5979	T5975	advcl	contain,modified
R1706	T5980	T5981	det	a,linker
R1707	T5981	T5979	dobj	linker,contain
R1708	T5982	T5981	nummod	4,linker
R1709	T5983	T5982	punct	×,4
R171	T1030	T1031	compound	gene,knockout
R1710	T5984	T5981	compound	alanine,linker
R1711	T5985	T5981	acl	followed,linker
R1712	T5986	T5985	agent	by,followed
R1713	T5987	T5988	det	a,tag
R1714	T5988	T5986	pobj	tag,by
R1715	T5989	T5990	compound	c,myc
R1716	T5990	T5988	compound	myc,tag
R1717	T5991	T5990	punct	-,myc
R1718	T5992	T5988	compound	epitope,tag
R1719	T5993	T5988	prep	at,tag
R172	T1031	T1029	compound	knockout,model
R1720	T5994	T5995	det	the,terminus
R1721	T5995	T5993	pobj	terminus,at
R1722	T5996	T5995	compound	carboxy,terminus
R1723	T5997	T5998	aux	to,facilitate
R1724	T5998	T5988	advcl	facilitate,tag
R1725	T5999	T5998	dobj	detection,facilitate
R1726	T6000	T5971	punct	", ",cDNA
R1727	T6001	T5971	cc	and,cDNA
R1728	T6002	T6003	det	an,sequence
R1729	T6003	T5971	conj	sequence,cDNA
R173	T1032	T1029	compound	mouse,model
R1730	T6004	T6003	nmod	SV40,sequence
R1731	T6005	T6003	amod	late,sequence
R1732	T6006	T6003	compound	polyadenylation,sequence
R1733	T6007	T6003	prep	from,sequence
R1734	T6008	T6007	pobj	pCI,from
R1735	T6009	T6003	prep	for,sequence
R1736	T6010	T6011	amod	enhanced,stability
R1737	T6011	T6009	pobj	stability,for
R1738	T6012	T6011	compound	mRNA,stability
R1739	T6013	T6011	cc	and,stability
R174	T1033	T1029	prep	of,model
R1740	T6014	T6011	conj	translation,stability
R1741	T6015	T5895	punct	.,contains
R1742	T6017	T6018	det	The,transgene
R1743	T6018	T6024	nsubj	transgene,purified
R1744	T6019	T6020	nummod	2.48,kb
R1745	T6020	T6018	compound	kb,transgene
R1746	T6021	T6022	compound	TRE,E2
R1747	T6022	T6018	compound	E2,transgene
R1748	T6023	T6022	punct	-,E2
R1749	T6025	T6024	aux	was,purified
R175	T1034	T1035	amod	classic,MSUD
R1750	T6026	T6024	prep	from,purified
R1751	T6027	T6028	compound	vector,sequences
R1752	T6028	T6026	pobj	sequences,from
R1753	T6029	T6024	prep	following,purified
R1754	T6030	T6029	pobj	digestion,following
R1755	T6031	T6030	prep	with,digestion
R1756	T6032	T6031	pobj	XhoI,with
R1757	T6033	T6032	cc	and,XhoI
R1758	T6034	T6032	conj	BamHI,XhoI
R1759	T6035	T6024	cc	and,purified
R176	T1035	T1033	pobj	MSUD,of
R1760	T6036	T6024	conj	injected,purified
R1761	T6037	T6036	prep	into,injected
R1762	T6038	T6039	nmod	C57BL,6J
R1763	T6039	T6041	nmod	6J,Strain
R1764	T6040	T6039	punct	/,6J
R1765	T6041	T6047	compound	Strain,embryos
R1766	T6042	T6039	cc	or,6J
R1767	T6043	T6044	compound	C57BL,6J
R1768	T6044	T6039	conj	6J,6J
R1769	T6045	T6044	punct	/,6J
R177	T1036	T1013	punct	.,created
R1770	T6046	T6041	punct	×,Strain
R1771	T6047	T6037	pobj	embryos,into
R1772	T6048	T6047	compound	129SvEv,embryos
R1773	T6049	T6047	compound	mouse,embryos
R1774	T6050	T6024	prep	at,purified
R1775	T6051	T6052	det	the,facilities
R1776	T6052	T6050	pobj	facilities,at
R1777	T6053	T6052	amod	transgenic,facilities
R1778	T6054	T6052	compound	core,facilities
R1779	T6055	T6052	prep	of,facilities
R178	T1038	T1039	det	The,mice
R1780	T6056	T6057	det	the,University
R1781	T6057	T6055	pobj	University,of
R1782	T6058	T6057	prep	of,University
R1783	T6059	T6058	pobj	Pittsburgh,of
R1784	T6060	T6057	cc	and,University
R1785	T6061	T6062	det	the,University
R1786	T6062	T6057	conj	University,University
R1787	T6063	T6062	prep	of,University
R1788	T6064	T6063	pobj	Cincinnati,of
R1789	T6065	T6024	punct	", ",purified
R179	T1039	T1042	nsubj	mice,lacked
R1790	T6066	T6024	advmod	respectively,purified
R1791	T6067	T6024	punct	.,purified
R1792	T6069	T6070	amod	Genomic,DNA
R1793	T6070	T6071	nsubjpass	DNA,screened
R1794	T6072	T6070	prep	from,DNA
R1795	T6073	T6074	det	the,tail
R1796	T6074	T6072	pobj	tail,from
R1797	T6075	T6074	prep	of,tail
R1798	T6076	T6075	pobj	mice,of
R1799	T6077	T6071	auxpass	was,screened
R18	T859	T855	appos	MSUD,Disease
R180	T1040	T1039	amod	homozygous,mice
R1800	T6078	T6071	prep	by,screened
R1801	T6079	T6080	compound	Southern,blot
R1802	T6080	T6081	compound	blot,analysis
R1803	T6081	T6078	pobj	analysis,by
R1804	T6082	T6071	prep	following,screened
R1805	T6083	T6082	pobj	digestion,following
R1806	T6084	T6083	prep	with,digestion
R1807	T6085	T6084	pobj	EcoRI,with
R1808	T6086	T6083	cc	and,digestion
R1809	T6087	T6083	conj	hybridization,digestion
R181	T1041	T1039	compound	knockout,mice
R1810	T6088	T6087	prep	to,hybridization
R1811	T6089	T6090	det	an,probe
R1812	T6090	T6088	pobj	probe,to
R1813	T6091	T6092	punct	~,400
R1814	T6092	T6093	nummod	400,bp
R1815	T6093	T6090	compound	bp,probe
R1816	T6094	T6090	acl	derived,probe
R1817	T6095	T6094	prep	from,derived
R1818	T6096	T6097	det	the,portion
R1819	T6097	T6095	pobj	portion,from
R182	T1043	T1044	amod	branched,chain
R1820	T6098	T6097	compound	SV40,portion
R1821	T6099	T6097	prep	of,portion
R1822	T6100	T6101	det	the,transgene
R1823	T6101	T6099	pobj	transgene,of
R1824	T6102	T6103	compound	TRE,E2
R1825	T6103	T6101	compound	E2,transgene
R1826	T6104	T6103	punct	-,E2
R1827	T6105	T6071	punct	.,screened
R1828	T6427	T6426	prep	of,Production
R1829	T6428	T6429	amod	intermediate,model
R183	T1044	T1046	compound	chain,dehydrogenase
R1830	T6429	T6427	pobj	model,of
R1831	T6430	T6429	nmod	MSUD,model
R1832	T6431	T6429	amod	murine,model
R1833	T6433	T6434	det	The,lines
R1834	T6434	T6440	nsubjpass	lines,mated
R1835	T6435	T6434	amod	various,lines
R1836	T6436	T6437	nmod	TRE,E2
R1837	T6437	T6434	nmod	E2,lines
R1838	T6438	T6437	punct	-,E2
R1839	T6439	T6434	amod	transgenic,lines
R184	T1045	T1044	punct	-,chain
R1840	T6441	T6434	acl	produced,lines
R1841	T6442	T6440	auxpass	were,mated
R1842	T6443	T6440	advmod	independently,mated
R1843	T6444	T6440	prep	to,mated
R1844	T6445	T6444	pobj	mice,to
R1845	T6446	T6447	dep	that,were
R1846	T6447	T6445	relcl	were,mice
R1847	T6448	T6447	acomp	heterozygous,were
R1848	T6449	T6448	prep	for,heterozygous
R1849	T6450	T6451	preconj	both,knockout
R185	T1046	T1049	compound	dehydrogenase,activity
R1850	T6451	T6449	pobj	knockout,for
R1851	T6452	T6451	det	the,knockout
R1852	T6453	T6451	compound	E2,knockout
R1853	T6454	T6451	cc	and,knockout
R1854	T6455	T6456	det	the,transgene
R1855	T6456	T6451	conj	transgene,knockout
R1856	T6457	T6458	compound	LAP,tTA
R1857	T6458	T6456	compound	tTA,transgene
R1858	T6459	T6458	punct	-,tTA
R1859	T6460	T6461	punct	[,background
R186	T1047	T1048	compound	keto,acid
R1860	T6461	T6440	parataxis	background,mated
R1861	T6462	T6463	nmod	Tg,Stock
R1862	T6463	T6461	dep	Stock,background
R1863	T6464	T6465	punct	(,tTALap
R1864	T6465	T6463	parataxis	tTALap,Stock
R1865	T6466	T6465	punct	),tTALap
R1866	T6467	T6468	nmod	Bjd,J
R1867	T6468	T6463	nmod	J,Stock
R1868	T6469	T6468	punct	/,J
R1869	T6470	T6463	punct	;,Stock
R187	T1048	T1046	compound	acid,dehydrogenase
R1870	T6471	T6463	nummod	3272,Stock
R1871	T6472	T6461	punct	;,background
R1872	T6473	T6474	det	The,Laboratory
R1873	T6474	T6461	dep	Laboratory,background
R1874	T6475	T6474	compound	Jackson,Laboratory
R1875	T6476	T6474	punct	", ",Laboratory
R1876	T6477	T6478	compound	Bar,Harbor
R1877	T6478	T6474	npadvmod	Harbor,Laboratory
R1878	T6479	T6474	punct	", ",Laboratory
R1879	T6480	T6474	npadvmod	ME,Laboratory
R188	T1049	T1042	dobj	activity,lacked
R1880	T6481	T6461	punct	;,background
R1881	T6482	T6461	nmod	NMRI,background
R1882	T6483	T6482	punct	×,NMRI
R1883	T6484	T6482	appos	FVB,NMRI
R1884	T6485	T6482	punct	×,NMRI
R1885	T6486	T6487	compound	C57BL,6J
R1886	T6487	T6482	appos	6J,NMRI
R1887	T6488	T6487	punct	/,6J
R1888	T6489	T6461	punct	],background
R1889	T6490	T6440	punct	.,mated
R189	T1050	T1049	punct	", ",activity
R1890	T6492	T6493	nsubj	Interbreeding,resulted
R1891	T6494	T6492	prep	of,Interbreeding
R1892	T6495	T6494	pobj	animals,of
R1893	T6496	T6497	dep	that,were
R1894	T6497	T6495	relcl	were,animals
R1895	T6498	T6497	acomp	heterozygous,were
R1896	T6499	T6498	prep	for,heterozygous
R1897	T6500	T6501	det	both,transgenes
R1898	T6501	T6499	pobj	transgenes,for
R1899	T6502	T6501	cc	and,transgenes
R19	T860	T857	punct	),is
R190	T1051	T1052	compound	E2,immunoreactivity
R1900	T6503	T6504	det	the,knockout
R1901	T6504	T6501	conj	knockout,transgenes
R1902	T6505	T6493	prep	in,resulted
R1903	T6506	T6507	det	the,production
R1904	T6507	T6505	pobj	production,in
R1905	T6508	T6507	prep	of,production
R1906	T6509	T6508	pobj	mice,of
R1907	T6510	T6509	prep	with,mice
R1908	T6511	T6512	det	a,variety
R1909	T6512	T6510	pobj	variety,with
R191	T1052	T1049	appos	immunoreactivity,activity
R1910	T6513	T6512	prep	of,variety
R1911	T6514	T6513	pobj	genotypes,of
R1912	T6515	T6509	prep	including,mice
R1913	T6516	T6517	det	some,animals
R1914	T6517	T6515	pobj	animals,including
R1915	T6518	T6519	dep	that,were
R1916	T6519	T6517	relcl	were,animals
R1917	T6520	T6519	acomp	homozygous,were
R1918	T6521	T6520	prep	for,homozygous
R1919	T6522	T6523	det	the,knockout
R192	T1053	T1042	punct	", ",lacked
R1920	T6523	T6521	pobj	knockout,for
R1921	T6524	T6519	cc	and,were
R1922	T6525	T6519	conj	were,were
R1923	T6526	T6525	acomp	positive,were
R1924	T6527	T6526	prep	for,positive
R1925	T6528	T6529	det	both,transgenes
R1926	T6529	T6527	pobj	transgenes,for
R1927	T6530	T6531	punct	(,refer
R1928	T6531	T6525	parataxis	refer,were
R1929	T6532	T6531	nsubj	we,refer
R193	T1054	T1042	cc	and,lacked
R1930	T6533	T6531	prep	to,refer
R1931	T6534	T6535	det	this,genotype
R1932	T6535	T6533	pobj	genotype,to
R1933	T6536	T6531	prep	as,refer
R1934	T6537	T6538	det	the,genotype
R1935	T6538	T6536	pobj	genotype,as
R1936	T6539	T6538	punct	"""",genotype
R1937	T6540	T6538	nmod	rescue,genotype
R1938	T6541	T6538	punct	"""",genotype
R1939	T6542	T6531	punct	),refer
R194	T1055	T1042	conj	had,lacked
R1940	T6543	T6493	punct	.,resulted
R1941	T6545	T6546	mark	If,were
R1942	T6546	T6558	advcl	were,survive
R1943	T6547	T6548	poss	our,strategy
R1944	T6548	T6546	nsubj	strategy,were
R1945	T6549	T6548	prep	for,strategy
R1946	T6550	T6549	pcomp	rescuing,for
R1947	T6551	T6552	det	the,phenotype
R1948	T6552	T6550	dobj	phenotype,rescuing
R1949	T6553	T6552	amod	neonatal,phenotype
R195	T1056	T1057	det	a,increase
R1950	T6554	T6552	amod	lethal,phenotype
R1951	T6555	T6552	prep	of,phenotype
R1952	T6556	T6557	det	the,knockout
R1953	T6557	T6555	pobj	knockout,of
R1954	T6559	T6546	acomp	successful,were
R1955	T6560	T6558	punct	", ",survive
R1956	T6561	T6558	advmod	then,survive
R1957	T6562	T6563	det	those,animals
R1958	T6563	T6558	nsubj	animals,survive
R1959	T6564	T6563	amod	homozygous,animals
R196	T1057	T1055	dobj	increase,had
R1960	T6565	T6563	compound	knockout,animals
R1961	T6566	T6567	dep	that,had
R1962	T6567	T6563	relcl	had,animals
R1963	T6568	T6569	det	both,transgenes
R1964	T6569	T6567	dobj	transgenes,had
R1965	T6570	T6558	aux	would,survive
R1966	T6571	T6558	prep	beyond,survive
R1967	T6572	T6573	det	the,period
R1968	T6573	T6571	pobj	period,beyond
R1969	T6574	T6573	amod	neonatal,period
R197	T1058	T1057	nummod	3,increase
R1970	T6575	T6558	punct	.,survive
R1971	T6672	T6673	det	All,mice
R1972	T6673	T6674	nsubjpass	mice,genotyped
R1973	T6675	T6674	auxpass	were,genotyped
R1974	T6676	T6674	prep	by,genotyped
R1975	T6677	T6678	compound	Southern,blot
R1976	T6678	T6679	compound	blot,analysis
R1977	T6679	T6676	pobj	analysis,by
R1978	T6680	T6674	punct	.,genotyped
R1979	T6682	T6683	amod	Genomic,DNAs
R198	T1059	T1058	punct	-,3
R1980	T6683	T6684	nsubjpass	DNAs,digested
R1981	T6685	T6683	acl	prepared,DNAs
R1982	T6686	T6685	prep	from,prepared
R1983	T6687	T6688	compound	tail,snips
R1984	T6688	T6686	pobj	snips,from
R1985	T6689	T6684	auxpass	were,digested
R1986	T6690	T6684	prep	with,digested
R1987	T6691	T6692	det	an,enzyme
R1988	T6692	T6690	pobj	enzyme,with
R1989	T6693	T6692	amod	appropriate,enzyme
R199	T1060	T1058	amod	fold,3
R1990	T6694	T6692	compound	restriction,enzyme
R1991	T6695	T6684	punct	", ",digested
R1992	T6696	T6697	dep	size,fractionated
R1993	T6697	T6684	conj	fractionated,digested
R1994	T6698	T6697	prep	by,fractionated
R1995	T6699	T6700	compound	agarose,gel
R1996	T6700	T6701	compound	gel,electrophoresis
R1997	T6701	T6698	pobj	electrophoresis,by
R1998	T6702	T6697	punct	", ",fractionated
R1999	T6703	T6697	conj	blotted,fractionated
R2	T837	T834	conj	characterization,Production
R20	T861	T862	det	an,error
R200	T1061	T1057	prep	in,increase
R2000	T6704	T6703	prep	to,blotted
R2001	T6705	T6704	pobj	nylon,to
R2002	T6706	T6703	punct	", ",blotted
R2003	T6707	T6703	cc	and,blotted
R2004	T6708	T6703	conj	probed,blotted
R2005	T6709	T6708	advcl	using,probed
R2006	T6710	T6711	amod	standard,procedures
R2007	T6711	T6709	dobj	procedures,using
R2008	T6712	T6684	punct	.,digested
R2009	T7185	T7186	amod	Primary,fibroblasts
R201	T1062	T1063	amod	circulating,acids
R2010	T7186	T7189	nsubjpass	fibroblasts,prepared
R2011	T7187	T7186	nmod	mouse,fibroblasts
R2012	T7188	T7186	amod	embryonic,fibroblasts
R2013	T7190	T7189	auxpass	were,prepared
R2014	T7191	T7189	prep	from,prepared
R2015	T7192	T7193	amod	embryonic,fetuses
R2016	T7193	T7191	pobj	fetuses,from
R2017	T7194	T7195	nmod	day,16.5
R2018	T7195	T7193	nummod	16.5,fetuses
R2019	T7196	T7195	punct	~,16.5
R202	T1063	T1061	pobj	acids,in
R2020	T7197	T7198	punct	–,18.5
R2021	T7198	T7195	prep	18.5,16.5
R2022	T7199	T7200	mark	as,described
R2023	T7200	T7189	advcl	described,prepared
R2024	T7201	T7202	punct	[,29
R2025	T7202	T7189	parataxis	29,prepared
R2026	T7203	T7202	punct	],29
R2027	T7204	T7189	punct	.,prepared
R2028	T7206	T7207	nsubjpass	Fibroblasts,passed
R2029	T7208	T7207	auxpass	were,passed
R203	T1064	T1065	amod	branched,chain
R2030	T7209	T7207	prep	onto,passed
R2031	T7210	T7209	pobj	glass,onto
R2032	T7211	T7207	punct	", ",passed
R2033	T7212	T7207	conj	fixed,passed
R2034	T7213	T7212	prep	in,fixed
R2035	T7214	T7215	nummod	2,%
R2036	T7215	T7216	compound	%,paraformaldehyde
R2037	T7216	T7213	pobj	paraformaldehyde,in
R2038	T7217	T7212	prep	in,fixed
R2039	T7218	T7217	pobj	PBS,in
R204	T1065	T1063	compound	chain,acids
R2040	T7219	T7212	prep	for,fixed
R2041	T7220	T7221	nummod	10,minutes
R2042	T7221	T7219	pobj	minutes,for
R2043	T7222	T7212	punct	", ",fixed
R2044	T7223	T7212	conj	permeabilized,fixed
R2045	T7224	T7223	prep	in,permeabilized
R2046	T7225	T7226	nummod	2,%
R2047	T7226	T7227	compound	%,paraformaldehyde
R2048	T7227	T7224	pobj	paraformaldehyde,in
R2049	T7228	T7227	acl	containing,paraformaldehyde
R205	T1066	T1065	punct	-,chain
R2050	T7229	T7230	nummod	0.1,%
R2051	T7230	T7231	compound	%,X100
R2052	T7231	T7228	dobj	X100,containing
R2053	T7232	T7231	compound	Triton,X100
R2054	T7233	T7223	prep	for,permeabilized
R2055	T7234	T7235	nummod	10,minutes
R2056	T7235	T7233	pobj	minutes,for
R2057	T7236	T7223	cc	and,permeabilized
R2058	T7237	T7223	conj	washed,permeabilized
R2059	T7238	T7239	nummod	three,times
R206	T1067	T1063	compound	amino,acids
R2060	T7239	T7237	npadvmod	times,washed
R2061	T7240	T7237	prep	in,washed
R2062	T7241	T7240	pobj	PBS,in
R2063	T7242	T7241	acl	containing,PBS
R2064	T7243	T7244	nummod	0.5,%
R2065	T7244	T7245	compound	%,BSA
R2066	T7245	T7242	dobj	BSA,containing
R2067	T7246	T7245	cc	and,BSA
R2068	T7247	T7248	nummod	0.15,%
R2069	T7248	T7249	compound	%,glycine
R207	T1068	T1042	punct	.,lacked
R2070	T7249	T7245	conj	glycine,BSA
R2071	T7250	T7242	punct	", ",containing
R2072	T7251	T7242	npadvmod	pH,containing
R2073	T7252	T7251	nummod	7.4,pH
R2074	T7253	T7254	punct	(,A
R2075	T7254	T7242	parataxis	A,containing
R2076	T7255	T7254	compound	Buffer,A
R2077	T7256	T7254	punct	),A
R2078	T7257	T7207	punct	.,passed
R2079	T7259	T7260	prep	Following,incubated
R208	T1070	T1071	det	These,derangements
R2080	T7261	T7262	det	a,incubation
R2081	T7262	T7259	pobj	incubation,Following
R2082	T7263	T7264	nummod	30,min
R2083	T7264	T7262	compound	min,incubation
R2084	T7265	T7262	prep	with,incubation
R2085	T7266	T7267	amod	purified,IgG
R2086	T7267	T7265	pobj	IgG,with
R2087	T7268	T7267	compound	goat,IgG
R2088	T7269	T7270	punct	(,μg
R2089	T7270	T7267	parataxis	μg,IgG
R209	T1071	T1073	nsubj	derangements,resulted
R2090	T7271	T7270	nummod	50,μg
R2091	T7272	T7270	punct	(,μg
R2092	T7273	T7274	punct	/,ml
R2093	T7274	T7270	prep	ml,μg
R2094	T7275	T7270	punct	),μg
R2095	T7276	T7262	prep	at,incubation
R2096	T7277	T7278	nummod	25,°C
R2097	T7278	T7276	pobj	°C,at
R2098	T7279	T7262	cc	and,incubation
R2099	T7280	T7281	nummod	three,washes
R21	T862	T857	attr	error,is
R210	T1072	T1071	amod	metabolic,derangements
R2100	T7281	T7262	conj	washes,incubation
R2101	T7282	T7281	amod	additional,washes
R2102	T7283	T7281	prep	with,washes
R2103	T7284	T7285	compound	Buffer,A
R2104	T7285	T7283	pobj	A,with
R2105	T7286	T7260	punct	", ",incubated
R2106	T7287	T7260	nsubjpass	cells,incubated
R2107	T7288	T7260	auxpass	were,incubated
R2108	T7289	T7260	prep	for,incubated
R2109	T7290	T7291	nummod	60,min
R211	T1074	T1073	prep	in,resulted
R2110	T7291	T7289	pobj	min,for
R2111	T7292	T7260	prep	with,incubated
R2112	T7293	T7294	npadvmod	E2,specific
R2113	T7294	T7296	amod	specific,antiserum
R2114	T7295	T7294	punct	-,specific
R2115	T7296	T7292	pobj	antiserum,with
R2116	T7297	T7298	punct	[,30
R2117	T7298	T7296	parataxis	30,antiserum
R2118	T7299	T7298	punct	],30
R2119	T7300	T7296	prep	at,antiserum
R212	T1075	T1076	amod	neonatal,lethality
R2120	T7301	T7302	nummod	1,μg
R2121	T7302	T7300	pobj	μg,at
R2122	T7303	T7304	punct	/,ml
R2123	T7304	T7302	prep	ml,μg
R2124	T7305	T7260	advcl	followed,incubated
R2125	T7306	T7305	agent	by,followed
R2126	T7307	T7308	nummod	three,washes
R2127	T7308	T7306	pobj	washes,by
R2128	T7309	T7308	prep	in,washes
R2129	T7310	T7311	compound	Buffer,A
R213	T1076	T1074	pobj	lethality,in
R2130	T7311	T7309	pobj	A,in
R2131	T7312	T7308	cc	and,washes
R2132	T7313	T7314	nummod	60,minute
R2133	T7314	T7315	compound	minute,incubation
R2134	T7315	T7308	conj	incubation,washes
R2135	T7316	T7315	prep	in,incubation
R2136	T7317	T7318	advmod	fluorescently,labeled
R2137	T7318	T7319	amod	labeled,antibody
R2138	T7319	T7316	pobj	antibody,in
R2139	T7320	T7319	amod	second,antibody
R214	T1077	T1073	punct	.,resulted
R2140	T7321	T7322	punct	(,μg
R2141	T7322	T7260	parataxis	μg,incubated
R2142	T7323	T7324	quantmod	1,2
R2143	T7324	T7322	nummod	2,μg
R2144	T7325	T7324	punct	–,2
R2145	T7326	T7327	punct	/,ml
R2146	T7327	T7322	prep	ml,μg
R2147	T7328	T7322	punct	),μg
R2148	T7329	T7260	punct	.,incubated
R2149	T7331	T7332	det	The,cells
R215	T1079	T1080	amod	Transgenic,expression
R2150	T7332	T7333	nsubjpass	cells,washed
R2151	T7334	T7333	auxpass	were,washed
R2152	T7335	T7333	advmod	then,washed
R2153	T7336	T7337	nummod	six,times
R2154	T7337	T7333	npadvmod	times,washed
R2155	T7338	T7339	punct	(,min
R2156	T7339	T7337	parataxis	min,times
R2157	T7340	T7339	nummod	5,min
R2158	T7341	T7342	punct	/,wash
R2159	T7342	T7339	prep	wash,min
R216	T1080	T1081	nsubj	expression,produced
R2160	T7343	T7339	punct	),min
R2161	T7344	T7333	prep	in,washed
R2162	T7345	T7346	compound	Buffer,A
R2163	T7346	T7344	pobj	A,in
R2164	T7347	T7333	cc	and,washed
R2165	T7348	T7349	advmod	then,mounted
R2166	T7349	T7333	conj	mounted,washed
R2167	T7350	T7349	prep	in,mounted
R2168	T7351	T7350	pobj	gelvatol,in
R2169	T7352	T7353	punct	(,Monsanto
R217	T1082	T1080	prep	of,expression
R2170	T7353	T7351	parataxis	Monsanto,gelvatol
R2171	T7354	T7353	punct	", ",Monsanto
R2172	T7355	T7356	compound	St,Louis
R2173	T7356	T7353	npadvmod	Louis,Monsanto
R2174	T7357	T7353	punct	),Monsanto
R2175	T7358	T7333	punct	.,washed
R2176	T7360	T7361	advmod	When,examined
R2177	T7361	T7367	advcl	examined,was
R2178	T7362	T7361	nsubjpass	livers,examined
R2179	T7363	T7362	prep	from,livers
R218	T1083	T1084	det	a,cDNA
R2180	T7364	T7365	amod	newborn,pups
R2181	T7365	T7363	pobj	pups,from
R2182	T7366	T7361	auxpass	were,examined
R2183	T7368	T7367	punct	", ",was
R2184	T7369	T7367	nsubj	fixation,was
R2185	T7370	T7367	prep	by,was
R2186	T7371	T7370	pobj	immersion,by
R2187	T7372	T7371	prep	in,immersion
R2188	T7373	T7374	nummod	2,%
R2189	T7374	T7375	compound	%,paraformaldehyde
R219	T1084	T1082	pobj	cDNA,of
R2190	T7375	T7372	pobj	paraformaldehyde,in
R2191	T7376	T7371	acl	followed,immersion
R2192	T7377	T7376	agent	by,followed
R2193	T7378	T7377	pobj	cryoprotection,by
R2194	T7379	T7378	cc	and,cryoprotection
R2195	T7380	T7381	compound	shock,freezing
R2196	T7381	T7378	conj	freezing,cryoprotection
R2197	T7382	T7378	prep	in,cryoprotection
R2198	T7383	T7384	amod	liquid,nitrogen
R2199	T7384	T7385	npadvmod	nitrogen,cooled
R22	T863	T862	amod	inborn,error
R220	T1085	T1084	amod	human,cDNA
R2200	T7385	T7386	amod	cooled,isopentane
R2201	T7386	T7382	pobj	isopentane,in
R2202	T7387	T7378	cc	and,cryoprotection
R2203	T7388	T7378	conj	sectioning,cryoprotection
R2204	T7389	T7390	punct	(,microns
R2205	T7390	T7388	parataxis	microns,sectioning
R2206	T7391	T7390	nummod	5,microns
R2207	T7392	T7390	punct	),microns
R2208	T7393	T7367	punct	.,was
R2209	T7395	T7396	advmod	Otherwise,was
R221	T1086	T1084	compound	E2,cDNA
R2210	T7397	T7396	nsubj	processing,was
R2211	T7398	T7396	prep	as,was
R2212	T7399	T7398	prep	for,as
R2213	T7400	T7401	det	the,cells
R2214	T7401	T7399	pobj	cells,for
R2215	T7402	T7401	advmod	above,cells
R2216	T7403	T7396	punct	(,was
R2217	T7404	T7396	prep	without,was
R2218	T7405	T7406	det	the,steps
R2219	T7406	T7404	pobj	steps,without
R222	T1087	T1080	prep	in,expression
R2220	T7407	T7406	nmod	fixation,steps
R2221	T7408	T7407	cc	and,fixation
R2222	T7409	T7407	conj	permabilization,fixation
R2223	T7410	T7396	punct	),was
R2224	T7411	T7396	punct	.,was
R2225	T7545	T7546	compound	Amino,acid
R2226	T7546	T7547	compound	acid,analysis
R2227	T7549	T7550	nsubjpass	Blood,collected
R2228	T7551	T7550	auxpass	was,collected
R2229	T7552	T7550	prep	from,collected
R223	T1088	T1089	det	the,liver
R2230	T7553	T7554	det	the,vein
R2231	T7554	T7552	pobj	vein,from
R2232	T7555	T7554	amod	retroorbital,vein
R2233	T7556	T7554	nmod	sinus,vein
R2234	T7557	T7556	cc	or,sinus
R2235	T7558	T7556	conj	tail,sinus
R2236	T7559	T7554	prep	of,vein
R2237	T7560	T7559	pobj	mice,of
R2238	T7561	T7550	cc	and,collected
R2239	T7562	T7550	conj	spotted,collected
R224	T1089	T1087	pobj	liver,in
R2240	T7563	T7562	prep	on,spotted
R2241	T7564	T7565	det	a,paper
R2242	T7565	T7563	pobj	paper,on
R2243	T7566	T7565	compound	filter,paper
R2244	T7567	T7568	advmod	routinely,used
R2245	T7568	T7565	acl	used,paper
R2246	T7569	T7568	prep	for,used
R2247	T7570	T7571	compound	blood,analysis
R2248	T7571	T7569	pobj	analysis,for
R2249	T7572	T7573	compound	amino,acid
R225	T1090	T1089	prep	of,liver
R2250	T7573	T7571	compound	acid,analysis
R2251	T7574	T7571	prep	for,analysis
R2252	T7575	T7576	amod	prenatal,screening
R2253	T7576	T7574	pobj	screening,for
R2254	T7577	T7550	punct	.,collected
R2255	T7579	T7580	nsubjpass	Concentrations,determined
R2256	T7581	T7579	prep	of,Concentrations
R2257	T7582	T7581	pobj	BCAA,of
R2258	T7583	T7582	cc	and,BCAA
R2259	T7584	T7585	amod	other,acids
R226	T1091	T1092	det	the,animals
R2260	T7585	T7582	conj	acids,BCAA
R2261	T7586	T7585	compound	amino,acids
R2262	T7587	T7579	prep	in,Concentrations
R2263	T7588	T7587	pobj	blood,in
R2264	T7589	T7580	auxpass	were,determined
R2265	T7590	T7580	prep	by,determined
R2266	T7591	T7592	amod	tandem,spectrometry
R2267	T7592	T7590	pobj	spectrometry,by
R2268	T7593	T7592	compound	mass,spectrometry
R2269	T7594	T7595	punct	(,Screening
R227	T1092	T1090	pobj	animals,of
R2270	T7595	T7592	parataxis	Screening,spectrometry
R2271	T7596	T7595	compound	Pediatrix,Screening
R2272	T7597	T7595	punct	", ",Screening
R2273	T7598	T7595	npadvmod	Bridgeville,Screening
R2274	T7599	T7595	punct	", ",Screening
R2275	T7600	T7595	npadvmod	PA,Screening
R2276	T7601	T7595	punct	),Screening
R2277	T7602	T7580	punct	.,determined
R228	T1093	T1092	compound	E2,animals
R2282	T8106	T8105	prep	of,Assay
R2283	T8107	T8108	compound	BCKDH,activity
R2284	T8108	T8106	pobj	activity,of
R2285	T8110	T8111	nsubjpass	Livers,removed
R2286	T8112	T8111	auxpass	were,removed
R2287	T8113	T8111	punct	", ",removed
R2288	T8114	T8111	conj	frozen,removed
R2289	T8115	T8114	prep	in,frozen
R229	T1094	T1092	compound	knockout,animals
R2290	T8116	T8117	amod	liquid,nitrogen
R2291	T8117	T8115	pobj	nitrogen,in
R2292	T8118	T8114	punct	", ",frozen
R2293	T8119	T8114	cc	and,frozen
R2294	T8120	T8114	conj	stored,frozen
R2295	T8121	T8120	prep	at,stored
R2296	T8122	T8123	punct	-,80
R2297	T8123	T8124	nummod	80,°C
R2298	T8124	T8121	pobj	°C,at
R2299	T8125	T8111	punct	.,removed
R23	T864	T862	prep	of,error
R230	T1095	T1096	det	a,model
R2300	T8127	T8128	prep	At,thawed
R2301	T8129	T8130	det	the,time
R2302	T8130	T8127	pobj	time,At
R2303	T8131	T8130	prep	of,time
R2304	T8132	T8133	compound	enzyme,assay
R2305	T8133	T8131	pobj	assay,of
R2306	T8134	T8128	punct	", ",thawed
R2307	T8135	T8128	nsubjpass	livers,thawed
R2308	T8136	T8128	auxpass	were,thawed
R2309	T8137	T8128	punct	", ",thawed
R231	T1096	T1081	dobj	model,produced
R2310	T8138	T8128	cc	and,thawed
R2311	T8139	T8128	conj	homogenized,thawed
R2312	T8140	T8141	punct	(,v
R2313	T8141	T8139	parataxis	v,homogenized
R2314	T8142	T8143	nummod	1,9
R2315	T8143	T8141	dep	9,v
R2316	T8144	T8143	punct	:,9
R2317	T8145	T8141	punct	", ",v
R2318	T8146	T8141	compound	w,v
R2319	T8147	T8141	punct	/,v
R232	T1097	T1096	prep	of,model
R2320	T8148	T8141	punct	),v
R2321	T8149	T8139	prep	in,homogenized
R2322	T8150	T8151	nummod	0.25,M
R2323	T8151	T8152	compound	M,sucrose
R2324	T8152	T8149	pobj	sucrose,in
R2325	T8153	T8152	punct	", ",sucrose
R2326	T8154	T8155	nummod	10,mM
R2327	T8155	T8156	compound	mM,HC1
R2328	T8156	T8152	appos	HC1,sucrose
R2329	T8157	T8156	compound	Tris,HC1
R233	T1098	T1099	amod	intermediate,MSUD
R2330	T8158	T8156	punct	-,HC1
R2331	T8159	T8156	punct	", ",HC1
R2332	T8160	T8156	npadvmod	pH,HC1
R2333	T8161	T8160	nummod	7.4,pH
R2334	T8162	T8128	punct	.,thawed
R2335	T8164	T8165	compound	Liver,homogenates
R2336	T8165	T8166	nsubjpass	homogenates,centrifuged
R2337	T8167	T8166	auxpass	were,centrifuged
R2338	T8168	T8166	prep	at,centrifuged
R2339	T8169	T8170	nummod	600,g
R234	T1099	T1097	pobj	MSUD,of
R2340	T8170	T8168	pobj	g,at
R2341	T8171	T8169	punct	×,600
R2342	T8172	T8166	prep	for,centrifuged
R2343	T8173	T8174	nummod	10,min
R2344	T8174	T8172	pobj	min,for
R2345	T8175	T8166	prep	at,centrifuged
R2346	T8176	T8177	nummod	4,°C
R2347	T8177	T8175	pobj	°C,at
R2348	T8178	T8166	cc	and,centrifuged
R2349	T8179	T8180	det	the,fraction
R235	T1100	T1081	punct	.,produced
R2350	T8180	T8182	nsubjpass	fraction,saved
R2351	T8181	T8180	compound	supernatant,fraction
R2352	T8182	T8166	conj	saved,centrifuged
R2353	T8183	T8182	auxpass	was,saved
R2354	T8184	T8185	aux	to,determine
R2355	T8185	T8182	advcl	determine,saved
R2356	T8186	T8187	det	the,activity
R2357	T8187	T8185	dobj	activity,determine
R2358	T8188	T8187	compound	BCKDH,activity
R2359	T8189	T8182	punct	.,saved
R236	T1102	T1103	amod	Branched,chain
R2360	T8191	T8192	det	The,use
R2361	T8192	T8193	nsubjpass	use,necessitated
R2362	T8194	T8192	prep	of,use
R2363	T8195	T8196	compound	tissue,homogenates
R2364	T8196	T8194	pobj	homogenates,of
R2365	T8197	T8193	auxpass	was,necessitated
R2366	T8198	T8193	agent	by,necessitated
R2367	T8199	T8200	det	the,availability
R2368	T8200	T8198	pobj	availability,by
R2369	T8201	T8200	amod	limited,availability
R237	T1103	T1105	compound	chain,dehydrogenase
R2370	T8202	T8200	prep	of,availability
R2371	T8203	T8204	compound	liver,tissue
R2372	T8204	T8202	pobj	tissue,of
R2373	T8205	T8204	punct	", ",tissue
R2374	T8206	T8207	advmod	particularly,from
R2375	T8207	T8204	prep	from,tissue
R2376	T8208	T8209	amod	newborn,pups
R2377	T8209	T8207	pobj	pups,from
R2378	T8210	T8193	punct	.,necessitated
R2379	T8212	T8213	compound	BCKDH,activity
R238	T1104	T1103	punct	-,chain
R2380	T8213	T8214	nsubjpass	activity,determined
R2381	T8215	T8214	auxpass	was,determined
R2382	T8216	T8214	prep	by,determined
R2383	T8217	T8216	pcomp	measuring,by
R2384	T8218	T8219	det	the,release
R2385	T8219	T8217	dobj	release,measuring
R2386	T8220	T8219	prep	of,release
R2387	T8221	T8220	pobj	14CO2,of
R2388	T8222	T8217	prep	from,measuring
R2389	T8223	T8224	nmod	α,keto
R239	T1105	T1108	compound	dehydrogenase,activity
R2390	T8224	T8226	nmod	keto,isocaproate
R2391	T8225	T8224	punct	-,keto
R2392	T8226	T8222	pobj	isocaproate,from
R2393	T8227	T8228	punct	[,1
R2394	T8228	T8224	parataxis	1,keto
R2395	T8229	T8230	punct	-,14C
R2396	T8230	T8228	prep	14C,1
R2397	T8231	T8228	punct	],1
R2398	T8232	T8233	mark	as,described
R2399	T8233	T8214	advcl	described,determined
R24	T865	T864	pobj	metabolism,of
R240	T1106	T1107	compound	keto,acid
R2400	T8234	T8233	advmod	previously,described
R2401	T8235	T8236	punct	[,31
R2402	T8236	T8233	parataxis	31,described
R2403	T8237	T8236	punct	],31
R2404	T8238	T8214	punct	.,determined
R2405	T8240	T8241	det	The,mixture
R2406	T8241	T8244	nsubj	mixture,contained
R2407	T8242	T8241	amod	complete,mixture
R2408	T8243	T8241	compound	reaction,mixture
R2409	T8245	T8246	punct	(,volume
R241	T1107	T1105	compound	acid,dehydrogenase
R2410	T8246	T8244	parataxis	volume,contained
R2411	T8247	T8246	amod	final,volume
R2412	T8248	T8249	nummod	1,ml
R2413	T8249	T8246	appos	ml,volume
R2414	T8250	T8246	punct	),volume
R2415	T8251	T8252	nummod	30,mM
R2416	T8252	T8253	compound	mM,buffer
R2417	T8253	T8244	dobj	buffer,contained
R2418	T8254	T8255	compound	potassium,phosphate
R2419	T8255	T8253	compound	phosphate,buffer
R242	T1108	T1109	nsubj	activity,was
R2420	T8256	T8253	punct	", ",buffer
R2421	T8257	T8253	npadvmod	pH,buffer
R2422	T8258	T8257	nummod	7.4,pH
R2423	T8259	T8253	punct	", ",buffer
R2424	T8260	T8261	nummod	0.20,mM
R2425	T8261	T8262	compound	mM,ketoisocaproate
R2426	T8262	T8253	conj	ketoisocaproate,buffer
R2427	T8263	T8262	compound	α,ketoisocaproate
R2428	T8264	T8262	punct	-,ketoisocaproate
R2429	T8265	T8262	punct	", ",ketoisocaproate
R243	T1110	T1111	quantmod	5,6
R2430	T8266	T8267	nummod	0.40,mM
R2431	T8267	T8268	compound	mM,CoASH
R2432	T8268	T8262	conj	CoASH,ketoisocaproate
R2433	T8269	T8268	punct	", ",CoASH
R2434	T8270	T8271	nummod	0.40,mM
R2435	T8271	T8272	compound	mM,pyrophosphate
R2436	T8272	T8268	conj	pyrophosphate,CoASH
R2437	T8273	T8272	compound	thiamin,pyrophosphate
R2438	T8274	T8272	punct	", ",pyrophosphate
R2439	T8275	T8276	nummod	2,mM
R244	T1111	T1113	nummod	6,%
R2440	T8276	T8277	compound	mM,NAD
R2441	T8277	T8272	conj	NAD,pyrophosphate
R2442	T8278	T8277	punct	+,NAD
R2443	T8279	T8277	punct	", ",NAD
R2444	T8280	T8281	nummod	2,mM
R2445	T8281	T8282	compound	mM,dithiothreitol
R2446	T8282	T8277	conj	dithiothreitol,NAD
R2447	T8283	T8282	punct	", ",dithiothreitol
R2448	T8284	T8285	nummod	5,mM
R2449	T8285	T8286	compound	mM,Mg2
R245	T1112	T1111	punct	–,6
R2450	T8286	T8282	conj	Mg2,dithiothreitol
R2451	T8287	T8286	punct	+,Mg2
R2452	T8288	T8286	punct	", ",Mg2
R2453	T8289	T8290	advmod	approximately,"250,000"
R2454	T8290	T8291	nummod	"250,000",DPM
R2455	T8291	T8286	conj	DPM,Mg2
R2456	T8292	T8291	prep	of,DPM
R2457	T8293	T8294	nmod	α,keto
R2458	T8294	T8296	nmod	keto,isocaproate
R2459	T8295	T8294	punct	-,keto
R246	T1113	T1109	attr	%,was
R2460	T8296	T8292	pobj	isocaproate,of
R2461	T8297	T8298	punct	[,1
R2462	T8298	T8294	parataxis	1,keto
R2463	T8299	T8300	punct	-,14C
R2464	T8300	T8298	prep	14C,1
R2465	T8301	T8298	punct	],1
R2466	T8302	T8291	punct	", ",DPM
R2467	T8303	T8291	cc	and,DPM
R2468	T8304	T8305	nummod	0.10,ml
R2469	T8305	T8291	conj	ml,DPM
R247	T1114	T1113	prep	of,%
R2470	T8306	T8305	prep	of,ml
R2471	T8307	T8308	compound	liver,homogenate
R2472	T8308	T8306	pobj	homogenate,of
R2473	T8309	T8310	punct	(,protein
R2474	T8310	T8308	parataxis	protein,homogenate
R2475	T8311	T8312	quantmod	2,3
R2476	T8312	T8314	nummod	3,mg
R2477	T8313	T8312	punct	–,3
R2478	T8314	T8310	compound	mg,protein
R2479	T8315	T8310	punct	),protein
R248	T1115	T1114	amod	normal,of
R2480	T8316	T8244	punct	.,contained
R2481	T8318	T8319	nsubjpass	Assays,carried
R2482	T8320	T8319	auxpass	were,carried
R2483	T8321	T8319	prt	out,carried
R2484	T8322	T8319	prep	for,carried
R2485	T8323	T8324	nummod	15,min
R2486	T8324	T8322	pobj	min,for
R2487	T8325	T8319	prep	at,carried
R2488	T8326	T8327	nummod	37,°C
R2489	T8327	T8325	pobj	°C,at
R249	T1116	T1109	cc	and,was
R2490	T8328	T8319	punct	", ",carried
R2491	T8329	T8330	nsubjpass	14CO2,trapped
R2492	T8330	T8319	conj	trapped,carried
R2493	T8331	T8330	auxpass	was,trapped
R2494	T8332	T8330	prep	in,trapped
R2495	T8333	T8332	pobj	hydroxide,in
R2496	T8334	T8333	prep	of,hydroxide
R2497	T8335	T8334	pobj	Hyamine,of
R2498	T8336	T8330	punct	", ",trapped
R2499	T8337	T8330	cc	and,trapped
R25	T866	T862	acl	caused,error
R250	T1117	T1109	conj	was,was
R2500	T8338	T8339	nsubjpass	radioactivity,determined
R2501	T8339	T8330	conj	determined,trapped
R2502	T8340	T8339	auxpass	was,determined
R2503	T8341	T8339	prep	by,determined
R2504	T8342	T8343	amod	liquid,scintillation
R2505	T8343	T8344	compound	scintillation,spectrometry
R2506	T8344	T8341	pobj	spectrometry,by
R2507	T8345	T8339	punct	.,determined
R2508	T8817	T8818	compound	Western,blot
R2509	T8818	T8819	compound	blot,analysis
R251	T1118	T1117	acomp	sufficient,was
R2510	T8821	T8822	compound	Protein,extracts
R2511	T8822	T8823	nsubjpass	extracts,isolated
R2512	T8824	T8823	auxpass	were,isolated
R2513	T8825	T8823	prep	from,isolated
R2514	T8826	T8827	amod	homogenized,liver
R2515	T8827	T8825	pobj	liver,from
R2516	T8828	T8827	punct	(,liver
R2517	T8829	T8830	advmod	freshly,harvested
R2518	T8830	T8827	acl	harvested,liver
R2519	T8831	T8830	cc	and,harvested
R252	T1119	T1120	aux	to,allow
R2520	T8832	T8833	dep	flash,frozen
R2521	T8833	T8830	conj	frozen,harvested
R2522	T8834	T8827	punct	),liver
R2523	T8835	T8827	prep	of,liver
R2524	T8836	T8837	npadvmod	wildtype,transgenic
R2525	T8837	T8845	amod	transgenic,mice
R2526	T8838	T8836	punct	", ",wildtype
R2527	T8839	T8840	compound	Line,525A
R2528	T8840	T8836	conj	525A,wildtype
R2529	T8841	T8840	punct	", ",525A
R253	T1120	T1118	xcomp	allow,sufficient
R2530	T8842	T8840	cc	and,525A
R2531	T8843	T8844	compound	Line,A
R2532	T8844	T8840	conj	A,525A
R2533	T8845	T8835	pobj	mice,of
R2534	T8846	T8823	punct	.,isolated
R2535	T8848	T8849	nsubjpass	Protein,analyzed
R2536	T8850	T8851	punct	(,μg
R2537	T8851	T8848	parataxis	μg,Protein
R2538	T8852	T8851	nummod	25,μg
R2539	T8853	T8851	prep	per,μg
R254	T1121	T1120	dobj	survival,allow
R2540	T8854	T8853	pobj	sample,per
R2541	T8855	T8851	punct	),μg
R2542	T8856	T8849	auxpass	was,analyzed
R2543	T8857	T8849	prep	by,analyzed
R2544	T8858	T8857	pobj	electrophoresis,by
R2545	T8859	T8849	prep	on,analyzed
R2546	T8860	T8861	det	a,Gel
R2547	T8861	T8859	pobj	Gel,on
R2548	T8862	T8863	nummod	10,%
R2549	T8863	T8861	nmod	%,Gel
R255	T1122	T1109	punct	", ",was
R2550	T8864	T8865	nmod	SDS,PAGE
R2551	T8865	T8861	nmod	PAGE,Gel
R2552	T8866	T8865	punct	-,PAGE
R2553	T8867	T8861	amod	Ready,Gel
R2554	T8868	T8869	punct	(,Rad
R2555	T8869	T8861	parataxis	Rad,Gel
R2556	T8870	T8869	compound	Bio,Rad
R2557	T8871	T8869	punct	-,Rad
R2558	T8872	T8869	punct	", ",Rad
R2559	T8873	T8869	npadvmod	Hercules,Rad
R256	T1123	T1109	cc	but,was
R2560	T8874	T8869	punct	", ",Rad
R2561	T8875	T8869	npadvmod	CA,Rad
R2562	T8876	T8869	punct	),Rad
R2563	T8877	T8849	cc	and,analyzed
R2564	T8878	T8849	conj	transferred,analyzed
R2565	T8879	T8878	prep	to,transferred
R2566	T8880	T8881	compound	PVDF,membrane
R2567	T8881	T8879	pobj	membrane,to
R2568	T8882	T8883	punct	(,Rad
R2569	T8883	T8881	parataxis	Rad,membrane
R257	T1124	T1109	conj	was,was
R2570	T8884	T8885	compound	Sequi,Blot
R2571	T8885	T8883	dep	Blot,Rad
R2572	T8886	T8885	punct	-,Blot
R2573	T8887	T8883	punct	;,Rad
R2574	T8888	T8883	compound	Bio,Rad
R2575	T8889	T8883	punct	-,Rad
R2576	T8890	T8883	punct	),Rad
R2577	T8891	T8878	prep	via,transferred
R2578	T8892	T8891	pobj	electroblotting,via
R2579	T8893	T8849	punct	.,analyzed
R258	T1125	T1124	acomp	insufficient,was
R2580	T8895	T8896	det	All,blots
R2581	T8896	T8897	nsubjpass	blots,probed
R2582	T8898	T8897	auxpass	were,probed
R2583	T8899	T8897	prep	for,probed
R2584	T8900	T8901	compound	E2,protein
R2585	T8901	T8899	pobj	protein,for
R2586	T8902	T8897	advcl	using,probed
R2587	T8903	T8904	amod	polyclonal,antisera
R2588	T8904	T8902	dobj	antisera,using
R2589	T8905	T8904	compound	rabbit,antisera
R259	T1126	T1127	aux	to,circulating
R2590	T8906	T8904	compound	E2,antisera
R2591	T8907	T8908	punct	(,"5,000"
R2592	T8908	T8904	parataxis	"5,000",antisera
R2593	T8909	T8908	nummod	1,"5,000"
R2594	T8910	T8908	punct	:,"5,000"
R2595	T8911	T8908	punct	),"5,000"
R2596	T8912	T8904	punct	", ",antisera
R2597	T8913	T8914	dep	which,detects
R2598	T8914	T8904	relcl	detects,antisera
R2599	T8915	T8916	preconj	both,subunits
R26	T867	T866	agent	by,caused
R260	T1127	T1125	xcomp	circulating,insufficient
R2600	T8916	T8914	dobj	subunits,detects
R2601	T8917	T8916	nmod	mouse,subunits
R2602	T8918	T8919	punct	(,kD
R2603	T8919	T8917	parataxis	kD,mouse
R2604	T8920	T8921	punct	~,47
R2605	T8921	T8919	nummod	47,kD
R2606	T8922	T8919	punct	),kD
R2607	T8923	T8917	cc	and,mouse
R2608	T8924	T8917	conj	human,mouse
R2609	T8925	T8926	punct	(,kD
R261	T1128	T1127	aux	normalize,circulating
R2610	T8926	T8924	parataxis	kD,human
R2611	T8927	T8928	punct	~,54
R2612	T8928	T8926	nummod	54,kD
R2613	T8929	T8926	punct	),kD
R2614	T8930	T8916	compound	E2,subunits
R2615	T8931	T8932	punct	(,gift
R2616	T8932	T8916	parataxis	gift,subunits
R2617	T8933	T8932	punct	[,gift
R2618	T8934	T8932	dep	30,gift
R2619	T8935	T8932	punct	],gift
R262	T1129	T1130	amod	branched,chain
R2620	T8936	T8932	punct	;,gift
R2621	T8937	T8932	prep	from,gift
R2622	T8938	T8939	compound	Dr.,Hutson
R2623	T8939	T8937	pobj	Hutson,from
R2624	T8940	T8939	compound	Susan,Hutson
R2625	T8941	T8939	punct	", ",Hutson
R2626	T8942	T8943	compound	Wake,Forest
R2627	T8943	T8944	compound	Forest,University
R2628	T8944	T8939	npadvmod	University,Hutson
R2629	T8945	T8932	punct	),gift
R263	T1130	T1132	compound	chain,acids
R2630	T8946	T8897	punct	.,probed
R2631	T8948	T8949	nsubjpass	Blots,re-probed
R2632	T8950	T8949	auxpass	were,re-probed
R2633	T8951	T8949	prep	with,re-probed
R2634	T8952	T8953	det	a,antibody
R2635	T8953	T8951	pobj	antibody,with
R2636	T8954	T8953	nmod	rabbit,antibody
R2637	T8955	T8956	amod	anti-c,myc
R2638	T8956	T8953	compound	myc,antibody
R2639	T8957	T8956	punct	-,myc
R264	T1131	T1130	punct	-,chain
R2640	T8958	T8953	compound	tag,antibody
R2641	T8959	T8960	punct	(,ab9106
R2642	T8960	T8953	parataxis	ab9106,antibody
R2643	T8961	T8962	nummod	1,"10,000"
R2644	T8962	T8960	dep	"10,000",ab9106
R2645	T8963	T8962	punct	:,"10,000"
R2646	T8964	T8960	punct	;,ab9106
R2647	T8965	T8960	dep	abCam,ab9106
R2648	T8966	T8965	punct	", ",abCam
R2649	T8967	T8965	npadvmod	Cambridge,abCam
R265	T1132	T1134	compound	acids,levels
R2650	T8968	T8965	punct	", ",abCam
R2651	T8969	T8965	npadvmod	MA,abCam
R2652	T8970	T8960	punct	;,ab9106
R2653	T8971	T8960	nmod	cat.,ab9106
R2654	T8972	T8960	punct	#,ab9106
R2655	T8973	T8960	punct	-,ab9106
R2656	T8974	T8960	nummod	100,ab9106
R2657	T8975	T8960	punct	),ab9106
R2658	T8976	T8949	punct	.,re-probed
R2659	T8978	T8979	nsubjpass	Blots,re-probed
R266	T1133	T1132	compound	amino,acids
R2660	T8980	T8979	auxpass	were,re-probed
R2661	T8981	T8979	advmod	also,re-probed
R2662	T8982	T8979	prep	with,re-probed
R2663	T8983	T8984	det	an,antibody
R2664	T8984	T8982	pobj	antibody,with
R2665	T8985	T8984	prep	for,antibody
R2666	T8986	T8987	compound	β,actin
R2667	T8987	T8985	pobj	actin,for
R2668	T8988	T8987	punct	-,actin
R2669	T8989	T8990	punct	(,ab8227
R267	T1134	T1127	dobj	levels,circulating
R2670	T8990	T8987	parataxis	ab8227,actin
R2671	T8991	T8992	nummod	43,kD
R2672	T8992	T8990	dep	kD,ab8227
R2673	T8993	T8990	punct	;,ab8227
R2674	T8994	T8995	nummod	1,"10,000"
R2675	T8995	T8990	dep	"10,000",ab8227
R2676	T8996	T8995	punct	:,"10,000"
R2677	T8997	T8990	punct	;,ab8227
R2678	T8998	T8990	dep	abCam,ab8227
R2679	T8999	T8990	punct	;,ab8227
R268	T1135	T1134	punct	", ",levels
R2680	T9000	T8990	nmod	cat.,ab8227
R2681	T9001	T8990	punct	#,ab8227
R2682	T9002	T8990	punct	-,ab8227
R2683	T9003	T8990	nummod	50,ab8227
R2684	T9004	T8990	punct	),ab8227
R2685	T9005	T9006	aux	to,allow
R2686	T9006	T8979	advcl	allow,re-probed
R2687	T9007	T9006	prep	for,allow
R2688	T9008	T9009	compound	loading,comparisons
R2689	T9009	T9007	pobj	comparisons,for
R269	T1136	T1137	dep	which,were
R2690	T9010	T8979	punct	.,re-probed
R2691	T9012	T9013	det	A,antibody
R2692	T9013	T9017	nsubjpass	antibody,used
R2693	T9014	T9013	nmod	goat,antibody
R2694	T9015	T9013	amod	anti-rabbit,antibody
R2695	T9016	T9013	amod	secondary,antibody
R2696	T9018	T9013	acl	conjugated,antibody
R2697	T9019	T9018	prep	to,conjugated
R2698	T9020	T9021	compound	horseradish,peroxidase
R2699	T9021	T9019	pobj	peroxidase,to
R27	T868	T869	det	a,deficiency
R270	T1137	T1134	relcl	were,levels
R2700	T9022	T9023	punct	(,H
R2701	T9023	T9021	parataxis	H,peroxidase
R2702	T9024	T9025	nummod	1,"10,000"
R2703	T9025	T9023	dep	"10,000",H
R2704	T9026	T9025	punct	:,"10,000"
R2705	T9027	T9023	punct	;,H
R2706	T9028	T9023	dep	Novus,H
R2707	T9029	T9028	punct	", ",Novus
R2708	T9030	T9028	npadvmod	Littleton,Novus
R2709	T9031	T9028	punct	", ",Novus
R271	T1138	T1137	acomp	intermediate,were
R2710	T9032	T9028	npadvmod	CO,Novus
R2711	T9033	T9023	punct	;,H
R2712	T9034	T9023	nmod	cat.,H
R2713	T9035	T9023	punct	#,H
R2714	T9036	T9023	compound	NB730,H
R2715	T9037	T9023	punct	-,H
R2716	T9038	T9023	punct	),H
R2717	T9039	T9017	auxpass	was,used
R2718	T9040	T9017	prep	for,used
R2719	T9041	T9040	pobj	detection,for
R272	T1139	T1137	prep	between,were
R2720	T9042	T9017	advcl	using,used
R2721	T9043	T9044	det	the,reagent
R2722	T9044	T9042	dobj	reagent,using
R2723	T9045	T9046	compound	Western,Lightning
R2724	T9046	T9044	compound	Lightning,reagent
R2725	T9047	T9044	compound	chemiluminescence,reagent
R2726	T9048	T9049	punct	(,Elmer
R2727	T9049	T9044	parataxis	Elmer,reagent
R2728	T9050	T9049	compound	Perkin,Elmer
R2729	T9051	T9049	punct	", ",Elmer
R273	T1140	T1139	pobj	wildtype,between
R2730	T9052	T9049	npadvmod	Boston,Elmer
R2731	T9053	T9049	punct	", ",Elmer
R2732	T9054	T9049	npadvmod	MA,Elmer
R2733	T9055	T9049	punct	),Elmer
R2734	T9056	T9017	cc	and,used
R2735	T9057	T9017	conj	exposed,used
R2736	T9058	T9057	prep	to,exposed
R2737	T9059	T9060	compound	X-ray,film
R2738	T9060	T9058	pobj	film,to
R2739	T9061	T9017	punct	.,used
R274	T1141	T1140	cc	and,wildtype
R2740	T9141	T9142	amod	Statistical,analyses
R2741	T9144	T9145	det	All,data
R2742	T9145	T9151	nsubjpass	data,presented
R2743	T9146	T9145	nmod	BCAA,data
R2744	T9147	T9146	cc	and,BCAA
R2745	T9148	T9146	conj	BCKDH,BCAA
R2746	T9149	T9145	compound	enzyme,data
R2747	T9150	T9145	compound	activity,data
R2748	T9152	T9151	auxpass	are,presented
R2749	T9153	T9151	prep	as,presented
R275	T1142	T1143	det	the,model
R2750	T9154	T9155	det	the,mean
R2751	T9155	T9153	pobj	mean,as
R2752	T9156	T9155	cc	+,mean
R2753	T9157	T9156	punct	/,+
R2754	T9158	T9156	punct	-,+
R2755	T9159	T9160	det	the,error
R2756	T9160	T9155	conj	error,mean
R2757	T9161	T9160	amod	standard,error
R2758	T9162	T9160	prep	of,error
R2759	T9163	T9164	det	the,mean
R276	T1143	T1140	conj	model,wildtype
R2760	T9164	T9162	pobj	mean,of
R2761	T9165	T9160	punct	(,error
R2762	T9166	T9160	appos	S.E.M.,error
R2763	T9167	T9151	punct	),presented
R2764	T9168	T9151	punct	.,presented
R2765	T9170	T9171	nsubjpass	Differences,compared
R2766	T9172	T9170	prep	between,Differences
R2767	T9173	T9172	pobj	genotypes,between
R2768	T9174	T9171	auxpass	were,compared
R2769	T9175	T9171	prep	by,compared
R277	T1144	T1143	amod	classic,model
R2770	T9176	T9177	poss	Student,test
R2771	T9177	T9175	pobj	test,by
R2772	T9178	T9176	case	's,Student
R2773	T9179	T9177	compound	t,test
R2774	T9180	T9181	punct	[,32
R2775	T9181	T9171	parataxis	32,compared
R2776	T9182	T9181	punct	],32
R2777	T9183	T9171	punct	.,compared
R2778	T10368	T10367	prep	of,Production
R2779	T10369	T10370	compound	E2,mice
R278	T1145	T1143	compound	MSUD,model
R2780	T10370	T10368	pobj	mice,of
R2781	T10371	T10372	compound	gene,knockout
R2782	T10372	T10370	compound	knockout,mice
R2783	T10374	T10375	aux	To,create
R2784	T10375	T10376	advcl	create,used
R2785	T10377	T10378	compound	E2,mice
R2786	T10378	T10375	dobj	mice,create
R2787	T10379	T10378	compound	knockout,mice
R2788	T10380	T10376	punct	", ",used
R2789	T10381	T10376	nsubj	we,used
R279	T1146	T1143	compound	mouse,model
R2790	T10382	T10383	compound	gene,targeting
R2791	T10383	T10376	dobj	targeting,used
R2792	T10384	T10376	prep	in,used
R2793	T10385	T10386	compound	mouse,cells
R2794	T10386	T10384	pobj	cells,in
R2795	T10387	T10386	compound	ES,cells
R2796	T10388	T10389	aux	to,disrupt
R2797	T10389	T10376	advcl	disrupt,used
R2798	T10390	T10391	det	the,gene
R2799	T10391	T10389	dobj	gene,disrupt
R28	T869	T867	pobj	deficiency,by
R280	T1147	T1109	punct	.,was
R2800	T10392	T10391	compound	E2,gene
R2801	T10393	T10376	punct	.,used
R2802	T10395	T10396	det	The,strategy
R2803	T10396	T10398	nsubjpass	strategy,illustrated
R2804	T10397	T10396	amod	overall,strategy
R2805	T10399	T10396	prep	for,strategy
R2806	T10400	T10399	pcomp	disrupting,for
R2807	T10401	T10402	det	the,gene
R2808	T10402	T10400	dobj	gene,disrupting
R2809	T10403	T10402	compound	E2,gene
R281	T1151	T1152	det	These,mice
R2810	T10404	T10398	auxpass	is,illustrated
R2811	T10405	T10398	prep	in,illustrated
R2812	T10406	T10407	compound	Fig.,1A
R2813	T10407	T10405	pobj	1A,in
R2814	T10408	T10398	punct	.,illustrated
R2815	T10410	T10411	det	The,construct
R2816	T10411	T10414	nsubjpass	construct,designed
R2817	T10412	T10413	compound	gene,targeting
R2818	T10413	T10411	compound	targeting,construct
R2819	T10415	T10414	auxpass	was,designed
R282	T1152	T1153	nsubj	mice,represent
R2820	T10416	T10417	aux	to,replace
R2821	T10417	T10414	advcl	replace,designed
R2822	T10418	T10419	det	a,fragment
R2823	T10419	T10417	dobj	fragment,replace
R2824	T10420	T10421	nummod	1.67,kb
R2825	T10421	T10419	nmod	kb,fragment
R2826	T10422	T10423	nmod	EcoRV,Smal
R2827	T10423	T10419	nmod	Smal,fragment
R2828	T10424	T10423	punct	/,Smal
R2829	T10425	T10419	amod	genomic,fragment
R283	T1154	T1155	amod	important,models
R2830	T10426	T10419	compound	DNA,fragment
R2831	T10427	T10419	acl	encompassing,fragment
R2832	T10428	T10427	dobj	part,encompassing
R2833	T10429	T10428	prep	of,part
R2834	T10430	T10429	pobj	Exon,of
R2835	T10431	T10430	nummod	4,Exon
R2836	T10432	T10428	cc	and,part
R2837	T10433	T10428	conj	all,part
R2838	T10434	T10433	prep	of,all
R2839	T10435	T10434	pobj	Exon,of
R284	T1155	T1153	dobj	models,represent
R2840	T10436	T10435	nummod	5,Exon
R2841	T10437	T10417	prep	with,replace
R2842	T10438	T10439	det	the,cassette
R2843	T10439	T10437	pobj	cassette,with
R2844	T10440	T10439	nmod	PGKneo,cassette
R2845	T10441	T10439	amod	selectable,cassette
R2846	T10442	T10439	compound	marker,cassette
R2847	T10443	T10414	punct	.,designed
R2848	T10445	T10446	prep	Of,displayed
R2849	T10447	T10448	nummod	522,clones
R285	T1156	T1155	compound	animal,models
R2850	T10448	T10445	pobj	clones,Of
R2851	T10449	T10450	compound	ES,cell
R2852	T10450	T10448	compound	cell,clones
R2853	T10451	T10448	acl	screened,clones
R2854	T10452	T10451	prep	for,screened
R2855	T10453	T10452	pobj	targeting,for
R2856	T10454	T10451	prep	by,screened
R2857	T10455	T10456	compound	Southern,blot
R2858	T10456	T10457	compound	blot,analysis
R2859	T10457	T10454	pobj	analysis,by
R286	T1157	T1158	dep	that,approximate
R2860	T10458	T10446	punct	", ",displayed
R2861	T10459	T10446	nsubj	29,displayed
R2862	T10460	T10461	punct	(,%
R2863	T10461	T10459	parataxis	%,29
R2864	T10462	T10461	nummod	5,%
R2865	T10463	T10461	punct	),%
R2866	T10464	T10465	det	the,polymorphisms
R2867	T10465	T10446	dobj	polymorphisms,displayed
R2868	T10466	T10465	amod	predicted,polymorphisms
R2869	T10467	T10468	compound	restriction,length
R287	T1158	T1155	relcl	approximate,models
R2870	T10468	T10465	compound	length,polymorphisms
R2871	T10469	T10468	compound	fragment,length
R2872	T10470	T10465	amod	indicative,polymorphisms
R2873	T10471	T10470	prep	of,indicative
R2874	T10472	T10471	pobj	gene,of
R2875	T10473	T10472	amod	targeting,gene
R2876	T10474	T10446	prep	at,displayed
R2877	T10475	T10476	det	the,locus
R2878	T10476	T10474	pobj	locus,at
R2879	T10477	T10476	compound	E2,locus
R288	T1159	T1158	advmod	closely,approximate
R2880	T10478	T10446	punct	.,displayed
R2881	T10480	T10481	mark	As,illustrated
R2882	T10481	T10482	advcl	illustrated,hybridizes
R2883	T10483	T10481	prep	in,illustrated
R2884	T10484	T10485	compound	Fig.,1A
R2885	T10485	T10483	pobj	1A,in
R2886	T10486	T10485	cc	and,1A
R2887	T10487	T10485	conj	1B,1A
R2888	T10488	T10482	punct	", ",hybridizes
R2889	T10489	T10490	det	an,probe
R289	T1160	T1161	det	the,phenotype
R2890	T10490	T10482	nsubj	probe,hybridizes
R2891	T10491	T10490	nmod	E2,probe
R2892	T10492	T10490	nmod	Exon,probe
R2893	T10493	T10492	nummod	6,Exon
R2894	T10494	T10490	amod	specific,probe
R2895	T10495	T10490	punct	", ",probe
R2896	T10496	T10497	dep	which,is
R2897	T10497	T10490	relcl	is,probe
R2898	T10498	T10497	acomp	external,is
R2899	T10499	T10498	prep	to,external
R29	T870	T869	prep	of,deficiency
R290	T1161	T1158	dobj	phenotype,approximate
R2900	T10500	T10501	det	the,construct
R2901	T10501	T10499	pobj	construct,to
R2902	T10502	T10501	compound	targeting,construct
R2903	T10503	T10482	punct	", ",hybridizes
R2904	T10504	T10482	prep	to,hybridizes
R2905	T10505	T10506	advmod	only,fragment
R2906	T10506	T10504	pobj	fragment,to
R2907	T10507	T10506	det	a,fragment
R2908	T10508	T10509	punct	~,16
R2909	T10509	T10510	nummod	16,kb
R291	T1162	T1161	prep	of,phenotype
R2910	T10510	T10506	compound	kb,fragment
R2911	T10511	T10506	compound	BglI,fragment
R2912	T10512	T10506	compound	restriction,fragment
R2913	T10513	T10506	prep	from,fragment
R2914	T10514	T10515	det	the,allele
R2915	T10515	T10513	pobj	allele,from
R2916	T10516	T10517	amod	wild,type
R2917	T10517	T10515	compound	type,allele
R2918	T10518	T10515	prep	in,allele
R2919	T10519	T10520	det	the,line
R292	T1163	T1162	pobj	humans,of
R2920	T10520	T10518	pobj	line,in
R2921	T10521	T10520	amod	parental,line
R2922	T10522	T10520	compound	R1,line
R2923	T10523	T10524	compound	ES,cell
R2924	T10524	T10520	compound	cell,line
R2925	T10525	T10482	punct	.,hybridizes
R2926	T10527	T10528	prep	In,hybridizes
R2927	T10529	T10530	advmod	correctly,targeted
R2928	T10530	T10531	amod	targeted,cells
R2929	T10531	T10527	pobj	cells,In
R293	T1164	T1163	prep	with,humans
R2930	T10532	T10531	compound	ES,cells
R2931	T10533	T10528	punct	", ",hybridizes
R2932	T10534	T10535	det	this,probe
R2933	T10535	T10528	nsubj	probe,hybridizes
R2934	T10536	T10528	advmod	also,hybridizes
R2935	T10537	T10528	prep	to,hybridizes
R2936	T10538	T10539	det	a,fragment
R2937	T10539	T10537	pobj	fragment,to
R2938	T10540	T10541	punct	~,11
R2939	T10541	T10542	nummod	11,kb
R294	T1165	T1166	det	the,forms
R2940	T10542	T10539	compound	kb,fragment
R2941	T10543	T10539	compound	BglI,fragment
R2942	T10544	T10539	compound	restriction,fragment
R2943	T10545	T10528	punct	.,hybridizes
R2944	T10547	T10548	nsubjpass	Targeting,confirmed
R2945	T10549	T10548	auxpass	was,confirmed
R2946	T10550	T10548	prep	with,confirmed
R2947	T10551	T10552	amod	several,enzymes
R2948	T10552	T10550	pobj	enzymes,with
R2949	T10553	T10552	amod	additional,enzymes
R295	T1166	T1164	pobj	forms,with
R2950	T10554	T10552	compound	restriction,enzymes
R2951	T10555	T10552	cc	and,enzymes
R2952	T10556	T10552	conj	probes,enzymes
R2953	T10557	T10558	punct	(,shown
R2954	T10558	T10548	parataxis	shown,confirmed
R2955	T10559	T10558	nsubj	data,shown
R2956	T10560	T10558	neg	not,shown
R2957	T10561	T10558	punct	),shown
R2958	T10562	T10548	punct	.,confirmed
R2959	T10564	T10565	advmod	Correctly,targeted
R296	T1167	T1166	amod	classic,forms
R2960	T10565	T10566	amod	targeted,cells
R2961	T10566	T10568	nsubjpass	cells,microinjected
R2962	T10567	T10566	compound	ES,cells
R2963	T10569	T10568	auxpass	were,microinjected
R2964	T10570	T10568	prep	into,microinjected
R2965	T10571	T10570	pobj	blastocysts,into
R2966	T10572	T10573	aux	to,produce
R2967	T10573	T10568	advcl	produce,microinjected
R2968	T10574	T10575	amod	chimeric,mice
R2969	T10575	T10573	dobj	mice,produce
R297	T1168	T1167	cc	and,classic
R2970	T10576	T10568	punct	.,microinjected
R2971	T10578	T10579	nsubjpass	Chimerics,bred
R2972	T10580	T10579	auxpass	were,bred
R2973	T10581	T10579	prep	to,bred
R2974	T10582	T10583	compound	C57BL,6J
R2975	T10583	T10585	compound	6J,mice
R2976	T10584	T10583	punct	/,6J
R2977	T10585	T10581	pobj	mice,to
R2978	T10586	T10579	punct	.,bred
R2979	T10588	T10589	prep	Following,interbred
R298	T1169	T1167	conj	intermediate,classic
R2980	T10590	T10591	compound	germline,transmission
R2981	T10591	T10588	pobj	transmission,Following
R2982	T10592	T10591	prep	of,transmission
R2983	T10593	T10594	det	the,allele
R2984	T10594	T10592	pobj	allele,of
R2985	T10595	T10594	amod	targeted,allele
R2986	T10596	T10589	punct	", ",interbred
R2987	T10597	T10598	amod	heterozygous,mice
R2988	T10598	T10589	nsubjpass	mice,interbred
R2989	T10599	T10589	auxpass	were,interbred
R299	T1170	T1166	prep	of,forms
R2990	T10600	T10601	aux	to,produce
R2991	T10601	T10589	advcl	produce,interbred
R2992	T10602	T10603	amod	wild,type
R2993	T10603	T10604	nmod	type,animals
R2994	T10604	T10601	dobj	animals,produce
R2995	T10605	T10606	punct	(,+
R2996	T10606	T10603	punct	+,type
R2997	T10607	T10606	punct	+,+
R2998	T10608	T10606	punct	/,+
R2999	T10609	T10606	punct	),+
R3	T838	T834	prep	of,Production
R30	T871	T872	amod	branched,chain
R300	T1171	T1170	pobj	MSUD,of
R3000	T10610	T10603	punct	", ",type
R3001	T10611	T10603	conj	heterozygous,type
R3002	T10612	T10613	punct	(,-
R3003	T10613	T10611	punct	-,heterozygous
R3004	T10614	T10613	punct	+,-
R3005	T10615	T10613	punct	/,-
R3006	T10616	T10613	punct	),-
R3007	T10617	T10611	cc	and,heterozygous
R3008	T10618	T10611	conj	homozygous,heterozygous
R3009	T10619	T10620	punct	(,-
R301	T1172	T1153	punct	.,represent
R3010	T10620	T10618	punct	-,homozygous
R3011	T10621	T10620	punct	-,-
R3012	T10622	T10620	punct	/,-
R3013	T10623	T10620	punct	),-
R3014	T10624	T10589	punct	.,interbred
R3015	T10626	T10627	nsubjpass	Mice,genotyped
R3016	T10628	T10627	auxpass	were,genotyped
R3017	T10629	T10627	prep	by,genotyped
R3018	T10630	T10631	compound	Southern,blot
R3019	T10631	T10632	compound	blot,analysis
R302	T1174	T1175	det	These,animals
R3020	T10632	T10629	pobj	analysis,by
R3021	T10633	T10627	punct	.,genotyped
R3022	T10635	T10636	det	The,probe
R3023	T10636	T10640	nsubj	probe,hybridized
R3024	T10637	T10638	npadvmod	Exon,specific
R3025	T10638	T10636	amod	specific,probe
R3026	T10639	T10637	nummod	6,Exon
R3027	T10641	T10640	prep	to,hybridized
R3028	T10642	T10643	advmod	only,fragment
R3029	T10643	T10641	pobj	fragment,to
R303	T1175	T1176	nsubj	animals,provide
R3030	T10644	T10643	det	a,fragment
R3031	T10645	T10646	punct	~,16
R3032	T10646	T10647	nummod	16,kb
R3033	T10647	T10643	compound	kb,fragment
R3034	T10648	T10643	compound	BglI,fragment
R3035	T10649	T10643	compound	restriction,fragment
R3036	T10650	T10643	prep	in,fragment
R3037	T10651	T10652	punct	+,+
R3038	T10652	T10654	punct	+,mice
R3039	T10653	T10652	punct	/,+
R304	T1177	T1178	amod	useful,models
R3040	T10654	T10650	pobj	mice,in
R3041	T10655	T10643	punct	", ",fragment
R3042	T10656	T10657	det	a,fragment
R3043	T10657	T10643	appos	fragment,fragment
R3044	T10658	T10659	punct	~,11
R3045	T10659	T10660	nummod	11,kb
R3046	T10660	T10657	compound	kb,fragment
R3047	T10661	T10657	compound	BglI,fragment
R3048	T10662	T10657	prep	in,fragment
R3049	T10663	T10664	punct	-,-
R305	T1178	T1176	dobj	models,provide
R3050	T10664	T10666	punct	-,mice
R3051	T10665	T10664	punct	/,-
R3052	T10666	T10662	pobj	mice,in
R3053	T10667	T10641	punct	", ",to
R3054	T10668	T10641	cc	and,to
R3055	T10669	T10641	conj	to,to
R3056	T10670	T10669	pobj	both,to
R3057	T10671	T10670	prep	of,both
R3058	T10672	T10673	det	these,fragments
R3059	T10673	T10671	pobj	fragments,of
R306	T1179	T1180	aux	to,characterize
R3060	T10674	T10670	prep	in,both
R3061	T10675	T10676	punct	+,-
R3062	T10676	T10678	punct	-,mice
R3063	T10677	T10676	punct	/,-
R3064	T10678	T10674	pobj	mice,in
R3065	T10679	T10680	punct	(,1B
R3066	T10680	T10640	parataxis	1B,hybridized
R3067	T10681	T10680	compound	Fig.,1B
R3068	T10682	T10680	punct	),1B
R3069	T10683	T10640	punct	.,hybridized
R307	T1180	T1178	advcl	characterize,models
R3070	T10685	T10686	nsubjpass	Mice,born
R3071	T10687	T10685	amod	homozygous,Mice
R3072	T10688	T10687	prep	for,homozygous
R3073	T10689	T10690	det	the,mutation
R3074	T10690	T10688	pobj	mutation,for
R3075	T10691	T10690	compound	E2,mutation
R3076	T10692	T10686	auxpass	were,born
R3077	T10693	T10686	prep	at,born
R3078	T10694	T10695	det	the,frequency
R3079	T10695	T10693	pobj	frequency,at
R308	T1181	T1180	advmod	further,characterize
R3080	T10696	T10695	amod	expected,frequency
R3081	T10697	T10686	punct	.,born
R3082	T10699	T10700	compound	Genotype,analysis
R3083	T10700	T10701	nsubj	analysis,revealed
R3084	T10702	T10700	prep	of,analysis
R3085	T10703	T10702	pobj	pups,of
R3086	T10704	T10703	acl	derived,pups
R3087	T10705	T10704	prep	from,derived
R3088	T10706	T10707	punct	+,-
R3089	T10707	T10709	punct	-,matings
R309	T1182	T1183	det	the,pathogenesis
R3090	T10708	T10707	punct	/,-
R3091	T10709	T10705	pobj	matings,from
R3092	T10710	T10707	prep	by,-
R3093	T10711	T10712	punct	+,-
R3094	T10712	T10710	punct	-,by
R3095	T10713	T10712	punct	/,-
R3096	T10714	T10715	mark	that,were
R3097	T10715	T10701	ccomp	were,revealed
R3098	T10716	T10717	punct	+,+
R3099	T10717	T10719	punct	+,mice
R31	T872	T874	compound	chain,dehydrogenase
R310	T1183	T1180	dobj	pathogenesis,characterize
R3100	T10718	T10717	punct	/,+
R3101	T10719	T10715	nsubj	mice,were
R3102	T10720	T10717	punct	", ",+
R3103	T10721	T10722	punct	+,-
R3104	T10722	T10717	conj	-,+
R3105	T10723	T10722	punct	/,-
R3106	T10724	T10722	punct	", ",-
R3107	T10725	T10722	cc	and,-
R3108	T10726	T10727	punct	-,-
R3109	T10727	T10722	conj	-,-
R311	T1184	T1183	prep	of,pathogenesis
R3110	T10728	T10727	punct	/,-
R3111	T10729	T10715	acomp	present,were
R3112	T10730	T10715	prep	at,were
R3113	T10731	T10732	advmod	nearly,frequency
R3114	T10732	T10730	pobj	frequency,at
R3115	T10733	T10732	det	the,frequency
R3116	T10734	T10732	amod	expected,frequency
R3117	T10735	T10736	nummod	1,1
R3118	T10736	T10732	nummod	1,frequency
R3119	T10737	T10736	punct	:,1
R312	T1185	T1184	pobj	MSUD,of
R3120	T10738	T10736	nummod	2,1
R3121	T10739	T10736	punct	:,1
R3122	T10740	T10701	punct	.,revealed
R3123	T10742	T10743	prep	Of,were
R3124	T10744	T10745	det	the,animals
R3125	T10745	T10742	pobj	animals,Of
R3126	T10746	T10745	amod	initial,animals
R3127	T10747	T10745	nummod	60,animals
R3128	T10748	T10745	acl	genotyped,animals
R3129	T10749	T10743	punct	", ",were
R313	T1186	T1178	punct	", ",models
R3130	T10750	T10743	nsubj	19,were
R3131	T10751	T10752	punct	(,%
R3132	T10752	T10750	parataxis	%,19
R3133	T10753	T10752	nummod	32,%
R3134	T10754	T10752	punct	),%
R3135	T10755	T10756	punct	+,+
R3136	T10756	T10743	attr	+,were
R3137	T10757	T10756	punct	/,+
R3138	T10758	T10743	punct	", ",were
R3139	T10759	T10760	nsubj	27,were
R314	T1187	T1188	advmod	as,as
R3140	T10760	T10743	conj	were,were
R3141	T10761	T10762	punct	(,%
R3142	T10762	T10759	parataxis	%,27
R3143	T10763	T10762	nummod	45,%
R3144	T10764	T10762	punct	),%
R3145	T10765	T10766	punct	+,-
R3146	T10766	T10760	attr	-,were
R3147	T10767	T10766	punct	/,-
R3148	T10768	T10760	punct	", ",were
R3149	T10769	T10760	cc	and,were
R315	T1188	T1178	cc	as,models
R3150	T10770	T10771	nsubj	14,were
R3151	T10771	T10760	conj	were,were
R3152	T10772	T10773	punct	(,%
R3153	T10773	T10770	parataxis	%,14
R3154	T10774	T10773	nummod	23,%
R3155	T10775	T10773	punct	),%
R3156	T10776	T10777	punct	-,-
R3157	T10777	T10771	attr	-,were
R3158	T10778	T10777	punct	/,-
R3159	T10779	T10743	punct	.,were
R316	T1189	T1188	advmod	well,as
R3160	T10781	T10782	advmod	Thus,was
R3161	T10783	T10782	punct	", ",was
R3162	T10784	T10785	det	the,gene
R3163	T10785	T10782	nsubj	gene,was
R3164	T10786	T10785	compound	E2,gene
R3165	T10787	T10782	acomp	dispensable,was
R3166	T10788	T10787	prep	for,dispensable
R3167	T10789	T10790	amod	normal,development
R3168	T10790	T10788	pobj	development,for
R3169	T10791	T10790	amod	embryonic,development
R317	T1190	T1178	conj	models,models
R3170	T10792	T10782	punct	.,was
R3171	T10794	T10795	advmod	However,died
R3172	T10796	T10795	punct	", ",died
R3173	T10797	T10798	mark	as,expected
R3174	T10798	T10795	advcl	expected,died
R3175	T10799	T10795	punct	", ",died
R3176	T10800	T10801	advmod	nearly,mice
R3177	T10801	T10795	nsubj	mice,died
R3178	T10802	T10801	det	all,mice
R3179	T10803	T10801	amod	homozygous,mice
R318	T1191	T1192	aux	to,test
R3180	T10804	T10795	prep	in,died
R3181	T10805	T10806	det	the,period
R3182	T10806	T10804	pobj	period,in
R3183	T10807	T10806	amod	perinatal,period
R3184	T10808	T10795	punct	.,died
R3185	T10810	T10811	advmod	Immediately,following
R3186	T10811	T10812	prep	following,were
R3187	T10812	T10817	ccomp	were,were
R3188	T10813	T10811	pobj	birth,following
R3189	T10814	T10812	punct	", ",were
R319	T1192	T1190	advcl	test,models
R3190	T10815	T10816	amod	homozygous,pups
R3191	T10816	T10812	nsubj	pups,were
R3192	T10818	T10812	acomp	indistinguishable,were
R3193	T10819	T10818	prep	from,indistinguishable
R3194	T10820	T10821	poss	their,littermates
R3195	T10821	T10819	pobj	littermates,from
R3196	T10822	T10823	punct	+,+
R3197	T10823	T10821	punct	+,littermates
R3198	T10824	T10823	punct	/,+
R3199	T10825	T10823	cc	and,+
R32	T873	T872	punct	-,chain
R320	T1193	T1194	amod	novel,strategies
R3200	T10826	T10827	punct	+,-
R3201	T10827	T10823	conj	-,+
R3202	T10828	T10827	punct	/,-
R3203	T10829	T10817	punct	;,were
R3204	T10830	T10817	nsubj	they,were
R3205	T10831	T10817	acomp	vigorous,were
R3206	T10832	T10831	punct	", ",vigorous
R3207	T10833	T10831	conj	active,vigorous
R3208	T10834	T10833	cc	and,active
R3209	T10835	T10833	conj	able,active
R321	T1194	T1192	dobj	strategies,test
R3210	T10836	T10837	aux	to,suckle
R3211	T10837	T10835	xcomp	suckle,able
R3212	T10838	T10817	punct	.,were
R3213	T10840	T10841	prep	By,became
R3214	T10842	T10843	amod	mid,day
R3215	T10843	T10840	pobj	day,By
R3216	T10844	T10842	prep	to,mid
R3217	T10845	T10844	amod	late,to
R3218	T10846	T10843	prep	on,day
R3219	T10847	T10848	amod	postnatal,day
R322	T1195	T1194	amod	therapeutic,strategies
R3220	T10848	T10846	pobj	day,on
R3221	T10849	T10848	nummod	one,day
R3222	T10850	T10841	punct	", ",became
R3223	T10851	T10852	advmod	most,pups
R3224	T10852	T10841	nsubj	pups,became
R3225	T10853	T10854	punct	-,-
R3226	T10854	T10852	punct	-,pups
R3227	T10855	T10854	punct	/,-
R3228	T10856	T10841	acomp	moribund,became
R3229	T10857	T10841	cc	and,became
R323	T1196	T1194	punct	", ",strategies
R3230	T10858	T10841	conj	were,became
R3231	T10859	T10860	advmod	readily,identifiable
R3232	T10860	T10858	acomp	identifiable,were
R3233	T10861	T10862	mark	as,were
R3234	T10862	T10860	advcl	were,identifiable
R3235	T10863	T10862	nsubj	they,were
R3236	T10864	T10865	amod	lethargic,pale
R3237	T10865	T10862	acomp	pale,were
R3238	T10866	T10865	punct	", ",pale
R3239	T10867	T10862	punct	", ",were
R324	T1197	T1198	amod	such,as
R3240	T10868	T10862	cc	and,were
R3241	T10869	T10862	conj	exhibited,were
R3242	T10870	T10871	amod	gasping,movements
R3243	T10871	T10869	dobj	movements,exhibited
R3244	T10872	T10871	amod	respiratory,movements
R3245	T10873	T10841	punct	.,became
R3246	T10875	T10876	prep	With,died
R3247	T10877	T10878	amod	few,exceptions
R3248	T10878	T10875	pobj	exceptions,With
R3249	T10879	T10876	punct	", ",died
R325	T1198	T1194	prep	as,strategies
R3250	T10880	T10881	punct	-,-
R3251	T10881	T10883	punct	-,pups
R3252	T10882	T10881	punct	/,-
R3253	T10883	T10876	nsubj	pups,died
R3254	T10884	T10876	prep	within,died
R3255	T10885	T10886	nummod	72,hours
R3256	T10886	T10884	pobj	hours,within
R3257	T10887	T10886	prep	of,hours
R3258	T10888	T10887	pobj	birth,of
R3259	T10889	T10876	punct	.,died
R326	T1199	T1200	nmod	gene,therapies
R3260	T10891	T10892	nsubj	We,observed
R3261	T10893	T10892	aux	have,observed
R3262	T10894	T10895	nummod	one,pup
R3263	T10895	T10892	dobj	pup,observed
R3264	T10896	T10895	amod	rare,pup
R3265	T10897	T10898	punct	-,-
R3266	T10898	T10895	punct	-,pup
R3267	T10899	T10898	punct	/,-
R3268	T10900	T10901	dep	that,survived
R3269	T10901	T10895	relcl	survived,pup
R327	T1200	T1198	pobj	therapies,as
R3270	T10902	T10901	prep	to,survived
R3271	T10903	T10904	amod	postnatal,day
R3272	T10904	T10902	pobj	day,to
R3273	T10905	T10904	nummod	13,day
R3274	T10906	T10892	punct	.,observed
R3275	T10908	T10909	det	The,reason
R3276	T10909	T10910	nsubj	reason,is
R3277	T10911	T10909	prep	for,reason
R3278	T10912	T10913	det	the,survival
R3279	T10913	T10911	pobj	survival,for
R328	T1201	T1199	cc	and,gene
R3280	T10914	T10913	amod	prolonged,survival
R3281	T10915	T10913	prep	of,survival
R3282	T10916	T10917	det	this,pup
R3283	T10917	T10915	pobj	pup,of
R3284	T10918	T10910	acomp	unknown,is
R3285	T10919	T10910	punct	.,is
R329	T1202	T1199	conj	cellular,gene
R3290	T12206	T12207	npadvmod	E2,deficient
R3291	T12207	T12208	amod	deficient,mice
R3292	T12208	T12209	nsubj	mice,model
R3293	T12210	T12209	advmod	accurately,model
R3294	T12211	T12212	amod	classic,MSUD
R3295	T12212	T12209	dobj	MSUD,model
R3296	T12214	T12215	aux	To,demonstrate
R3297	T12215	T12216	advcl	demonstrate,determined
R3298	T12217	T12218	mark	that,created
R3299	T12218	T12215	ccomp	created,demonstrate
R33	T874	T870	pobj	dehydrogenase,of
R330	T1203	T1200	punct	", ",therapies
R3300	T12219	T12220	det	the,event
R3301	T12220	T12218	nsubj	event,created
R3302	T12221	T12222	compound	gene,targeting
R3303	T12222	T12220	compound	targeting,event
R3304	T12223	T12224	det	a,allele
R3305	T12224	T12218	dobj	allele,created
R3306	T12225	T12224	amod	true,allele
R3307	T12226	T12227	npadvmod	E2,null
R3308	T12227	T12224	amod	null,allele
R3309	T12228	T12216	punct	", ",determined
R331	T1204	T1205	aux	to,treat
R3310	T12229	T12216	nsubj	we,determined
R3311	T12230	T12231	compound	BCKDH,activity
R3312	T12231	T12216	dobj	activity,determined
R3313	T12232	T12216	prep	in,determined
R3314	T12233	T12234	compound	liver,homogenates
R3315	T12234	T12232	pobj	homogenates,in
R3316	T12235	T12234	prep	of,homogenates
R3317	T12236	T12237	amod	postnatal,day
R3318	T12237	T12238	nmod	day,pups
R3319	T12238	T12235	pobj	pups,of
R332	T1205	T1200	advcl	treat,therapies
R3320	T12239	T12237	nummod	1,day
R3321	T12240	T12238	compound	mouse,pups
R3322	T12241	T12238	acl	derived,pups
R3323	T12242	T12241	prep	from,derived
R3324	T12243	T12244	punct	+,-
R3325	T12244	T12246	punct	-,pairs
R3326	T12245	T12244	punct	/,-
R3327	T12246	T12242	pobj	pairs,from
R3328	T12247	T12244	prep	by,-
R3329	T12248	T12249	punct	+,-
R333	T1206	T1207	det	this,disease
R3330	T12249	T12247	punct	-,by
R3331	T12250	T12249	punct	/,-
R3332	T12251	T12246	compound	mating,pairs
R3333	T12252	T12216	punct	.,determined
R3334	T12254	T12255	mark	As,shown
R3335	T12255	T12256	advcl	shown,was
R3336	T12257	T12255	prep	in,shown
R3337	T12258	T12259	compound	Figure,2A
R3338	T12259	T12257	pobj	2A,in
R3339	T12260	T12256	punct	", ",was
R334	T1207	T1205	dobj	disease,treat
R3340	T12261	T12262	det	the,activity
R3341	T12262	T12256	nsubj	activity,was
R3342	T12263	T12262	compound	BCKDH,activity
R3343	T12264	T12262	prep	in,activity
R3344	T12265	T12266	punct	+,+
R3345	T12266	T12268	punct	+,mice
R3346	T12267	T12266	punct	/,+
R3347	T12268	T12264	pobj	mice,in
R3348	T12269	T12270	advmod	readily,detectable
R3349	T12270	T12256	acomp	detectable,was
R335	T1208	T1207	amod	devastating,disease
R3350	T12271	T12256	punct	.,was
R3351	T12273	T12274	prep	In,was
R3352	T12275	T12276	amod	marked,contrast
R3353	T12276	T12273	pobj	contrast,In
R3354	T12277	T12274	punct	", ",was
R3355	T12278	T12279	compound	BCKDH,activity
R3356	T12279	T12274	nsubj	activity,was
R3357	T12280	T12281	advmod	completely,absent
R3358	T12281	T12274	acomp	absent,was
R3359	T12282	T12274	prep	in,was
R336	T1209	T1207	amod	metabolic,disease
R3360	T12283	T12284	punct	-,-
R3361	T12284	T12286	punct	-,livers
R3362	T12285	T12284	punct	/,-
R3363	T12286	T12282	pobj	livers,in
R3364	T12287	T12286	compound	mouse,livers
R3365	T12288	T12274	punct	.,was
R3366	T12290	T12291	mark	As,expected
R3367	T12291	T12292	advcl	expected,had
R3368	T12293	T12292	punct	", ",had
R3369	T12294	T12292	nsubj	homogenates,had
R337	T1210	T1176	punct	.,provide
R3370	T12295	T12294	prep	from,homogenates
R3371	T12296	T12297	punct	+,-
R3372	T12297	T12299	punct	-,mice
R3373	T12298	T12297	punct	/,-
R3374	T12299	T12295	pobj	mice,from
R3375	T12300	T12301	advmod	approximately,half
R3376	T12301	T12302	nummod	half,activity
R3377	T12302	T12292	dobj	activity,had
R3378	T12303	T12302	det	the,activity
R3379	T12304	T12302	prep	of,activity
R338	T847	T843	compound	syrup,disease
R3380	T12305	T12306	poss	their,littermates
R3381	T12306	T12304	pobj	littermates,of
R3382	T12307	T12308	punct	+,+
R3383	T12308	T12306	punct	+,littermates
R3384	T12309	T12308	punct	/,+
R3385	T12310	T12311	punct	(,2A
R3386	T12311	T12292	parataxis	2A,had
R3387	T12312	T12311	compound	Figure,2A
R3388	T12313	T12311	punct	),2A
R3389	T12314	T12292	punct	.,had
R3390	T12316	T12317	det	The,results
R3391	T12317	T12318	nsubj	results,are
R3392	T12319	T12317	prep	from,results
R3393	T12320	T12321	punct	-,-
R3394	T12321	T12323	punct	-,mice
R3395	T12322	T12321	punct	/,-
R3396	T12323	T12319	pobj	mice,from
R3397	T12324	T12318	acomp	similar,are
R3398	T12325	T12324	prep	to,similar
R3399	T12326	T12325	pobj	that,to
R34	T875	T876	compound	keto,acid
R3400	T12327	T12326	acl	observed,that
R3401	T12328	T12327	prep	in,observed
R3402	T12329	T12328	pobj	humans,in
R3403	T12330	T12329	prep	with,humans
R3404	T12331	T12332	amod	classic,MSUD
R3405	T12332	T12330	pobj	MSUD,with
R3406	T12333	T12334	punct	[,1
R3407	T12334	T12318	parataxis	1,are
R3408	T12335	T12334	punct	],1
R3409	T12336	T12318	punct	.,are
R3410	T12339	T12340	nsubjpass	Immunohistochemistry,used
R3411	T12341	T12339	prep	with,Immunohistochemistry
R3412	T12342	T12343	det	an,antibody
R3413	T12343	T12341	pobj	antibody,with
R3414	T12344	T12345	npadvmod	E2,specific
R3415	T12345	T12343	amod	specific,antibody
R3416	T12346	T12340	auxpass	was,used
R3417	T12347	T12348	aux	to,examine
R3418	T12348	T12340	advcl	examine,used
R3419	T12349	T12350	compound	E2,protein
R3420	T12350	T12348	dobj	protein,examine
R3421	T12351	T12348	prep	in,examine
R3422	T12352	T12353	det	the,mice
R3423	T12353	T12351	pobj	mice,in
R3424	T12354	T12340	punct	.,used
R3425	T12356	T12357	mark	As,shown
R3426	T12357	T12358	advcl	shown,was
R3427	T12359	T12357	prep	in,shown
R3428	T12360	T12361	compound	Figure,1C
R3429	T12361	T12359	pobj	1C,in
R3430	T12362	T12361	cc	and,1C
R3431	T12363	T12361	conj	1D,1C
R3432	T12364	T12358	punct	", ",was
R3433	T12365	T12366	amod	immunoreactive,protein
R3434	T12366	T12358	nsubj	protein,was
R3435	T12367	T12366	compound	E2,protein
R3436	T12368	T12358	acomp	abundant,was
R3437	T12369	T12358	prep	in,was
R3438	T12370	T12369	pobj	liver,in
R3439	T12371	T12370	cc	and,liver
R3440	T12372	T12373	amod	embryonic,fibroblasts
R3441	T12373	T12370	conj	fibroblasts,liver
R3442	T12374	T12370	prep	of,liver
R3443	T12375	T12376	punct	+,+
R3444	T12376	T12378	punct	+,mice
R3445	T12377	T12376	punct	/,+
R3446	T12378	T12374	pobj	mice,of
R3447	T12379	T12358	punct	.,was
R3448	T12381	T12382	prep	In,was
R3449	T12383	T12384	amod	marked,contrast
R3450	T12384	T12381	pobj	contrast,In
R3451	T12385	T12382	punct	", ",was
R3452	T12386	T12387	amod	immunoreactive,protein
R3453	T12387	T12382	nsubj	protein,was
R3454	T12388	T12387	compound	E2,protein
R3455	T12389	T12382	acomp	absent,was
R3456	T12390	T12382	prep	in,was
R3457	T12391	T12392	det	these,tissues
R3458	T12392	T12390	pobj	tissues,in
R3459	T12393	T12392	amod	same,tissues
R3460	T12394	T12392	prep	of,tissues
R3461	T12395	T12396	punct	-,-
R3462	T12396	T12398	punct	-,mice
R3463	T12397	T12396	punct	/,-
R3464	T12398	T12394	pobj	mice,of
R3465	T12399	T12382	punct	.,was
R3466	T12401	T12402	amod	Homozygous,knockout
R3467	T12402	T12404	compound	knockout,mice
R3468	T12403	T12402	compound	E2,knockout
R3469	T12404	T12405	nsubj	mice,had
R3470	T12406	T12407	det	a,increase
R3471	T12407	T12405	dobj	increase,had
R3472	T12408	T12409	advmod	nearly,3
R3473	T12409	T12410	quantmod	3,fold
R3474	T12410	T12407	nummod	fold,increase
R3475	T12411	T12410	punct	-,fold
R3476	T12412	T12407	prep	in,increase
R3477	T12413	T12414	nmod	blood,levels
R3478	T12414	T12412	pobj	levels,in
R3479	T12415	T12416	punct	(,2B
R348	T3304	T3305	compound	Maple,Syrup
R3480	T12416	T12413	parataxis	2B,blood
R3481	T12417	T12416	compound	Figure,2B
R3482	T12418	T12416	punct	),2B
R3483	T12419	T12413	cc	and,blood
R3484	T12420	T12413	conj	urine,blood
R3485	T12421	T12422	punct	(,shown
R3486	T12422	T12420	parataxis	shown,urine
R3487	T12423	T12422	nsubj	data,shown
R3488	T12424	T12422	neg	not,shown
R3489	T12425	T12422	punct	),shown
R349	T3305	T3306	compound	Syrup,Disease
R3490	T12426	T12414	prep	of,levels
R3491	T12427	T12426	pobj	BCAA,of
R3492	T12428	T12429	punct	(,sum
R3493	T12429	T12427	parataxis	sum,BCAA
R3494	T12430	T12429	prep	of,sum
R3495	T12431	T12430	pobj	leucine,of
R3496	T12432	T12431	punct	", ",leucine
R3497	T12433	T12431	conj	isoleucine,leucine
R3498	T12434	T12433	punct	", ",isoleucine
R3499	T12435	T12433	cc	and,isoleucine
R35	T876	T874	compound	acid,dehydrogenase
R350	T3306	T3308	nsubj	Disease,is
R3500	T12436	T12433	conj	valine,isoleucine
R3501	T12437	T12429	punct	),sum
R3502	T12438	T12407	prep	as,increase
R3503	T12439	T12438	prep	compared,as
R3504	T12440	T12439	prep	to,compared
R3505	T12441	T12442	poss	their,littermates
R3506	T12442	T12440	pobj	littermates,to
R3507	T12443	T12444	punct	+,+
R3508	T12444	T12442	punct	+,littermates
R3509	T12445	T12444	punct	/,+
R351	T3307	T3306	compound	Urine,Disease
R3510	T12446	T12405	punct	.,had
R3511	T12448	T12449	mark	Because,analyzed
R3512	T12449	T12453	advcl	analyzed,include
R3513	T12450	T12451	compound	amino,acids
R3514	T12451	T12449	nsubjpass	acids,analyzed
R3515	T12452	T12449	auxpass	were,analyzed
R3516	T12454	T12449	prep	by,analyzed
R3517	T12455	T12456	compound	tandem,spectrometry
R3518	T12456	T12454	pobj	spectrometry,by
R3519	T12457	T12456	compound	mass,spectrometry
R352	T3309	T3306	punct	(,Disease
R3520	T12458	T12453	punct	", ",include
R3521	T12459	T12460	det	the,sum
R3522	T12460	T12453	nsubj	sum,include
R3523	T12461	T12460	prep	of,sum
R3524	T12462	T12461	pobj	BCAA,of
R3525	T12463	T12460	acl	shown,sum
R3526	T12464	T12463	prep	in,shown
R3527	T12465	T12466	compound	Figure,2B
R3528	T12466	T12464	pobj	2B,in
R3529	T12467	T12453	advmod	also,include
R353	T3310	T3306	appos	MSUD,Disease
R3530	T12468	T12453	aux	may,include
R3531	T12469	T12453	dobj	alloisoleucine,include
R3532	T12470	T12471	dep	that,produced
R3533	T12471	T12469	relcl	produced,alloisoleucine
R3534	T12472	T12471	aux	may,produced
R3535	T12473	T12471	aux	have,produced
R3536	T12474	T12471	auxpass	been,produced
R3537	T12475	T12471	prep	in,produced
R3538	T12476	T12477	punct	-,-
R3539	T12477	T12479	punct	-,mice
R354	T3311	T3308	punct	),is
R3540	T12478	T12477	punct	/,-
R3541	T12479	T12475	pobj	mice,in
R3542	T12480	T12479	compound	MSUD,mice
R3543	T12481	T12453	punct	.,include
R3544	T12483	T12484	det	The,metabolism
R3545	T12484	T12485	nsubjpass	metabolism,linked
R3546	T12486	T12484	prep	of,metabolism
R3547	T12487	T12486	pobj	BCAA,of
R3548	T12488	T12485	auxpass	is,linked
R3549	T12489	T12485	prep	with,linked
R355	T3312	T3313	det	a,disorder
R3550	T12490	T12491	det	the,synthesis
R3551	T12491	T12489	pobj	synthesis,with
R3552	T12492	T12491	prep	of,synthesis
R3553	T12493	T12492	pobj	alanine,of
R3554	T12494	T12493	punct	", ",alanine
R3555	T12495	T12493	conj	glutamate,alanine
R3556	T12496	T12495	punct	", ",glutamate
R3557	T12497	T12495	cc	and,glutamate
R3558	T12498	T12495	conj	glutamine,glutamate
R3559	T12499	T12500	punct	[,33
R356	T3313	T3308	attr	disorder,is
R3560	T12500	T12485	parataxis	33,linked
R3561	T12501	T12500	punct	],33
R3562	T12502	T12485	punct	.,linked
R3563	T12504	T12505	prep	Because,analyzed
R3564	T12506	T12504	pcomp	of,Because
R3565	T12507	T12508	amod	impaired,metabolism
R3566	T12508	T12504	pobj	metabolism,Because
R3567	T12509	T12508	prep	of,metabolism
R3568	T12510	T12509	pobj	BCAA,of
R3569	T12511	T12508	prep	in,metabolism
R357	T3314	T3313	amod	genetic,disorder
R3570	T12512	T12513	compound	MSUD,mice
R3571	T12513	T12511	pobj	mice,in
R3572	T12514	T12504	punct	", ",Because
R3573	T12515	T12504	cc	and,Because
R3574	T12516	T12517	aux	to,characterize
R3575	T12517	T12504	conj	characterize,Because
R3576	T12518	T12517	advmod	further,characterize
R3577	T12519	T12520	det	the,biochemistry
R3578	T12520	T12517	dobj	biochemistry,characterize
R3579	T12521	T12520	amod	abnormal,biochemistry
R358	T3315	T3313	acl	caused,disorder
R3580	T12522	T12517	prep	in,characterize
R3581	T12523	T12524	det	this,model
R3582	T12524	T12522	pobj	model,in
R3583	T12525	T12505	punct	", ",analyzed
R3584	T12526	T12505	nsubj	we,analyzed
R3585	T12527	T12528	det	the,levels
R3586	T12528	T12505	dobj	levels,analyzed
R3587	T12529	T12528	compound	blood,levels
R3588	T12530	T12528	prep	of,levels
R3589	T12531	T12532	det	the,alanine
R359	T3316	T3315	agent	by,caused
R3590	T12532	T12530	pobj	alanine,of
R3591	T12533	T12532	punct	", ",alanine
R3592	T12534	T12532	conj	glutamate,alanine
R3593	T12535	T12534	punct	", ",glutamate
R3594	T12536	T12534	cc	and,glutamate
R3595	T12537	T12534	conj	glutamine,glutamate
R3596	T12538	T12505	punct	.,analyzed
R3597	T12540	T12541	mark	As,shown
R3598	T12541	T12542	advcl	shown,were
R3599	T12543	T12541	prep	in,shown
R36	T877	T857	punct	.,is
R360	T3317	T3318	det	a,deficiency
R3600	T12544	T12543	pobj	Table,in
R3601	T12545	T12544	nummod	1,Table
R3602	T12546	T12542	punct	", ",were
R3603	T12547	T12548	det	the,levels
R3604	T12548	T12542	nsubj	levels,were
R3605	T12549	T12548	prep	of,levels
R3606	T12550	T12551	det	all,acids
R3607	T12551	T12549	pobj	acids,of
R3608	T12552	T12551	nummod	three,acids
R3609	T12553	T12551	compound	amino,acids
R361	T3318	T3316	pobj	deficiency,by
R3610	T12554	T12548	prep	in,levels
R3611	T12555	T12556	amod	homozygous,mice
R3612	T12556	T12554	pobj	mice,in
R3613	T12557	T12558	advmod	markedly,lower
R3614	T12558	T12542	acomp	lower,were
R3615	T12559	T12558	prep	than,lower
R3616	T12560	T12561	det	the,levels
R3617	T12561	T12559	pobj	levels,than
R3618	T12562	T12561	prep	in,levels
R3619	T12563	T12564	punct	+,+
R362	T3319	T3318	prep	of,deficiency
R3620	T12564	T12566	punct	+,mice
R3621	T12565	T12564	punct	/,+
R3622	T12566	T12562	pobj	mice,in
R3623	T12567	T12564	cc	or,+
R3624	T12568	T12569	punct	+,-
R3625	T12569	T12564	conj	-,+
R3626	T12570	T12569	punct	/,-
R3627	T12571	T12542	punct	.,were
R3628	T12573	T12574	det	The,levels
R3629	T12574	T12575	nsubj	levels,were
R363	T3320	T3321	amod	branched,chain
R3630	T12576	T12574	prep	of,levels
R3631	T12577	T12578	det	these,acids
R3632	T12578	T12576	pobj	acids,of
R3633	T12579	T12578	compound	amino,acids
R3634	T12580	T12574	prep	in,levels
R3635	T12581	T12582	det	the,mice
R3636	T12582	T12580	pobj	mice,in
R3637	T12583	T12584	punct	+,-
R3638	T12584	T12582	punct	-,mice
R3639	T12585	T12584	punct	/,-
R364	T3321	T3323	compound	chain,dehydrogenase
R3640	T12586	T12575	acomp	comparable,were
R3641	T12587	T12586	prep	to,comparable
R3642	T12588	T12587	pobj	those,to
R3643	T12589	T12588	prep	in,those
R3644	T12590	T12591	punct	+,+
R3645	T12591	T12593	punct	+,mice
R3646	T12592	T12591	punct	/,+
R3647	T12593	T12589	pobj	mice,in
R3648	T12594	T12595	punct	(,Table
R3649	T12595	T12575	parataxis	Table,were
R365	T3322	T3321	punct	-,chain
R3650	T12596	T12595	nummod	1,Table
R3651	T12597	T12595	punct	),Table
R3652	T12598	T12575	punct	.,were
R3653	T12600	T12601	prep	Because,suggested
R3654	T12602	T12600	pcomp	of,Because
R3655	T12603	T12604	det	the,decrease
R3656	T12604	T12600	pobj	decrease,Because
R3657	T12605	T12604	amod	abnormal,decrease
R3658	T12606	T12604	prep	in,decrease
R3659	T12607	T12608	det	the,level
R366	T3323	T3319	pobj	dehydrogenase,of
R3660	T12608	T12606	pobj	level,in
R3661	T12609	T12608	compound	blood,level
R3662	T12610	T12608	compound	alanine,level
R3663	T12611	T12604	cc	and,decrease
R3664	T12612	T12613	amod	marked,rise
R3665	T12613	T12604	conj	rise,decrease
R3666	T12614	T12613	prep	in,rise
R3667	T12615	T12616	compound	blood,levels
R3668	T12616	T12614	pobj	levels,in
R3669	T12617	T12616	compound	BCAA,levels
R367	T3324	T3325	compound	keto,acid
R3670	T12618	T12619	dep	that,are
R3671	T12619	T12604	relcl	are,decrease
R3672	T12620	T12619	acomp	characteristic,are
R3673	T12621	T12620	prep	of,characteristic
R3674	T12622	T12621	pobj	MSUD,of
R3675	T12623	T12601	punct	", ",suggested
R3676	T12624	T12625	det	a,report
R3677	T12625	T12601	nsubj	report,suggested
R3678	T12626	T12625	amod	recent,report
R3679	T12627	T12625	prep	on,report
R368	T3325	T3323	compound	acid,dehydrogenase
R3680	T12628	T12629	compound	MSUD,patients
R3681	T12629	T12627	pobj	patients,on
R3682	T12630	T12601	aux	has,suggested
R3683	T12631	T12632	mark	that,provides
R3684	T12632	T12601	ccomp	provides,suggested
R3685	T12633	T12634	det	the,ratio
R3686	T12634	T12632	nsubj	ratio,provides
R3687	T12635	T12634	prep	of,ratio
R3688	T12636	T12637	compound	BCAA,alanine
R3689	T12637	T12635	pobj	alanine,of
R369	T3326	T3323	punct	(,dehydrogenase
R3690	T12638	T12637	punct	/,alanine
R3691	T12639	T12640	det	a,measure
R3692	T12640	T12632	dobj	measure,provides
R3693	T12641	T12642	advmod	more,sensitive
R3694	T12642	T12640	amod	sensitive,measure
R3695	T12643	T12640	prep	of,measure
R3696	T12644	T12645	det	the,biochemistry
R3697	T12645	T12643	pobj	biochemistry,of
R3698	T12646	T12645	amod	abnormal,biochemistry
R3699	T12647	T12645	prep	of,biochemistry
R37	T879	T880	nsubj	MSUD,has
R370	T3327	T3323	appos	BCKDH,dehydrogenase
R3700	T12648	T12647	pobj	MSUD,of
R3701	T12649	T12640	prep	than,measure
R3702	T12650	T12651	compound	BCAA,level
R3703	T12651	T12649	pobj	level,than
R3704	T12652	T12651	advmod	alone,level
R3705	T12653	T12654	punct	[,8
R3706	T12654	T12601	parataxis	8,suggested
R3707	T12655	T12654	punct	],8
R3708	T12656	T12601	punct	.,suggested
R3709	T12658	T12659	advmod	Therefore,expressed
R371	T3328	T3318	punct	),deficiency
R3710	T12660	T12659	punct	", ",expressed
R3711	T12661	T12659	nsubj	we,expressed
R3712	T12662	T12659	advmod	also,expressed
R3713	T12663	T12664	det	the,results
R3714	T12664	T12659	dobj	results,expressed
R3715	T12665	T12666	compound	amino,acid
R3716	T12666	T12664	compound	acid,results
R3717	T12667	T12659	prep	as,expressed
R3718	T12668	T12669	compound	BCAA,alanine
R3719	T12669	T12671	compound	alanine,ratio
R372	T3329	T3318	punct	", ",deficiency
R3720	T12670	T12669	punct	/,alanine
R3721	T12671	T12667	pobj	ratio,as
R3722	T12672	T12659	punct	.,expressed
R3723	T12674	T12675	mark	As,shown
R3724	T12675	T12676	advcl	shown,was
R3725	T12677	T12675	prep	in,shown
R3726	T12678	T12679	compound	Figure,2C
R3727	T12679	T12677	pobj	2C,in
R3728	T12680	T12676	punct	", ",was
R3729	T12681	T12682	det	this,ratio
R373	T3330	T3331	det	a,complex
R3730	T12682	T12676	nsubj	ratio,was
R3731	T12683	T12682	prep	in,ratio
R3732	T12684	T12685	punct	-,-
R3733	T12685	T12687	punct	-,pups
R3734	T12686	T12685	punct	/,-
R3735	T12687	T12683	pobj	pups,in
R3736	T12688	T12689	advmod	more,6
R3737	T12689	T12691	quantmod	6,fold
R3738	T12690	T12689	quantmod	than,6
R3739	T12691	T12693	advmod	fold,higher
R374	T3331	T3318	appos	complex,deficiency
R3740	T12692	T12691	punct	-,fold
R3741	T12693	T12676	acomp	higher,was
R3742	T12694	T12693	prep	than,higher
R3743	T12695	T12696	punct	+,+
R3744	T12696	T12698	punct	+,littermates
R3745	T12697	T12696	punct	/,+
R3746	T12698	T12694	pobj	littermates,than
R3747	T12699	T12696	cc	or,+
R3748	T12700	T12701	punct	+,-
R3749	T12701	T12696	conj	-,+
R375	T3332	T3331	amod	mitochondrial,complex
R3750	T12702	T12701	punct	/,-
R3751	T12703	T12676	punct	.,was
R3752	T12705	T12706	det	These,results
R3753	T12706	T12710	nsubj	results,are
R3754	T12707	T12706	compound	blood,results
R3755	T12708	T12709	compound	amino,acid
R3756	T12709	T12706	compound	acid,results
R3757	T12711	T12710	acomp	consistent,are
R3758	T12712	T12711	prep	with,consistent
R3759	T12713	T12714	det	the,concentrations
R376	T3333	T3331	compound	multienzyme,complex
R3760	T12714	T12712	pobj	concentrations,with
R3761	T12715	T12714	acl	seen,concentrations
R3762	T12716	T12715	prep	in,seen
R3763	T12717	T12716	pobj	patients,in
R3764	T12718	T12717	prep	with,patients
R3765	T12719	T12718	pobj	MSUD,with
R3766	T12720	T12721	punct	[,15
R3767	T12721	T12710	parataxis	15,are
R3768	T12722	T12721	nummod	1,15
R3769	T12723	T12721	punct	",",15
R377	T3334	T3331	amod	responsible,complex
R3770	T12724	T12721	nummod	8,15
R3771	T12725	T12721	punct	",",15
R3772	T12726	T12721	punct	],15
R3773	T12727	T12710	punct	.,are
R3774	T12729	T12730	prep	In,lack
R3775	T12731	T12729	pobj	summary,In
R3776	T12732	T12730	punct	", ",lack
R3777	T12733	T12734	compound	E2,mice
R3778	T12734	T12730	nsubj	mice,lack
R3779	T12735	T12734	compound	knockout,mice
R378	T3335	T3334	prep	for,responsible
R3780	T12736	T12737	nmod	BCKDH,activity
R3781	T12737	T12730	dobj	activity,lack
R3782	T12738	T12737	amod	enzymatic,activity
R3783	T12739	T12737	punct	", ",activity
R3784	T12740	T12741	compound	E2,immunoreactivity
R3785	T12741	T12737	appos	immunoreactivity,activity
R3786	T12742	T12730	punct	", ",lack
R3787	T12743	T12730	cc	and,lack
R3788	T12744	T12730	conj	have,lack
R3789	T12745	T12746	advmod	markedly,elevated
R379	T3336	T3337	det	the,decarboxylation
R3790	T12746	T12747	amod	elevated,levels
R3791	T12747	T12744	dobj	levels,have
R3792	T12748	T12747	prep	of,levels
R3793	T12749	T12748	pobj	BCAA,of
R3794	T12750	T12744	prep	in,have
R3795	T12751	T12752	det	the,blood
R3796	T12752	T12750	pobj	blood,in
R3797	T12753	T12752	cc	and,blood
R3798	T12754	T12752	conj	urine,blood
R3799	T12755	T12730	punct	.,lack
R38	T881	T882	amod	several,phenotypes
R380	T3337	T3335	pobj	decarboxylation,for
R3800	T12757	T12758	det	These,derangements
R3801	T12758	T12760	nsubj	derangements,result
R3802	T12759	T12758	amod	metabolic,derangements
R3803	T12761	T12760	advmod	ultimately,result
R3804	T12762	T12760	prep	in,result
R3805	T12763	T12764	amod	neonatal,lethality
R3806	T12764	T12762	pobj	lethality,in
R3807	T12765	T12760	punct	.,result
R3808	T12767	T12768	det	These,phenotypes
R3809	T12768	T12769	nsubj	phenotypes,are
R381	T3338	T3337	amod	oxidative,decarboxylation
R3810	T12770	T12771	advmod	remarkably,similar
R3811	T12771	T12769	acomp	similar,are
R3812	T12772	T12771	prep	to,similar
R3813	T12773	T12772	pobj	that,to
R3814	T12774	T12773	acl	observed,that
R3815	T12775	T12774	prep	in,observed
R3816	T12776	T12775	pobj	humans,in
R3817	T12777	T12776	prep	with,humans
R3818	T12778	T12779	det	the,form
R3819	T12779	T12777	pobj	form,with
R382	T3339	T3337	prep	of,decarboxylation
R3820	T12780	T12779	amod	classic,form
R3821	T12781	T12779	prep	of,form
R3822	T12782	T12781	pobj	MSUD,of
R3823	T12783	T12769	punct	.,are
R3824	T12785	T12786	advmod	Thus,model
R3825	T12787	T12786	punct	", ",model
R3826	T12788	T12789	compound	E2,mice
R3827	T12789	T12786	nsubj	mice,model
R3828	T12790	T12789	compound	knockout,mice
R3829	T12791	T12786	advmod	closely,model
R383	T3423	T3422	prep	of,%
R3830	T12792	T12793	amod	classic,MSUD
R3831	T12793	T12786	dobj	MSUD,model
R3832	T12794	T12786	punct	.,model
R3835	T15726	T15727	compound	Knockout,mice
R3836	T15727	T15728	nsubj	mice,model
R3837	T15729	T15727	acl	expressing,mice
R3838	T15730	T15731	det	a,transgene
R3839	T15731	T15729	dobj	transgene,expressing
R384	T3340	T3341	amod	branched,chain
R3840	T15732	T15731	amod	human,transgene
R3841	T15733	T15731	compound	E2,transgene
R3842	T15734	T15735	det	a,form
R3843	T15735	T15728	dobj	form,model
R3844	T15736	T15735	amod	variant,form
R3845	T15737	T15735	prep	of,form
R3846	T15738	T15737	pobj	MSUD,of
R3847	T15739	T15740	dep	that,mimics
R3848	T15740	T15735	relcl	mimics,form
R3849	T15741	T15742	amod	intermediate,MSUD
R385	T3424	T3423	pobj	normal,of
R3850	T15742	T15740	dobj	MSUD,mimics
R3851	T15744	T15745	aux	To,create
R3852	T15745	T15746	advcl	create,used
R3853	T15747	T15748	det	a,model
R3854	T15748	T15745	dobj	model,create
R3855	T15749	T15748	amod	murine,model
R3856	T15750	T15748	prep	of,model
R3857	T15751	T15752	det	the,form
R3858	T15752	T15750	pobj	form,of
R3859	T15753	T15752	amod	intermediate,form
R386	T3425	T3426	punct	[,1
R3860	T15754	T15752	compound	variant,form
R3861	T15755	T15752	prep	of,form
R3862	T15756	T15755	pobj	MSUD,of
R3863	T15757	T15746	punct	", ",used
R3864	T15758	T15746	nsubj	we,used
R3865	T15759	T15760	det	a,strategy
R3866	T15760	T15746	dobj	strategy,used
R3867	T15761	T15760	amod	transgenic,strategy
R3868	T15762	T15763	aux	to,rescue
R3869	T15763	T15746	advcl	rescue,used
R387	T3341	T3343	compound	chain,acids
R3870	T15764	T15765	det	the,elevation
R3871	T15765	T15763	dobj	elevation,rescue
R3872	T15766	T15765	amod	severe,elevation
R3873	T15767	T15765	prep	of,elevation
R3874	T15768	T15767	pobj	BCAA,of
R3875	T15769	T15765	cc	and,elevation
R3876	T15770	T15771	amod	neonatal,lethality
R3877	T15771	T15765	conj	lethality,elevation
R3878	T15772	T15773	dep	that,occurs
R3879	T15773	T15771	relcl	occurs,lethality
R388	T3426	T3402	parataxis	1,have
R3880	T15774	T15773	prep	in,occurs
R3881	T15775	T15776	det	the,model
R3882	T15776	T15774	pobj	model,in
R3883	T15777	T15776	amod	classic,model
R3884	T15778	T15776	compound	MSUD,model
R3885	T15779	T15776	compound	mouse,model
R3886	T15780	T15746	punct	.,used
R3887	T15782	T15783	poss	Our,strategy
R3888	T15783	T15784	nsubjpass	strategy,composed
R3889	T15785	T15784	auxpass	was,composed
R389	T3427	T3426	punct	],1
R3890	T15786	T15784	prep	of,composed
R3891	T15787	T15788	det	a,system
R3892	T15788	T15786	pobj	system,of
R3893	T15789	T15790	nummod	two,part
R3894	T15790	T15788	nmod	part,system
R3895	T15791	T15788	amod	transgenic,system
R3896	T15792	T15793	aux	to,express
R3897	T15793	T15788	advcl	express,system
R3898	T15794	T15795	amod	human,E2
R3899	T15795	T15793	dobj	E2,express
R39	T882	T880	dobj	phenotypes,has
R390	T3428	T3402	punct	.,have
R3900	T15796	T15793	prep	in,express
R3901	T15797	T15796	pobj	liver,in
R3902	T15798	T15793	prep	on,express
R3903	T15799	T15800	det	the,background
R3904	T15800	T15798	pobj	background,on
R3905	T15801	T15802	compound	E2,knockout
R3906	T15802	T15800	compound	knockout,background
R3907	T15803	T15784	punct	.,composed
R3908	T15805	T15806	det	This,system
R3909	T15806	T15808	nsubj	system,consisted
R391	T3430	T3431	det	These,patients
R3910	T15807	T15806	amod	bi-transgenic,system
R3911	T15809	T15808	prep	of,consisted
R3912	T15810	T15811	det	a,transgene
R3913	T15811	T15809	pobj	transgene,of
R3914	T15812	T15813	compound	LAP,tTA
R3915	T15813	T15811	compound	tTA,transgene
R3916	T15814	T15813	punct	-,tTA
R3917	T15815	T15811	cc	and,transgene
R3918	T15816	T15817	det	a,transgene
R3919	T15817	T15811	conj	transgene,transgene
R392	T3431	T3432	nsubj	patients,show
R3920	T15818	T15819	compound	TRE,E2
R3921	T15819	T15817	compound	E2,transgene
R3922	T15820	T15819	punct	-,E2
R3923	T15821	T15822	punct	(,3A
R3924	T15822	T15808	parataxis	3A,consisted
R3925	T15823	T15822	compound	Fig.,3A
R3926	T15824	T15822	punct	),3A
R3927	T15825	T15808	punct	.,consisted
R3928	T15827	T15828	det	The,transgene
R3929	T15828	T15832	nsubj	transgene,directs
R393	T3433	T3434	advmod	markedly,elevated
R3930	T15829	T15830	compound	LAP,tTA
R3931	T15830	T15828	compound	tTA,transgene
R3932	T15831	T15830	punct	-,tTA
R3933	T15833	T15834	punct	[,34
R3934	T15834	T15828	parataxis	34,transgene
R3935	T15835	T15834	punct	],34
R3936	T15836	T15837	amod	high,levels
R3937	T15837	T15832	dobj	levels,directs
R3938	T15838	T15837	prep	of,levels
R3939	T15839	T15840	npadvmod	liver,specific
R394	T3434	T3435	amod	elevated,levels
R3940	T15840	T15841	amod	specific,expression
R3941	T15841	T15838	pobj	expression,of
R3942	T15842	T15841	prep	of,expression
R3943	T15843	T15844	det	the,transactivator
R3944	T15844	T15842	pobj	transactivator,of
R3945	T15845	T15846	npadvmod	tetracycline,controlled
R3946	T15846	T15844	amod	controlled,transactivator
R3947	T15847	T15846	punct	-,controlled
R3948	T15848	T15844	punct	(,transactivator
R3949	T15849	T15844	appos	tTA,transactivator
R395	T3435	T3432	dobj	levels,show
R3950	T15850	T15844	punct	),transactivator
R3951	T15851	T15844	punct	", ",transactivator
R3952	T15852	T15853	det	a,factor
R3953	T15853	T15844	appos	factor,transactivator
R3954	T15854	T15853	compound	transcription,factor
R3955	T15855	T15856	dep	that,stimulates
R3956	T15856	T15853	relcl	stimulates,factor
R3957	T15857	T15856	dobj	expression,stimulates
R3958	T15858	T15857	prep	of,expression
R3959	T15859	T15858	pobj	promoters,of
R396	T3342	T3341	punct	-,chain
R3960	T15860	T15861	dep	that,harbor
R3961	T15861	T15859	relcl	harbor,promoters
R3962	T15862	T15863	det	a,element
R3963	T15863	T15861	dobj	element,harbor
R3964	T15864	T15863	compound	transactivator,element
R3965	T15865	T15863	compound	response,element
R3966	T15866	T15863	punct	(,element
R3967	T15867	T15863	appos	TRE,element
R3968	T15868	T15832	punct	),directs
R3969	T15869	T15832	punct	.,directs
R397	T3436	T3435	prep	of,levels
R3970	T15871	T15872	det	The,transgene
R3971	T15872	T15876	nsubjpass	transgene,designed
R3972	T15873	T15874	compound	TRE,E2
R3973	T15874	T15872	compound	E2,transgene
R3974	T15875	T15874	punct	-,E2
R3975	T15877	T15876	auxpass	was,designed
R3976	T15878	T15879	aux	to,express
R3977	T15879	T15876	advcl	express,designed
R3978	T15880	T15881	det	a,cDNA
R3979	T15881	T15879	dobj	cDNA,express
R398	T3437	T3436	pobj	BCAA,of
R3980	T15882	T15881	amod	human,cDNA
R3981	T15883	T15881	compound	E2,cDNA
R3982	T15884	T15879	prep	from,express
R3983	T15885	T15886	det	a,TRE
R3984	T15886	T15884	pobj	TRE,from
R3985	T15887	T15886	acl	containing,TRE
R3986	T15888	T15889	amod	minimal,promoter
R3987	T15889	T15887	dobj	promoter,containing
R3988	T15890	T15879	prep	upon,express
R3989	T15891	T15890	pobj	stimulation,upon
R399	T3438	T3432	prep	in,show
R3990	T15892	T15891	prep	by,stimulation
R3991	T15893	T15892	pobj	tTA,by
R3992	T15894	T15876	punct	.,designed
R3993	T15897	T15898	nmod	LAP,tTA
R3994	T15898	T15900	nmod	tTA,mice
R3995	T15899	T15898	punct	-,tTA
R3996	T15903	T15902	auxpass	w,e
R3997	T15904	T15902	advcl	re,e
R3998	T15919	T15920	amod	c,e
R3999	T15923	T15922	cc	and,d
R4	T839	T840	amod	murine,models
R40	T883	T882	amod	clinical,phenotypes
R400	T3439	T3438	pobj	blood,in
R4000	T15924	T15922	conj	c,d
R4001	T15930	T15929	acl	a,r
R4002	T15932	T15928	aux	t,r
R4003	T15933	T15928	auxpass	e,r
R4004	T15937	T15936	agent	by,zed
R4005	T15938	T15939	det	the,laboratory
R4006	T15939	T15937	pobj	laboratory,by
R4007	T15940	T15939	compound	Bujard,laboratory
R4008	T15941	T15942	punct	[,34
R4009	T15942	T15936	parataxis	34,zed
R401	T3343	T3339	pobj	acids,of
R4010	T15943	T15942	punct	],34
R4011	T15946	T15947	amod	Pronuclear,microinjection
R4012	T15947	T15948	nsubjpass	microinjection,used
R4013	T15949	T15948	auxpass	was,used
R4014	T15950	T15951	aux	to,produce
R4015	T15951	T15948	advcl	produce,used
R4016	T15952	T15953	amod	transgenic,mice
R4017	T15953	T15951	dobj	mice,produce
R4018	T15954	T15955	dep	that,harbored
R4019	T15955	T15953	relcl	harbored,mice
R402	T3440	T3439	cc	and,blood
R4020	T15956	T15957	det	the,transgene
R4021	T15957	T15955	dobj	transgene,harbored
R4022	T15958	T15959	compound	TRE,E2
R4023	T15959	T15957	compound	E2,transgene
R4024	T15960	T15959	punct	-,E2
R4025	T15961	T15948	punct	.,used
R4026	T15963	T15964	prep	From,identified
R4027	T15965	T15963	pobj	injections,From
R4028	T15966	T15965	prep	at,injections
R4029	T15967	T15968	det	the,University
R403	T3441	T3442	amod	other,fluids
R4030	T15968	T15966	pobj	University,at
R4031	T15969	T15968	prep	of,University
R4032	T15970	T15969	pobj	Pittsburgh,of
R4033	T15971	T15964	punct	", ",identified
R4034	T15972	T15973	nummod	2,founders
R4035	T15973	T15964	nsubjpass	founders,identified
R4036	T15974	T15975	npadvmod	transgene,positive
R4037	T15975	T15973	amod	positive,founders
R4038	T15976	T15964	auxpass	were,identified
R4039	T15977	T15978	dep	that,led
R404	T3344	T3343	compound	keto,acids
R4040	T15978	T15964	ccomp	led,identified
R4041	T15979	T15978	prep	to,led
R4042	T15980	T15981	det	the,generation
R4043	T15981	T15979	pobj	generation,to
R4044	T15982	T15981	prep	of,generation
R4045	T15983	T15984	nummod	3,lines
R4046	T15984	T15982	pobj	lines,of
R4047	T15985	T15984	amod	different,lines
R4048	T15986	T15984	amod	transgenic,lines
R4049	T15987	T15988	punct	(,A
R405	T3442	T3439	conj	fluids,blood
R4050	T15988	T15984	parataxis	A,lines
R4051	T15989	T15988	compound	Lines,A
R4052	T15990	T15988	punct	", ",A
R4053	T15991	T15988	conj	B,A
R4054	T15992	T15991	punct	", ",B
R4055	T15993	T15991	cc	&,B
R4056	T15994	T15991	conj	D,B
R4057	T15995	T15988	punct	),A
R4058	T15996	T15964	punct	.,identified
R4059	T15998	T15999	prep	From,obtained
R406	T3443	T3442	compound	body,fluids
R4060	T16000	T16001	det	the,injections
R4061	T16001	T15998	pobj	injections,From
R4062	T16002	T16001	prep	at,injections
R4063	T16003	T16004	det	the,University
R4064	T16004	T16002	pobj	University,at
R4065	T16005	T16004	prep	of,University
R4066	T16006	T16005	pobj	Cincinnati,of
R4067	T16007	T15999	punct	", ",obtained
R4068	T16008	T15999	nsubj	we,obtained
R4069	T16009	T16010	nummod	8,founders
R407	T3444	T3445	punct	[,1
R4070	T16010	T15999	dobj	founders,obtained
R4071	T16011	T16012	npadvmod	transgene,positive
R4072	T16012	T16010	amod	positive,founders
R4073	T16013	T15999	punct	.,obtained
R4074	T16015	T16016	amod	Subsequent,breeding
R4075	T16016	T16017	nsubj	breeding,revealed
R4076	T16018	T16016	prep	of,breeding
R4077	T16019	T16020	det	these,founders
R4078	T16020	T16018	pobj	founders,of
R4079	T16021	T16022	mark	that,had
R408	T3345	T3343	acl	derived,acids
R4080	T16022	T16017	ccomp	had,revealed
R4081	T16023	T16022	nsubj	many,had
R4082	T16024	T16023	prep	of,many
R4083	T16025	T16026	det	the,animals
R4084	T16026	T16024	pobj	animals,of
R4085	T16027	T16028	amod	multiple,sites
R4086	T16028	T16022	dobj	sites,had
R4087	T16029	T16028	compound	transgene,sites
R4088	T16030	T16028	compound	insertion,sites
R4089	T16031	T16032	dep	that,segregated
R409	T3445	T3432	parataxis	1,show
R4090	T16032	T16028	relcl	segregated,sites
R4091	T16033	T16017	punct	.,revealed
R4092	T16035	T16036	nsubj	We,established
R4093	T16037	T16038	det	a,total
R4094	T16038	T16036	dobj	total,established
R4095	T16039	T16038	prep	of,total
R4096	T16040	T16041	nummod	15,lines
R4097	T16041	T16039	pobj	lines,of
R4098	T16042	T16041	amod	different,lines
R4099	T16043	T16041	amod	transgenic,lines
R41	T884	T880	prep	depending,has
R410	T3446	T3445	punct	],1
R4100	T16044	T16036	prep	from,established
R4101	T16045	T16046	det	these,founders
R4102	T16046	T16044	pobj	founders,from
R4103	T16047	T16036	punct	.,established
R4104	T16049	T16050	amod	Limited,resources
R4105	T16050	T16051	nsubj	resources,allowed
R4106	T16052	T16053	nsubj	us,focus
R4107	T16053	T16051	ccomp	focus,allowed
R4108	T16054	T16053	aux	to,focus
R4109	T16055	T16053	advmod	only,focus
R411	T3447	T3432	punct	.,show
R4110	T16056	T16053	prep	on,focus
R4111	T16057	T16058	det	a,total
R4112	T16058	T16056	pobj	total,on
R4113	T16059	T16058	prep	of,total
R4114	T16060	T16059	pobj	9,of
R4115	T16061	T16060	prep	of,9
R4116	T16062	T16063	det	these,lines
R4117	T16063	T16061	pobj	lines,of
R4118	T16064	T16051	punct	.,allowed
R4119	T16066	T16067	det	These,lines
R412	T3346	T3345	prep	from,derived
R4120	T16067	T16068	nsubj	lines,differed
R4121	T16069	T16068	advmod	substantially,differed
R4122	T16070	T16068	prep	in,differed
R4123	T16071	T16072	compound	transgene,number
R4124	T16072	T16070	pobj	number,in
R4125	T16073	T16072	compound	copy,number
R4126	T16074	T16075	mark	as,compared
R4127	T16075	T16068	advcl	compared,differed
R4128	T16076	T16075	prep	by,compared
R4129	T16077	T16078	compound	Southern,blot
R413	T3449	T3450	prep	Besides,are
R4130	T16078	T16079	compound	blot,analysis
R4131	T16079	T16076	pobj	analysis,by
R4132	T16080	T16079	prep	of,analysis
R4133	T16081	T16082	compound	tail,DNA
R4134	T16082	T16080	pobj	DNA,of
R4135	T16083	T16084	punct	(,shown
R4136	T16084	T16068	parataxis	shown,differed
R4137	T16085	T16084	nsubj	data,shown
R4138	T16086	T16084	neg	not,shown
R4139	T16087	T16084	punct	),shown
R414	T3347	T3348	amod	branched,chain
R4140	T16088	T16068	punct	.,differed
R4141	T16090	T16091	amod	Transgenic,mice
R4142	T16091	T16095	nsubjpass	mice,crossed
R4143	T16092	T16093	compound	TRE,E2
R4144	T16093	T16091	compound	E2,mice
R4145	T16094	T16093	punct	-,E2
R4146	T16096	T16091	prep	from,mice
R4147	T16097	T16098	det	each,line
R4148	T16098	T16096	pobj	line,from
R4149	T16099	T16100	punct	(,offspring
R415	T3451	T3452	det	the,form
R4150	T16100	T16098	parataxis	offspring,line
R4151	T16101	T16100	preconj	either,offspring
R4152	T16102	T16100	nmod	founders,offspring
R4153	T16103	T16102	cc	or,founders
R4154	T16104	T16102	conj	F1,founders
R4155	T16105	T16100	punct	),offspring
R4156	T16106	T16095	auxpass	were,crossed
R4157	T16107	T16095	prep	to,crossed
R4158	T16108	T16107	pobj	mice,to
R4159	T16109	T16110	dep	that,were
R416	T3452	T3449	pobj	form,Besides
R4160	T16110	T16108	relcl	were,mice
R4161	T16111	T16110	acomp	positive,were
R4162	T16112	T16111	prep	for,positive
R4163	T16113	T16114	det	the,transgene
R4164	T16114	T16112	pobj	transgene,for
R4165	T16115	T16116	compound	LAP,tTA
R4166	T16116	T16114	compound	tTA,transgene
R4167	T16117	T16116	punct	-,tTA
R4168	T16118	T16110	cc	and,were
R4169	T16119	T16110	conj	were,were
R417	T3348	T3350	compound	chain,acids
R4170	T16120	T16119	acomp	heterozygous,were
R4171	T16121	T16120	prep	for,heterozygous
R4172	T16122	T16123	det	the,knockout
R4173	T16123	T16121	pobj	knockout,for
R4174	T16124	T16123	compound	E2,knockout
R4175	T16125	T16095	punct	.,crossed
R4176	T16127	T16128	advmod	Ultimately,used
R4177	T16129	T16128	punct	", ",used
R4178	T16130	T16131	amod	breeding,pairs
R4179	T16131	T16128	nsubjpass	pairs,used
R418	T3453	T3452	amod	classic,form
R4180	T16132	T16128	auxpass	were,used
R4181	T16133	T16134	prep	in,were
R4182	T16134	T16128	ccomp	were,used
R4183	T16135	T16133	pobj	which,in
R4184	T16136	T16137	det	the,mice
R4185	T16137	T16134	nsubj	mice,were
R4186	T16138	T16134	acomp	heterozygous,were
R4187	T16139	T16138	prep	for,heterozygous
R4188	T16140	T16141	det	the,transgene
R4189	T16141	T16139	pobj	transgene,for
R419	T3454	T3450	punct	", ",are
R4190	T16142	T16143	compound	LAP,tTA
R4191	T16143	T16141	compound	tTA,transgene
R4192	T16144	T16143	punct	-,tTA
R4193	T16145	T16141	punct	", ",transgene
R4194	T16146	T16147	det	the,transgene
R4195	T16147	T16141	conj	transgene,transgene
R4196	T16148	T16149	compound	TRE,E2
R4197	T16149	T16147	compound	E2,transgene
R4198	T16150	T16149	punct	-,E2
R4199	T16151	T16147	punct	", ",transgene
R42	T885	T884	prep	on,depending
R420	T3455	T3450	expl	there,are
R4200	T16152	T16147	cc	and,transgene
R4201	T16153	T16154	det	the,knockout
R4202	T16154	T16147	conj	knockout,transgene
R4203	T16155	T16154	compound	E2,knockout
R4204	T16156	T16128	punct	.,used
R4205	T16158	T16159	aux	To,screen
R4206	T16159	T16161	advcl	screen,genotyped
R4207	T16160	T16159	advmod	efficiently,screen
R4208	T16162	T16159	prep	for,screen
R4209	T16163	T16164	det	the,ability
R421	T3349	T3348	punct	-,chain
R4210	T16164	T16162	pobj	ability,for
R4211	T16165	T16164	prep	of,ability
R4212	T16166	T16167	det	the,transgenes
R4213	T16167	T16165	pobj	transgenes,of
R4214	T16168	T16169	aux	to,rescue
R4215	T16169	T16164	acl	rescue,ability
R4216	T16170	T16171	det	the,knockout
R4217	T16171	T16169	dobj	knockout,rescue
R4218	T16172	T16169	prep	from,rescue
R4219	T16173	T16174	amod	neonatal,lethality
R422	T3456	T3457	amod	other,variants
R4220	T16174	T16172	pobj	lethality,from
R4221	T16175	T16161	punct	", ",genotyped
R4222	T16176	T16161	nsubj	we,genotyped
R4223	T16177	T16161	dobj	litters,genotyped
R4224	T16178	T16161	prep	at,genotyped
R4225	T16179	T16178	pobj	weaning,at
R4226	T16180	T16161	punct	.,genotyped
R4227	T16182	T16183	det	This,strategy
R4228	T16183	T16185	nsubjpass	strategy,expected
R4229	T16184	T16183	amod	breeding,strategy
R423	T3350	T3346	pobj	acids,from
R4230	T16186	T16185	auxpass	is,expected
R4231	T16187	T16188	aux	to,result
R4232	T16188	T16185	xcomp	result,expected
R4233	T16189	T16188	prep	in,result
R4234	T16190	T16191	det	a,maximum
R4235	T16191	T16189	pobj	maximum,in
R4236	T16192	T16191	amod	theoretical,maximum
R4237	T16193	T16191	prep	of,maximum
R4238	T16194	T16193	pobj	animals,of
R4239	T16195	T16194	prep	with,animals
R424	T3457	T3450	attr	variants,are
R4240	T16196	T16197	det	the,genotype
R4241	T16197	T16195	pobj	genotype,with
R4242	T16198	T16197	compound	rescue,genotype
R4243	T16199	T16200	punct	(,knockout
R4244	T16200	T16197	parataxis	knockout,genotype
R4245	T16201	T16200	advmod	i.e.,knockout
R4246	T16202	T16200	punct	", ",knockout
R4247	T16203	T16200	amod	homozygous,knockout
R4248	T16204	T16200	compound	E2,knockout
R4249	T16205	T16200	cc	and,knockout
R425	T3458	T3457	prep	of,variants
R4250	T16206	T16200	conj	positive,knockout
R4251	T16207	T16206	prep	for,positive
R4252	T16208	T16209	det	both,transgenes
R4253	T16209	T16207	pobj	transgenes,for
R4254	T16210	T16200	punct	),knockout
R4255	T16211	T16191	prep	of,maximum
R4256	T16212	T16213	nummod	14,%
R4257	T16213	T16211	pobj	%,of
R4258	T16214	T16185	punct	.,expected
R4259	T16216	T16217	compound	Fig.,4A
R426	T3459	T3460	det	the,disease
R4260	T16217	T16218	nsubj	4A,shows
R4261	T16219	T16220	det	the,percentage
R4262	T16220	T16218	dobj	percentage,shows
R4263	T16221	T16220	prep	of,percentage
R4264	T16222	T16221	pobj	mice,of
R4265	T16223	T16222	amod	alive,mice
R4266	T16224	T16223	prep	at,alive
R4267	T16225	T16224	pobj	weaning,at
R4268	T16226	T16220	prep	with,percentage
R4269	T16227	T16228	det	the,genotype
R427	T3460	T3458	pobj	disease,of
R4270	T16228	T16226	pobj	genotype,with
R4271	T16229	T16228	compound	rescue,genotype
R4272	T16230	T16228	prep	from,genotype
R4273	T16231	T16232	det	each,line
R4274	T16232	T16230	pobj	line,from
R4275	T16279	T16280	advmod	Thus,appears
R4276	T16233	T16232	amod	transgenic,line
R4277	T16281	T16280	punct	", ",appears
R4278	T16234	T16232	acl	tested,line
R4279	T16282	T16283	det	the,transgene
R428	T3351	T3350	compound	amino,acids
R4280	T16283	T16280	nsubj	transgene,appears
R4281	T16235	T16218	punct	.,shows
R4282	T16284	T16283	compound	525A,transgene
R4283	T16285	T16286	aux	to,be
R4284	T16237	T16238	amod	Considerable,variability
R4285	T16286	T16280	xcomp	be,appears
R4286	T16287	T16288	advmod	highly,effective
R4287	T16288	T16286	acomp	effective,be
R4288	T16289	T16288	prep	at,effective
R4289	T16238	T16239	nsubjpass	variability,observed
R429	T3461	T3450	punct	.,are
R4290	T16240	T16239	auxpass	was,observed
R4291	T16290	T16289	pcomp	rescuing,at
R4292	T16291	T16292	det	the,phenotype
R4293	T16241	T16239	prep	between,observed
R4294	T16292	T16290	dobj	phenotype,rescuing
R4295	T16293	T16292	amod	neonatal,phenotype
R4296	T16294	T16292	amod	lethal,phenotype
R4297	T16242	T16241	pobj	lines,between
R4298	T16295	T16292	prep	of,phenotype
R4299	T16296	T16297	det	the,knockout
R43	T886	T887	det	the,degree
R430	T3463	T3464	nsubj	Patients,show
R4300	T16297	T16295	pobj	knockout,of
R4301	T16298	T16297	compound	E2,knockout
R4302	T16243	T16239	punct	.,observed
R4303	T16299	T16280	punct	.,appears
R4304	T16301	T16302	prep	In,failed
R4305	T16245	T16246	det	The,line
R4306	T16303	T16301	pobj	contrast,In
R4307	T16304	T16302	punct	", ",failed
R4308	T16305	T16306	compound	line,520B
R4309	T16246	T16247	nsubj	line,was
R431	T3352	T3350	punct	(,acids
R4310	T16306	T16302	nsubj	520B,failed
R4311	T16248	T16246	prep	with,line
R4312	T16307	T16302	advmod	completely,failed
R4313	T16308	T16309	aux	to,rescue
R4314	T16309	T16302	xcomp	rescue,failed
R4315	T16310	T16311	det	the,phenotype
R4316	T16249	T16250	det	the,percentage
R4317	T16311	T16309	dobj	phenotype,rescue
R4318	T16250	T16248	pobj	percentage,with
R4319	T16312	T16311	amod	neonatal,phenotype
R432	T3465	T3463	prep	with,Patients
R4320	T16313	T16311	amod	lethal,phenotype
R4321	T16314	T16302	punct	.,failed
R4322	T16251	T16250	amod	highest,percentage
R4323	T16316	T16317	det	All,lines
R4324	T16317	T16319	nsubj	lines,produced
R4325	T16318	T16317	amod	other,lines
R4326	T16252	T16250	prep	of,percentage
R4327	T16320	T16317	acl	tested,lines
R4328	T16253	T16254	compound	rescue,mice
R4329	T16321	T16322	amod	surviving,animals
R433	T3466	T3467	det	the,form
R4330	T16322	T16319	dobj	animals,produced
R4331	T16323	T16322	compound	rescue,animals
R4332	T16254	T16252	pobj	mice,of
R4333	T16324	T16319	prep	at,produced
R4334	T16325	T16326	det	a,frequency
R4335	T16326	T16324	pobj	frequency,at
R4336	T16255	T16250	prep	at,percentage
R4337	T16327	T16326	prep	between,frequency
R4338	T16328	T16329	nummod	1,%
R4339	T16329	T16327	pobj	%,between
R434	T3353	T3350	appos	BCAA,acids
R4340	T16256	T16255	pobj	weaning,at
R4341	T16330	T16328	cc	and,1
R4342	T16331	T16328	conj	5,1
R4343	T16332	T16329	prep	of,%
R4344	T16257	T16258	nmod	line,A
R4345	T16333	T16334	amod	weaned,pups
R4346	T16334	T16332	pobj	pups,of
R4347	T16335	T16319	punct	.,produced
R4348	T16258	T16247	attr	A,was
R4349	T16259	T16258	nummod	525,A
R435	T3354	T3350	punct	),acids
R4350	T16260	T16247	punct	.,was
R4351	T16262	T16263	advmod	Approximately,10
R4352	T16263	T16264	nummod	10,%
R4353	T16341	T16340	nsubj	n,i
R4354	T16264	T16265	nsubj	%,had
R4355	T16266	T16264	prep	of,%
R4356	T16267	T16268	amod	weaned,pups
R4357	T16268	T16266	pobj	pups,of
R4358	T16269	T16268	punct	", ",pups
R4359	T16270	T16268	amod	surviving,pups
R436	T3467	T3465	pobj	form,with
R4360	T16352	T16353	advmod	s,e
R4361	T16353	T16351	acomp	e,were
R4362	T16271	T16264	prep	from,%
R4363	T16272	T16273	det	this,line
R4364	T16273	T16271	pobj	line,from
R4365	T16274	T16275	det	the,genotype
R4366	T16275	T16265	dobj	genotype,had
R4367	T16276	T16275	compound	rescue,genotype
R4368	T16277	T16265	punct	.,had
R4369	T16380	T16381	compound	Fig.,4B
R437	T3468	T3467	amod	intermediate,form
R4370	T16381	T16382	nsubj	4B,shows
R4371	T16383	T16384	det	the,results
R4372	T16363	T16362	cc	d,or
R4373	T16384	T16382	dobj	results,shows
R4374	T16385	T16384	prep	of,results
R4375	T16367	T16366	pobj	ai,t
R4376	T16386	T16387	compound	blood,analysis
R4377	T16371	T16370	conj	c,d
R4378	T16387	T16385	pobj	analysis,of
R4379	T16372	T16373	poss	h,a
R438	T3355	T3350	punct	", ",acids
R4380	T16388	T16389	compound	amino,acid
R4381	T16389	T16387	compound	acid,analysis
R4382	T16376	T16377	amod	a,cterization
R4383	T16390	T16384	acl	presented,results
R4384	T16391	T16390	prep	as,presented
R4385	T16392	T16393	det	a,ratio
R4386	T16393	T16391	pobj	ratio,as
R4387	T16394	T16393	prep	of,ratio
R4388	T16484	T16485	mark	Because,demonstrated
R4389	T16395	T16396	amod	total,BCAA
R439	T3469	T3467	prep	of,form
R4390	T16485	T16503	advcl	demonstrated,examined
R4391	T16486	T16487	det	the,mice
R4392	T16396	T16394	pobj	BCAA,of
R4393	T16487	T16485	nsubjpass	mice,demonstrated
R4394	T16488	T16489	compound	LAP,tTA
R4395	T16489	T16487	compound	tTA,mice
R4396	T16397	T16393	prep	to,ratio
R4397	T16490	T16489	punct	-,tTA
R4398	T16491	T16492	dep	that,used
R4399	T16492	T16487	relcl	used,mice
R44	T887	T885	pobj	degree,on
R440	T3470	T3471	det	the,disease
R4400	T16398	T16397	pobj	alanine,to
R4401	T16493	T16492	auxpass	were,used
R4402	T16494	T16495	aux	to,drive
R4403	T16495	T16492	advcl	drive,used
R4404	T16496	T16495	dobj	expression,drive
R4405	T16497	T16496	prep	of,expression
R4406	T16399	T16382	punct	.,shows
R4407	T16498	T16499	amod	transgenic,E2
R4408	T16499	T16497	pobj	E2,of
R4409	T16500	T16499	amod	human,E2
R441	T3471	T3469	pobj	disease,of
R4410	T16401	T16402	det	The,values
R4411	T16501	T16485	aux	have,demonstrated
R4412	T16402	T16406	nsubj	values,were
R4413	T16403	T16404	compound	BCAA,alanine
R4414	T16404	T16402	compound	alanine,values
R4415	T16502	T16485	auxpass	been,demonstrated
R4416	T16405	T16404	punct	/,alanine
R4417	T16504	T16505	aux	to,produce
R4418	T16505	T16485	xcomp	produce,demonstrated
R4419	T16506	T16507	npadvmod	liver,specific
R442	T3472	T3473	compound	BCKDH,activity
R4420	T16407	T16402	prep	for,values
R4421	T16507	T16508	amod	specific,expression
R4422	T16508	T16505	dobj	expression,produce
R4423	T16509	T16510	punct	[,34
R4424	T16408	T16407	pobj	mice,for
R4425	T16510	T16485	parataxis	34,demonstrated
R4426	T16511	T16510	punct	],34
R4427	T16512	T16503	punct	", ",examined
R4428	T16409	T16408	prep	from,mice
R4429	T16513	T16514	compound	BCKDH,activity
R443	T3473	T3464	dobj	activity,show
R4430	T16514	T16503	nsubjpass	activity,examined
R4431	T16515	T16514	compound	enzyme,activity
R4432	T16410	T16411	det	both,lines
R4433	T16516	T16514	cc	and,activity
R4434	T16517	T16514	conj	production,activity
R4435	T16518	T16517	prep	of,production
R4436	T16411	T16409	pobj	lines,from
R4437	T16519	T16520	amod	human,protein
R4438	T16520	T16518	pobj	protein,of
R4439	T16521	T16520	compound	E2,protein
R444	T3474	T3473	prep	in,activity
R4440	T16412	T16413	advmod	significantly,elevated
R4441	T16522	T16514	prep	in,activity
R4442	T16523	T16522	pobj	liver,in
R4443	T16524	T16523	prep	of,liver
R4444	T16413	T16406	acomp	elevated,were
R4445	T16525	T16526	compound	Lines,A
R4446	T16526	T16524	pobj	A,of
R4447	T16527	T16526	cc	and,A
R4448	T16414	T16406	prep	compared,were
R4449	T16528	T16526	conj	525A,A
R445	T3356	T3350	conj	leucine,acids
R4450	T16529	T16503	auxpass	was,examined
R4451	T16415	T16414	prep	to,compared
R4452	T16530	T16503	punct	.,examined
R4453	T16416	T16415	pobj	controls,to
R4454	T16417	T16416	prep	for,controls
R4455	T16532	T16533	nmod	BCKDH,activity
R4456	T16533	T16535	nsubj	activity,was
R4457	T16534	T16533	amod	enzymatic,activity
R4458	T16418	T16417	pobj	all,for
R4459	T16536	T16533	prep	in,activity
R446	T3475	T3476	det	the,range
R4460	T16537	T16536	pobj	liver,in
R4461	T16538	T16537	prep	from,liver
R4462	T16539	T16540	compound	Lines,A
R4463	T16419	T16418	prep	of,all
R4464	T16540	T16538	pobj	A,from
R4465	T16541	T16540	cc	and,A
R4466	T16420	T16421	det	the,points
R4467	T16542	T16540	conj	525A,A
R4468	T16543	T16544	punct	~,6
R4469	T16544	T16545	nummod	6,%
R447	T3476	T3474	pobj	range,in
R4470	T16421	T16419	pobj	points,of
R4471	T16545	T16535	attr	%,was
R4472	T16546	T16544	cc	and,6
R4473	T16422	T16421	compound	time,points
R4474	T16547	T16544	conj	5,6
R4475	T16548	T16535	punct	", ",was
R4476	T16549	T16535	advmod	respectively,was
R4477	T16423	T16421	acl	analyzed,points
R4478	T16550	T16535	punct	", ",was
R4479	T16551	T16535	prep	of,was
R448	T3477	T3476	prep	of,range
R4480	T16552	T16553	det	the,activity
R4481	T16424	T16406	punct	.,were
R4482	T16553	T16551	pobj	activity,of
R4483	T16554	T16553	amod	enzymatic,activity
R4484	T16555	T16553	amod	present,activity
R4485	T16427	T16428	mark	that,are
R4486	T16556	T16555	prep	in,present
R4487	T16557	T16558	compound	control,liver
R4488	T16558	T16556	pobj	liver,in
R4489	T16559	T16560	punct	(,4C
R449	T3357	T3356	punct	", ",leucine
R4490	T16560	T16535	parataxis	4C,was
R4491	T16428	T16426	ccomp	are,Note
R4492	T16561	T16560	compound	Fig.,4C
R4493	T16562	T16560	punct	),4C
R4494	T16563	T16535	punct	.,was
R4495	T16429	T16430	det	the,values
R4496	T16565	T16566	mark	As,shown
R4497	T16430	T16428	nsubj	values,are
R4498	T16566	T16567	advcl	shown,was
R4499	T16568	T16566	prep	in,shown
R45	T888	T887	prep	of,degree
R450	T3478	T3479	quantmod	3,30
R4500	T16431	T16432	compound	BCAA,alanine
R4501	T16569	T16570	compound	Fig.,3B
R4502	T16570	T16568	pobj	3B,in
R4503	T16571	T16567	punct	", ",was
R4504	T16432	T16430	compound	alanine,values
R4505	T16572	T16573	det	the,amount
R4506	T16573	T16567	nsubj	amount,was
R4507	T16574	T16573	prep	of,amount
R4508	T16575	T16576	amod	human,E2
R4509	T16576	T16574	pobj	E2,of
R451	T3479	T3481	nummod	30,%
R4510	T16433	T16432	punct	/,alanine
R4511	T16577	T16578	punct	(,Kd
R4512	T16578	T16576	parataxis	Kd,E2
R4513	T16579	T16580	amod	predicted,MW
R4514	T16580	T16578	nsubj	MW,Kd
R4515	T16434	T16430	prep	for,values
R4516	T16581	T16582	punct	~,54
R4517	T16582	T16578	nummod	54,Kd
R4518	T16583	T16578	punct	),Kd
R4519	T16435	T16436	det	the,mice
R452	T3480	T3479	punct	–,30
R4520	T16584	T16573	prep	in,amount
R4521	T16585	T16586	det	these,mice
R4522	T16586	T16584	pobj	mice,in
R4523	T16436	T16434	pobj	mice,for
R4524	T16587	T16586	amod	transgenic,mice
R4525	T16588	T16589	advmod	quite,variable
R4526	T16589	T16567	acomp	variable,was
R4527	T16437	T16436	amod	transgenic,mice
R4528	T16590	T16589	prep	between,variable
R4529	T16438	T16428	acomp	intermediate,are
R453	T3358	T3356	conj	isoleucine,leucine
R4530	T16439	T16428	prep	between,are
R4531	T16591	T16590	pobj	mice,between
R4532	T16592	T16567	punct	.,was
R4533	T16440	T16439	pobj	controls,between
R4534	T16594	T16595	prep	In,was
R4535	T16596	T16594	pobj	some,In
R4536	T16597	T16596	prep	of,some
R4537	T16441	T16440	cc	and,controls
R4538	T16598	T16599	det	these,mice
R4539	T16599	T16597	pobj	mice,of
R454	T3481	T3477	pobj	%,of
R4540	T16442	T16440	conj	knockouts,controls
R4541	T16600	T16599	amod	transgenic,mice
R4542	T16601	T16595	punct	", ",was
R4543	T16602	T16603	det	the,level
R4544	T16443	T16444	punct	(,see
R4545	T16603	T16595	nsubj	level,was
R4546	T16604	T16603	prep	of,level
R4547	T16605	T16606	amod	human,E2
R4548	T16444	T16428	parataxis	see,are
R4549	T16606	T16604	pobj	E2,of
R455	T3482	T3481	prep	of,%
R4550	T16607	T16608	advmod	approximately,equal
R4551	T16445	T16446	compound	Figure,2C
R4552	T16446	T16444	dobj	2C,see
R4553	T16447	T16444	punct	),see
R4554	T16608	T16595	acomp	equal,was
R4555	T16609	T16608	prep	to,equal
R4556	T16448	T16426	punct	.,Note
R4557	T16610	T16611	det	the,amount
R4558	T16611	T16609	pobj	amount,to
R4559	T16612	T16611	prep	of,amount
R456	T3483	T3482	pobj	normal,of
R4560	T16613	T16614	compound	mouse,E2
R4561	T16614	T16612	pobj	E2,of
R4562	T16615	T16616	punct	(,Kd
R4563	T16450	T16451	compound	Blood,levels
R4564	T16616	T16614	parataxis	Kd,E2
R4565	T16451	T16452	nsubjpass	levels,reduced
R4566	T16617	T16618	punct	~,47
R4567	T16618	T16616	nummod	47,Kd
R4568	T16453	T16451	prep	of,levels
R4569	T16619	T16616	punct	),Kd
R457	T3359	T3358	cc	and,isoleucine
R4570	T16620	T16611	acl	produced,amount
R4571	T16621	T16620	prep	in,produced
R4572	T16622	T16621	pobj	liver,in
R4573	T16623	T16622	prep	of,liver
R4574	T16454	T16453	pobj	alanine,of
R4575	T16624	T16625	amod	nontransgenic,mice
R4576	T16625	T16623	pobj	mice,of
R4577	T16455	T16454	punct	", ",alanine
R4578	T16626	T16625	compound	control,mice
R4579	T16627	T16595	punct	.,was
R458	T3484	T3464	punct	.,show
R4580	T16456	T16454	conj	glutamate,alanine
R4581	T16629	T16630	det	The,observations
R4582	T16630	T16631	nsubj	observations,suggest
R4583	T16457	T16456	punct	", ",glutamate
R4584	T16632	T16633	mark	that,result
R4585	T16633	T16630	acl	result,observations
R4586	T16458	T16456	cc	and,glutamate
R4587	T16634	T16635	det	these,amounts
R4588	T16635	T16633	nsubj	amounts,result
R4589	T16636	T16637	advmod	near,normal
R459	T3486	T3487	prep	In,delayed
R4590	T16459	T16456	conj	glutamine,glutamate
R4591	T16637	T16635	amod	normal,amounts
R4592	T16638	T16635	prep	of,amounts
R4593	T16460	T16452	auxpass	were,reduced
R4594	T16639	T16640	compound	E2,protein
R4595	T16640	T16638	pobj	protein,of
R4596	T16641	T16633	prep	in,result
R4597	T16461	T16452	prep	in,reduced
R4598	T16642	T16643	advmod	only,6
R4599	T16643	T16647	nummod	6,%
R46	T889	T890	compound	enzyme,deficiency
R460	T3360	T3358	conj	valine,isoleucine
R4600	T16644	T16643	punct	~,6
R4601	T16462	T16463	compound	Line,A
R4602	T16645	T16643	quantmod	5,6
R4603	T16646	T16643	punct	–,6
R4604	T16647	T16641	pobj	%,in
R4605	T16463	T16464	compound	A,mice
R4606	T16648	T16647	prep	of,%
R4607	T16649	T16650	amod	normal,activity
R4608	T16650	T16648	pobj	activity,of
R4609	T16464	T16461	pobj	mice,in
R461	T3488	T3489	amod	such,patients
R4610	T16651	T16650	compound	BCKDH,activity
R4611	T16652	T16650	compound	enzyme,activity
R4612	T16653	T16654	mark	that,functioning
R4613	T16654	T16631	ccomp	functioning,suggest
R4614	T16655	T16656	nmod	transgene,E2
R4615	T16656	T16654	nsubj	E2,functioning
R4616	T16465	T16452	punct	", ",reduced
R4617	T16657	T16656	amod	derived,E2
R4618	T16658	T16654	aux	is,functioning
R4619	T16659	T16654	prep	at,functioning
R462	T3489	T3486	pobj	patients,In
R4620	T16466	T16467	mark	as,was
R4621	T16660	T16661	det	a,level
R4622	T16661	T16659	pobj	level,at
R4623	T16467	T16452	advcl	was,reduced
R4624	T16662	T16661	amod	suboptimal,level
R4625	T16663	T16631	punct	.,suggest
R4626	T16468	T16467	nsubj	glutamate,was
R4627	T16665	T16666	nsubj	It,is
R4628	T16667	T16666	acomp	probable,is
R4629	T16469	T16468	prep	in,glutamate
R463	T3361	T3362	punct	(,see
R4630	T16668	T16669	mark	that,interfered
R4631	T16669	T16666	ccomp	interfered,is
R4632	T16670	T16671	det	the,tag
R4633	T16470	T16471	compound	Line,525A
R4634	T16671	T16669	nsubj	tag,interfered
R4635	T16672	T16673	compound	c,myc
R4636	T16471	T16472	compound	525A,mice
R4637	T16673	T16671	compound	myc,tag
R4638	T16674	T16673	punct	-,myc
R4639	T16675	T16671	prep	at,tag
R464	T3490	T3491	det	the,onset
R4640	T16472	T16469	pobj	mice,in
R4641	T16676	T16677	det	the,terminus
R4642	T16677	T16675	pobj	terminus,at
R4643	T16678	T16677	compound	carboxy,terminus
R4644	T16473	T16468	punct	", ",glutamate
R4645	T16679	T16677	prep	of,terminus
R4646	T16680	T16681	det	the,E2
R4647	T16681	T16679	pobj	E2,of
R4648	T16474	T16468	prep	compared,glutamate
R4649	T16682	T16683	npadvmod	transgene,derived
R465	T3491	T3487	nsubjpass	onset,delayed
R4650	T16683	T16681	amod	derived,E2
R4651	T16684	T16681	amod	human,E2
R4652	T16475	T16474	prep	to,compared
R4653	T16685	T16669	prep	with,interfered
R4654	T16686	T16687	amod	enzymatic,activity
R4655	T16687	T16685	pobj	activity,with
R4656	T16476	T16475	pobj	controls,to
R4657	T16688	T16666	punct	.,is
R4658	T16477	T16478	punct	(,Table
R4659	T16690	T16691	det	This,interpretation
R466	T3362	T3308	parataxis	see,is
R4660	T16691	T16692	nsubj	interpretation,is
R4661	T16478	T16467	parataxis	Table,was
R4662	T16693	T16692	acomp	consistent,is
R4663	T16694	T16693	prep	with,consistent
R4664	T16695	T16696	amod	previous,studies
R4665	T16696	T16694	pobj	studies,with
R4666	T16697	T16698	dep	which,revealed
R4667	T16479	T16478	nummod	2,Table
R4668	T16698	T16696	relcl	revealed,studies
R4669	T16699	T16698	aux	have,revealed
R467	T3492	T3491	prep	of,onset
R4670	T16700	T16701	mark	that,is
R4671	T16480	T16478	punct	),Table
R4672	T16481	T16452	punct	.,reduced
R4673	T16701	T16698	ccomp	is,revealed
R4674	T16702	T16703	det	the,terminus
R4675	T16714	T16701	acomp	essential,is
R4676	T16703	T16701	nsubj	terminus,is
R4677	T16704	T16703	compound	carboxy,terminus
R4678	T16705	T16703	prep	of,terminus
R4679	T16706	T16707	det	an,subunit
R468	T3493	T3494	det	the,disease
R4680	T16707	T16705	pobj	subunit,of
R4681	T16708	T16707	amod	analogous,subunit
R4682	T16715	T16714	prep	for,essential
R4683	T16709	T16707	prep	of,subunit
R4684	T16710	T16711	det	the,complex
R4685	T16711	T16709	pobj	complex,of
R4686	T16716	T16717	compound	subunit,interactions
R4687	T16712	T16711	compound	pyruvate,complex
R4688	T16713	T16711	compound	dehydrogenase,complex
R4689	T16717	T16715	pobj	interactions,for
R469	T3494	T3492	pobj	disease,of
R4690	T16718	T16719	punct	[,35
R4691	T16719	T16701	parataxis	35,is
R4692	T16820	T16819	pobj	E2,of
R4693	T16821	T16808	acomp	negligible,is
R4694	T16720	T16719	punct	],35
R4695	T16822	T16808	prep	in,is
R4696	T16823	T16824	det	those,tissues
R4697	T16824	T16822	pobj	tissues,in
R4698	T16721	T16701	cc	and,is
R4699	T16825	T16808	punct	.,is
R47	T890	T888	pobj	deficiency,of
R470	T3495	T3487	auxpass	is,delayed
R4700	T16722	T16723	nsubj	insertion,interfered
R4701	T16827	T16828	amod	Long,term
R4702	T16828	T16830	compound	term,survival
R4703	T16829	T16828	punct	-,term
R4704	T16723	T16701	conj	interfered,is
R4705	T16830	T16831	nsubjpass	survival,plotted
R4706	T16724	T16722	prep	of,insertion
R4707	T16832	T16830	prep	of,survival
R4708	T16833	T16834	det	the,mice
R4709	T16834	T16832	pobj	mice,of
R471	T3363	T3362	prep	for,see
R4710	T16725	T16726	det	a,tag
R4711	T16835	T16834	prep	from,mice
R4712	T16836	T16837	det	these,lines
R4713	T16837	T16835	pobj	lines,from
R4714	T16838	T16837	nummod	two,lines
R4715	T16839	T16837	amod	transgenic,lines
R4716	T16840	T16841	dep	that,survived
R4717	T16726	T16724	pobj	tag,of
R4718	T16841	T16837	relcl	survived,lines
R4719	T16842	T16841	prep	beyond,survived
R472	T3496	T3487	punct	", ",delayed
R4720	T16843	T16842	pobj	weaning,beyond
R4721	T16727	T16726	compound	Hisx6,tag
R4722	T16844	T16831	auxpass	is,plotted
R4723	T16845	T16831	prep	in,plotted
R4724	T16846	T16847	compound	Fig.,4D
R4725	T16728	T16722	prep	on,insertion
R4726	T16847	T16845	pobj	4D,in
R4727	T16848	T16831	punct	.,plotted
R4728	T16729	T16730	det	the,terminus
R4729	T16850	T16851	mark	Although,collected
R473	T3497	T3487	cc	but,delayed
R4730	T16730	T16728	pobj	terminus,on
R4731	T16851	T16859	advcl	collected,is
R4732	T16852	T16853	compound	survival,data
R4733	T16853	T16851	nsubjpass	data,collected
R4734	T16854	T16853	prep	for,data
R4735	T16731	T16730	compound	carboxy,terminus
R4736	T16855	T16856	compound	control,animals
R4737	T16856	T16854	pobj	animals,for
R4738	T16857	T16851	auxpass	was,collected
R4739	T16732	T16730	prep	of,terminus
R474	T3498	T3499	expl	there,are
R4740	T16858	T16851	neg	not,collected
R4741	T16860	T16859	punct	", ",is
R4742	T16733	T16734	det	an,subunit
R4743	T16861	T16859	nsubj	it,is
R4744	T16862	T16863	advmod	readily,apparent
R4745	T16734	T16732	pobj	subunit,of
R4746	T16863	T16859	acomp	apparent,is
R4747	T16864	T16865	mark	that,compromised
R4748	T16865	T16859	ccomp	compromised,is
R4749	T16735	T16734	amod	analogous,subunit
R475	T3364	T3363	pobj	review,for
R4750	T16866	T16865	nsubjpass	survival,compromised
R4751	T16867	T16866	prep	of,survival
R4752	T16868	T16869	det	the,mice
R4753	T16736	T16737	nmod	E.,coli
R4754	T16869	T16867	pobj	mice,of
R4755	T16870	T16869	compound	rescue,mice
R4756	T16871	T16865	auxpass	was,compromised
R4757	T16737	T16734	nmod	coli,subunit
R4758	T16872	T16859	punct	.,is
R4759	T16874	T16875	prep	By,were
R476	T3499	T3487	conj	are,delayed
R4760	T16738	T16723	prep	with,interfered
R4761	T16876	T16877	nummod	16,weeks
R4762	T16739	T16740	amod	normal,assembly
R4763	T16877	T16874	pobj	weeks,By
R4764	T16878	T16875	punct	", ",were
R4765	T16879	T16880	det	all,mice
R4766	T16740	T16738	pobj	assembly,with
R4767	T16880	T16875	nsubj	mice,were
R4768	T16881	T16880	prep	of,mice
R4769	T16741	T16740	compound	subunit,assembly
R477	T3365	T3362	punct	", ",see
R4770	T16882	T16883	compound	Line,A
R4771	T16883	T16881	pobj	A,of
R4772	T16884	T16875	acomp	moribund,were
R4773	T16742	T16743	punct	[,36
R4774	T16885	T16884	cc	and,moribund
R4775	T16886	T16887	advmod	humanely,sacrificed
R4776	T16887	T16884	conj	sacrificed,moribund
R4777	T16743	T16723	parataxis	36,interfered
R4778	T16888	T16875	punct	", ",were
R4779	T16889	T16875	cc	or,were
R478	T3500	T3501	advmod	persistently,elevated
R4780	T16744	T16743	punct	],36
R4781	T16890	T16891	auxpass	were,found
R4782	T16891	T16875	conj	found,were
R4783	T16892	T16891	advcl	dead,found
R4784	T16745	T16692	punct	.,is
R4785	T16893	T16891	prep	in,found
R4786	T16894	T16895	poss	their,cage
R4787	T16747	T16748	nsubj	It,is
R4788	T16895	T16893	pobj	cage,in
R4789	T16896	T16875	punct	.,were
R479	T3501	T3502	amod	elevated,levels
R4790	T16898	T16899	nsubj	Survival,appeared
R4791	T16749	T16748	advmod	also,is
R4792	T16900	T16898	prep	of,Survival
R4793	T16901	T16902	compound	Line,525A
R4794	T16902	T16900	pobj	525A,of
R4795	T16750	T16748	acomp	possible,is
R4796	T16903	T16904	aux	to,be
R4797	T16904	T16899	xcomp	be,appeared
R4798	T16905	T16906	advmod	somewhat,better
R4799	T16751	T16752	mark	that,was
R48	T891	T880	punct	.,has
R480	T3502	T3499	attr	levels,are
R4800	T16906	T16904	acomp	better,be
R4801	T16907	T16899	punct	.,appeared
R4802	T16752	T16748	ccomp	was,is
R4803	T16909	T16910	prep	At,were
R4804	T16753	T16754	amod	human,E2
R4805	T16911	T16912	nummod	20,weeks
R4806	T16912	T16909	pobj	weeks,At
R4807	T16913	T16912	prep	of,weeks
R4808	T16754	T16752	nsubj	E2,was
R4809	T16914	T16913	pobj	age,of
R481	T3503	T3502	prep	of,levels
R4810	T16915	T16910	punct	", ",were
R4811	T16916	T16917	punct	~,12
R4812	T16755	T16752	neg	not,was
R4813	T16917	T16918	nummod	12,%
R4814	T16918	T16910	nsubj	%,were
R4815	T16919	T16918	prep	of,%
R4816	T16756	T16757	advmod	fully,functional
R4817	T16920	T16919	pobj	mice,of
R4818	T16921	T16922	punct	(,2
R4819	T16922	T16920	parataxis	2,mice
R482	T3366	T3362	punct	: ,see
R4820	T16923	T16922	prep	of,2
R4821	T16924	T16923	pobj	16,of
R4822	T16757	T16752	acomp	functional,was
R4823	T16925	T16922	punct	),2
R4824	T16758	T16759	advmod	when,complexed
R4825	T16759	T16752	advcl	complexed,was
R4826	T16760	T16759	prep	with,complexed
R4827	T16926	T16910	advmod	still,were
R4828	T16927	T16910	acomp	alive,were
R4829	T16761	T16762	compound	mouse,E1
R483	T3504	T3503	pobj	BCAA,of
R4830	T16928	T16910	punct	.,were
R4831	T16762	T16760	pobj	E1,with
R4832	T16930	T16931	det	These,survivors
R4833	T16931	T16934	nsubj	survivors,died
R4834	T16932	T16931	nummod	two,survivors
R4835	T16763	T16762	cc	and,E1
R4836	T16933	T16931	amod	rare,survivors
R4837	T16764	T16765	compound	E3,subunits
R4838	T16935	T16934	prep	at,died
R4839	T16936	T16937	nummod	40,weeks
R484	T3505	T3506	punct	[,1
R4840	T16937	T16935	pobj	weeks,at
R4841	T16765	T16762	conj	subunits,E1
R4842	T16938	T16936	cc	and,40
R4843	T16766	T16748	punct	.,is
R4844	T16939	T16936	conj	60,40
R4845	T16940	T16937	prep	of,weeks
R4846	T16941	T16940	pobj	age,of
R4847	T16942	T16934	punct	.,died
R4848	T16768	T16769	amod	Human,E2
R4849	T16944	T16945	prep	In,had
R485	T3367	T3362	punct	[,see
R4850	T16946	T16944	pobj	summary,In
R4851	T16769	T16770	nsubj	E2,shares
R4852	T16947	T16945	punct	", ",had
R4853	T16948	T16949	det	these,mice
R4854	T16771	T16772	punct	~,88
R4855	T16949	T16945	nsubj	mice,had
R4856	T16950	T16949	amod	surviving,mice
R4857	T16951	T16952	compound	BCAA,alanine
R4858	T16952	T16954	compound	alanine,ratios
R4859	T16772	T16773	nummod	88,%
R486	T3506	T3499	parataxis	1,are
R4860	T16953	T16952	punct	/,alanine
R4861	T16954	T16945	dobj	ratios,had
R4862	T16955	T16956	dep	that,were
R4863	T16773	T16774	compound	%,identity
R4864	T16956	T16954	relcl	were,ratios
R4865	T16957	T16956	acomp	intermediate,were
R4866	T16958	T16957	prep	between,intermediate
R4867	T16774	T16770	dobj	identity,shares
R4868	T16959	T16958	pobj	controls,between
R4869	T16960	T16959	cc	and,controls
R487	T3507	T3506	punct	],1
R4870	T16961	T16959	conj	knockouts,controls
R4871	T16962	T16945	cc	and,had
R4872	T16963	T16964	nsubj	they,expressed
R4873	T16775	T16774	prep	to,identity
R4874	T16964	T16945	conj	expressed,had
R4875	T16965	T16966	punct	~,6
R4876	T16776	T16777	compound	mouse,E2
R4877	T16777	T16775	pobj	E2,to
R4878	T16966	T16969	nummod	6,%
R4879	T16778	T16770	prep	at,shares
R488	T3508	T3499	punct	.,are
R4880	T16967	T16966	quantmod	5,6
R4881	T16968	T16966	punct	–,6
R4882	T16969	T16964	dobj	%,expressed
R4883	T16779	T16780	det	the,level
R4884	T16970	T16969	prep	of,%
R4885	T16971	T16972	amod	normal,activity
R4886	T16780	T16778	pobj	level,at
R4887	T16972	T16970	pobj	activity,of
R4888	T16973	T16972	compound	BCKDH,activity
R4889	T16974	T16972	compound	enzyme,activity
R489	T3368	T3362	dobj	1,see
R4890	T16781	T16782	compound	amino,acid
R4891	T16975	T16972	prep	in,activity
R4892	T16976	T16977	det	the,liver
R4893	T16782	T16780	compound	acid,level
R4894	T16977	T16975	pobj	liver,in
R4895	T16978	T16945	punct	.,had
R4896	T16783	T16770	punct	.,shares
R4897	T16980	T16981	det	These,observations
R4898	T16785	T16786	nsubj	We,used
R4899	T16981	T16983	nsubj	observations,are
R49	T893	T894	amod	Current,treatments
R490	T3510	T3511	nsubj	Patients,show
R4900	T16982	T16981	amod	phenotypic,observations
R4901	T16984	T16985	advmod	remarkably,similar
R4902	T16985	T16983	acomp	similar,are
R4903	T16787	T16786	advmod	also,used
R4904	T16986	T16985	prep	to,similar
R4905	T16987	T16988	det	the,phenotype
R4906	T16988	T16986	pobj	phenotype,to
R4907	T16788	T16789	compound	western,blot
R4908	T16789	T16790	compound	blot,analysis
R4909	T16989	T16988	amod	clinical,phenotype
R491	T3369	T3362	punct	],see
R4910	T16790	T16786	dobj	analysis,used
R4911	T16990	T16988	acl	observed,phenotype
R4912	T16991	T16990	prep	in,observed
R4913	T16992	T16993	compound	MSUD,patients
R4914	T16791	T16792	aux	to,analyze
R4915	T16993	T16991	pobj	patients,in
R4916	T16994	T16993	prep	with,patients
R4917	T16995	T16996	det	the,form
R4918	T16792	T16786	advcl	analyze,used
R4919	T16996	T16994	pobj	form,with
R492	T3512	T3510	prep	with,Patients
R4920	T16997	T16996	amod	intermediate,form
R4921	T16998	T16996	prep	of,form
R4922	T16999	T17000	det	the,disease
R4923	T17000	T16998	pobj	disease,of
R4924	T17001	T17002	punct	[,1
R4925	T16793	T16792	dobj	expression,analyze
R4926	T17002	T16983	parataxis	1,are
R4927	T17003	T17002	punct	],1
R4928	T17004	T16983	punct	.,are
R4929	T16794	T16793	prep	of,expression
R493	T3513	T3514	det	the,form
R4930	T17006	T17007	advmod	Thus,represent
R4931	T16795	T16796	det	the,transgene
R4932	T17008	T17007	punct	", ",represent
R4933	T17009	T17010	det	these,mice
R4934	T17010	T17007	nsubj	mice,represent
R4935	T16796	T16794	pobj	transgene,of
R4936	T17011	T17010	compound	rescue,mice
R4937	T17012	T17013	det	a,model
R4938	T17013	T17007	dobj	model,represent
R4939	T16797	T16796	compound	E2,transgene
R494	T3514	T3512	pobj	form,with
R4940	T17014	T17015	advmod	very,useful
R4941	T17015	T17013	amod	useful,model
R4942	T16798	T16792	prep	in,analyze
R4943	T17016	T17013	prep	of,model
R4944	T17017	T17018	det	the,phenotype
R4945	T17018	T17016	pobj	phenotype,of
R4946	T16799	T16798	pobj	brain,in
R4947	T17019	T17018	amod	intermediate,phenotype
R4948	T17020	T17018	compound	MSUD,phenotype
R4949	T17021	T17007	punct	.,represent
R495	T3515	T3514	amod	intermittent,form
R4950	T16800	T16799	punct	", ",brain
R4951	T16801	T16799	conj	kidney,brain
R4952	T16802	T16801	cc	and,kidney
R4953	T16803	T16801	conj	muscle,kidney
R4954	T16804	T16786	punct	.,used
R4955	T16806	T16807	mark	As,shown
R4956	T16807	T16808	advcl	shown,is
R4957	T16809	T16807	prep	in,shown
R4958	T16810	T16811	compound	Figure,3C
R4959	T16811	T16809	pobj	3C,in
R496	T3516	T3514	prep	of,form
R4960	T16812	T16811	punct	", ",3C
R4961	T16813	T16811	conj	D,3C
R4962	T16814	T16813	cc	and,D
R4963	T16815	T16813	conj	3E,D
R4964	T16816	T16807	advmod	respectively,shown
R4965	T16817	T16808	punct	", ",is
R4966	T16818	T16808	nsubj	expression,is
R4967	T16819	T16818	prep	of,expression
R4968	T19826	T19827	det	A,hurdle
R4969	T19827	T19829	nsubj	hurdle,been
R497	T3370	T3362	punct	),see
R4970	T19828	T19827	amod	major,hurdle
R4971	T19830	T19827	prep	in,hurdle
R4972	T19831	T19830	pcomp	developing,in
R4973	T19832	T19833	amod	new,treatments
R4974	T19833	T19831	dobj	treatments,developing
R4975	T19834	T19833	prep	for,treatments
R4976	T19835	T19834	pobj	MSUD,for
R4977	T19836	T19829	aux	has,been
R4978	T19837	T19838	det	the,lack
R4979	T19838	T19829	attr	lack,been
R498	T3517	T3516	pobj	MSUD,of
R4980	T19839	T19838	prep	of,lack
R4981	T19840	T19841	det	a,model
R4982	T19841	T19839	pobj	model,of
R4983	T19842	T19841	amod	practical,model
R4984	T19843	T19841	punct	", ",model
R4985	T19844	T19841	amod	accurate,model
R4986	T19845	T19841	compound	animal,model
R4987	T19846	T19841	prep	of,model
R4988	T19847	T19848	det	the,disease
R4989	T19848	T19846	pobj	disease,of
R499	T3518	T3519	compound	BCKDH,activity
R4990	T19849	T19829	punct	.,been
R4991	T19851	T19852	det	This,hurdle
R4992	T19852	T19853	nsubjpass	hurdle,overcome
R4993	T19854	T19853	aux	has,overcome
R4994	T19855	T19853	advmod	now,overcome
R4995	T19856	T19853	auxpass	been,overcome
R4996	T19857	T19853	punct	.,overcome
R4997	T19859	T19860	prep	In,describe
R4998	T19861	T19862	det	this,report
R4999	T19862	T19859	pobj	report,In
R5	T840	T838	pobj	models,of
R50	T894	T895	nsubj	treatments,are
R500	T3519	T3511	dobj	activity,show
R5000	T19863	T19860	punct	", ",describe
R5001	T19864	T19860	nsubj	we,describe
R5002	T19865	T19866	det	the,development
R5003	T19866	T19860	dobj	development,describe
R5004	T19867	T19866	cc	and,development
R5005	T19868	T19866	conj	characterization,development
R5006	T19869	T19866	prep	of,development
R5007	T19870	T19871	nummod	two,models
R5008	T19871	T19869	pobj	models,of
R5009	T19872	T19873	advmod	genetically,engineered
R501	T3520	T3511	prep	in,show
R5010	T19873	T19871	amod	engineered,models
R5011	T19874	T19871	compound	mouse,models
R5012	T19875	T19876	dep	that,are
R5013	T19876	T19871	relcl	are,models
R5014	T19877	T19876	advmod	phenotypically,are
R5015	T19878	T19879	advmod	very,similar
R5016	T19879	T19876	acomp	similar,are
R5017	T19880	T19879	prep	to,similar
R5018	T19881	T19882	compound	MSUD,patients
R5019	T19882	T19880	pobj	patients,to
R502	T3371	T3308	punct	.,is
R5020	T19883	T19882	prep	with,patients
R5021	T19884	T19885	det	the,forms
R5022	T19885	T19883	pobj	forms,with
R5023	T19886	T19885	amod	classic,forms
R5024	T19887	T19886	cc	and,classic
R5025	T19888	T19886	conj	intermediate,classic
R5026	T19889	T19885	prep	of,forms
R5027	T19890	T19891	det	the,disease
R5028	T19891	T19889	pobj	disease,of
R5029	T19892	T19860	punct	.,describe
R503	T3521	T3522	det	the,range
R5030	T19894	T19895	poss	Our,model
R5031	T19895	T19896	nsubjpass	model,created
R5032	T19897	T19895	prep	of,model
R5033	T19898	T19899	amod	classic,MSUD
R5034	T19899	T19897	pobj	MSUD,of
R5035	T19900	T19896	auxpass	was,created
R5036	T19901	T19896	prep	by,created
R5037	T19902	T19903	compound	gene,knockout
R5038	T19903	T19901	pobj	knockout,by
R5039	T19904	T19903	prep	of,knockout
R504	T3522	T3520	pobj	range,in
R5040	T19905	T19906	det	the,subunit
R5041	T19906	T19904	pobj	subunit,of
R5042	T19907	T19906	compound	E2,subunit
R5043	T19908	T19906	prep	of,subunit
R5044	T19909	T19908	pobj	BCKDH,of
R5045	T19910	T19896	punct	.,created
R5046	T19912	T19913	det	The,phenotype
R5047	T19913	T19914	nsubj	phenotype,is
R5048	T19915	T19913	prep	of,phenotype
R5049	T19916	T19917	det	these,animals
R505	T3523	T3522	prep	of,range
R5050	T19917	T19915	pobj	animals,of
R5051	T19918	T19917	compound	knockout,animals
R5052	T19919	T19920	advmod	strikingly,similar
R5053	T19920	T19914	acomp	similar,is
R5054	T19921	T19920	prep	to,similar
R5055	T19922	T19921	pobj	humans,to
R5056	T19923	T19922	prep	with,humans
R5057	T19924	T19925	amod	classic,MSUD
R5058	T19925	T19923	pobj	MSUD,with
R5059	T19926	T19914	punct	.,is
R506	T3373	T3374	nsubj	Patients,show
R5060	T19928	T19929	compound	Knockout,mice
R5061	T19929	T19930	nsubjpass	mice,born
R5062	T19931	T19930	auxpass	were,born
R5063	T19932	T19930	prep	at,born
R5064	T19933	T19934	det	the,frequency
R5065	T19934	T19932	pobj	frequency,at
R5066	T19935	T19934	amod	expected,frequency
R5067	T19936	T19930	cc	and,born
R5068	T19937	T19930	conj	appeared,born
R5069	T19938	T19937	oprd	normal,appeared
R507	T3524	T3525	quantmod	5,20
R5070	T19939	T19937	prep	at,appeared
R5071	T19940	T19939	pobj	birth,at
R5072	T19941	T19930	punct	.,born
R5073	T19943	T19944	prep	Within,were
R5074	T19945	T19946	det	a,day
R5075	T19946	T19943	pobj	day,Within
R5076	T19947	T19946	prep	of,day
R5077	T19948	T19947	pobj	birth,of
R5078	T19949	T19943	cc	and,Within
R5079	T19950	T19943	conj	following,Within
R508	T3525	T3527	nummod	20,%
R5080	T19951	T19950	pobj	suckling,following
R5081	T19952	T19944	punct	", ",were
R5082	T19953	T19954	compound	blood,levels
R5083	T19954	T19944	nsubj	levels,were
R5084	T19955	T19954	prep	of,levels
R5085	T19956	T19955	pobj	BCAA,of
R5086	T19957	T19958	advmod	markedly,elevated
R5087	T19958	T19944	acomp	elevated,were
R5088	T19959	T19944	punct	.,were
R5089	T19961	T19962	advmod	Concomitantly,decreased
R509	T3526	T3525	punct	–,20
R5090	T19963	T19962	punct	", ",decreased
R5091	T19964	T19962	nsubjpass	levels,decreased
R5092	T19965	T19964	prep	of,levels
R5093	T19966	T19967	det	the,acids
R5094	T19967	T19965	pobj	acids,of
R5095	T19968	T19967	compound	amino,acids
R5096	T19969	T19967	appos	alanine,acids
R5097	T19970	T19969	punct	", ",alanine
R5098	T19971	T19969	conj	glutamate,alanine
R5099	T19972	T19971	punct	", ",glutamate
R51	T896	T895	neg	not,are
R510	T3527	T3523	pobj	%,of
R5100	T19973	T19971	cc	and,glutamate
R5101	T19974	T19971	conj	glutamine,glutamate
R5102	T19975	T19967	punct	", ",acids
R5103	T19976	T19977	poss	whose,synthesis
R5104	T19977	T19978	dep	synthesis,linked
R5105	T19978	T19967	relcl	linked,acids
R5106	T19979	T19978	auxpass	is,linked
R5107	T19980	T19978	prep	with,linked
R5108	T19981	T19982	amod	normal,metabolism
R5109	T19982	T19980	pobj	metabolism,with
R511	T3375	T3373	prep	with,Patients
R5110	T19983	T19982	prep	of,metabolism
R5111	T19984	T19983	pobj	BCAA,of
R5112	T19985	T19962	punct	", ",decreased
R5113	T19986	T19962	auxpass	were,decreased
R5114	T19987	T19988	punct	(,Table
R5115	T19988	T19962	parataxis	Table,decreased
R5116	T19989	T19988	nummod	1,Table
R5117	T19990	T19988	punct	),Table
R5118	T19991	T19962	punct	.,decreased
R5119	T19993	T19994	det	The,activity
R512	T3528	T3511	cc	and,show
R5120	T19994	T19995	nsubj	activity,was
R5121	T19996	T19994	prep	of,activity
R5122	T19997	T19996	pobj	BCKDH,of
R5123	T19998	T19994	prep	in,activity
R5124	T19999	T19998	pobj	livers,in
R5125	T20000	T19999	prep	of,livers
R5126	T20001	T20002	amod	homozygous,knockout
R5127	T20002	T20003	compound	knockout,pups
R5128	T20003	T20000	pobj	pups,of
R5129	T20004	T20003	compound	mouse,pups
R513	T3376	T3375	pobj	MSUD,with
R5130	T20005	T19995	acomp	undetectable,was
R5131	T20006	T19995	punct	", ",was
R5132	T20007	T19995	advcl	accounting,was
R5133	T20008	T20007	prep	for,accounting
R5134	T20009	T20010	det	the,accumulation
R5135	T20010	T20008	pobj	accumulation,for
R5136	T20011	T20010	prep	of,accumulation
R5137	T20012	T20013	amod	unmetabolized,BCAA
R5138	T20013	T20011	pobj	BCAA,of
R5139	T20014	T19995	punct	.,was
R514	T3377	T3374	punct	", ",show
R5140	T20016	T20017	amod	Immunoreactive,protein
R5141	T20017	T20019	nsubj	protein,was
R5142	T20018	T20017	compound	E2,protein
R5143	T20020	T20019	acomp	absent,was
R5144	T20021	T20019	prep	in,was
R5145	T20022	T20021	pobj	liver,in
R5146	T20023	T20022	cc	and,liver
R5147	T20024	T20022	conj	fibroblasts,liver
R5148	T20025	T20022	prep	of,liver
R5149	T20026	T20027	amod	homozygous,pups
R515	T3378	T3374	prep	depending,show
R5150	T20027	T20025	pobj	pups,of
R5151	T20028	T20019	punct	.,was
R5152	T20030	T20031	det	The,behavior
R5153	T20031	T20033	nsubj	behavior,resembles
R5154	T20032	T20031	amod	phenotypic,behavior
R5155	T20034	T20031	prep	of,behavior
R5156	T20035	T20036	amod	homozygous,pups
R5157	T20036	T20034	pobj	pups,of
R5158	T20037	T20036	compound	mouse,pups
R5159	T20038	T20033	dobj	symptoms,resembles
R516	T3529	T3530	prep	during,are
R5160	T20039	T20038	acl	seen,symptoms
R5161	T20040	T20039	prep	in,seen
R5162	T20041	T20042	amod	newborn,patients
R5163	T20042	T20040	pobj	patients,in
R5164	T20043	T20042	amod	classic,patients
R5165	T20044	T20042	compound	MSUD,patients
R5166	T20045	T20033	punct	.,resembles
R5167	T20047	T20048	nsubj	These,include
R5168	T20049	T20048	dobj	signs,include
R5169	T20050	T20049	prep	of,signs
R517	T3530	T3511	conj	are,show
R5170	T20051	T20052	amod	neurologic,dysfunction
R5171	T20052	T20050	pobj	dysfunction,of
R5172	T20053	T20054	amod	such,as
R5173	T20054	T20049	prep	as,signs
R5174	T20055	T20054	pobj	seizures,as
R5175	T20056	T20055	punct	", ",seizures
R5176	T20057	T20055	conj	stupor,seizures
R5177	T20058	T20057	punct	", ",stupor
R5178	T20059	T20057	conj	lethargy,stupor
R5179	T20060	T20059	punct	", ",lethargy
R518	T3379	T3378	prep	on,depending
R5180	T20061	T20059	conj	loss,lethargy
R5181	T20062	T20061	prep	of,loss
R5182	T20063	T20064	compound	motor,activity
R5183	T20064	T20062	pobj	activity,of
R5184	T20065	T20061	punct	", ",loss
R5185	T20066	T20061	cc	and,loss
R5186	T20067	T20068	amod	respiratory,difficulties
R5187	T20068	T20061	conj	difficulties,loss
R5188	T20069	T20070	punct	[,1
R5189	T20070	T20048	parataxis	1,include
R519	T3531	T3532	det	the,phase
R5190	T20071	T20070	punct	],1
R5191	T20072	T20048	punct	.,include
R5192	T20074	T20075	det	These,symptoms
R5193	T20075	T20077	nsubj	symptoms,result
R5194	T20076	T20075	amod	neurologic,symptoms
R5195	T20078	T20077	aux	may,result
R5196	T20079	T20077	prep	from,result
R5197	T20080	T20081	amod	reduced,levels
R5198	T20081	T20079	pobj	levels,from
R5199	T20082	T20081	prep	of,levels
R52	T897	T895	acomp	satisfactory,are
R520	T3532	T3529	pobj	phase,during
R5200	T20083	T20082	pobj	glutamate,of
R5201	T20084	T20083	punct	", ",glutamate
R5202	T20085	T20083	conj	glutamine,glutamate
R5203	T20086	T20085	punct	", ",glutamine
R5204	T20087	T20085	conj	alanine,glutamine
R5205	T20088	T20087	punct	", ",alanine
R5206	T20089	T20087	cc	and,alanine
R5207	T20090	T20091	amod	other,acids
R5208	T20091	T20087	conj	acids,alanine
R5209	T20092	T20091	amod	similar,acids
R521	T3380	T3381	det	the,mutation
R5210	T20093	T20091	amod	neuroactive,acids
R5211	T20094	T20091	compound	amino,acids
R5212	T20095	T20083	punct	", ",glutamate
R5213	T20096	T20097	dep	which,considered
R5214	T20097	T20083	relcl	considered,glutamate
R5215	T20098	T20097	auxpass	are,considered
R5216	T20099	T20100	det	the,culprit
R5217	T20100	T20097	oprd	culprit,considered
R5218	T20101	T20100	prep	for,culprit
R5219	T20102	T20103	compound	MSUD,encephalopathies
R522	T3533	T3532	amod	asymptomatic,phase
R5220	T20103	T20101	pobj	encephalopathies,for
R5221	T20104	T20100	prep	in,culprit
R5222	T20105	T20104	pobj	patients,in
R5223	T20106	T20107	punct	[,37
R5224	T20107	T20077	parataxis	37,result
R5225	T20108	T20107	nummod	1,37
R5226	T20109	T20107	punct	",",37
R5227	T20110	T20107	punct	],37
R5228	T20111	T20077	punct	.,result
R5229	T20113	T20114	advmod	Finally,died
R523	T3534	T3535	det	the,levels
R5230	T20115	T20114	punct	", ",died
R5231	T20116	T20117	advmod	nearly,all
R5232	T20117	T20114	nsubj	all,died
R5233	T20118	T20117	prep	of,all
R5234	T20119	T20120	det	the,pups
R5235	T20120	T20118	pobj	pups,of
R5236	T20121	T20120	amod	homozygous,pups
R5237	T20122	T20114	prep	within,died
R5238	T20123	T20124	nummod	72,hours
R5239	T20124	T20122	pobj	hours,within
R524	T3535	T3530	nsubj	levels,are
R5240	T20125	T20124	prep	of,hours
R5241	T20126	T20125	pobj	birth,of
R5242	T20127	T20114	punct	.,died
R5243	T20129	T20130	det	This,lethality
R5244	T20130	T20132	nsubj	lethality,is
R5245	T20131	T20130	amod	neonatal,lethality
R5246	T20133	T20134	advmod	likely,due
R5247	T20134	T20132	prep	due,is
R5248	T20135	T20134	pcomp	to,due
R5249	T20136	T20137	det	the,accumulation
R525	T3381	T3379	pobj	mutation,on
R5250	T20137	T20134	pobj	accumulation,due
R5251	T20138	T20137	prep	of,accumulation
R5252	T20139	T20138	pobj	BCAA,of
R5253	T20140	T20137	prep	to,accumulation
R5254	T20141	T20142	amod	neurotoxic,levels
R5255	T20142	T20140	pobj	levels,to
R5256	T20143	T20142	punct	", ",levels
R5257	T20144	T20142	conj	ketoacidosis,levels
R5258	T20145	T20144	punct	", ",ketoacidosis
R5259	T20146	T20147	compound	brain,edema
R526	T3536	T3535	compound	blood,levels
R5260	T20147	T20144	conj	edema,ketoacidosis
R5261	T20148	T20147	punct	", ",edema
R5262	T20149	T20147	conj	dehydration,edema
R5263	T20150	T20149	punct	", ",dehydration
R5264	T20151	T20149	cc	and,dehydration
R5265	T20152	T20149	conj	malnutrition,dehydration
R5266	T20153	T20154	mark	as,observed
R5267	T20154	T20132	advcl	observed,is
R5268	T20155	T20154	prep	in,observed
R5269	T20156	T20157	det	the,calf
R527	T3537	T3535	compound	BCAA,levels
R5270	T20157	T20155	pobj	calf,in
R5271	T20158	T20157	compound	MSUD,calf
R5272	T20159	T20160	punct	[,17
R5273	T20160	T20157	parataxis	17,calf
R5274	T20161	T20160	punct	],17
R5275	T20162	T20155	cc	and,in
R5276	T20163	T20155	conj	in,in
R5277	T20164	T20165	amod	classic,patients
R5278	T20165	T20163	pobj	patients,in
R5279	T20166	T20165	compound	MSUD,patients
R528	T3538	T3530	acomp	normal,are
R5280	T20167	T20168	punct	[,1
R5281	T20168	T20165	parataxis	1,patients
R5282	T20169	T20168	punct	],1
R5283	T20170	T20132	punct	.,is
R5284	T20172	T20173	amod	Heterozygous,knockouts
R5285	T20173	T20174	nsubj	knockouts,were
R5286	T20175	T20174	acomp	normal,were
R5287	T20176	T20174	cc	and,were
R5288	T20177	T20174	conj	had,were
R5289	T20178	T20179	amod	normal,levels
R529	T3382	T3374	punct	", ",show
R5290	T20179	T20177	dobj	levels,had
R5291	T20180	T20179	prep	of,levels
R5292	T20181	T20180	pobj	BCAA,of
R5293	T20182	T20177	prep	despite,had
R5294	T20183	T20182	pcomp	having,despite
R5295	T20184	T20185	advmod	approximately,half
R5296	T20185	T20183	dobj	half,having
R5297	T20186	T20185	prep	of,half
R5298	T20187	T20188	nmod	BCKDH,activity
R5299	T20188	T20186	pobj	activity,of
R53	T898	T895	cc	and,are
R530	T3539	T3540	punct	[,1
R5300	T20189	T20188	amod	enzymatic,activity
R5301	T20190	T20174	punct	.,were
R5302	T20192	T20193	prep	From,represents
R5303	T20194	T20195	det	the,studies
R5304	T20195	T20192	pobj	studies,From
R5305	T20196	T20195	compound	characterization,studies
R5306	T20197	T20195	acl	completed,studies
R5307	T20198	T20199	advmod	thus,far
R5308	T20199	T20197	advmod	far,completed
R5309	T20200	T20193	punct	", ",represents
R531	T3540	T3530	parataxis	1,are
R5310	T20201	T20202	det	the,mouse
R5311	T20202	T20193	nsubj	mouse,represents
R5312	T20203	T20204	amod	null,mutation
R5313	T20204	T20202	compound	mutation,mouse
R5314	T20205	T20193	advmod	accurately,represents
R5315	T20206	T20207	det	a,model
R5316	T20207	T20193	dobj	model,represents
R5317	T20208	T20207	prep	of,model
R5318	T20209	T20210	amod	classic,MSUD
R5319	T20210	T20208	pobj	MSUD,of
R532	T3383	T3384	amod	variable,degrees
R5320	T20211	T20193	cc	and,represents
R5321	T20212	T20193	conj	appears,represents
R5322	T20213	T20214	aux	to,be
R5323	T20214	T20212	xcomp	be,appears
R5324	T20215	T20216	det	a,model
R5325	T20216	T20214	attr	model,be
R5326	T20217	T20216	amod	faithful,model
R5327	T20218	T20216	prep	of,model
R5328	T20219	T20220	det	the,disease
R5329	T20220	T20218	pobj	disease,of
R533	T3541	T3540	punct	],1
R5330	T20221	T20220	amod	human,disease
R5331	T20222	T20214	prep	with,be
R5332	T20223	T20222	pobj	respect,with
R5333	T20224	T20223	prep	to,respect
R5334	T20225	T20226	amod	several,phenotypes
R5335	T20226	T20224	pobj	phenotypes,to
R5336	T20227	T20226	amod	biochemical,phenotypes
R5337	T20228	T20229	punct	[,1
R5338	T20229	T20212	parataxis	1,appears
R5339	T20230	T20229	punct	],1
R534	T3542	T3530	punct	.,are
R5340	T20231	T20193	punct	.,represents
R5341	T20233	T20234	aux	To,create
R5342	T20234	T20235	advcl	create,used
R5343	T20236	T20237	det	a,model
R5344	T20237	T20234	dobj	model,create
R5345	T20238	T20237	prep	of,model
R5346	T20239	T20240	amod	intermediate,MSUD
R5347	T20240	T20238	pobj	MSUD,of
R5348	T20241	T20235	punct	", ",used
R5349	T20242	T20235	nsubj	we,used
R535	T3384	T3374	dobj	degrees,show
R5350	T20243	T20244	det	a,strategy
R5351	T20244	T20235	dobj	strategy,used
R5352	T20245	T20244	amod	transgenic,strategy
R5353	T20246	T20247	aux	to,express
R5354	T20247	T20235	advcl	express,used
R5355	T20248	T20249	amod	human,E2
R5356	T20249	T20247	dobj	E2,express
R5357	T20250	T20247	prep	in,express
R5358	T20251	T20252	det	the,liver
R5359	T20252	T20250	pobj	liver,in
R536	T3544	T3545	det	The,incidence
R5360	T20253	T20252	prep	of,liver
R5361	T20254	T20255	compound	E2,mice
R5362	T20255	T20253	pobj	mice,of
R5363	T20256	T20255	compound	knockout,mice
R5364	T20257	T20235	punct	.,used
R5365	T20259	T20260	mark	As,mentioned
R5366	T20260	T20261	advcl	mentioned,die
R5367	T20262	T20260	advmod	above,mentioned
R5368	T20263	T20261	punct	", ",die
R5369	T20264	T20265	compound	E2,knockouts
R537	T3545	T3547	nsubj	incidence,is
R5370	T20265	T20261	nsubj	knockouts,die
R5371	T20266	T20265	prep	without,knockouts
R5372	T20267	T20268	npadvmod	transgene,derived
R5373	T20268	T20269	amod	derived,E2
R5374	T20269	T20266	pobj	E2,without
R5375	T20270	T20261	prep	during,die
R5376	T20271	T20272	det	the,period
R5377	T20272	T20270	pobj	period,during
R5378	T20273	T20274	amod	early,neonatal
R5379	T20274	T20272	amod	neonatal,period
R538	T3546	T3545	amod	overall,incidence
R5380	T20275	T20261	punct	.,die
R5381	T20277	T20278	nsubj	We,show
R5382	T20279	T20280	mark	that,is
R5383	T20280	T20278	ccomp	is,show
R5384	T20281	T20280	nsubj	expression,is
R5385	T20282	T20281	prep	of,expression
R5386	T20283	T20284	det	a,transgene
R5387	T20284	T20282	pobj	transgene,of
R5388	T20285	T20284	amod	human,transgene
R5389	T20286	T20284	compound	E2,transgene
R539	T3385	T3384	prep	of,degrees
R5390	T20287	T20281	prep	in,expression
R5391	T20288	T20289	det	the,liver
R5392	T20289	T20287	pobj	liver,in
R5393	T20290	T20289	prep	of,liver
R5394	T20291	T20292	det	these,mice
R5395	T20292	T20290	pobj	mice,of
R5396	T20293	T20292	compound	knockout,mice
R5397	T20294	T20280	acomp	able,is
R5398	T20295	T20296	aux	to,rescue
R5399	T20296	T20294	xcomp	rescue,able
R54	T899	T895	conj	require,are
R540	T3548	T3545	prep	of,incidence
R5400	T20297	T20298	det	the,lethality
R5401	T20298	T20296	dobj	lethality,rescue
R5402	T20299	T20298	amod	neonatal,lethality
R5403	T20300	T20278	punct	.,show
R5404	T20302	T20303	nsubj	Many,survived
R5405	T20304	T20302	prep	of,Many
R5406	T20305	T20306	det	the,mice
R5407	T20306	T20304	pobj	mice,of
R5408	T20307	T20306	compound	rescue,mice
R5409	T20308	T20303	prep	to,survived
R541	T3386	T3387	compound	enzyme,deficiency
R5410	T20309	T20308	pobj	adulthood,to
R5411	T20310	T20303	punct	.,survived
R5412	T20312	T20313	nsubj	It,is
R5413	T20314	T20313	acomp	interesting,is
R5414	T20315	T20316	aux	to,note
R5415	T20316	T20313	xcomp	note,is
R5416	T20317	T20318	mark	that,was
R5417	T20318	T20316	ccomp	was,note
R5418	T20319	T20318	prep	in,was
R5419	T20320	T20321	det	these,lines
R542	T3549	T3548	pobj	MSUD,of
R5420	T20321	T20319	pobj	lines,in
R5421	T20322	T20321	compound	mouse,lines
R5422	T20323	T20324	advmod	only,6
R5423	T20324	T20328	nummod	6,%
R5424	T20325	T20324	punct	~,6
R5425	T20326	T20324	quantmod	5,6
R5426	T20327	T20324	punct	–,6
R5427	T20328	T20318	nsubj	%,was
R5428	T20329	T20328	prep	of,%
R5429	T20330	T20331	amod	normal,activity
R543	T3550	T3545	prep	in,incidence
R5430	T20331	T20329	pobj	activity,of
R5431	T20332	T20331	compound	BCKDH,activity
R5432	T20333	T20331	prep	in,activity
R5433	T20334	T20335	det	the,liver
R5434	T20335	T20333	pobj	liver,in
R5435	T20336	T20318	acomp	sufficient,was
R5436	T20337	T20338	aux	to,allow
R5437	T20338	T20336	xcomp	allow,sufficient
R5438	T20339	T20338	dobj	survival,allow
R5439	T20340	T20313	punct	.,is
R544	T3551	T3552	det	the,population
R5440	T20342	T20343	nsubj	We,demonstrated
R5441	T20344	T20343	advmod	also,demonstrated
R5442	T20345	T20346	mark	that,were
R5443	T20346	T20343	ccomp	were,demonstrated
R5444	T20347	T20348	compound	BCAA,levels
R5445	T20348	T20346	nsubj	levels,were
R5446	T20349	T20348	prep	in,levels
R5447	T20350	T20349	pobj	blood,in
R5448	T20351	T20350	prep	of,blood
R5449	T20352	T20353	det	these,mice
R545	T3552	T3550	pobj	population,in
R5450	T20353	T20351	pobj	mice,of
R5451	T20354	T20353	compound	rescue,mice
R5452	T20355	T20346	acomp	intermediate,were
R5453	T20356	T20346	prep	between,were
R5454	T20357	T20356	pobj	controls,between
R5455	T20358	T20357	cc	and,controls
R5456	T20359	T20357	conj	knockouts,controls
R5457	T20360	T20343	punct	.,demonstrated
R5458	T20362	T20363	det	The,hallmarks
R5459	T20363	T20364	nsubj	hallmarks,are
R546	T3553	T3552	amod	general,population
R5460	T20365	T20363	prep	of,hallmarks
R5461	T20366	T20367	amod	intermediate,patients
R5462	T20367	T20365	pobj	patients,of
R5463	T20368	T20367	nmod	MSUD,patients
R5464	T20369	T20367	amod	human,patients
R5465	T20370	T20371	advmod	persistently,increased
R5466	T20371	T20372	amod	increased,levels
R5467	T20372	T20364	attr	levels,are
R5468	T20373	T20372	prep	of,levels
R5469	T20374	T20375	nmod	BCAA,activity
R547	T3387	T3385	pobj	deficiency,of
R5470	T20375	T20373	pobj	activity,of
R5471	T20376	T20374	cc	and,BCAA
R5472	T20377	T20374	conj	BCKDH,BCAA
R5473	T20378	T20372	prep	in,levels
R5474	T20379	T20380	det	the,range
R5475	T20380	T20378	pobj	range,in
R5476	T20381	T20380	prep	of,range
R5477	T20382	T20383	quantmod	3,30
R5478	T20383	T20385	nummod	30,%
R5479	T20384	T20383	punct	–,30
R548	T3554	T3555	nummod	1,"185,000"
R5480	T20385	T20381	pobj	%,of
R5481	T20386	T20385	prep	of,%
R5482	T20387	T20386	pobj	normal,of
R5483	T20388	T20389	punct	[,1
R5484	T20389	T20364	parataxis	1,are
R5485	T20390	T20389	punct	],1
R5486	T20391	T20364	punct	.,are
R5487	T20393	T20394	prep	Because,represent
R5488	T20395	T20393	pcomp	of,Because
R5489	T20396	T20397	det	the,similarities
R549	T3555	T3547	attr	"185,000",is
R5490	T20397	T20393	pobj	similarities,Because
R5491	T20398	T20397	amod	phenotypic,similarities
R5492	T20399	T20397	prep	of,similarities
R5493	T20400	T20401	det	the,mice
R5494	T20401	T20399	pobj	mice,of
R5495	T20402	T20401	compound	rescue,mice
R5496	T20403	T20397	prep	to,similarities
R5497	T20404	T20405	det	the,patients
R5498	T20405	T20403	pobj	patients,to
R5499	T20406	T20405	amod	human,patients
R55	T900	T901	amod	new,approaches
R550	T3556	T3555	punct	:,"185,000"
R5500	T20407	T20394	punct	", ",represent
R5501	T20408	T20409	det	these,mice
R5502	T20409	T20394	nsubj	mice,represent
R5503	T20410	T20411	advmod	genetically,engineered
R5504	T20411	T20409	amod	engineered,mice
R5505	T20412	T20413	det	a,model
R5506	T20413	T20394	dobj	model,represent
R5507	T20414	T20413	amod	useful,model
R5508	T20415	T20416	amod	small,animal
R5509	T20416	T20413	compound	animal,model
R551	T3388	T3387	acl	leading,deficiency
R5510	T20417	T20413	prep	of,model
R5511	T20418	T20419	det	the,form
R5512	T20419	T20417	pobj	form,of
R5513	T20420	T20419	amod	intermediate,form
R5514	T20421	T20419	prep	of,form
R5515	T20422	T20421	pobj	MSUD,of
R5516	T20423	T20394	punct	.,represent
R5517	T20425	T20426	det	The,approach
R5518	T20426	T20428	nsubjpass	approach,based
R5519	T20427	T20426	amod	transgenic,approach
R552	T3557	T3558	punct	[,1
R5520	T20429	T20430	dep	that,used
R5521	T20430	T20426	relcl	used,approach
R5522	T20431	T20430	auxpass	was,used
R5523	T20432	T20433	aux	to,create
R5524	T20433	T20430	advcl	create,used
R5525	T20434	T20435	det	the,model
R5526	T20435	T20433	dobj	model,create
R5527	T20436	T20435	amod	intermediate,model
R5528	T20437	T20435	compound	MSUD,model
R5529	T20438	T20428	auxpass	was,based
R553	T3558	T3547	parataxis	1,is
R5530	T20439	T20428	prep	on,based
R5531	T20440	T20441	det	the,system
R5532	T20441	T20439	pobj	system,on
R5533	T20442	T20443	npadvmod	tetracycline,regulated
R5534	T20443	T20441	amod	regulated,system
R5535	T20444	T20445	compound	gene,switch
R5536	T20445	T20441	compound	switch,system
R5537	T20446	T20447	dep	that,used
R5538	T20447	T20441	relcl	used,system
R5539	T20448	T20447	aux	has,used
R554	T3559	T3558	punct	],1
R5540	T20449	T20447	auxpass	been,used
R5541	T20450	T20447	prep	with,used
R5542	T20451	T20452	amod	great,success
R5543	T20452	T20450	pobj	success,with
R5544	T20453	T20447	prep	in,used
R5545	T20454	T20455	amod	other,studies
R5546	T20455	T20453	pobj	studies,in
R5547	T20456	T20447	punct	", ",used
R5548	T20457	T20447	prep	for,used
R5549	T20458	T20457	pobj	example,for
R555	T3389	T3388	prep	to,leading
R5550	T20459	T20460	punct	[,34
R5551	T20460	T20428	parataxis	34,based
R5552	T20461	T20460	punct	],34
R5553	T20462	T20428	punct	.,based
R5554	T20464	T20465	det	The,strategy
R5555	T20465	T20466	nsubj	strategy,was
R5556	T20467	T20465	prep	behind,strategy
R5557	T20468	T20469	poss	our,approach
R5558	T20469	T20467	pobj	approach,behind
R5559	T20470	T20471	aux	to,create
R556	T3560	T3547	punct	", ",is
R5560	T20471	T20466	xcomp	create,was
R5561	T20472	T20471	dobj	mice,create
R5562	T20473	T20472	prep	with,mice
R5563	T20474	T20475	det	an,phenotype
R5564	T20475	T20473	pobj	phenotype,with
R5565	T20476	T20475	amod	intermediate,phenotype
R5566	T20477	T20475	compound	MSUD,phenotype
R5567	T20478	T20479	dep	that,converted
R5568	T20479	T20475	relcl	converted,phenotype
R5569	T20480	T20479	aux	could,converted
R557	T3561	T3547	cc	and,is
R5570	T20481	T20479	auxpass	be,converted
R5571	T20482	T20479	prep	to,converted
R5572	T20483	T20484	det	the,phenotype
R5573	T20484	T20482	pobj	phenotype,to
R5574	T20485	T20484	amod	classic,phenotype
R5575	T20486	T20479	prep	at,converted
R5576	T20487	T20488	det	the,investigator
R5577	T20488	T20489	poss	investigator,discretion
R5578	T20489	T20486	pobj	discretion,at
R5579	T20490	T20488	case	's,investigator
R558	T3562	T3563	prep	in,estimated
R5580	T20491	T20479	prep	by,converted
R5581	T20492	T20491	pcomp	turning,by
R5582	T20493	T20492	prt	off,turning
R5583	T20494	T20495	det	the,transgene
R5584	T20495	T20492	dobj	transgene,turning
R5585	T20496	T20495	amod	rescuing,transgene
R5586	T20497	T20466	punct	.,was
R5587	T20499	T20500	advmod	However,were
R5588	T20501	T20500	punct	", ",were
R5589	T20502	T20500	nsubj	we,were
R559	T3390	T3391	amod	several,phenotypes
R5590	T20503	T20500	acomp	unable,were
R5591	T20504	T20505	aux	to,turn
R5592	T20505	T20503	xcomp	turn,unable
R5593	T20506	T20505	advmod	consistently,turn
R5594	T20507	T20505	prt	off,turn
R5595	T20508	T20509	det	the,cassette
R5596	T20509	T20505	dobj	cassette,turn
R5597	T20510	T20509	amod	human,cassette
R5598	T20511	T20509	compound	E2,cassette
R5599	T20512	T20509	compound	expression,cassette
R56	T901	T899	dobj	approaches,require
R560	T3563	T3547	conj	estimated,is
R5600	T20513	T20505	prep	by,turn
R5601	T20514	T20515	det	the,analogue
R5602	T20515	T20513	pobj	analogue,by
R5603	T20516	T20515	amod	potent,analogue
R5604	T20517	T20515	compound	tetracycline,analogue
R5605	T20518	T20515	punct	", ",analogue
R5606	T20519	T20515	appos	doxycycline,analogue
R5607	T20520	T20505	prep	in,turn
R5608	T20521	T20520	pobj	either,in
R5609	T20522	T20521	prep	of,either
R561	T3391	T3389	pobj	phenotypes,to
R5610	T20523	T20524	det	the,lines
R5611	T20524	T20522	pobj	lines,of
R5612	T20525	T20524	nummod	two,lines
R5613	T20526	T20524	amod	transgenic,lines
R5614	T20527	T20524	acl	tested,lines
R5615	T20528	T20529	punct	(,shown
R5616	T20529	T20500	parataxis	shown,were
R5617	T20530	T20529	nsubj	data,shown
R5618	T20531	T20529	neg	not,shown
R5619	T20532	T20529	punct	),shown
R562	T3564	T3565	amod	certain,groups
R5620	T20533	T20500	punct	.,were
R5621	T20535	T20536	det	The,reason
R5622	T20536	T20537	nsubj	reason,is
R5623	T20538	T20539	det	these,lines
R5624	T20539	T20541	nsubj	lines,were
R5625	T20540	T20539	nummod	two,lines
R5626	T20541	T20536	advcl	were,reason
R5627	T20542	T20541	acomp	unresponsive,were
R5628	T20543	T20542	prep	to,unresponsive
R5629	T20544	T20543	pobj	doxycycline,to
R563	T3565	T3562	pobj	groups,in
R5630	T20545	T20537	acomp	unknown,is
R5631	T20546	T20537	punct	.,is
R5632	T20548	T20549	nsubj	It,is
R5633	T20550	T20549	acomp	conceivable,is
R5634	T20551	T20552	mark	that,was
R5635	T20552	T20549	ccomp	was,is
R5636	T20553	T20554	det	the,site
R5637	T20554	T20552	nsubj	site,was
R5638	T20555	T20554	compound	integration,site
R5639	T20556	T20554	prep	of,site
R564	T3566	T3565	compound	population,groups
R5640	T20557	T20558	det	the,transgenes
R5641	T20558	T20556	pobj	transgenes,of
R5642	T20559	T20552	neg	not,was
R5643	T20560	T20552	acomp	permissive,was
R5644	T20561	T20560	prep	for,permissive
R5645	T20562	T20563	amod	regulated,expression
R5646	T20563	T20561	pobj	expression,for
R5647	T20564	T20549	punct	.,is
R5648	T20566	T20567	nsubjpass	Screening,needed
R5649	T20568	T20566	prep	of,Screening
R565	T3392	T3391	amod	different,phenotypes
R5650	T20569	T20570	amod	additional,lines
R5651	T20570	T20568	pobj	lines,of
R5652	T20571	T20567	aux	may,needed
R5653	T20572	T20567	auxpass	be,needed
R5654	T20573	T20574	aux	to,find
R5655	T20574	T20567	advcl	find,needed
R5656	T20575	T20574	dobj	lines,find
R5657	T20576	T20577	dep	that,allow
R5658	T20577	T20575	relcl	allow,lines
R5659	T20578	T20577	prep	for,allow
R566	T3567	T3565	punct	", ",groups
R5660	T20579	T20578	pobj	survival,for
R5661	T20580	T20577	cc	and,allow
R5662	T20581	T20582	aux	can,regulated
R5663	T20582	T20577	conj	regulated,allow
R5664	T20583	T20582	advmod	also,regulated
R5665	T20584	T20582	auxpass	be,regulated
R5666	T20585	T20567	punct	.,needed
R5667	T20587	T20588	det	The,models
R5668	T20588	T20589	nsubj	models,are
R5669	T20590	T20588	acl	described,models
R567	T3568	T3569	amod	such,as
R5670	T20591	T20590	prep	in,described
R5671	T20592	T20593	det	this,communication
R5672	T20593	T20591	pobj	communication,in
R5673	T20594	T20595	amod	important,advances
R5674	T20595	T20589	attr	advances,are
R5675	T20596	T20595	prep	over,advances
R5676	T20597	T20598	det	the,models
R5677	T20598	T20596	pobj	models,over
R5678	T20599	T20600	advmod	previously,described
R5679	T20600	T20598	amod	described,models
R568	T3569	T3565	prep	as,groups
R5680	T20601	T20589	punct	.,are
R5681	T20603	T20604	det	An,mouse
R5682	T20604	T20606	nsubjpass	mouse,created
R5683	T20605	T20604	compound	ENU,mouse
R5684	T20607	T20604	acl	resembling,mouse
R5685	T20608	T20609	amod	human,MSUD
R5686	T20609	T20607	dobj	MSUD,resembling
R5687	T20610	T20606	auxpass	was,created
R5688	T20611	T20606	advmod	previously,created
R5689	T20612	T20606	prep	by,created
R569	T3570	T3569	pobj	Mennonites,as
R5690	T20613	T20612	pcomp	disrupting,by
R5691	T20614	T20615	det	the,gene
R5692	T20615	T20613	dobj	gene,disrupting
R5693	T20616	T20615	amod	mitochondrial,gene
R5694	T20617	T20615	compound	BCAT,gene
R5695	T20618	T20619	punct	[,20
R5696	T20619	T20606	parataxis	20,created
R5697	T20620	T20619	punct	],20
R5698	T20621	T20606	punct	.,created
R5699	T20623	T20624	det	The,activity
R57	T902	T903	aux	to,combat
R570	T3393	T3391	amod	clinical,phenotypes
R5700	T20624	T20626	nsubj	activity,was
R5701	T20625	T20624	compound	BCKDH,activity
R5702	T20627	T20624	prep	in,activity
R5703	T20628	T20629	det	the,liver
R5704	T20629	T20627	pobj	liver,in
R5705	T20630	T20629	cc	and,liver
R5706	T20631	T20629	conj	muscle,liver
R5707	T20632	T20629	prep	of,liver
R5708	T20633	T20634	det	this,mouse
R5709	T20634	T20632	pobj	mouse,of
R571	T3571	T3570	prep	of,Mennonites
R5710	T20635	T20626	acomp	normal,was
R5711	T20636	T20637	advmod	even,were
R5712	T20637	T20626	advcl	were,was
R5713	T20638	T20637	mark	though,were
R5714	T20639	T20640	det	the,levels
R5715	T20640	T20637	nsubj	levels,were
R5716	T20641	T20640	compound	blood,levels
R5717	T20642	T20640	compound	BCAA,levels
R5718	T20643	T20644	advmod	markedly,elevated
R5719	T20644	T20637	acomp	elevated,were
R572	T3572	T3571	pobj	Pennsylvania,of
R5720	T20645	T20626	punct	.,was
R5721	T20647	T20648	mark	While,is
R5722	T20648	T20650	advcl	is,is
R5723	T20649	T20648	nsubj	this,is
R5724	T20651	T20652	det	an,model
R5725	T20652	T20648	attr	model,is
R5726	T20653	T20652	amod	interesting,model
R5727	T20654	T20650	punct	", ",is
R5728	T20655	T20650	nsubj	it,is
R5729	T20656	T20650	neg	not,is
R573	T3394	T3395	punct	[,1
R5730	T20657	T20658	det	a,model
R5731	T20658	T20650	attr	model,is
R5732	T20659	T20658	amod	true,model
R5733	T20660	T20658	prep	of,model
R5734	T20661	T20660	pobj	MSUD,of
R5735	T20662	T20663	mark	because,is
R5736	T20663	T20650	advcl	is,is
R5737	T20664	T20665	det	the,mutation
R5738	T20665	T20663	nsubj	mutation,is
R5739	T20666	T20663	neg	not,is
R574	T3573	T3563	punct	", ",estimated
R5740	T20667	T20663	prep	in,is
R5741	T20668	T20667	pobj	BCKDH,in
R5742	T20669	T20663	cc	and,is
R5743	T20670	T20671	det	the,levels
R5744	T20671	T20672	nsubj	levels,are
R5745	T20672	T20663	conj	are,is
R5746	T20673	T20671	prep	of,levels
R5747	T20674	T20673	pobj	BCKDH,of
R5748	T20675	T20672	acomp	normal,are
R5749	T20676	T20650	punct	.,is
R575	T3574	T3575	det	the,incidence
R5750	T20678	T20679	advmod	Furthermore,described
R5751	T20680	T20679	punct	", ",described
R5752	T20681	T20682	det	no,case
R5753	T20682	T20679	nsubjpass	case,described
R5754	T20683	T20682	prep	of,case
R5755	T20790	T20789	compound	MSUD,models
R5756	T20791	T20789	compound	mouse,models
R5757	T20792	T20793	aux	to,test
R5758	T20793	T20785	xcomp	test,is
R5759	T20794	T20795	amod	novel,treatments
R576	T3575	T3563	nsubjpass	incidence,estimated
R5760	T20795	T20793	dobj	treatments,test
R5761	T20796	T20797	amod	such,as
R5762	T20797	T20795	prep	as,treatments
R5763	T20798	T20799	npadvmod	gene,based
R5764	T20799	T20807	amod	based,therapies
R5765	T20800	T20801	punct	[,38
R5766	T20801	T20798	parataxis	38,gene
R5767	T20802	T20803	punct	-,41
R5768	T20803	T20801	prep	41,38
R5769	T20804	T20801	punct	],38
R577	T3576	T3563	auxpass	is,estimated
R5770	T20805	T20798	cc	and,gene
R5771	T20806	T20798	conj	cell,gene
R5772	T20684	T20683	pobj	MSUD,of
R5773	T20807	T20797	pobj	therapies,as
R5774	T20685	T20679	aux	has,described
R5775	T20686	T20679	auxpass	been,described
R5776	T20808	T20809	punct	(,hepatocytes
R5777	T20687	T20679	advcl	attributing,described
R5778	T20688	T20689	det	this,disease
R5779	T20809	T20807	parataxis	hepatocytes,therapies
R578	T3395	T3374	parataxis	1,show
R5780	T20689	T20687	dobj	disease,attributing
R5781	T20690	T20687	prep	to,attributing
R5782	T20691	T20692	compound	BCAT,deficiency
R5783	T20810	T20809	advmod	e.g.,hepatocytes
R5784	T20692	T20690	pobj	deficiency,to
R5785	T20693	T20679	punct	.,described
R5786	T20811	T20809	punct	", ",hepatocytes
R5787	T20695	T20696	prep	In,described
R5788	T20812	T20813	punct	[,43
R5789	T20697	T20695	pobj	addition,In
R579	T3577	T3578	aux	to,be
R5790	T20698	T20697	prep	to,addition
R5791	T20813	T20809	parataxis	43,hepatocytes
R5792	T20699	T20700	det	this,model
R5793	T20700	T20698	pobj	model,to
R5794	T20701	T20700	compound	mouse,model
R5795	T20702	T20696	punct	", ",described
R5796	T20814	T20813	nummod	42,43
R5797	T20703	T20704	det	a,model
R5798	T20704	T20696	nsubjpass	model,described
R5799	T20705	T20704	compound	cow,model
R58	T903	T899	advcl	combat,require
R580	T3578	T3563	xcomp	be,estimated
R5800	T20706	T20704	prep	of,model
R5801	T20707	T20706	pobj	MSUD,of
R5802	T20708	T20696	aux	has,described
R5803	T20815	T20813	punct	",",43
R5804	T20709	T20696	advmod	also,described
R5805	T20710	T20696	auxpass	been,described
R5806	T20711	T20696	advmod	previously,described
R5807	T20816	T20813	punct	],43
R5808	T20712	T20713	punct	[,17
R5809	T20713	T20696	parataxis	17,described
R581	T3396	T3395	punct	],1
R5810	T20714	T20715	punct	-,19
R5811	T20817	T20809	cc	or,hepatocytes
R5812	T20715	T20713	prep	19,17
R5813	T20716	T20713	punct	],17
R5814	T20717	T20696	punct	.,described
R5815	T20818	T20819	amod	embryonic,cells
R5816	T20719	T20720	advmod	However,is
R5817	T20819	T20809	conj	cells,hepatocytes
R5818	T20721	T20720	punct	", ",is
R5819	T20722	T20720	prep	due,is
R582	T3579	T3578	prep	as,be
R5820	T20723	T20722	pcomp	to,due
R5821	T20724	T20725	amod	practical,constraints
R5822	T20820	T20819	compound	stem,cells
R5823	T20725	T20722	pobj	constraints,due
R5824	T20726	T20725	acl	imposed,constraints
R5825	T20821	T20822	punct	[,44
R5826	T20727	T20726	agent	by,imposed
R5827	T20728	T20729	amod	such,model
R5828	T20729	T20727	pobj	model,by
R5829	T20822	T20819	parataxis	44,cells
R583	T3580	T3579	amod	high,as
R5830	T20730	T20729	det	a,model
R5831	T20731	T20729	amod	large,model
R5832	T20732	T20729	compound	animal,model
R5833	T20823	T20822	punct	],44
R5834	T20733	T20722	cc	and,due
R5835	T20734	T20722	conj	due,due
R5836	T20735	T20734	pcomp	to,due
R5837	T20824	T20809	punct	),hepatocytes
R5838	T20736	T20737	det	the,observation
R5839	T20737	T20734	pobj	observation,due
R584	T3581	T3580	prep	as,high
R5840	T20738	T20737	prep	of,observation
R5841	T20825	T20785	punct	.,is
R5842	T20739	T20738	pobj	differences,of
R5843	T20740	T20739	prep	between,differences
R5844	T20827	T20828	advcl	Relevant,highlight
R5845	T20741	T20742	det	the,model
R5846	T20742	T20740	pobj	model,between
R5847	T20829	T20827	prep	to,Relevant
R5848	T20743	T20742	compound	MSUD,model
R5849	T20744	T20742	compound	cow,model
R585	T3397	T3374	punct	.,show
R5850	T20745	T20742	cc	and,model
R5851	T20746	T20747	compound	MSUD,humans
R5852	T20747	T20742	conj	humans,model
R5853	T20748	T20749	punct	[,1
R5854	T20749	T20734	parataxis	1,due
R5855	T20830	T20831	compound	gene,therapy
R5856	T20750	T20749	punct	],1
R5857	T20751	T20720	punct	", ",is
R5858	T20752	T20753	det	this,model
R5859	T20831	T20829	pobj	therapy,to
R586	T3582	T3583	nummod	1,176
R5860	T20753	T20720	nsubj	model,is
R5861	T20754	T20720	advmod	also,is
R5862	T20755	T20720	prep	of,is
R5863	T20832	T20828	punct	", ",highlight
R5864	T20756	T20757	amod	limited,utility
R5865	T20833	T20834	amod	recent,problems
R5866	T20757	T20755	pobj	utility,of
R5867	T20758	T20720	punct	.,is
R5868	T20760	T20761	prep	In,are
R5869	T20834	T20828	nsubj	problems,highlight
R587	T3583	T3581	pobj	176,as
R5870	T20762	T20760	pobj	contrast,In
R5871	T20763	T20761	punct	", ",are
R5872	T20835	T20834	prep	with,problems
R5873	T20764	T20765	det	the,models
R5874	T20765	T20761	nsubj	models,are
R5875	T20766	T20765	amod	murine,models
R5876	T20836	T20837	compound	gene,therapy
R5877	T20767	T20765	acl	described,models
R5878	T20768	T20767	prep	in,described
R5879	T20769	T20770	det	this,report
R588	T3584	T3583	punct	:,176
R5880	T20837	T20835	pobj	therapy,with
R5881	T20770	T20768	pobj	report,in
R5882	T20771	T20772	advmod	more,valid
R5883	T20772	T20761	acomp	valid,are
R5884	T20838	T20834	prep	in,problems
R5885	T20773	T20772	cc	and,valid
R5886	T20774	T20772	conj	appropriate,valid
R5887	T20775	T20774	prep	for,appropriate
R5888	T20839	T20838	pobj	humans,in
R5889	T20776	T20777	amod	modeling,MSUD
R589	T3399	T3400	advmod	Approximately,75
R5890	T20777	T20775	pobj	MSUD,for
R5891	T20778	T20777	amod	human,MSUD
R5892	T20840	T20841	det	the,importance
R5893	T20779	T20761	punct	.,are
R5894	T20841	T20828	dobj	importance,highlight
R5895	T20781	T20782	det	The,opportunity
R5896	T20782	T20785	nsubj	opportunity,is
R5897	T20783	T20784	advmod	most,significant
R5898	T20784	T20782	amod	significant,opportunity
R5899	T20842	T20841	cc	and,importance
R59	T904	T905	det	this,disease
R590	T3585	T3586	punct	[,2
R5900	T20786	T20782	acl	presented,opportunity
R5901	T20787	T20786	agent	by,presented
R5902	T20788	T20789	det	the,models
R5903	T20789	T20787	pobj	models,by
R5904	T20843	T20844	amod	critical,need
R5905	T20844	T20841	conj	need,importance
R5906	T20845	T20841	prep	of,importance
R5907	T20846	T20847	compound	animal,models
R5908	T20847	T20845	pobj	models,of
R5909	T20848	T20847	prep	of,models
R591	T3586	T3563	parataxis	2,estimated
R5910	T20896	T20892	auxpass	is,expected
R5911	T20897	T20898	aux	to,advance
R5912	T20849	T20850	amod	genetic,diseases
R5913	T20898	T20892	xcomp	advance,expected
R5914	T20899	T20900	amod	such,approaches
R5915	T20900	T20898	dobj	approaches,advance
R5916	T20850	T20848	pobj	diseases,of
R5917	T20901	T20900	amod	therapeutic,approaches
R5918	T20902	T20892	cc	and,expected
R5919	T20903	T20904	aux	could,pave
R592	T3587	T3586	punct	],2
R5920	T20904	T20892	conj	pave,expected
R5921	T20851	T20841	prep	for,importance
R5922	T20905	T20906	det	the,way
R5923	T20906	T20904	dobj	way,pave
R5924	T20852	T20853	amod	preclinical,studies
R5925	T20907	T20904	prep	for,pave
R5926	T20908	T20907	pobj	studies,for
R5927	T20853	T20851	pobj	studies,for
R5928	T20909	T20908	prep	in,studies
R5929	T20910	T20909	pobj	humans,in
R593	T3588	T3563	punct	.,estimated
R5930	T20911	T20892	punct	.,expected
R5931	T20854	T20828	punct	.,highlight
R5932	T20913	T20914	prep	In,be
R5933	T20856	T20857	det	The,testing
R5934	T20857	T20858	nsubj	testing,provides
R5935	T20915	T20913	pobj	addition,In
R5936	T20916	T20915	prep	to,addition
R5937	T20859	T20857	prep	of,testing
R5938	T20917	T20916	pcomp	testing,to
R5939	T20918	T20919	amod	various,treatments
R594	T3400	T3401	nummod	75,%
R5940	T20919	T20917	dobj	treatments,testing
R5941	T20920	T20914	punct	", ",be
R5942	T20860	T20861	npadvmod	gene,based
R5943	T20921	T20922	det	these,models
R5944	T20922	T20914	nsubj	models,be
R5945	T20923	T20914	aux	should,be
R5946	T20861	T20864	amod	based,therapies
R5947	T20924	T20914	advmod	also,be
R5948	T20925	T20926	advmod	very,useful
R5949	T20862	T20860	cc	and,gene
R595	T3590	T3591	det	The,complex
R5950	T20926	T20914	acomp	useful,be
R5951	T20927	T20926	prep	for,useful
R5952	T20863	T20860	conj	cell,gene
R5953	T20928	T20927	pcomp	investigating,for
R5954	T20929	T20930	det	the,consequences
R5955	T20930	T20928	dobj	consequences,investigating
R5956	T20931	T20930	amod	underlying,consequences
R5957	T20932	T20930	amod	pathophysiologic,consequences
R5958	T20933	T20930	prep	of,consequences
R5959	T20864	T20859	pobj	therapies,of
R596	T3401	T3402	nsubj	%,have
R5960	T20934	T20935	det	the,disease
R5961	T20935	T20933	pobj	disease,of
R5962	T20936	T20914	punct	.,be
R5963	T20865	T20857	prep	on,testing
R5964	T20938	T20939	amod	Such,studies
R5965	T20939	T20940	nsubj	studies,are
R5966	T20866	T20867	amod	appropriate,systems
R5967	T20941	T20942	advmod	very,difficult
R5968	T20942	T20943	amod	difficult,impossible
R5969	T20867	T20865	pobj	systems,on
R597	T3591	T3593	nsubj	complex,consists
R5970	T20943	T20940	acomp	impossible,are
R5971	T20944	T20943	punct	/,impossible
R5972	T20945	T20946	aux	to,do
R5973	T20868	T20867	compound	animal,systems
R5974	T20946	T20943	advcl	do,impossible
R5975	T20947	T20946	prep	in,do
R5976	T20948	T20949	amod	human,patients
R5977	T20869	T20867	compound	model,systems
R5978	T20949	T20947	pobj	patients,in
R5979	T20950	T20946	prep	for,do
R598	T3592	T3591	compound	BCKDH,complex
R5980	T20951	T20952	amod	obvious,reasons
R5981	T20870	T20871	det	a,method
R5982	T20952	T20950	pobj	reasons,for
R5983	T20953	T20952	amod	ethical,reasons
R5984	T20954	T20940	punct	", ",are
R5985	T20871	T20858	dobj	method,provides
R5986	T20955	T20940	cc	and,are
R5987	T20956	T20957	advmod	often,rely
R5988	T20957	T20940	conj	rely,are
R5989	T20872	T20871	amod	preliminary,method
R599	T3594	T3591	punct	", ",complex
R5990	T20958	T20957	aux	must,rely
R5991	T20959	T20957	prep	on,rely
R5992	T20960	T20961	compound	autopsy,tissue
R5993	T20873	T20871	prep	of,method
R5994	T20961	T20959	pobj	tissue,on
R5995	T20962	T20940	punct	.,are
R5996	T20874	T20873	pcomp	establishing,of
R5997	T20964	T20965	advmod	Additionally,be
R5998	T20875	T20876	preconj	not,efficacy
R5999	T20966	T20965	punct	", ",be
R6	T841	T840	prep	of,models
R60	T905	T903	dobj	disease,combat
R600	T3595	T3596	det	the,enzyme
R6000	T20967	T20968	det	the,model
R6001	T20968	T20965	nsubj	model,be
R6002	T20969	T20968	amod	intermediate,model
R6003	T20876	T20874	dobj	efficacy,establishing
R6004	T20970	T20968	compound	MSUD,model
R6005	T20971	T20965	aux	may,be
R6006	T20877	T20875	advmod	only,not
R6007	T20972	T20965	advmod	also,be
R6008	T20973	T20965	acomp	useful,be
R6009	T20974	T20973	prep	for,useful
R601	T3596	T3591	appos	enzyme,complex
R6010	T20975	T20974	pobj	studies,for
R6011	T20976	T20977	aux	to,test
R6012	T20977	T20975	advcl	test,studies
R6013	T20878	T20876	det	the,efficacy
R6014	T20978	T20979	det	the,effect
R6015	T20979	T20977	dobj	effect,test
R6016	T20980	T20979	prep	of,effect
R6017	T20879	T20876	punct	", ",efficacy
R6018	T20981	T20980	pobj	thiamin,of
R6019	T20982	T20965	punct	.,be
R602	T3403	T3401	prep	of,%
R6020	T20880	T20876	cc	but,efficacy
R6021	T20984	T20985	det	A,form
R6022	T20881	T20880	advmod	also,but
R6023	T20985	T20989	nsubjpass	form,described
R6024	T20882	T20883	amod	short,term
R6025	T20986	T20987	npadvmod	thiamin,responsive
R6026	T20987	T20985	amod	responsive,form
R6027	T20883	T20888	compound	term,safety
R6028	T20988	T20987	punct	-,responsive
R6029	T20990	T20985	prep	of,form
R603	T3597	T3596	amod	deficient,enzyme
R6030	T20884	T20882	punct	-,short
R6031	T20991	T20990	pobj	MSUD,of
R6032	T20992	T20989	aux	has,described
R6033	T20885	T20882	cc	and,short
R6034	T20993	T20989	auxpass	been,described
R6035	T20994	T20995	punct	[,1
R6036	T20886	T20882	conj	long,short
R6037	T20995	T20989	parataxis	1,described
R6038	T20996	T20995	punct	],1
R6039	T20997	T20989	punct	.,described
R604	T3598	T3596	prep	in,enzyme
R6040	T20887	T20883	punct	-,term
R6041	T20999	T21000	det	The,phenotype
R6042	T21000	T21001	nsubj	phenotype,is
R6043	T20888	T20876	conj	safety,efficacy
R6044	T20889	T20858	punct	.,provides
R6045	T20891	T20892	nsubjpass	Success,expected
R6046	T20893	T20891	prep	with,Success
R6047	T21002	T21000	prep	of,phenotype
R6048	T21003	T21004	det	these,patients
R6049	T20894	T20895	compound	animal,studies
R605	T3599	T3598	pobj	MSUD,in
R6050	T21004	T21002	pobj	patients,of
R6051	T20895	T20893	pobj	studies,with
R6052	T21005	T21001	acomp	heterogeneous,is
R6053	T21006	T21001	cc	and,is
R6054	T21108	T21109	punct	[,46
R6055	T21007	T21008	nsubj	treatment,produced
R6056	T21008	T21001	conj	produced,is
R6057	T21009	T21007	prep	with,treatment
R6058	T21010	T21011	det	a,range
R6059	T21011	T21009	pobj	range,with
R606	T3600	T3593	punct	", ",consists
R6060	T21012	T21011	amod	wide,range
R6061	T21013	T21011	prep	of,range
R6062	T21014	T21015	compound	thiamin,doses
R6063	T21109	T21097	parataxis	46,Because
R6064	T21015	T21013	pobj	doses,of
R6065	T21016	T21008	aux	has,produced
R6066	T21017	T21018	amod	limited,success
R6067	T21018	T21008	dobj	success,produced
R6068	T21110	T21109	nummod	35,46
R6069	T21019	T21020	punct	[,1
R607	T3601	T3593	prep	of,consists
R6070	T21020	T21008	parataxis	1,produced
R6071	T21111	T21109	punct	",",46
R6072	T21021	T21020	punct	],1
R6073	T21022	T21008	punct	.,produced
R6074	T21112	T21109	nummod	36,46
R6075	T21024	T21025	amod	Recent,studies
R6076	T21025	T21028	nsubj	studies,suggested
R6077	T21026	T21027	compound	cell,culture
R6078	T21113	T21109	punct	",",46
R6079	T21027	T21025	compound	culture,studies
R608	T3404	T3405	compound	MSUD,patients
R6080	T21029	T21025	prep	with,studies
R6081	T21114	T21109	nummod	45,46
R6082	T21030	T21031	compound	MSUD,cells
R6083	T21031	T21029	pobj	cells,with
R6084	T21032	T21028	aux	have,suggested
R6085	T21115	T21109	punct	",",46
R6086	T21033	T21034	mark	that,is
R6087	T21034	T21028	ccomp	is,suggested
R6088	T21116	T21109	punct	],46
R6089	T21035	T21036	det	the,phenotype
R609	T3602	T3603	nummod	three,proteins
R6090	T21036	T21034	nsubj	phenotype,is
R6091	T21117	T21098	punct	", ",provide
R6092	T21037	T21038	npadvmod	thiamin,responsive
R6093	T21038	T21036	amod	responsive,phenotype
R6094	T21118	T21119	det	the,model
R6095	T21039	T21038	punct	-,responsive
R6096	T21040	T21034	acomp	dependent,is
R6097	T21041	T21040	prep	upon,dependent
R6098	T21119	T21098	nsubj	model,provide
R6099	T21042	T21043	det	the,presence
R61	T906	T895	punct	.,are
R610	T3603	T3601	pobj	proteins,of
R6100	T21043	T21041	pobj	presence,upon
R6101	T21044	T21043	prep	of,presence
R6102	T21045	T21046	advmod	at,one
R6103	T21120	T21119	amod	intermediate,model
R6104	T21046	T21048	nummod	one,allele
R6105	T21047	T21046	advmod	least,one
R6106	T21048	T21044	pobj	allele,of
R6107	T21121	T21119	compound	MSUD,model
R6108	T21049	T21050	npadvmod	E2,expressing
R6109	T21050	T21048	amod	expressing,allele
R611	T3604	T3603	amod	catalytic,proteins
R6110	T21051	T21052	punct	[,46
R6111	T21052	T21028	parataxis	46,suggested
R6112	T21053	T21052	nummod	45,46
R6113	T21054	T21052	punct	",",46
R6114	T21055	T21052	punct	],46
R6115	T21122	T21098	aux	may,provide
R6116	T21056	T21028	punct	.,suggested
R6117	T21058	T21059	prep	In,provides
R6118	T21123	T21124	det	a,resource
R6119	T21060	T21058	pobj	light,In
R612	T3405	T3403	pobj	patients,of
R6120	T21061	T21060	prep	of,light
R6121	T21124	T21098	dobj	resource,provide
R6122	T21062	T21063	det	these,findings
R6123	T21063	T21061	pobj	findings,of
R6124	T21064	T21063	amod	newer,findings
R6125	T21125	T21124	amod	valuable,resource
R6126	T21065	T21059	punct	", ",provides
R6127	T21066	T21067	poss	our,mouse
R6128	T21126	T21124	prep	for,resource
R6129	T21067	T21059	nsubj	mouse,provides
R613	T3605	T3603	punct	", ",proteins
R6130	T21068	T21067	amod	intermediate,mouse
R6131	T21069	T21067	compound	MSUD,mouse
R6132	T21127	T21126	pobj	studies,for
R6133	T21070	T21071	det	a,model
R6134	T21071	T21059	dobj	model,provides
R6135	T21072	T21073	aux	to,test
R6136	T21073	T21071	advcl	test,model
R6137	T21128	T21127	prep	of,studies
R6138	T21074	T21075	det	the,effect
R6139	T21075	T21073	dobj	effect,test
R614	T3606	T3607	det	a,decarboxylase
R6140	T21076	T21075	prep	of,effect
R6141	T21129	T21130	amod	structural,biology
R6142	T21077	T21076	pobj	thiamin,of
R6143	T21130	T21128	pobj	biology,of
R6144	T21078	T21073	prep	with,test
R6145	T21079	T21078	pobj	respect,with
R6146	T21080	T21079	prep	to,respect
R6147	T21081	T21082	compound	BCKDH,activity
R6148	T21131	T21098	punct	.,provide
R6149	T21082	T21080	pobj	activity,to
R615	T3607	T3603	appos	decarboxylase,proteins
R6150	T21083	T21082	cc	and,activity
R6151	T21084	T21085	compound	blood,levels
R6152	T21133	T21134	advmod	Lastly,be
R6153	T21085	T21082	conj	levels,activity
R6154	T21086	T21087	compound	amino,acid
R6155	T21087	T21085	compound	acid,levels
R6156	T21088	T21073	prep	at,test
R6157	T21135	T21134	punct	", ",be
R6158	T21089	T21090	det	the,level
R6159	T21090	T21088	pobj	level,at
R616	T3406	T3407	det	the,form
R6160	T21136	T21137	det	the,information
R6161	T21091	T21090	prep	of,level
R6162	T21092	T21093	det	the,animal
R6163	T21093	T21091	pobj	animal,of
R6164	T21094	T21093	amod	whole,animal
R6165	T21095	T21059	punct	.,provides
R6166	T21137	T21134	nsubj	information,be
R6167	T21097	T21098	prep	Because,provide
R6168	T21138	T21137	cc	and,information
R6169	T21099	T21097	pcomp	of,Because
R617	T3608	T3609	punct	(,E1
R6170	T21100	T21101	amod	recent,interest
R6171	T21101	T21097	pobj	interest,Because
R6172	T21139	T21137	conj	knowledge,information
R6173	T21102	T21101	prep	in,interest
R6174	T21103	T21104	amod	structural,analysis
R6175	T21104	T21102	pobj	analysis,in
R6176	T21140	T21137	acl	gained,information
R6177	T21105	T21104	prep	of,analysis
R6178	T21106	T21107	compound	multienzyme,complexes
R6179	T21107	T21105	pobj	complexes,of
R618	T3609	T3607	parataxis	E1,decarboxylase
R6180	T21141	T21140	prep	from,gained
R6181	T21142	T21141	pobj	studies,from
R6182	T21143	T21142	prep	with,studies
R6183	T21144	T21145	det	the,models
R6184	T21145	T21143	pobj	models,with
R6185	T21146	T21145	compound	MSUD,models
R6186	T21147	T21145	acl	described,models
R6187	T21148	T21147	advmod	here,described
R6188	T21149	T21134	aux	will,be
R6189	T21150	T21134	advmod	also,be
R619	T3610	T3609	punct	),E1
R6190	T21151	T21134	acomp	applicable,be
R6191	T21152	T21151	cc	and,applicable
R6192	T21153	T21151	conj	transferable,applicable
R6193	T21154	T21153	prep	to,transferable
R6194	T21155	T21156	amod	other,disorders
R6195	T21156	T21154	pobj	disorders,to
R6196	T21157	T21156	amod	mitochondrial,disorders
R6197	T21158	T21134	prep	due,be
R6198	T21159	T21158	pcomp	to,due
R6199	T21160	T21158	pobj	defects,due
R62	T908	T909	det	A,hurdle
R620	T3407	T3402	dobj	form,have
R6200	T21161	T21160	prep	in,defects
R6201	T21162	T21163	amod	multisubunit,enzymes
R6202	T21163	T21161	pobj	enzymes,in
R6203	T21164	T21134	punct	.,be
R6204	T21291	T21292	prep	In,describes
R6205	T21293	T21291	pobj	summary,In
R6206	T21294	T21292	punct	", ",describes
R6207	T21295	T21296	det	this,report
R6208	T21296	T21292	nsubj	report,describes
R6209	T21297	T21298	det	the,development
R621	T3611	T3607	punct	", ",decarboxylase
R6210	T21298	T21292	dobj	development,describes
R6211	T21299	T21298	cc	and,development
R6212	T21300	T21298	conj	characterization,development
R6213	T21301	T21298	prep	of,development
R6214	T21302	T21303	advmod	genetically,engineered
R6215	T21303	T21304	amod	engineered,models
R6216	T21304	T21301	pobj	models,of
R6217	T21305	T21304	compound	mouse,models
R6218	T21306	T21304	prep	of,models
R6219	T21307	T21308	amod	classic,MSUD
R622	T3612	T3613	det	a,transacylase
R6220	T21308	T21306	pobj	MSUD,of
R6221	T21309	T21310	advmod	as,as
R6222	T21310	T21307	cc	as,classic
R6223	T21311	T21310	advmod	well,as
R6224	T21312	T21307	conj	intermediate,classic
R6225	T21313	T21292	punct	.,describes
R6226	T21315	T21316	det	These,animals
R6227	T21316	T21317	nsubj	animals,provide
R6228	T21318	T21319	amod	useful,models
R6229	T21319	T21317	dobj	models,provide
R623	T3408	T3407	amod	classic,form
R6230	T21320	T21321	aux	to,characterize
R6231	T21321	T21319	advcl	characterize,models
R6232	T21322	T21321	advmod	further,characterize
R6233	T21323	T21324	det	the,pathogenesis
R6234	T21324	T21321	dobj	pathogenesis,characterize
R6235	T21325	T21324	prep	of,pathogenesis
R6236	T21326	T21325	pobj	MSUD,of
R6237	T21327	T21317	punct	", ",provide
R6238	T21328	T21329	advmod	as,as
R6239	T21329	T21317	cc	as,provide
R624	T3613	T3607	conj	transacylase,decarboxylase
R6240	T21330	T21329	advmod	well,as
R6241	T21331	T21317	conj	models,provide
R6242	T21332	T21333	aux	to,test
R6243	T21333	T21331	advcl	test,models
R6244	T21334	T21335	amod	novel,strategies
R6245	T21335	T21333	dobj	strategies,test
R6246	T21336	T21335	amod	therapeutic,strategies
R6247	T21337	T21335	punct	", ",strategies
R6248	T21338	T21339	amod	such,as
R6249	T21339	T21335	prep	as,strategies
R625	T3614	T3613	compound	dihydrolipoyl,transacylase
R6250	T21340	T21341	nmod	gene,therapies
R6251	T21341	T21339	pobj	therapies,as
R6252	T21342	T21340	cc	and,gene
R6253	T21343	T21340	conj	cellular,gene
R6254	T21344	T21333	punct	", ",test
R6255	T21345	T21346	aux	to,treat
R6256	T21346	T21333	advcl	treat,test
R6257	T21347	T21348	det	this,disease
R6258	T21348	T21346	dobj	disease,treat
R6259	T21349	T21348	amod	devastating,disease
R626	T3615	T3616	punct	(,E2
R6260	T21350	T21348	amod	metabolic,disease
R6261	T21351	T21317	punct	.,provide
R6262	T21373	T21374	det	The,history
R6263	T21374	T21376	nsubjpass	history,accessed
R6264	T21375	T21374	amod	pre-publication,history
R6265	T21377	T21374	prep	for,history
R6266	T21378	T21379	det	this,paper
R6267	T21379	T21377	pobj	paper,for
R6268	T21380	T21376	aux	can,accessed
R6269	T21381	T21376	auxpass	be,accessed
R627	T3409	T3407	prep	of,form
R6270	T21382	T21376	advmod	here,accessed
R6271	T21383	T21376	punct	:,accessed
R6272	T22055	T22056	compound	E2,production
R6273	T22057	T22058	compound	gene,knockout
R6274	T22058	T22056	compound	knockout,production
R6275	T22059	T22056	compound	mouse,production
R6276	T22060	T22056	punct	.,production
R6277	T22062	T22063	meta	A,strategy
R6278	T22064	T22063	punct	", ",strategy
R6279	T22065	T22066	compound	Gene,targeting
R628	T3616	T3613	parataxis	E2,transacylase
R6280	T22066	T22063	compound	targeting,strategy
R6281	T22067	T22063	acl	used,strategy
R6282	T22068	T22067	prep	for,used
R6283	T22069	T22068	pcomp	targeting,for
R6284	T22070	T22071	det	the,locus
R6285	T22071	T22069	dobj	locus,targeting
R6286	T22072	T22071	compound	E2,locus
R6287	T22073	T22067	prep	in,used
R6288	T22074	T22075	compound	mouse,cells
R6289	T22075	T22073	pobj	cells,in
R629	T3617	T3616	punct	),E2
R6290	T22076	T22075	compound	ES,cells
R6291	T22077	T22063	punct	.,strategy
R6292	T22079	T22080	det	The,construct
R6293	T22080	T22082	nsubjpass	construct,designed
R6294	T22081	T22080	compound	targeting,construct
R6295	T22083	T22082	auxpass	was,designed
R6296	T22084	T22085	aux	to,delete
R6297	T22085	T22082	advcl	delete,designed
R6298	T22086	T22087	nummod	1.67,kb
R6299	T22087	T22085	dobj	kb,delete
R63	T909	T911	nsubj	hurdle,been
R630	T3618	T3613	punct	", ",transacylase
R6300	T22088	T22087	prep	of,kb
R6301	T22089	T22088	pobj	sequence,of
R6302	T22090	T22085	prep	between,delete
R6303	T22091	T22092	det	an,site
R6304	T22092	T22090	pobj	site,between
R6305	T22093	T22092	compound	EcoRV,site
R6306	T22094	T22092	prep	in,site
R6307	T22095	T22094	pobj	Exon,in
R6308	T22096	T22095	nummod	4,Exon
R6309	T22097	T22092	cc	and,site
R631	T3410	T3411	det	the,disease
R6310	T22098	T22099	det	a,site
R6311	T22099	T22092	conj	site,site
R6312	T22100	T22099	compound	Smal,site
R6313	T22101	T22099	prep	in,site
R6314	T22102	T22101	pobj	intron,in
R6315	T22103	T22102	nummod	5,intron
R6316	T22104	T22082	punct	.,designed
R6317	T22106	T22107	det	The,gene
R6318	T22107	T22111	nsubj	gene,contains
R6319	T22108	T22109	amod	wild,type
R632	T3619	T3613	cc	and,transacylase
R6320	T22109	T22107	compound	type,gene
R6321	T22110	T22107	compound	E2,gene
R6322	T22112	T22113	det	an,fragment
R6323	T22113	T22111	dobj	fragment,contains
R6324	T22114	T22115	punct	~,16
R6325	T22115	T22116	nummod	16,kb
R6326	T22116	T22113	compound	kb,fragment
R6327	T22117	T22113	compound	BglI,fragment
R6328	T22118	T22113	compound	restriction,fragment
R6329	T22119	T22120	dep	that,hybridizes
R633	T3620	T3621	det	a,dehydrogenase
R6330	T22120	T22113	relcl	hybridizes,fragment
R6331	T22121	T22120	prep	to,hybridizes
R6332	T22122	T22123	det	the,probe
R6333	T22123	T22121	pobj	probe,to
R6334	T22124	T22125	npadvmod	Exon,specific
R6335	T22125	T22123	amod	specific,probe
R6336	T22126	T22124	nummod	6,Exon
R6337	T22127	T22111	punct	.,contains
R6338	T22129	T22130	det	A,locus
R6339	T22130	T22134	nsubj	locus,harbors
R634	T3621	T3613	conj	dehydrogenase,transacylase
R6340	T22131	T22132	advmod	correctly,targeted
R6341	T22132	T22130	amod	targeted,locus
R6342	T22133	T22130	compound	E2,locus
R6343	T22135	T22136	det	an,fragment
R6344	T22136	T22134	dobj	fragment,harbors
R6345	T22137	T22138	punct	~,11
R6346	T22138	T22139	nummod	11,kb
R6347	T22139	T22136	compound	kb,fragment
R6348	T22140	T22136	compound	BglI,fragment
R6349	T22141	T22136	compound	restriction,fragment
R635	T3622	T3621	compound	dihydrolipoyl,dehydrogenase
R6350	T22142	T22143	dep	that,hybridizes
R6351	T22143	T22136	relcl	hybridizes,fragment
R6352	T22144	T22143	prep	to,hybridizes
R6353	T22145	T22146	det	the,probe
R6354	T22146	T22144	pobj	probe,to
R6355	T22147	T22146	amod	same,probe
R6356	T22148	T22134	punct	.,harbors
R6357	T22151	T22152	mark	that,detect
R6358	T22152	T22150	ccomp	detect,Note
R6359	T22153	T22154	det	the,probe
R636	T3411	T3409	pobj	disease,of
R6360	T22154	T22152	nsubj	probe,detect
R6361	T22155	T22152	aux	will,detect
R6362	T22156	T22152	neg	not,detect
R6363	T22157	T22158	amod	random,integration
R6364	T22158	T22152	dobj	integration,detect
R6365	T22159	T22158	prep	of,integration
R6366	T22160	T22161	det	the,vector
R6367	T22161	T22159	pobj	vector,of
R6368	T22162	T22161	compound	targeting,vector
R6369	T22163	T22164	mark	because,is
R637	T3623	T3624	punct	(,E3
R6370	T22164	T22152	advcl	is,detect
R6371	T22165	T22164	nsubj	it,is
R6372	T22166	T22164	acomp	external,is
R6373	T22167	T22166	prep	to,external
R6374	T22168	T22169	det	the,vector
R6375	T22169	T22167	pobj	vector,to
R6376	T22170	T22169	compound	targeting,vector
R6377	T22171	T22150	punct	.,Note
R6378	T22173	T22174	meta	B,analysis
R6379	T22175	T22174	punct	", ",analysis
R638	T3624	T3621	parataxis	E3,dehydrogenase
R6380	T22176	T22177	compound	Southern,blot
R6381	T22177	T22174	compound	blot,analysis
R6382	T22178	T22174	prep	of,analysis
R6383	T22179	T22180	npadvmod	Bgll,digested
R6384	T22180	T22181	amod	digested,DNA
R6385	T22181	T22178	pobj	DNA,of
R6386	T22182	T22181	amod	genomic,DNA
R6387	T22183	T22181	acl	derived,DNA
R6388	T22184	T22183	prep	from,derived
R6389	T22185	T22186	det	the,line
R639	T3625	T3624	punct	),E3
R6390	T22186	T22184	pobj	line,from
R6391	T22187	T22186	amod	parental,line
R6392	T22188	T22189	amod	wild,type
R6393	T22189	T22186	compound	type,line
R6394	T22190	T22191	compound	ES,cell
R6395	T22191	T22186	compound	cell,line
R6396	T22192	T22193	punct	(,R1
R6397	T22193	T22186	parataxis	R1,line
R6398	T22194	T22193	punct	),R1
R6399	T22195	T22186	punct	", ",line
R64	T910	T909	amod	major,hurdle
R640	T3412	T3407	prep	with,form
R6400	T22196	T22197	det	a,line
R6401	T22197	T22186	appos	line,line
R6402	T22198	T22197	amod	heterozygous,line
R6403	T22199	T22197	amod	targeted,line
R6404	T22200	T22201	compound	ES,cell
R6405	T22201	T22197	compound	cell,line
R6406	T22202	T22203	punct	(,362
R6407	T22203	T22197	parataxis	362,line
R6408	T22204	T22203	punct	),362
R6409	T22205	T22184	punct	", ",from
R641	T3626	T3593	punct	.,consists
R6410	T22206	T22184	cc	and,from
R6411	T22207	T22184	conj	from,from
R6412	T22208	T22209	amod	wild,type
R6413	T22209	T22210	nmod	type,knockout
R6414	T22210	T22225	nmod	knockout,mice
R6415	T22211	T22212	punct	(,+
R6416	T22212	T22209	punct	+,type
R6417	T22213	T22212	punct	+,+
R6418	T22214	T22212	punct	/,+
R6419	T22215	T22212	punct	),+
R642	T3413	T3414	compound	BCKDH,activity
R6420	T22216	T22209	punct	", ",type
R6421	T22217	T22209	conj	heterozygous,type
R6422	T22218	T22219	punct	(,-
R6423	T22219	T22217	punct	-,heterozygous
R6424	T22220	T22219	punct	+,-
R6425	T22221	T22219	punct	/,-
R6426	T22222	T22219	punct	),-
R6427	T22223	T22217	cc	and,heterozygous
R6428	T22224	T22217	conj	homozygous,heterozygous
R6429	T22225	T22207	pobj	mice,from
R643	T3628	T3629	det	The,component
R6430	T22226	T22227	punct	(,-
R6431	T22227	T22210	punct	-,knockout
R6432	T22228	T22227	punct	-,-
R6433	T22229	T22227	punct	/,-
R6434	T22230	T22227	punct	),-
R6435	T22231	T22174	punct	.,analysis
R6436	T22233	T22234	det	The,blot
R6437	T22234	T22235	nsubjpass	blot,hybridized
R6438	T22236	T22235	auxpass	was,hybridized
R6439	T22237	T22235	prep	with,hybridized
R644	T3629	T3631	nsubj	component,is
R6440	T22238	T22239	det	an,probe
R6441	T22239	T22237	pobj	probe,with
R6442	T22240	T22241	npadvmod	Exon,specific
R6443	T22241	T22239	amod	specific,probe
R6444	T22242	T22240	nummod	6,Exon
R6445	T22243	T22235	punct	.,hybridized
R6446	T22245	T22246	meta	C,analysis
R6447	T22247	T22246	punct	", ",analysis
R6448	T22248	T22246	amod	Immunohistochemical,analysis
R6449	T22249	T22246	prep	of,analysis
R645	T3630	T3629	compound	E1,component
R6450	T22250	T22251	amod	fresh,frozen
R6451	T22251	T22252	amod	frozen,sections
R6452	T22252	T22249	pobj	sections,of
R6453	T22253	T22252	compound	liver,sections
R6454	T22254	T22252	prep	from,sections
R6455	T22255	T22256	nmod	control,knockout
R6456	T22256	T22264	nmod	knockout,pups
R6457	T22257	T22258	punct	(,+
R6458	T22258	T22255	punct	+,control
R6459	T22259	T22258	punct	+,+
R646	T3414	T3412	pobj	activity,with
R6460	T22260	T22258	punct	/,+
R6461	T22261	T22258	punct	),+
R6462	T22262	T22255	cc	and,control
R6463	T22263	T22255	conj	E2,control
R6464	T22264	T22254	pobj	pups,from
R6465	T22265	T22266	punct	(,-
R6466	T22266	T22256	punct	-,knockout
R6467	T22267	T22266	punct	-,-
R6468	T22268	T22266	punct	/,-
R6469	T22269	T22266	punct	),-
R647	T3632	T3633	det	a,heterotetramer
R6470	T22270	T22271	amod	postnatal,day
R6471	T22271	T22264	nmod	day,pups
R6472	T22272	T22271	nummod	1,day
R6473	T22273	T22264	compound	mouse,pups
R6474	T22274	T22246	punct	.,analysis
R6475	T22276	T22277	nsubjpass	Sections,stained
R6476	T22278	T22277	auxpass	were,stained
R6477	T22279	T22277	prep	for,stained
R6478	T22280	T22279	pobj	E2,for
R6479	T22281	T22277	advcl	using,stained
R648	T3633	T3631	attr	heterotetramer,is
R6480	T22282	T22283	det	an,antibody
R6481	T22283	T22281	dobj	antibody,using
R6482	T22284	T22285	npadvmod	E2,specific
R6483	T22285	T22283	amod	specific,antibody
R6484	T22286	T22287	punct	(,green
R6485	T22287	T22283	parataxis	green,antibody
R6486	T22288	T22287	punct	),green
R6487	T22289	T22283	cc	and,antibody
R6488	T22290	T22291	det	a,stain
R6489	T22291	T22283	conj	stain,antibody
R649	T3415	T3414	prep	in,activity
R6490	T22292	T22291	amod	nuclear,stain
R6491	T22293	T22294	punct	(,blue
R6492	T22294	T22291	parataxis	blue,stain
R6493	T22295	T22294	punct	),blue
R6494	T22296	T22277	punct	.,stained
R6495	T22299	T22300	det	the,absence
R6496	T22300	T22298	dobj	absence,Note
R6497	T22301	T22300	amod	complete,absence
R6498	T22302	T22300	prep	of,absence
R6499	T22303	T22304	compound	E2,immunoreactivity
R65	T912	T909	prep	in,hurdle
R650	T3416	T3417	det	the,range
R6500	T22304	T22302	pobj	immunoreactivity,of
R6501	T22305	T22298	prep	in,Note
R6502	T22306	T22307	det	the,section
R6503	T22307	T22305	pobj	section,in
R6504	T22308	T22307	prep	from,section
R6505	T22309	T22310	det	the,mouse
R6506	T22310	T22308	pobj	mouse,from
R6507	T22311	T22310	compound	knockout,mouse
R6508	T22312	T22298	punct	.,Note
R6509	T22314	T22315	meta	D,observed
R651	T3634	T3633	acl	composed,heterotetramer
R6510	T22316	T22315	punct	", ",observed
R6511	T22317	T22318	amod	Similar,results
R6512	T22318	T22315	nsubjpass	results,observed
R6513	T22319	T22315	auxpass	were,observed
R6514	T22320	T22315	prep	upon,observed
R6515	T22321	T22322	amod	immunohistochemical,analysis
R6516	T22322	T22320	pobj	analysis,upon
R6517	T22323	T22322	prep	of,analysis
R6518	T22324	T22325	amod	primary,fibroblasts
R6519	T22325	T22323	pobj	fibroblasts,of
R652	T3417	T3415	pobj	range,in
R6520	T22326	T22325	nmod	mouse,fibroblasts
R6521	T22327	T22325	amod	embryonic,fibroblasts
R6522	T22328	T22325	punct	(,fibroblasts
R6523	T22329	T22325	appos	MEFs,fibroblasts
R6524	T22330	T22315	punct	),observed
R6525	T22331	T22315	punct	.,observed
R6526	T22334	T22335	mark	that,was
R6527	T22335	T22333	ccomp	was,Note
R6528	T22336	T22337	det	the,signal
R6529	T22337	T22335	nsubj	signal,was
R653	T3635	T3634	prep	of,composed
R6530	T22338	T22339	advmod	readily,detectable
R6531	T22339	T22337	amod	detectable,signal
R6532	T22340	T22337	prep	for,signal
R6533	T22341	T22340	pobj	E2,for
R6534	T22342	T22337	prep	in,signal
R6535	T22343	T22344	det	the,cells
R6536	T22344	T22342	pobj	cells,in
R6537	T22345	T22344	compound	control,cells
R6538	T22346	T22335	acomp	present,was
R6539	T22347	T22346	prep	in,present
R654	T3636	T3637	nummod	two,α
R6540	T22348	T22349	det	a,pattern
R6541	T22349	T22347	pobj	pattern,in
R6542	T22350	T22349	amod	characteristic,pattern
R6543	T22351	T22350	prep	of,characteristic
R6544	T22352	T22351	pobj	mitochondria,of
R6545	T22353	T22352	punct	", ",mitochondria
R6546	T22354	T22355	det	the,location
R6547	T22355	T22352	appos	location,mitochondria
R6548	T22356	T22355	amod	subcellular,location
R6549	T22357	T22355	prep	of,location
R655	T3637	T3638	nmod	α,subunits
R6550	T22358	T22357	pobj	BCKDH,of
R6551	T22359	T22333	punct	.,Note
R6552	T22807	T22808	amod	Biochemical,characterization
R6553	T22809	T22808	prep	of,characterization
R6554	T22810	T22811	det	the,model
R6555	T22811	T22809	pobj	model,of
R6556	T22812	T22811	amod	classic,model
R6557	T22813	T22811	nmod	MSUD,model
R6558	T22814	T22811	amod	murine,model
R6559	T22815	T22808	punct	.,characterization
R656	T3418	T3417	prep	of,range
R6560	T22817	T22818	meta	A,activity
R6561	T22819	T22818	punct	", ",activity
R6562	T22820	T22818	compound	BCKDH,activity
R6563	T22821	T22818	compound	enzyme,activity
R6564	T22822	T22818	prep	in,activity
R6565	T22823	T22822	pobj	liver,in
R6566	T22824	T22823	prep	of,liver
R6567	T22825	T22826	amod	newborn,mice
R6568	T22826	T22824	pobj	mice,of
R6569	T22827	T22828	amod	wild,type
R657	T3638	T3635	pobj	subunits,of
R6570	T22828	T22829	nmod	type,control
R6571	T22829	T22830	nmod	control,knockout
R6572	T22830	T22826	compound	knockout,mice
R6573	T22831	T22832	punct	(,+
R6574	T22832	T22829	punct	+,control
R6575	T22833	T22832	punct	+,+
R6576	T22834	T22832	punct	/,+
R6577	T22835	T22832	punct	),+
R6578	T22836	T22829	punct	", ",control
R6579	T22837	T22829	conj	heterozygous,control
R658	T3639	T3637	cc	and,α
R6580	T22838	T22839	punct	(,-
R6581	T22839	T22837	punct	-,heterozygous
R6582	T22840	T22839	punct	+,-
R6583	T22841	T22839	punct	/,-
R6584	T22842	T22839	punct	),-
R6585	T22843	T22837	punct	", ",heterozygous
R6586	T22844	T22837	cc	and,heterozygous
R6587	T22845	T22837	conj	homozygous,heterozygous
R6588	T22846	T22847	punct	(,-
R6589	T22847	T22845	punct	-,homozygous
R659	T3419	T3420	quantmod	0,2
R6590	T22848	T22847	punct	-,-
R6591	T22849	T22847	punct	/,-
R6592	T22850	T22847	punct	),-
R6593	T22851	T22818	punct	.,activity
R6594	T22853	T22854	compound	Enzyme,activity
R6595	T22854	T22855	nsubj	activity,was
R6596	T22856	T22857	advmod	significantly,reduced
R6597	T22857	T22855	acomp	reduced,was
R6598	T22858	T22855	prep	in,was
R6599	T22859	T22860	punct	+,-
R66	T913	T912	pcomp	developing,in
R660	T3640	T3641	nummod	two,β
R6600	T22860	T22862	punct	-,liver
R6601	T22861	T22860	punct	/,-
R6602	T22862	T22858	pobj	liver,in
R6603	T22863	T22855	prep	compared,was
R6604	T22864	T22863	prep	to,compared
R6605	T22865	T22866	punct	+,+
R6606	T22866	T22864	punct	+,to
R6607	T22867	T22866	punct	/,+
R6608	T22868	T22855	punct	", ",was
R6609	T22869	T22855	cc	and,was
R661	T3641	T3637	conj	β,α
R6610	T22870	T22855	conj	was,was
R6611	T22871	T22870	prep	below,was
R6612	T22872	T22873	det	the,level
R6613	T22873	T22871	pobj	level,below
R6614	T22874	T22873	prep	of,level
R6615	T22875	T22874	pobj	detection,of
R6616	T22876	T22870	prep	in,was
R6617	T22877	T22878	punct	-,-
R6618	T22878	T22880	punct	-,liver
R6619	T22879	T22878	punct	/,-
R662	T3642	T3643	punct	[,3
R6620	T22880	T22876	pobj	liver,in
R6621	T22881	T22855	punct	.,was
R6622	T22883	T22884	meta	B,concentrations
R6623	T22885	T22884	punct	", ",concentrations
R6624	T22886	T22884	amod	Total,concentrations
R6625	T22887	T22884	compound	BCAA,concentrations
R6626	T22888	T22884	prep	in,concentrations
R6627	T22889	T22888	pobj	blood,in
R6628	T22890	T22889	prep	of,blood
R6629	T22891	T22890	pobj	mice,of
R663	T3420	T3422	nummod	2,%
R6630	T22892	T22884	punct	.,concentrations
R6631	T22894	T22895	amod	Total,BCAA
R6632	T22895	T22896	nsubj	BCAA,represent
R6633	T22897	T22898	det	the,sum
R6634	T22898	T22896	dobj	sum,represent
R6635	T22899	T22898	prep	of,sum
R6636	T22900	T22899	pobj	leucine,of
R6637	T22901	T22900	punct	", ",leucine
R6638	T22902	T22900	conj	isoleucine,leucine
R6639	T22903	T22902	punct	", ",isoleucine
R664	T3643	T3631	parataxis	3,is
R6640	T22904	T22902	cc	and,isoleucine
R6641	T22905	T22902	conj	valine,isoleucine
R6642	T22906	T22896	punct	.,represent
R6643	T22908	T22909	amod	Total,concentrations
R6644	T22909	T22911	nsubj	concentrations,were
R6645	T22910	T22909	compound	BCAA,concentrations
R6646	T22912	T22909	prep	in,concentrations
R6647	T22913	T22912	pobj	blood,in
R6648	T22914	T22913	prep	from,blood
R6649	T22915	T22916	punct	-,-
R665	T3644	T3643	punct	],3
R6650	T22916	T22918	punct	-,mice
R6651	T22917	T22916	punct	/,-
R6652	T22918	T22914	pobj	mice,from
R6653	T22919	T22920	advmod	significantly,elevated
R6654	T22920	T22911	acomp	elevated,were
R6655	T22921	T22911	prep	compared,were
R6656	T22922	T22921	prep	to,compared
R6657	T22923	T22924	punct	+,+
R6658	T22924	T22922	punct	+,to
R6659	T22925	T22924	punct	/,+
R666	T3645	T3631	punct	.,is
R6660	T22926	T22924	cc	and,+
R6661	T22927	T22928	punct	+,-
R6662	T22928	T22924	conj	-,+
R6663	T22929	T22928	punct	/,-
R6664	T22930	T22911	punct	.,were
R6665	T22932	T22933	meta	C,Ratio
R6666	T22934	T22933	punct	", ",Ratio
R6667	T22935	T22933	prep	of,Ratio
R6668	T22936	T22937	amod	total,BCAA
R6669	T22937	T22935	pobj	BCAA,of
R667	T3421	T3420	punct	–,2
R6670	T22938	T22933	prep	to,Ratio
R6671	T22939	T22938	pobj	alanine,to
R6672	T22940	T22933	prep	in,Ratio
R6673	T22941	T22940	pobj	blood,in
R6674	T22942	T22941	prep	of,blood
R6675	T22943	T22942	pobj	mice,of
R6676	T22944	T22933	punct	.,Ratio
R6677	T22946	T22947	det	This,ratio
R6678	T22947	T22948	nsubj	ratio,was
R6679	T22949	T22950	advmod	significantly,elevated
R668	T3647	T3648	det	The,components
R6680	T22950	T22948	acomp	elevated,was
R6681	T22951	T22948	prep	in,was
R6682	T22952	T22953	punct	-,-
R6683	T22953	T22955	punct	-,mice
R6684	T22954	T22953	punct	/,-
R6685	T22955	T22951	pobj	mice,in
R6686	T22956	T22948	prep	compared,was
R6687	T22957	T22956	prep	to,compared
R6688	T22958	T22959	punct	+,+
R6689	T22959	T22961	punct	+,mice
R669	T3422	T3418	pobj	%,of
R6690	T22960	T22959	punct	/,+
R6691	T22961	T22957	pobj	mice,to
R6692	T22962	T22959	cc	and,+
R6693	T22963	T22964	punct	+,-
R6694	T22964	T22959	conj	-,+
R6695	T22965	T22964	punct	/,-
R6696	T22966	T22948	punct	.,was
R6697	T22968	T22969	det	The,numbers
R6698	T22969	T22970	nsubj	numbers,indicates
R6699	T22971	T22969	prep	on,numbers
R67	T914	T915	amod	new,treatments
R670	T3648	T3652	nsubj	components,are
R6700	T22972	T22973	det	the,bars
R6701	T22973	T22971	pobj	bars,on
R6702	T22974	T22975	det	the,number
R6703	T22975	T22970	dobj	number,indicates
R6704	T22976	T22975	prep	of,number
R6705	T22977	T22976	pobj	mice,of
R6706	T22978	T22977	acl	analyzed,mice
R6707	T22979	T22970	punct	.,indicates
R6708	T22981	T22982	punct	,*
R6709	T22983	T22981	punct	", ",*
R671	T3649	T3648	nmod	E1,components
R6710	T22984	T22985	advmod	Significantly,different
R6711	T22985	T22981	amod	different,*
R6712	T22986	T22985	prep	from,different
R6713	T22987	T22988	punct	+,+
R6714	T22988	T22986	punct	+,from
R6715	T22989	T22988	punct	/,+
R6716	T22990	T22991	punct	(,0.001
R6717	T22991	T22985	parataxis	0.001,different
R6718	T22992	T22991	nsubj	P,0.001
R6719	T22993	T22991	punct	<,0.001
R672	T3650	T3649	cc	and,E1
R6720	T22994	T22991	punct	),0.001
R6721	T22995	T22982	punct	;,**
R6722	T22996	T22982	punct	", ",**
R6723	T22997	T22998	advmod	significantly,different
R6724	T22998	T22982	amod	different,**
R6725	T22999	T22998	prep	from,different
R6726	T23000	T23001	punct	+,+
R6727	T23001	T22999	punct	+,from
R6728	T23002	T23001	punct	/,+
R6729	T23003	T23001	cc	and,+
R673	T3739	T3737	conj	E2,E1
R6730	T23004	T23005	punct	+,-
R6731	T23005	T23001	conj	-,+
R6732	T23006	T23005	punct	/,-
R6733	T23007	T23008	punct	(,0.001
R6734	T23008	T22998	parataxis	0.001,different
R6735	T23009	T23008	nsubj	P,0.001
R6736	T23010	T23008	punct	<,0.001
R6737	T23011	T23008	punct	),0.001
R6738	T23012	T22982	punct	.,**
R6739	T23655	T23656	amod	Transgenic,production
R674	T3651	T3649	conj	E2,E1
R6740	T23657	T23656	compound	mouse,production
R6741	T23658	T23656	cc	and,production
R6742	T23659	T23656	conj	characterization,production
R6743	T23660	T23656	punct	.,production
R6744	T23662	T23663	meta	A,strategy
R6745	T23664	T23663	punct	", ",strategy
R6746	T23665	T23663	amod	Transgenic,strategy
R6747	T23666	T23663	acl	used,strategy
R6748	T23667	T23668	aux	to,produce
R6749	T23668	T23666	advcl	produce,used
R675	T3653	T3652	acomp	specific,are
R6750	T23669	T23668	dobj	mice,produce
R6751	T23670	T23671	dep	that,express
R6752	T23671	T23669	relcl	express,mice
R6753	T23672	T23673	amod	human,E2
R6754	T23673	T23671	dobj	E2,express
R6755	T23674	T23663	punct	.,strategy
R6756	T23676	T23677	nmod	LAP,tTA
R6757	T23677	T23679	nmod	tTA,mice
R6758	T23678	T23677	punct	-,tTA
R6759	T23679	T23681	nsubjpass	mice,described
R676	T3654	T3653	prep	to,specific
R6760	T23680	T23679	amod	transgenic,mice
R6761	T23682	T23681	aux	have,described
R6762	T23683	T23681	auxpass	been,described
R6763	T23684	T23681	advmod	previously,described
R6764	T23685	T23686	punct	[,34
R6765	T23686	T23681	parataxis	34,described
R6766	T23687	T23686	punct	],34
R6767	T23688	T23681	punct	.,described
R6768	T23690	T23691	det	These,mice
R6769	T23691	T23692	nsubj	mice,express
R677	T3655	T3654	pobj	BCKDH,to
R6770	T23693	T23694	det	the,transactivator
R6771	T23694	T23692	dobj	transactivator,express
R6772	T23695	T23696	npadvmod	tetracycline,controlled
R6773	T23696	T23694	amod	controlled,transactivator
R6774	T23697	T23696	punct	-,controlled
R6775	T23698	T23694	punct	(,transactivator
R6776	T23699	T23694	appos	tTA,transactivator
R6777	T23700	T23694	punct	),transactivator
R6778	T23701	T23694	prep	from,transactivator
R6779	T23702	T23703	det	the,promoter
R678	T3740	T3736	prep	of,subunits
R6780	T23703	T23701	pobj	promoter,from
R6781	T23704	T23705	npadvmod	liver,specific
R6782	T23705	T23703	amod	specific,promoter
R6783	T23706	T23703	compound	LAP,promoter
R6784	T23707	T23692	punct	.,express
R6785	T23709	T23710	det	The,transgene
R6786	T23710	T23714	nsubj	transgene,contains
R6787	T23711	T23712	compound	TRE,E2
R6788	T23712	T23710	compound	E2,transgene
R6789	T23713	T23712	punct	-,E2
R679	T3656	T3652	punct	", ",are
R6790	T23715	T23716	det	the,element
R6791	T23716	T23714	dobj	element,contains
R6792	T23717	T23716	compound	tetracycline,element
R6793	T23718	T23716	compound	response,element
R6794	T23719	T23716	punct	(,element
R6795	T23720	T23716	appos	TRE,element
R6796	T23721	T23716	punct	),element
R6797	T23722	T23716	prep	as,element
R6798	T23723	T23722	pobj	part,as
R6799	T23724	T23723	prep	of,part
R68	T915	T913	dobj	treatments,developing
R680	T3657	T3658	mark	whereas,used
R6800	T23725	T23726	det	the,promoter
R6801	T23726	T23724	pobj	promoter,of
R6802	T23727	T23716	punct	", ",element
R6803	T23728	T23729	det	a,intron
R6804	T23729	T23716	conj	intron,element
R6805	T23730	T23729	amod	synthetic,intron
R6806	T23731	T23732	punct	(,line
R6807	T23732	T23729	parataxis	line,intron
R6808	T23733	T23732	amod	thin,line
R6809	T23734	T23732	punct	),line
R681	T3658	T3652	advcl	used,are
R6810	T23735	T23729	punct	", ",intron
R6811	T23736	T23737	det	the,cDNA
R6812	T23737	T23729	conj	cDNA,intron
R6813	T23738	T23737	amod	human,cDNA
R6814	T23739	T23737	compound	E2,cDNA
R6815	T23740	T23737	punct	", ",cDNA
R6816	T23741	T23742	det	an,spacer
R6817	T23742	T23737	conj	spacer,cDNA
R6818	T23743	T23742	compound	alanine,spacer
R6819	T23744	T23742	punct	", ",spacer
R682	T3741	T3740	pobj	BCKDH,of
R6820	T23745	T23746	det	a,tag
R6821	T23746	T23742	conj	tag,spacer
R6822	T23747	T23748	compound	c,myc
R6823	T23748	T23746	compound	myc,tag
R6824	T23749	T23748	punct	-,myc
R6825	T23750	T23746	compound	epitope,tag
R6826	T23751	T23746	punct	", ",tag
R6827	T23752	T23746	cc	and,tag
R6828	T23753	T23754	npadvmod	SV40,derived
R6829	T23754	T23755	amod	derived,sequence
R683	T3659	T3658	nsubjpass	E3,used
R6830	T23755	T23746	conj	sequence,tag
R6831	T23756	T23755	compound	polyadenylation,sequence
R6832	T23757	T23714	punct	.,contains
R6833	T23759	T23760	det	This,construct
R6834	T23760	T23761	nsubjpass	construct,used
R6835	T23762	T23761	auxpass	was,used
R6836	T23763	T23764	aux	to,create
R6837	T23764	T23761	advcl	create,used
R6838	T23765	T23766	amod	several,lines
R6839	T23766	T23764	dobj	lines,create
R684	T3660	T3658	auxpass	is,used
R6840	T23767	T23766	prep	of,lines
R6841	T23768	T23769	amod	transgenic,mice
R6842	T23769	T23767	pobj	mice,of
R6843	T23770	T23761	punct	.,used
R6844	T23772	T23773	meta	B,analysis
R6845	T23774	T23773	punct	", ",analysis
R6846	T23775	T23776	compound	Western,blot
R6847	T23776	T23773	compound	blot,analysis
R6848	T23777	T23773	prep	of,analysis
R6849	T23778	T23779	compound	E2,protein
R685	T3661	T3658	advmod	also,used
R6850	T23779	T23777	pobj	protein,of
R6851	T23780	T23779	prep	in,protein
R6852	T23781	T23780	pobj	liver,in
R6853	T23782	T23781	prep	of,liver
R6854	T23783	T23784	nmod	control,mice
R6855	T23784	T23782	pobj	mice,of
R6856	T23785	T23783	cc	and,control
R6857	T23786	T23783	conj	intermediate,control
R6858	T23787	T23784	compound	MSUD,mice
R6859	T23788	T23773	punct	.,analysis
R686	T3742	T3730	aux	have,described
R6860	T23791	T23792	mark	that,was
R6861	T23792	T23790	ccomp	was,Note
R6862	T23793	T23794	det	the,amount
R6863	T23794	T23792	nsubj	amount,was
R6864	T23795	T23794	prep	of,amount
R6865	T23796	T23797	amod	human,protein
R6866	T23797	T23795	pobj	protein,of
R6867	T23798	T23797	compound	E2,protein
R6868	T23799	T23800	punct	(,MW
R6869	T23800	T23797	parataxis	MW,protein
R687	T3662	T3658	agent	by,used
R6870	T23801	T23800	amod	predicted,MW
R6871	T23802	T23803	punct	~,54
R6872	T23803	T23804	nummod	54,Kd
R6873	T23804	T23800	appos	Kd,MW
R6874	T23805	T23800	punct	),MW
R6875	T23806	T23794	prep	in,amount
R6876	T23807	T23806	pobj	mice,in
R6877	T23808	T23807	prep	from,mice
R6878	T23809	T23810	compound	Lines,A
R6879	T23810	T23808	pobj	A,from
R688	T3663	T3664	nmod	pyruvate,complexes
R6880	T23811	T23810	cc	and,A
R6881	T23812	T23813	nummod	525,A
R6882	T23813	T23810	conj	A,A
R6883	T23814	T23792	acomp	variable,was
R6884	T23815	T23792	cc	but,was
R6885	T23816	T23817	prep	in,was
R6886	T23817	T23792	conj	was,was
R6887	T23818	T23816	pobj	many,in
R6888	T23819	T23818	prep	of,many
R6889	T23820	T23821	det	the,animals
R689	T3664	T3662	pobj	complexes,by
R6890	T23821	T23819	pobj	animals,of
R6891	T23822	T23823	det	the,amount
R6892	T23823	T23817	nsubj	amount,was
R6893	T23824	T23817	acomp	similar,was
R6894	T23825	T23824	prep	to,similar
R6895	T23826	T23827	det	the,amount
R6896	T23827	T23825	pobj	amount,to
R6897	T23828	T23827	prep	of,amount
R6898	T23829	T23830	compound	mouse,E2
R6899	T23830	T23828	pobj	E2,of
R69	T916	T911	aux	has,been
R690	T3743	T3730	auxpass	been,described
R6900	T23831	T23832	punct	(,Kd
R6901	T23832	T23830	parataxis	Kd,E2
R6902	T23833	T23832	nsubj	MW,Kd
R6903	T23834	T23832	punct	=,Kd
R6904	T23835	T23836	punct	~,47
R6905	T23836	T23832	nummod	47,Kd
R6906	T23837	T23832	punct	),Kd
R6907	T23838	T23827	prep	in,amount
R6908	T23839	T23840	compound	control,animals
R6909	T23840	T23838	pobj	animals,in
R691	T3665	T3663	cc	and,pyruvate
R6910	T23841	T23790	punct	.,Note
R6911	T23843	T23844	compound	Re,probing
R6912	T23844	T23846	nsubj	probing,confirmed
R6913	T23845	T23844	punct	-,probing
R6914	T23847	T23844	prep	with,probing
R6915	T23848	T23849	det	a,antibody
R6916	T23849	T23847	pobj	antibody,with
R6917	T23850	T23851	compound	c,myc
R6918	T23851	T23849	compound	myc,antibody
R6919	T23852	T23851	punct	-,myc
R692	T3666	T3667	compound	α,ketoglutarate
R6920	T23853	T23849	compound	tag,antibody
R6921	T23854	T23855	det	the,presence
R6922	T23855	T23846	dobj	presence,confirmed
R6923	T23856	T23855	prep	of,presence
R6924	T23857	T23858	det	the,E2
R6925	T23858	T23856	pobj	E2,of
R6926	T23859	T23860	npadvmod	transgene,derived
R6927	T23860	T23858	amod	derived,E2
R6928	T23861	T23858	punct	", ",E2
R6929	T23862	T23863	compound	c,myc
R693	T3745	T3744	punct	", ",are
R6930	T23863	T23865	npadvmod	myc,tagged
R6931	T23864	T23863	punct	-,myc
R6932	T23865	T23858	amod	tagged,E2
R6933	T23866	T23858	punct	", ",E2
R6934	T23867	T23858	amod	human,E2
R6935	T23868	T23846	prep	in,confirmed
R6936	T23869	T23870	amod	transgenic,mice
R6937	T23870	T23868	pobj	mice,in
R6938	T23871	T23868	cc	but,in
R6939	T23872	T23871	neg	not,but
R694	T3667	T3663	conj	ketoglutarate,pyruvate
R6940	T23873	T23868	conj	in,in
R6941	T23874	T23873	pobj	controls,in
R6942	T23875	T23846	punct	.,confirmed
R6943	T23877	T23878	compound	Western,blot
R6944	T23878	T23879	compound	blot,analysis
R6945	T23879	T23880	nsubj	analysis,revealed
R6946	T23881	T23879	prep	of,analysis
R6947	T23882	T23881	pobj	brain,of
R6948	T23883	T23884	punct	(,C
R6949	T23884	T23882	parataxis	C,brain
R695	T3668	T3667	punct	-,ketoglutarate
R6950	T23885	T23884	punct	),C
R6951	T23886	T23882	punct	", ",brain
R6952	T23887	T23882	conj	kidney,brain
R6953	T23888	T23889	punct	(,D
R6954	T23889	T23887	parataxis	D,kidney
R6955	T23890	T23889	punct	),D
R6956	T23891	T23887	punct	", ",kidney
R6957	T23892	T23887	cc	and,kidney
R6958	T23893	T23887	conj	muscle,kidney
R6959	T23894	T23895	punct	(,E
R696	T3669	T3664	compound	dehydrogenase,complexes
R6960	T23895	T23893	parataxis	E,muscle
R6961	T23896	T23895	punct	),E
R6962	T23897	T23898	amod	negligible,amounts
R6963	T23898	T23880	dobj	amounts,revealed
R6964	T23899	T23898	prep	of,amounts
R6965	T23900	T23901	npadvmod	transgene,derived
R6966	T23901	T23902	amod	derived,E2
R6967	T23902	T23899	pobj	E2,of
R6968	T23903	T23880	prep	in,revealed
R6969	T23904	T23905	det	those,tissues
R697	T3670	T3671	punct	[,3
R6970	T23905	T23903	pobj	tissues,in
R6971	T23906	T23880	punct	.,revealed
R6972	T23908	T23909	det	All,blots
R6973	T23909	T23910	nsubjpass	blots,re-probed
R6974	T23911	T23910	auxpass	were,re-probed
R6975	T23912	T23910	prep	with,re-probed
R6976	T23913	T23914	det	an,antibody
R6977	T23914	T23912	pobj	antibody,with
R6978	T23915	T23914	compound	actin,antibody
R6979	T23916	T23917	aux	to,allow
R698	T3671	T3664	parataxis	3,complexes
R6980	T23917	T23910	advcl	allow,re-probed
R6981	T23918	T23919	nsubjpass	amount,compared
R6982	T23919	T23917	ccomp	compared,allow
R6983	T23920	T23918	prep	of,amount
R6984	T23921	T23920	pobj	protein,of
R6985	T23922	T23921	acl	loaded,protein
R6986	T23923	T23922	prep	in,loaded
R6987	T23924	T23925	det	each,lane
R6988	T23925	T23923	pobj	lane,in
R6989	T23926	T23919	aux	to,compared
R699	T3746	T3744	advmod	however,are
R6990	T23927	T23919	auxpass	be,compared
R6991	T23928	T23910	punct	.,re-probed
R6992	T24446	T24445	prep	of,Characterization
R6993	T24447	T24448	det	the,model
R6994	T24448	T24446	pobj	model,of
R6995	T24449	T24448	amod	intermediate,model
R6996	T24450	T24448	nmod	MSUD,model
R6997	T24451	T24448	amod	murine,model
R6998	T24452	T24445	punct	.,Characterization
R6999	T24454	T24455	meta	A,analysis
R7	T842	T843	amod	classic,disease
R70	T917	T918	det	the,lack
R700	T3672	T3671	punct	],3
R7000	T24456	T24455	punct	", ",analysis
R7001	T24457	T24455	compound	Survival,analysis
R7002	T24458	T24455	prep	of,analysis
R7003	T24459	T24460	amod	transgenic,lines
R7004	T24460	T24458	pobj	lines,of
R7005	T24461	T24460	compound	rescue,lines
R7006	T24462	T24455	punct	.,analysis
R7007	T24464	T24465	dep	Presented,are
R7008	T24466	T24465	nsubj	percentages,are
R7009	T24467	T24466	prep	of,percentages
R701	T3673	T3664	cc	and,complexes
R7010	T24468	T24467	pobj	mice,of
R7011	T24469	T24468	amod	alive,mice
R7012	T24470	T24469	prep	at,alive
R7013	T24471	T24470	pobj	weaning,at
R7014	T24472	T24468	prep	from,mice
R7015	T24473	T24474	det	each,line
R7016	T24474	T24472	pobj	line,from
R7017	T24475	T24474	amod	transgenic,line
R7018	T24476	T24474	acl	tested,line
R7019	T24477	T24478	dep	that,had
R702	T3674	T3675	det	the,system
R7020	T24478	T24476	ccomp	had,tested
R7021	T24479	T24480	det	the,genotype
R7022	T24480	T24478	dobj	genotype,had
R7023	T24481	T24480	compound	rescue,genotype
R7024	T24482	T24483	punct	(,homozygous
R7025	T24483	T24478	parataxis	homozygous,had
R7026	T24484	T24483	advmod	i.e.,homozygous
R7027	T24485	T24483	punct	", ",homozygous
R7028	T24486	T24483	prep	for,homozygous
R7029	T24487	T24486	pobj	knockout,for
R703	T3675	T3664	conj	system,complexes
R7030	T24488	T24487	prep	of,knockout
R7031	T24489	T24490	amod	endogenous,E2
R7032	T24490	T24488	pobj	E2,of
R7033	T24491	T24483	cc	and,homozygous
R7034	T24492	T24483	conj	positive,homozygous
R7035	T24493	T24492	prep	for,positive
R7036	T24494	T24495	preconj	both,transgenes
R7037	T24495	T24493	pobj	transgenes,for
R7038	T24496	T24495	det	the,transgenes
R7039	T24497	T24498	nmod	LAP,tTA
R704	T3676	T3677	compound	glycine,cleavage
R7040	T24498	T24495	nmod	tTA,transgenes
R7041	T24499	T24498	punct	-,tTA
R7042	T24500	T24498	cc	and,tTA
R7043	T24501	T24502	compound	TRE,E2
R7044	T24502	T24498	conj	E2,tTA
R7045	T24503	T24502	punct	-,E2
R7046	T24504	T24483	punct	),homozygous
R7047	T24505	T24465	punct	.,are
R7048	T24507	T24508	advmod	Also,is
R7049	T24509	T24508	dep	plotted,is
R705	T3677	T3675	compound	cleavage,system
R7050	T24510	T24511	det	the,frequency
R7051	T24511	T24508	nsubj	frequency,is
R7052	T24512	T24511	amod	theoretical,frequency
R7053	T24513	T24511	amod	maximum,frequency
R7054	T24514	T24515	prep	at,expected
R7055	T24515	T24511	relcl	expected,frequency
R7056	T24516	T24514	pobj	which,at
R7057	T24517	T24518	det	this,genotype
R7058	T24518	T24515	nsubjpass	genotype,expected
R7059	T24519	T24515	auxpass	is,expected
R706	T3678	T3679	punct	[,4
R7060	T24520	T24515	prep	in,expected
R7061	T24521	T24522	det	this,population
R7062	T24522	T24520	pobj	population,in
R7063	T24523	T24522	prep	of,population
R7064	T24524	T24523	pobj	animals,of
R7065	T24525	T24508	punct	.,is
R7066	T24527	T24528	det	The,numbers
R7067	T24528	T24529	nsubj	numbers,indicates
R7068	T24530	T24528	prep	on,numbers
R7069	T24531	T24532	det	the,bars
R707	T3747	T3744	punct	", ",are
R7070	T24532	T24530	pobj	bars,on
R7071	T24533	T24534	det	the,numbers
R7072	T24534	T24529	dobj	numbers,indicates
R7073	T24535	T24534	prep	of,numbers
R7074	T24536	T24535	pobj	observations,of
R7075	T24537	T24534	prep	for,numbers
R7076	T24538	T24539	det	each,line
R7077	T24539	T24537	pobj	line,for
R7078	T24540	T24529	punct	.,indicates
R7079	T24542	T24543	meta	B,Ratio
R708	T3679	T3675	parataxis	4,system
R7080	T24544	T24543	punct	", ",Ratio
R7081	T24545	T24543	prep	of,Ratio
R7082	T24546	T24547	amod	total,BCAA
R7083	T24547	T24545	pobj	BCAA,of
R7084	T24548	T24543	prep	to,Ratio
R7085	T24549	T24548	pobj	alanine,to
R7086	T24550	T24543	prep	in,Ratio
R7087	T24551	T24550	pobj	blood,in
R7088	T24552	T24551	prep	of,blood
R7089	T24553	T24552	pobj	mice,of
R709	T3680	T3679	punct	],4
R7090	T24554	T24543	prep	from,Ratio
R7091	T24555	T24554	pobj	controls,from
R7092	T24556	T24555	cc	and,controls
R7093	T24557	T24558	compound	Lines,A
R7094	T24558	T24555	conj	A,controls
R7095	T24559	T24558	cc	and,A
R7096	T24560	T24558	conj	525A,A
R7097	T24561	T24543	punct	.,Ratio
R7098	T24563	T24564	amod	Total,BCAA
R7099	T24564	T24565	nsubj	BCAA,represent
R71	T918	T911	attr	lack,been
R710	T3681	T3652	punct	.,are
R7100	T24566	T24567	det	the,sum
R7101	T24567	T24565	dobj	sum,represent
R7102	T24568	T24567	prep	of,sum
R7103	T24569	T24568	pobj	leucine,of
R7104	T24570	T24569	punct	", ",leucine
R7105	T24571	T24569	conj	isoleucine,leucine
R7106	T24572	T24571	punct	", ",isoleucine
R7107	T24573	T24571	cc	and,isoleucine
R7108	T24574	T24571	conj	valine,isoleucine
R7109	T24575	T24565	punct	.,represent
R711	T3748	T3749	det	the,majority
R7110	T24577	T24578	compound	BCAA,alanine
R7111	T24578	T24580	compound	alanine,values
R7112	T24579	T24578	punct	/,alanine
R7113	T24580	T24581	nsubj	values,were
R7114	T24582	T24583	advmod	significantly,greater
R7115	T24583	T24581	acomp	greater,were
R7116	T24584	T24583	prep	for,greater
R7117	T24585	T24586	compound	Lines,A
R7118	T24586	T24584	pobj	A,for
R7119	T24587	T24586	cc	and,A
R712	T3683	T3684	nsubjpass	BCKDH,associated
R7120	T24588	T24586	conj	525A,A
R7121	T24589	T24583	prep	compared,greater
R7122	T24590	T24589	prep	to,compared
R7123	T24591	T24590	pobj	controls,to
R7124	T24592	T24581	prep	at,were
R7125	T24593	T24594	det	all,ages
R7126	T24594	T24592	pobj	ages,at
R7127	T24595	T24594	acl	tested,ages
R7128	T24596	T24581	punct	.,were
R7129	T24598	T24599	det	The,numbers
R713	T3749	T3744	nsubj	majority,are
R7130	T24599	T24600	nsubj	numbers,indicates
R7131	T24601	T24599	prep	on,numbers
R7132	T24602	T24603	det	the,bars
R7133	T24603	T24601	pobj	bars,on
R7134	T24604	T24605	det	the,numbers
R7135	T24605	T24600	dobj	numbers,indicates
R7136	T24606	T24605	prep	of,numbers
R7137	T24607	T24606	pobj	samples,of
R7138	T24608	T24607	acl	analyzed,samples
R7139	T24609	T24600	punct	.,indicates
R714	T3685	T3684	auxpass	is,associated
R7140	T24611	T24612	punct	*,0.01
R7141	T24612	T24616	ccomp	0.01,0.005
R7142	T24613	T24612	punct	", ",0.01
R7143	T24614	T24612	nsubj	P,0.01
R7144	T24615	T24612	punct	≤,0.01
R7145	T24617	T24616	punct	;,0.005
R7146	T24618	T24616	punct	**,0.005
R7147	T24619	T24616	punct	", ",0.005
R7148	T24620	T24616	nsubj	P,0.005
R7149	T24621	T24616	punct	<,0.005
R715	T3686	T3684	advmod	also,associated
R7150	T24622	T24616	punct	.,0.005
R7151	T24624	T24625	meta	C,activity
R7152	T24626	T24625	punct	", ",activity
R7153	T24627	T24625	compound	BCKDH,activity
R7154	T24628	T24625	compound	enzyme,activity
R7155	T24629	T24625	prep	in,activity
R7156	T24630	T24629	pobj	liver,in
R7157	T24631	T24630	prep	of,liver
R7158	T24632	T24633	nmod	control,mice
R7159	T24633	T24631	pobj	mice,of
R716	T3750	T3749	prep	of,majority
R7160	T24634	T24632	cc	and,control
R7161	T24635	T24636	compound	Lines,A
R7162	T24636	T24632	conj	A,control
R7163	T24637	T24636	cc	and,A
R7164	T24638	T24636	conj	525A,A
R7165	T24639	T24625	punct	.,activity
R7166	T24641	T24642	punct	*,0.001
R7167	T24642	T24646	ccomp	0.001,0.0001
R7168	T24643	T24642	punct	", ",0.001
R7169	T24644	T24642	nsubj	P,0.001
R717	T3687	T3684	prep	with,associated
R7170	T24645	T24642	punct	≤,0.001
R7171	T24647	T24646	punct	;,0.0001
R7172	T24648	T24646	punct	**,0.0001
R7173	T24649	T24646	punct	", ",0.0001
R7174	T24650	T24646	nsubj	P,0.0001
R7175	T24651	T24646	punct	≤,0.0001
R7176	T24652	T24646	punct	.,0.0001
R7177	T24654	T24655	meta	D,curves
R7178	T24656	T24655	punct	", ",curves
R7179	T24657	T24655	compound	Survival,curves
R718	T3688	T3689	nummod	two,proteins
R7180	T24658	T24655	prep	for,curves
R7181	T24659	T24658	pobj	mice,for
R7182	T24660	T24659	prep	from,mice
R7183	T24661	T24662	compound	Lines,A
R7184	T24662	T24660	pobj	A,from
R7185	T24663	T24662	cc	and,A
R7186	T24664	T24662	conj	525A,A
R7187	T24665	T24655	punct	.,curves
R7188	T24667	T24668	compound	Rescue,mice
R7189	T24668	T24669	nsubjpass	mice,monitored
R719	T3689	T3687	pobj	proteins,with
R7190	T24670	T24668	amod	alive,mice
R7191	T24671	T24670	prep	at,alive
R7192	T24672	T24671	pobj	weaning,at
R7193	T24673	T24669	auxpass	were,monitored
R7194	T24674	T24675	mark	until,were
R7195	T24675	T24669	advcl	were,monitored
R7196	T24676	T24675	nsubj	they,were
R7197	T24677	T24675	acomp	moribund,were
R7198	T24678	T24677	cc	and,moribund
R7199	T24679	T24680	advmod	subsequently,sacrificed
R72	T919	T918	prep	of,lack
R720	T3751	T3752	compound	MSUD,mutations
R7200	T24680	T24677	conj	sacrificed,moribund
R7201	T24681	T24675	cc	or,were
R7202	T24682	T24683	auxpass	were,found
R7203	T24683	T24675	conj	found,were
R7204	T24684	T24683	advcl	dead,found
R7205	T24685	T24683	prep	in,found
R7206	T24686	T24687	poss	their,cages
R7207	T24687	T24685	pobj	cages,in
R7208	T24688	T24669	punct	.,monitored
R7209	T24775	T24774	prep	of,Summary
R721	T3690	T3689	amod	regulatory,proteins
R7210	T24776	T24777	amod	additional,levels
R7211	T24777	T24775	pobj	levels,of
R7212	T24778	T24777	compound	blood,levels
R7213	T24779	T24780	compound	amino,acid
R7214	T24780	T24777	compound	acid,levels
R7215	T24781	T24777	prep	in,levels
R7216	T24782	T24783	det	the,model
R7217	T24783	T24781	pobj	model,in
R7218	T24784	T24783	amod	classic,model
R7219	T24785	T24783	compound	MSUD,model
R722	T3691	T3689	punct	", ",proteins
R7220	T24786	T24783	cc	and,model
R7221	T24787	T24788	compound	control,littermates
R7222	T24788	T24783	conj	littermates,model
R7223	T24789	T24790	mark	as,determined
R7224	T24790	T24777	advcl	determined,levels
R7225	T24791	T24790	prep	by,determined
R7226	T24792	T24793	amod	tandem,spectrometry
R7227	T24793	T24791	pobj	spectrometry,by
R7228	T24794	T24793	compound	mass,spectrometry
R7229	T24795	T24774	punct	.,Summary
R723	T3692	T3693	det	a,kinase
R7230	T24797	T24798	det	All,samples
R7231	T24798	T24799	nsubjpass	samples,collected
R7232	T24800	T24799	auxpass	were,collected
R7233	T24801	T24799	prep	on,collected
R7234	T24802	T24803	det	the,day
R7235	T24803	T24801	pobj	day,on
R7236	T24804	T24803	prep	of,day
R7237	T24805	T24804	pobj	birth,of
R7238	T24806	T24799	punct	.,collected
R7239	T24808	T24809	det	All,values
R724	T3693	T3689	appos	kinase,proteins
R7240	T24809	T24810	nsubj	values,are
R7241	T24811	T24810	attr	mean,are
R7242	T24812	T24811	cc	+,mean
R7243	T24813	T24812	punct	/,+
R7244	T24814	T24812	punct	-,+
R7245	T24815	T24811	conj	SEM,mean
R7246	T24816	T24810	punct	.,are
R7247	T24915	T24914	prep	of,Summary
R7248	T24916	T24917	amod	additional,levels
R7249	T24917	T24915	pobj	levels,of
R725	T3752	T3750	pobj	mutations,of
R7250	T24918	T24917	compound	blood,levels
R7251	T24919	T24920	compound	amino,acid
R7252	T24920	T24917	compound	acid,levels
R7253	T24921	T24917	prep	in,levels
R7254	T24922	T24923	det	the,model
R7255	T24923	T24921	pobj	model,in
R7256	T24924	T24923	amod	intermediate,model
R7257	T24925	T24923	compound	MSUD,model
R7258	T24926	T24923	cc	and,model
R7259	T24927	T24928	compound	littermate,controls
R726	T3694	T3693	amod	specific,kinase
R7260	T24928	T24923	conj	controls,model
R7261	T24929	T24930	mark	as,determined
R7262	T24930	T24917	advcl	determined,levels
R7263	T24931	T24930	prep	by,determined
R7264	T24932	T24933	amod	tandem,spectrometry
R7265	T24933	T24931	pobj	spectrometry,by
R7266	T24934	T24933	compound	mass,spectrometry
R7267	T24935	T24914	punct	.,Summary
R7268	T24937	T24938	nsubjpass	Samples,collected
R7269	T24939	T24938	auxpass	were,collected
R727	T3695	T3693	cc	and,kinase
R7270	T24940	T24938	prep	from,collected
R7271	T24941	T24940	pobj	mice,from
R7272	T24942	T24943	dep	that,were
R7273	T24943	T24941	relcl	were,mice
R7274	T24944	T24945	quantmod	4,6
R7275	T24945	T24947	nummod	6,weeks
R7276	T24946	T24945	punct	–,6
R7277	T24947	T24943	attr	weeks,were
R7278	T24948	T24947	prep	of,weeks
R7279	T24949	T24948	pobj	age,of
R728	T3753	T3752	acl	identified,mutations
R7280	T24950	T24938	punct	.,collected
R7281	T24952	T24953	det	All,values
R7282	T24953	T24954	nsubj	values,are
R7283	T24955	T24954	attr	mean,are
R7284	T24956	T24955	cc	+,mean
R7285	T24957	T24956	punct	/,+
R7286	T24958	T24956	punct	-,+
R7287	T24959	T24955	conj	SEM,mean
R7288	T24960	T24954	punct	.,are
R729	T3696	T3697	det	a,phosphatase
R73	T920	T921	det	a,model
R730	T3697	T3693	conj	phosphatase,kinase
R731	T3698	T3699	dep	which,regulate
R732	T3699	T3689	relcl	regulate,proteins
R733	T3754	T3755	advmod	thus,far
R734	T3700	T3701	det	the,activity
R735	T3701	T3699	dobj	activity,regulate
R736	T3702	T3701	prep	of,activity
R737	T3755	T3753	advmod	far,identified
R738	T3703	T3704	det	this,enzyme
R739	T3704	T3702	pobj	enzyme,of
R74	T921	T919	pobj	model,of
R740	T3705	T3699	prep	through,regulate
R741	T3756	T3744	prep	in,are
R742	T3706	T3707	det	a,cycle
R743	T3707	T3705	pobj	cycle,through
R744	T3757	T3758	det	the,subunit
R745	T3708	T3707	nmod	phosphorylation,cycle
R746	T3709	T3710	punct	(,inactivation
R747	T3710	T3708	parataxis	inactivation,phosphorylation
R748	T3758	T3756	pobj	subunit,in
R749	T3711	T3710	punct	),inactivation
R75	T922	T921	amod	suitable,model
R750	T3712	T3708	cc	and,phosphorylation
R751	T3713	T3708	conj	dephosphorylation,phosphorylation
R752	T3759	T3758	compound	E2,subunit
R753	T3714	T3715	punct	(,activation
R754	T3715	T3713	parataxis	activation,dephosphorylation
R755	T3716	T3715	punct	),activation
R756	T3717	T3707	prep	of,cycle
R757	T3718	T3719	det	the,subunit
R758	T3719	T3717	pobj	subunit,of
R759	T3760	T3761	punct	[,7
R76	T923	T921	compound	animal,model
R760	T3720	T3719	compound	E1,subunit
R761	T3721	T3719	compound	α,subunit
R762	T3722	T3723	punct	[,6
R763	T3761	T3744	parataxis	7,are
R764	T3723	T3684	parataxis	6,associated
R765	T3762	T3761	nummod	1,7
R766	T3724	T3723	nummod	5,6
R767	T3725	T3723	punct	",",6
R768	T3726	T3723	punct	],6
R769	T3763	T3761	punct	",",7
R77	T924	T911	punct	.,been
R770	T3727	T3684	punct	.,associated
R771	T3764	T3761	punct	],7
R772	T3765	T3744	punct	.,are
R773	T3767	T3768	prep	To,associated
R774	T3729	T3730	nsubjpass	Mutations,described
R775	T3730	T3744	ccomp	described,are
R776	T3731	T3729	prep	in,Mutations
R777	T3769	T3767	pobj	date,To
R778	T3732	T3733	det	the,genes
R779	T3733	T3731	pobj	genes,in
R78	T928	T929	aux	To,create
R780	T3734	T3733	prep	of,genes
R781	T3735	T3736	det	the,subunits
R782	T3770	T3768	punct	", ",associated
R783	T3736	T3734	pobj	subunits,of
R784	T3737	T3736	nmod	E1,subunits
R785	T3738	T3737	cc	and,E1
R786	T3771	T3768	nsubjpass	cases,associated
R787	T3772	T3771	prep	of,cases
R788	T3773	T3772	pobj	MSUD,of
R789	T3845	T3843	pobj	spite,In
R79	T929	T930	advcl	create,used
R790	T3774	T3768	aux	have,associated
R791	T3846	T3845	prep	of,spite
R792	T3847	T3848	amod	dietary,intervention
R793	T3775	T3768	neg	not,associated
R794	T3848	T3846	pobj	intervention,of
R795	T3849	T3844	punct	", ",is
R796	T3850	T3844	expl	there,is
R797	T3776	T3768	auxpass	been,associated
R798	T3851	T3852	amod	significant,mortality
R799	T3852	T3844	attr	mortality,is
R8	T843	T841	pobj	disease,of
R80	T931	T932	det	a,model
R800	T3777	T3768	prep	with,associated
R801	T3853	T3852	acl	associated,mortality
R802	T3854	T3853	prep	with,associated
R803	T3778	T3777	pobj	defects,with
R804	T3855	T3854	pobj	MSUD,with
R805	T3856	T3844	cc	and,is
R806	T3857	T3858	expl	there,is
R807	T3858	T3844	conj	is,is
R808	T3859	T3860	det	a,incidence
R809	T3860	T3858	attr	incidence,is
R81	T932	T929	dobj	model,create
R810	T3779	T3778	prep	in,defects
R811	T3861	T3860	amod	high,incidence
R812	T3862	T3860	prep	of,incidence
R813	T3863	T3864	amod	mental,retardation
R814	T3864	T3862	pobj	retardation,of
R815	T3780	T3781	det	the,kinase
R816	T3865	T3858	prep	in,is
R817	T3866	T3865	pobj	survivors,in
R818	T3867	T3868	punct	[,9
R819	T3781	T3779	pobj	kinase,in
R82	T933	T932	amod	murine,model
R820	T3868	T3858	parataxis	9,is
R821	T3869	T3868	punct	],9
R822	T3870	T3844	punct	.,is
R823	T3782	T3781	amod	regulatory,kinase
R824	T3872	T3873	prep	Because,treated
R825	T3783	T3781	cc	and,kinase
R826	T3874	T3872	pcomp	of,Because
R827	T3875	T3876	det	the,role
R828	T3784	T3781	conj	phosphatase,kinase
R829	T3876	T3872	pobj	role,Because
R83	T934	T932	prep	of,model
R830	T3877	T3876	amod	central,role
R831	T3878	T3876	prep	of,role
R832	T3785	T3786	punct	[,1
R833	T3879	T3880	det	the,liver
R834	T3880	T3878	pobj	liver,of
R835	T3881	T3876	prep	in,role
R836	T3786	T3768	parataxis	1,associated
R837	T3882	T3883	compound	amino,acid
R838	T3883	T3884	compound	acid,metabolism
R839	T3884	T3881	pobj	metabolism,in
R84	T935	T936	amod	classic,MSUD
R840	T3787	T3786	punct	],1
R841	T3885	T3876	cc	and,role
R842	T3886	T3887	amod	moderate,high
R843	T3887	T3889	amod	high,levels
R844	T3788	T3768	punct	.,associated
R845	T3888	T3887	punct	/,high
R846	T3889	T3876	conj	levels,role
R847	T3890	T3889	prep	of,levels
R848	T3790	T3791	amod	Current,management
R849	T3891	T3892	compound	BCKDH,activity
R85	T936	T934	pobj	MSUD,of
R850	T3892	T3890	pobj	activity,of
R851	T3893	T3892	prep	in,activity
R852	T3894	T3895	amod	human,liver
R853	T3791	T3792	nsubj	management,relies
R854	T3895	T3893	pobj	liver,in
R855	T3793	T3791	prep	of,management
R856	T3896	T3897	punct	[,10
R857	T3794	T3795	compound	MSUD,patients
R858	T3897	T3889	parataxis	10,levels
R859	T3898	T3899	punct	-,12
R86	T937	T930	punct	", ",used
R860	T3899	T3897	prep	12,10
R861	T3900	T3897	punct	],10
R862	T3901	T3873	punct	", ",treated
R863	T3902	T3903	det	a,cases
R864	T3795	T3793	pobj	patients,of
R865	T3903	T3873	nsubjpass	cases,treated
R866	T3904	T3903	amod	few,cases
R867	T3796	T3792	prep	on,relies
R868	T3905	T3903	prep	of,cases
R869	T3906	T3905	pobj	MSUD,of
R87	T938	T930	nsubj	we,used
R870	T3907	T3873	aux	have,treated
R871	T3797	T3798	det	a,restriction
R872	T3908	T3873	advmod	recently,treated
R873	T3909	T3873	auxpass	been,treated
R874	T3910	T3873	prep	by,treated
R875	T3798	T3796	pobj	restriction,on
R876	T3911	T3912	compound	liver,transplantation
R877	T3799	T3798	amod	strict,restriction
R878	T3912	T3910	pobj	transplantation,by
R879	T3800	T3798	amod	lifelong,restriction
R88	T939	T940	compound	gene,targeting
R880	T3913	T3914	punct	[,13
R881	T3801	T3798	amod	dietary,restriction
R882	T3914	T3873	parataxis	13,treated
R883	T3915	T3916	punct	-,16
R884	T3916	T3914	prep	16,13
R885	T3802	T3798	prep	of,restriction
R886	T3917	T3914	punct	],13
R887	T3918	T3873	punct	.,treated
R888	T3803	T3802	pobj	protein,of
R889	T3920	T3921	mark	While,is
R89	T940	T930	dobj	targeting,used
R890	T3921	T3930	advcl	is,known
R891	T3922	T3923	det	the,outcome
R892	T3804	T3803	cc	or,protein
R893	T3923	T3921	nsubj	outcome,is
R894	T3924	T3925	amod	short,term
R895	T3925	T3923	compound	term,outcome
R896	T3805	T3803	conj	BCAA,protein
R897	T3926	T3925	punct	-,term
R898	T3927	T3923	prep	of,outcome
R899	T3928	T3929	compound	liver,transplantation
R9	T844	T842	cc	and,classic
R90	T941	T940	cc	and,targeting
R900	T3806	T3807	punct	[,8
R901	T3929	T3927	pobj	transplantation,of
R902	T3931	T3921	acomp	encouraging,is
R903	T3807	T3792	parataxis	8,relies
R904	T3932	T3930	punct	", ",known
R905	T3933	T3934	amod	long,term
R906	T3934	T3936	compound	term,effects
R907	T3808	T3807	nummod	1,8
R908	T3935	T3934	punct	-,term
R909	T3936	T3930	nsubjpass	effects,known
R91	T942	T943	amod	embryonic,cell
R910	T3809	T3807	punct	",",8
R911	T3937	T3936	prep	of,effects
R912	T3938	T3939	det	this,approach
R913	T3939	T3937	pobj	approach,of
R914	T3940	T3930	auxpass	are,known
R915	T3941	T3930	neg	not,known
R916	T3942	T3930	punct	.,known
R917	T3810	T3807	punct	],8
R918	T3944	T3945	advmod	However,required
R919	T3946	T3945	punct	", ",required
R92	T943	T945	compound	cell,technologies
R920	T3811	T3792	punct	.,relies
R921	T3947	T3948	det	these,patients
R922	T3948	T3945	nsubjpass	patients,required
R923	T3949	T3945	auxpass	are,required
R924	T3813	T3814	predet	Such,management
R925	T3950	T3945	advmod	now,required
R926	T3814	T3817	nsubj	management,is
R927	T3815	T3814	det	a,management
R928	T3951	T3952	aux	to,take
R929	T3952	T3945	xcomp	take,required
R93	T944	T943	compound	stem,cell
R930	T3953	T3954	compound	immunosuppressant,drugs
R931	T3816	T3814	amod	dietary,management
R932	T3954	T3952	dobj	drugs,take
R933	T3955	T3952	prep	for,take
R934	T3818	T3814	prep	of,management
R935	T3956	T3957	det	the,rest
R936	T3957	T3955	pobj	rest,for
R937	T3958	T3957	prep	of,rest
R938	T3959	T3960	poss	their,lives
R939	T3819	T3820	det	the,disease
R94	T945	T940	conj	technologies,targeting
R940	T3960	T3958	pobj	lives,of
R941	T3961	T3952	punct	", ",take
R942	T3962	T3963	advmod	often,with
R943	T3820	T3818	pobj	disease,of
R944	T3963	T3952	prep	with,take
R945	T3964	T3965	amod	undesirable,effects
R946	T3965	T3963	pobj	effects,with
R947	T3821	T3817	punct	", ",is
R948	T3966	T3965	compound	side,effects
R949	T3967	T3945	punct	.,required
R95	T946	T947	aux	to,create
R950	T3822	T3817	advmod	however,is
R951	T3969	T3970	advmod	Moreover,are
R952	T3823	T3817	punct	", ",is
R953	T3971	T3970	punct	", ",are
R954	T3972	T3973	det	the,cost
R955	T3973	T3970	nsubj	cost,are
R956	T3824	T3817	neg	not,is
R957	T3974	T3973	acl	associated,cost
R958	T3975	T3974	prep	with,associated
R959	T3976	T3977	compound	liver,transplantation
R96	T947	T930	advcl	create,used
R960	T3825	T3826	advmod	entirely,satisfactory
R961	T3977	T3975	pobj	transplantation,with
R962	T3978	T3973	cc	and,cost
R963	T3979	T3980	det	the,availability
R964	T3980	T3973	conj	availability,cost
R965	T3981	T3980	prep	of,availability
R966	T3826	T3817	acomp	satisfactory,is
R967	T3827	T3828	advmod	especially,in
R968	T3982	T3983	compound	donor,livers
R969	T3828	T3817	prep	in,is
R97	T948	T949	det	a,line
R970	T3983	T3981	pobj	livers,of
R971	T3984	T3985	amod	additional,factors
R972	T3829	T3828	pobj	times,in
R973	T3985	T3970	attr	factors,are
R974	T3986	T3985	amod	limiting,factors
R975	T3987	T3985	prep	for,factors
R976	T3830	T3829	prep	of,times
R977	T3988	T3989	det	the,practicality
R978	T3989	T3987	pobj	practicality,for
R979	T3990	T3989	prep	of,practicality
R98	T949	T947	dobj	line,create
R980	T3831	T3832	amod	metabolic,decompensation
R981	T3991	T3990	pobj	treatment,of
R982	T3992	T3991	prep	of,treatment
R983	T3832	T3830	pobj	decompensation,of
R984	T3993	T3994	det	this,disease
R985	T3994	T3992	pobj	disease,of
R986	T3833	T3832	prep	due,decompensation
R987	T3995	T3970	punct	.,are
R988	T3997	T3998	prep	Because,is
R989	T3834	T3833	pcomp	to,due
R99	T950	T949	compound	mouse,line
R990	T3999	T3997	pcomp	of,Because
R991	T3835	T3833	pobj	infection,due
R992	T4000	T4001	det	the,options
R993	T4001	T3997	pobj	options,Because
R994	T3836	T3835	punct	", ",infection
R995	T4002	T4001	amod	current,options
R996	T4003	T4001	amod	unsatisfactory,options
R997	T4004	T4001	prep	for,options
R998	T4005	T4006	det	the,treatment
R999	T3837	T3835	conj	injuries,infection

PMC:1448208 JSONTXT 7 Projects

Annnotations TAB TSV DIC JSON TextAE

PMC:1448208 JSON TXT 7 Projects