;PMID: 9592195 ;source_file_1403.mrg ;Release 0.9 of PennBioIE ;Funded by NSF ITR EIA-0205448 ;0)section:[e:0..34] = [t:0..34] ;1)sentence:[e:39..199] = [t:39..199] ;2)section:[e:203..243] = [t:203..243] ;3)section:[e:247..339] = [t:247..339] ;4)sentence:[e:343..617] = [t:343..617] ;5)sentence:[e:618..906] = [t:618..906] ;6)sentence:[e:907..973] = [t:907..973] ;7)sentence:[e:974..1155] = [t:974..1155] ;8)sentence:[e:1156..1478] = [t:1156..1478] ;9)sentence:[e:1479..1632] = [t:1479..1632] ;10)section:[e:1636..1680] = [t:1636..1680] ;section 0 Span:0..34 ;Int J Oncol 1998 Jun;12(6):1333-8 (SEC (FRAG (NNP:[0..3] Int) (DT:[4..5] J) (NNP:[6..11] Oncol) (CD:[13..17] 1998) (.:[18..25] Jun;12-LRB-) (CD:[25..27] 6-RRB-) (CD:[27..32] :1333) (HYPH:[32..33] -) (CD:[33..34] 8))) ;sentence 1 Span:39..199 ;Detection of ras gene mutations in peripheral blood of carcinoma patients ;using CD45 immunomagnetic separation and nested mutant allele specific ;amplification. ;[52..55]:gene-rna:"ras" ;[94..103]:malignancy:"carcinoma" ;[120..124]:gene-protein:"CD45" (SENT (NP-HLN (NP (NN:[39..48] Detection)) (PP (IN:[49..51] of) (NP (NN:[52..55] ras) (NN:[56..60] gene) (NNS:[61..70] mutations))) (PP-LOC (IN:[71..73] in) (NP (NP (JJ:[74..84] peripheral) (NN:[85..90] blood)) (PP (IN:[91..93] of) (NP (NN:[94..103] carcinoma) (NNS:[104..112] patients))))) (S-MNR (NP-SBJ (-NONE-:[112..112] *)) (VP (VBG:[113..118] using) (NP (NP (NN:[120..124] CD45) (JJ:[125..139] immunomagnetic) (NN:[140..150] separation)) (CC:[151..154] and) (NP (ADJP (NML (VBN:[155..161] nested) (JJ:[162..168] mutant) (NN:[169..175] allele)) (JJ:[176..184] specific)) (NN:[185..198] amplification))))) (.:[198..199] .))) ;section 2 Span:203..243 ;Shibata K, Mori M, Kitano S, Akiyoshi T. (SEC (FRAG (NNP:[203..210] Shibata) (NNP:[211..212] K) (,:[212..213] ,) (NNP:[214..218] Mori) (NNP:[219..221] M,) (NNP:[222..228] Kitano) (NNP:[229..231] S,) (NNP:[232..240] Akiyoshi) (NNP:[241..243] T.))) ;section 3 Span:247..339 ;Department of Surgery, Medical Institute of Bioregulation, Kyushu University, ; Beppu, Japan. (SEC (FRAG (NNP:[247..257] Department) (IN:[258..260] of) (NNP:[261..268] Surgery) (NNP:[268..269] ,) (NNP:[270..277] Medical) (NNP:[278..287] Institute) (IN:[288..290] of) (NNP:[291..304] Bioregulation) (,:[304..305] ,) (NNP:[306..312] Kyushu) (NNP:[313..323] University) (,:[323..324] ,) (NNP:[326..331] Beppu) (,:[331..332] ,) (NNP:[333..338] Japan) (.:[338..339] .))) ;sentence 4 Span:343..617 ;We developed a sensitive technique of detecting circulating tumor cells in ;carcinoma patients, using CD45 immunomagnetic separation to isolate ;epithelial cells in blood samples and specific polymerase chain reaction ;analysis to identify point mutations of the K-ras gene. ;[403..414]:malignancy:"tumor cells" ;[419..428]:malignancy:"carcinoma" ;[445..449]:gene-protein:"CD45" ;[535..545]:gene-protein:"polymerase" ;[583..598]:variation-type:"point mutations" ;[606..611]:gene-rna:"K-ras" (SENT (S (NP-SBJ (PRP:[343..345] We)) (VP (VBD:[346..355] developed) (NP (NP (DT:[356..357] a) (JJ:[358..367] sensitive) (NN:[368..377] technique)) (PP (IN:[378..380] of) (S-NOM (NP-SBJ (-NONE-:[380..380] *)) (VP (VBG:[381..390] detecting) (NP (NP (VBG:[391..402] circulating) (NN:[403..408] tumor) (NNS:[409..414] cells)) (PP-LOC (IN:[415..417] in) (NP (NN:[419..428] carcinoma) (NNS:[429..437] patients))))))))) (,:[437..438] ,) (S-MNR (NP-SBJ (-NONE-:[438..438] *)) (VP (VP (VBG:[439..444] using) (NP (NN:[445..449] CD45) (JJ:[450..464] immunomagnetic) (NN:[465..475] separation)) (S-PRP (NP-SBJ (-NONE-:[475..475] *)) (VP (TO:[476..478] to) (VP (VB:[479..486] isolate) (NP (JJ:[487..497] epithelial) (NNS:[499..504] cells)) (PP-LOC (IN:[505..507] in) (NP (NN:[508..513] blood) (NNS:[514..521] samples))))))) (CC:[522..525] and) (VP (NP (NML (JJ:[526..534] specific) (NN:[535..545] polymerase) (NN:[546..551] chain) (NN:[552..560] reaction)) (NN:[561..569] analysis)) (S-PRP (NP-SBJ (-NONE-:[569..569] *)) (VP (TO:[570..572] to) (VP (VB:[574..582] identify) (NP (NP (NN:[583..588] point) (NNS:[589..598] mutations)) (PP (IN:[599..601] of) (NP (DT:[602..605] the) (NN:[606..611] K-ras) (NN:[612..616] gene)))))))))) (.:[616..617] .))) ;sentence 5 Span:618..906 ;The method is based on the fact that the peripheral blood mononuclear cells ;(PBMC) that express CD45 antigen are trapped with anti-CD45 conjugated ;supramagnetic microbeads while the carcinoma cells that do not express CD45 ;antigen are not trapped and pass through the magnetic fields. ;[715..727]:gene-protein:"CD45 antigen" ;[751..755]:gene-protein:"CD45" ;[802..818]:malignancy:"carcinoma cells" ;[839..851]:gene-protein:"CD45 antigen" (SENT (S (NP-SBJ-1 (DT:[618..621] The) (NN:[622..628] method)) (VP (VBZ:[629..631] is) (VP (VBN:[632..637] based) (NP-1 (-NONE-:[637..637] *)) (PP-CLR (IN:[638..640] on) (NP (DT:[641..644] the) (NN:[645..649] fact) (SBAR (IN:[650..654] that) (S (NP-SBJ-4 (NP (DT:[656..659] the) (NML (NML (JJ:[660..670] peripheral) (NN:[671..676] blood) (JJ:[677..688] mononuclear) (NNS:[689..694] cells)) (NML (-LRB-:[695..696] -LRB-) (NN:[696..700] PBMC) (-RRB-:[700..701] -RRB-)))) (SBAR (WHNP-5 (WDT:[702..706] that)) (S (NP-SBJ-5 (-NONE-:[706..706] *T*)) (VP (VBP:[707..714] express) (NP (NN:[715..719] CD45) (NN:[720..727] antigen)))))) (VP (VBP:[728..731] are) (VP (VBN:[733..740] trapped) (NP-4 (-NONE-:[740..740] *)) (PP (IN:[741..745] with) (NP (NML (AFX:[746..750] anti) (HYPH:[750..751] -) (NN:[751..755] CD45)) (VBN:[756..766] conjugated) (JJ:[767..780] supramagnetic) (NNS:[781..791] microbeads))) (SBAR-ADV (IN:[792..797] while) (S (NP-SBJ-3 (NP (DT:[798..801] the) (NN:[802..811] carcinoma) (NNS:[813..818] cells)) (SBAR (WHNP-2 (WDT:[819..823] that)) (S (NP-SBJ-2 (-NONE-:[823..823] *T*)) (VP (VBP:[824..826] do) (RB:[827..830] not) (VP (VB:[831..838] express) (NP (NN:[839..843] CD45) (NN:[844..851] antigen))))))) (VP (VP (VBP:[852..855] are) (RB:[856..859] not) (VP (VBN:[860..867] trapped) (NP-3 (-NONE-:[867..867] *)))) (CC:[868..871] and) (VP (VBP:[872..876] pass) (PP (IN:[877..884] through) (NP (DT:[885..888] the) (JJ:[890..898] magnetic) (NNS:[899..905] fields))))))))))))))) (.:[905..906] .))) ;sentence 6 Span:907..973 ;This method concentrated the number of carcinoma cells 3.3 times. ;[946..961]:malignancy:"carcinoma cells" (SENT (S (NP-SBJ (DT:[907..911] This) (NN:[912..918] method)) (VP (VBD:[919..931] concentrated) (NP (NP (DT:[932..935] the) (NN:[936..942] number)) (PP (IN:[943..945] of) (NP (NN:[946..955] carcinoma) (NNS:[956..961] cells)))) (NP-EXT (QP (CD:[962..965] 3.3) (NNS:[967..972] times)))) (.:[972..973] .))) ;sentence 7 Span:974..1155 ;After this separation, the modified method of mutant allele specific ;amplification was applied and this method was able to ten control carcinoma ;cells in a background of 107 PBMC. ;[1110..1126]:malignancy:"carcinoma cells" (SENT (S (S (PP-TMP (IN:[974..979] After) (NP (DT:[980..984] this) (NN:[985..995] separation))) (,:[995..996] ,) (NP-SBJ-1 (NP (DT:[997..1000] the) (VBN:[1001..1009] modified) (NN:[1010..1016] method)) (PP (IN:[1017..1019] of) (NP (ADJP (NML (JJ:[1020..1026] mutant) (NN:[1027..1033] allele)) (JJ:[1034..1042] specific)) (NN:[1044..1057] amplification)))) (VP (VBD:[1058..1061] was) (VP (VBN:[1062..1069] applied) (NP-1 (-NONE-:[1069..1069] *))))) (CC:[1070..1073] and) (S (NP-SBJ (DT:[1074..1078] this) (NN:[1079..1085] method)) (VP (VBD:[1086..1089] was) (ADJP-PRD (JJ:[1090..1094] able) (PP (TO:[1095..1097] to) (NP (CD:[1098..1101] ten) (NN:[1102..1109] control) (NN:[1110..1119] carcinoma) (NNS:[1121..1126] cells)))) (PP-LOC (IN:[1127..1129] in) (NP (NP (DT:[1130..1131] a) (NN:[1132..1142] background)) (PP (IN:[1143..1145] of) (NP (CD:[1146..1149] 107) (NN:[1150..1154] PBMC))))))) (.:[1154..1155] .))) ;sentence 8 Span:1156..1478 ;A preliminary clinical study demonstrated that six cases of end-stage ;carcinoma with K-ras mutations in the primary tumor showed the same ;mutations in the peripheral blood samples, while two cases without K-ras ;mutation in the primary tumor and 10 healthy volunteers showed no mutation ;in the peripheral blood samples. ;[1217..1236]:malignancy:"end-stage carcinoma" ;[1242..1247]:gene-rna:"K-ras" ;[1296..1305]:variation-event:"mutations" ;[1364..1369]:gene-rna:"K-ras" ;[1437..1445]:variation-event:"mutation" (SENT (S (NP-SBJ (DT:[1156..1157] A) (JJ:[1158..1169] preliminary) (JJ:[1170..1178] clinical) (NN:[1179..1184] study)) (VP (VBD:[1185..1197] demonstrated) (SBAR (IN:[1199..1203] that) (S (NP-SBJ (NP (CD:[1204..1207] six) (NNS:[1208..1213] cases)) (PP (IN:[1214..1216] of) (NP (NP (NML (NN:[1217..1220] end) (HYPH:[1220..1221] -) (NN:[1221..1226] stage)) (NN:[1227..1236] carcinoma)) (PP (IN:[1237..1241] with) (NP (NP (NN:[1242..1247] K-ras) (NNS:[1248..1257] mutations)) (PP-LOC (IN:[1258..1260] in) (NP (DT:[1261..1264] the) (JJ:[1265..1272] primary) (NN:[1273..1278] tumor)))))))) (VP (VBD:[1280..1286] showed) (NP (DT:[1287..1290] the) (JJ:[1291..1295] same) (NNS:[1296..1305] mutations)) (PP-LOC (IN:[1306..1308] in) (NP (DT:[1309..1312] the) (JJ:[1313..1323] peripheral) (NN:[1324..1329] blood) (NNS:[1330..1337] samples))) (,:[1337..1338] ,) (SBAR-ADV (IN:[1339..1344] while) (S (NP-SBJ (NP (NP (CD:[1345..1348] two) (NNS:[1349..1354] cases)) (PP (IN:[1356..1363] without) (NP (NP (NN:[1364..1369] K-ras) (NN:[1370..1378] mutation)) (PP (IN:[1379..1381] in) (NP (DT:[1382..1385] the) (JJ:[1386..1393] primary) (NN:[1394..1399] tumor)))))) (CC:[1400..1403] and) (NP (CD:[1404..1406] 10) (JJ:[1407..1414] healthy) (NNS:[1415..1425] volunteers))) (VP (VBD:[1426..1432] showed) (NP (DT:[1433..1435] no) (NN:[1437..1445] mutation)) (PP-LOC (IN:[1446..1448] in) (NP (DT:[1449..1452] the) (JJ:[1453..1463] peripheral) (NN:[1464..1469] blood) (NNS:[1470..1477] samples)))))))))) (.:[1477..1478] .))) ;sentence 9 Span:1479..1632 ;The results suggest that this method may be very useful to detect ;circulating carcinoma cells in the patient whose primary tumor shows K-ras ;mutations. ;[1558..1573]:malignancy:"carcinoma cells" ;[1616..1621]:gene-rna:"K-ras" (SENT (S (NP-SBJ (DT:[1479..1482] The) (NNS:[1483..1490] results)) (VP (VBP:[1491..1498] suggest) (SBAR (IN:[1499..1503] that) (S (NP-SBJ (DT:[1504..1508] this) (NN:[1509..1515] method)) (VP (MD:[1517..1520] may) (VP (VB:[1521..1523] be) (ADJP-PRD (RB:[1524..1528] very) (JJ:[1529..1535] useful)) (S-PRP (NP-SBJ (-NONE-:[1535..1535] *)) (VP (TO:[1536..1538] to) (VP (VB:[1539..1545] detect) (NP (VBG:[1546..1557] circulating) (NN:[1558..1567] carcinoma) (NNS:[1568..1573] cells)) (PP-LOC (IN:[1574..1576] in) (NP (NP (DT:[1577..1580] the) (NN:[1581..1588] patient)) (SBAR (WHNP-1 (WP$:[1589..1594] whose) (JJ:[1596..1603] primary) (NN:[1604..1609] tumor)) (S (NP-SBJ-1 (-NONE-:[1609..1609] *T*)) (VP (VBZ:[1610..1615] shows) (NP (NN:[1616..1621] K-ras) (NNS:[1622..1631] mutations))))))))))))))) (.:[1631..1632] .))) ;section 10 Span:1636..1680 ;PMID: 9592195 [PubMed - indexed for MEDLINE] (SEC (FRAG (NNP:[1636..1640] PMID) (::[1640..1641] :) (CD:[1642..1649] 9592195) (IN:[1650..1651] -LSB-) (NNP:[1651..1657] PubMed) (HYPH:[1658..1659] -) (JJ:[1660..1667] indexed) (IN:[1668..1671] for) (NNP:[1672..1680] MEDLINE-RSB-)))