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PMC:2800179 / 874-17737 JSONTXT

Annnotations TAB JSON ListView MergeView

ICD10

Id Subject Object Predicate Lexical cue
T7121 15821-15829 http://purl.bioontology.org/ontology/ICD10/T14.9 denotes injuries

GO-CC

Id Subject Object Predicate Lexical cue
T601 20-24 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T602 63-68 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T603 150-155 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T604 399-404 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T606 311-319 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T607 311-319 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T1992 1810-1814 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T1993 1946-1950 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T1994 2350-2354 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T1995 2386-2390 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T1996 2870-2874 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T1997 3226-3230 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T2807 5679-5683 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T3977 7788-7792 http://purl.obolibrary.org/obo/GO_0005623 denotes Cell
T3978 8345-8349 http://purl.obolibrary.org/obo/GO_0005623 denotes Cell
T3979 8503-8511 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T3980 8503-8511 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T4227 8584-8588 http://purl.obolibrary.org/obo/GO_0005623 denotes Cell
T4228 8885-8889 http://purl.obolibrary.org/obo/GO_0005623 denotes Cell
T4229 9031-9036 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4547 9280-9284 http://purl.obolibrary.org/obo/GO_0005623 denotes Cell
T4548 9357-9362 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4549 9494-9499 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4550 9616-9621 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4551 9864-9869 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4552 10081-10086 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4553 9280-9292 http://purl.obolibrary.org/obo/GO_0009986 denotes Cell Surface
T4554 9650-9660 http://purl.obolibrary.org/obo/GO_0019815 denotes antibodies
T4555 9750-9758 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T4556 9650-9660 http://purl.obolibrary.org/obo/GO_0042571 denotes antibodies
T4557 9750-9758 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T4715 10130-10135 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T4716 10365-10369 http://purl.obolibrary.org/obo/GO_0005623 denotes cell
T5447 11873-11878 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T6420 12154-12159 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T6421 13171-13176 http://purl.obolibrary.org/obo/GO_0005623 denotes cells
T6422 12711-12719 http://purl.obolibrary.org/obo/GO_0016020 denotes membrane
T6423 13536-13544 http://purl.obolibrary.org/obo/GO_0016020 denotes membrane
T6424 13584-13593 http://purl.obolibrary.org/obo/GO_0016020 denotes Membranes
T6425 13680-13689 http://purl.obolibrary.org/obo/GO_0016020 denotes Membranes
T6426 13917-13926 http://purl.obolibrary.org/obo/GO_0016020 denotes Membranes
T6427 14450-14459 http://purl.obolibrary.org/obo/GO_0016020 denotes membranes
T6428 12774-12782 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T6429 12875-12883 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T6430 13738-13746 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T6431 13796-13804 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T6432 14219-14227 http://purl.obolibrary.org/obo/GO_0019815 denotes antibody
T6433 12774-12782 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T6434 12875-12883 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T6435 13738-13746 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T6436 13796-13804 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody
T6437 14219-14227 http://purl.obolibrary.org/obo/GO_0042571 denotes antibody

GO-MF

Id Subject Object Predicate Lexical cue
T599 311-319 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T1991 1895-1899 http://purl.obolibrary.org/obo/GO_0005153 denotes IL-8
T3641 7517-7520 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3642 7576-7579 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3643 7701-7704 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3976 8503-8511 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T4226 9191-9193 http://purl.obolibrary.org/obo/GO_0033968 denotes CA
T4544 9579-9586 http://purl.obolibrary.org/obo/GO_0005488 denotes binding
T4545 9650-9660 http://purl.obolibrary.org/obo/GO_0003823 denotes antibodies
T4546 9750-9758 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T5200 10434-10436 http://purl.obolibrary.org/obo/GO_0003964 denotes RT
T5201 10615-10628 http://purl.obolibrary.org/obo/GO_0003899 denotes transcriptase
T5202 10615-10628 http://purl.obolibrary.org/obo/GO_0003968 denotes transcriptase
T5203 10615-10628 http://purl.obolibrary.org/obo/GO_0034062 denotes transcriptase
T5442 12025-12044 http://purl.obolibrary.org/obo/GO_0045289 denotes Luciferase activity
T5443 12025-12044 http://purl.obolibrary.org/obo/GO_0047077 denotes Luciferase activity
T5444 12025-12044 http://purl.obolibrary.org/obo/GO_0047712 denotes Luciferase activity
T5445 12025-12044 http://purl.obolibrary.org/obo/GO_0050248 denotes Luciferase activity
T5446 12025-12044 http://purl.obolibrary.org/obo/GO_0050397 denotes Luciferase activity
T6413 12774-12782 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T6414 12875-12883 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T6415 13738-13746 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T6416 13796-13804 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T6417 14219-14227 http://purl.obolibrary.org/obo/GO_0003823 denotes antibody
T6418 12810-12812 http://purl.obolibrary.org/obo/GO_0033968 denotes CA
T6419 14490-14493 http://purl.obolibrary.org/obo/GO_0004707 denotes ERK
T7120 15356-15380 http://purl.obolibrary.org/obo/GO_0004601 denotes myeloperoxidase activity
T15792 16557-16560 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO

GO-BP

Id Subject Object Predicate Lexical cue
T586 494-502 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T589 367-376 http://purl.obolibrary.org/obo/GO_0046903 denotes secretion
T595 20-38 http://purl.obolibrary.org/obo/GO_0008283 denotes cell proliferation
T596 72-80 http://purl.obolibrary.org/obo/GO_0051318 denotes G1 phase
T597 231-251 http://purl.obolibrary.org/obo/GO_0070487 denotes monocyte aggregation
T598 439-455 http://purl.obolibrary.org/obo/GO_0051092 denotes NF-κB activation
T1955 659-668 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphated
T1956 2492-2501 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphated
T1957 2727-2736 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphated
T1958 1168-1176 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T1959 3647-3655 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T1960 1208-1220 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1961 1699-1711 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1962 2105-2117 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1963 2548-2560 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1964 2627-2639 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1965 2664-2676 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1966 2831-2843 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T1967 2391-2418 http://purl.obolibrary.org/obo/GO_0006954 denotes response to an inflammatory
T1968 3576-3597 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammatory response
T1969 1574-1583 http://purl.obolibrary.org/obo/GO_0007586 denotes digestion
T1970 1895-1910 http://purl.obolibrary.org/obo/GO_0032677 denotes IL-8 production
T1971 1895-1910 http://purl.obolibrary.org/obo/GO_0032637 denotes IL-8 production
T1972 1924-1950 http://purl.obolibrary.org/obo/GO_0061582 denotes intestinal epithelial cell
T1973 1924-1950 http://purl.obolibrary.org/obo/GO_0060574 denotes intestinal epithelial cell
T1974 1924-1950 http://purl.obolibrary.org/obo/GO_0060575 denotes intestinal epithelial cell
T1975 1924-1950 http://purl.obolibrary.org/obo/GO_0060576 denotes intestinal epithelial cell
T1976 1984-2005 http://purl.obolibrary.org/obo/GO_0051092 denotes κB (NF-κB) activation
T1977 3464-3480 http://purl.obolibrary.org/obo/GO_0051092 denotes NF-κB activation
T1978 2029-2042 http://purl.obolibrary.org/obo/GO_0006351 denotes transcription
T1979 2069-2088 http://purl.obolibrary.org/obo/GO_0010467 denotes expression of genes
T1980 2350-2363 http://purl.obolibrary.org/obo/GO_0007155 denotes cell-adhesion
T1981 2350-2373 http://purl.obolibrary.org/obo/GO_0060352 denotes cell-adhesion molecules
T1982 2379-2399 http://purl.obolibrary.org/obo/GO_0002418 denotes immune cell response
T1983 2379-2399 http://purl.obolibrary.org/obo/GO_0002443 denotes immune cell response
T1984 2391-2427 http://purl.obolibrary.org/obo/GO_0002437 denotes response to an inflammatory stimulus
T1985 2850-2869 http://purl.obolibrary.org/obo/GO_0001865 denotes T-lymphocyte and NK
T1986 2850-2869 http://purl.obolibrary.org/obo/GO_0001866 denotes T-lymphocyte and NK
T1987 2850-2869 http://purl.obolibrary.org/obo/GO_0051132 denotes T-lymphocyte and NK
T1988 3226-3244 http://purl.obolibrary.org/obo/GO_0008283 denotes cell proliferation
T1989 3312-3332 http://purl.obolibrary.org/obo/GO_0070487 denotes monocyte aggregation
T1990 3370-3379 http://purl.obolibrary.org/obo/GO_0046903 denotes secretion
T2804 3786-3794 http://purl.obolibrary.org/obo/GO_0009056 denotes Degraded
T2805 3827-3835 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T2806 5296-5304 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphate
T3085 5932-5940 http://purl.obolibrary.org/obo/GO_0009056 denotes Degraded
T3086 5984-5992 http://purl.obolibrary.org/obo/GO_0007631 denotes drinking
T3087 6350-6358 http://purl.obolibrary.org/obo/GO_0007631 denotes drinking
T3634 7141-7153 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T3635 7246-7258 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T3636 7368-7377 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphated
T3637 7517-7520 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3638 7576-7579 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3639 7701-7704 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO
T3640 7733-7744 http://purl.obolibrary.org/obo/GO_0009056 denotes degradation
T3974 8252-8268 http://purl.obolibrary.org/obo/GO_0098743 denotes cell aggregation
T3975 8257-8279 http://purl.obolibrary.org/obo/GO_0070487 denotes aggregation, monocytes
T4223 8584-8594 http://purl.obolibrary.org/obo/GO_0007049 denotes Cell Cycle
T4224 8631-8637 http://purl.obolibrary.org/obo/GO_0040007 denotes growth
T4225 9191-9193 http://purl.obolibrary.org/obo/GO_0033968 denotes CA
T4714 10365-10377 http://purl.obolibrary.org/obo/GO_0001906 denotes cell killing
T5196 10434-10436 http://purl.obolibrary.org/obo/GO_0003964 denotes RT
T5197 10615-10628 http://purl.obolibrary.org/obo/GO_0003899 denotes transcriptase
T5198 10615-10628 http://purl.obolibrary.org/obo/GO_0003968 denotes transcriptase
T5199 10615-10628 http://purl.obolibrary.org/obo/GO_0034062 denotes transcriptase
T5436 11577-11590 http://purl.obolibrary.org/obo/GO_0006351 denotes Transcription
T5437 12025-12044 http://purl.obolibrary.org/obo/GO_0045289 denotes Luciferase activity
T5438 12025-12044 http://purl.obolibrary.org/obo/GO_0047077 denotes Luciferase activity
T5439 12025-12044 http://purl.obolibrary.org/obo/GO_0047712 denotes Luciferase activity
T5440 12025-12044 http://purl.obolibrary.org/obo/GO_0050248 denotes Luciferase activity
T5441 12025-12044 http://purl.obolibrary.org/obo/GO_0050397 denotes Luciferase activity
T6409 12300-12305 http://purl.obolibrary.org/obo/GO_0019835 denotes lysis
T6410 12460-12468 http://purl.obolibrary.org/obo/GO_0007349 denotes cellular
T6411 12810-12812 http://purl.obolibrary.org/obo/GO_0033968 denotes CA
T6412 14490-14493 http://purl.obolibrary.org/obo/GO_0004707 denotes ERK
T7112 14892-14900 http://purl.obolibrary.org/obo/GO_0009056 denotes Degraded
T7113 14968-14976 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T7114 14920-14932 http://purl.obolibrary.org/obo/GO_0006954 denotes Inflammation
T7115 15319-15331 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T7116 15507-15519 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T7117 15596-15608 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T7118 15356-15380 http://purl.obolibrary.org/obo/GO_0004601 denotes myeloperoxidase activity
T7119 15992-16001 http://purl.obolibrary.org/obo/GO_0051923 denotes sulphated
T15785 16357-16365 http://purl.obolibrary.org/obo/GO_0009056 denotes Degraded
T15786 16439-16447 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T15787 16610-16618 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T15788 16656-16664 http://purl.obolibrary.org/obo/GO_0009056 denotes degraded
T15789 16384-16396 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T15790 16511-16523 http://purl.obolibrary.org/obo/GO_0006954 denotes inflammation
T15791 16557-16560 http://purl.obolibrary.org/obo/GO_0004601 denotes MPO

2_test

Id Subject Object Predicate Lexical cue
20072622-3943102-93233346 1317-1318 3943102 denotes 4
20072622-2570843-93233347 1337-1338 2570843 denotes 5
20072622-3943102-93233348 1409-1410 3943102 denotes 4
20072622-11675262-93233349 1414-1415 11675262 denotes 6
20072622-17034924-93233350 1418-1419 17034924 denotes 8
20072622-8849638-93233351 1635-1637 8849638 denotes 11
20072622-9178362-93233352 1641-1643 9178362 denotes 12
20072622-17095757-93233353 2007-2009 17095757 denotes 13
20072622-3140380-93233354 2119-2121 3140380 denotes 15
20072622-8011280-93233355 2125-2127 8011280 denotes 16
20072622-9136827-93233356 2222-2224 9136827 denotes 17
20072622-1316857-93233357 2647-2649 1316857 denotes 18
20072622-7557106-93233358 2653-2655 7557106 denotes 19
20072622-8741008-93233359 2801-2803 8741008 denotes 20
20072622-8741008-93233360 2886-2888 8741008 denotes 20
20072622-4212170-93233361 6403-6405 4212170 denotes 23
20072622-1688816-93233362 6760-6762 1688816 denotes 24
20072622-9824605-93233363 7298-7300 9824605 denotes 25
20072622-10940278-93233364 7304-7306 10940278 denotes 26
20072622-6092199-93233365 7558-7560 6092199 denotes 27
20072622-8482919-93233366 8243-8245 8482919 denotes 28
20072622-3782828-93233367 10385-10387 3782828 denotes 29
20072622-8625304-93233368 11933-11935 8625304 denotes 30
20072622-19342628-93233369 14508-14510 19342628 denotes 32

pmc-enju-pas

Id Subject Object Predicate Lexical cue
T221 0-3 NN denotes CGN
T222 4-13 VB denotes inhibited
T223 14-19 NN denotes THP-1
T224 20-24 NN denotes cell
T225 25-38 NN denotes proliferation
T226 39-41 FW denotes in
T227 42-47 FW denotes vitro
T228 47-48 -COMMA- denotes ,
T229 49-58 VB denotes arresting
T230 59-62 DT denotes the
T231 63-68 NN denotes cells
T232 69-71 IN denotes in
T233 72-74 NN denotes G1
T234 75-80 NN denotes phase
T235 82-84 IN denotes In
T236 85-93 NN denotes addition
T237 93-94 -COMMA- denotes ,
T238 95-99 NN denotes dCGN
T239 100-109 VB denotes increased
T240 110-116 NN denotes ICAM-1
T241 117-127 NN denotes expression
T242 128-130 IN denotes in
T243 131-135 CC denotes both
T244 136-139 NN denotes PBM
T245 140-143 CC denotes and
T246 144-149 NN denotes THP-1
T247 150-155 NN denotes cells
T248 156-160 IN denotes with
T249 161-162 DT denotes a
T250 163-168 JJ denotes major
T251 169-175 NN denotes effect
T252 176-180 VB denotes seen
T253 181-186 IN denotes after
T254 187-189 CD denotes 40
T255 190-193 NN denotes kDa
T256 194-198 NN denotes dCGN
T257 199-207 NN denotes exposure
T258 209-213 RB denotes Also
T259 213-214 -COMMA- denotes ,
T260 215-219 NN denotes dCGN
T261 220-230 VB denotes stimulated
T262 231-239 NN denotes monocyte
T263 240-251 NN denotes aggregation
T264 252-254 FW denotes in
T265 255-260 FW denotes vitro
T266 261-265 WDT denotes that
T267 266-269 VB denotes was
T268 270-279 VB denotes prevented
T269 280-282 IN denotes by
T270 283-293 NN denotes incubation
T271 294-298 IN denotes with
T272 299-310 JJ denotes anti-ICAM-1
T273 311-319 NN denotes antibody
T274 321-328 RB denotes Finally
T275 328-329 -COMMA- denotes ,
T276 330-334 NN denotes dCGN
T277 335-345 VB denotes stimulated
T278 346-351 NN denotes TNF-α
T279 352-362 NN denotes expression
T280 363-366 CC denotes and
T281 367-376 NN denotes secretion
T282 377-379 IN denotes by
T283 380-384 CC denotes both
T284 385-388 NN denotes PBM
T285 389-392 CC denotes and
T286 393-398 NN denotes THP-1
T287 399-404 NN denotes cells
T288 406-409 PDT denotes All
T289 410-415 DT denotes these
T290 416-423 NN denotes effects
T291 424-428 VB denotes were
T292 429-435 VB denotes linked
T293 436-438 TO denotes to
T294 439-444 NN denotes NF-κB
T295 445-455 NN denotes activation
T296 457-462 DT denotes These
T297 463-467 NN denotes data
T298 468-476 RB denotes strongly
T299 477-484 VB denotes suggest
T300 485-489 IN denotes that
T301 490-493 DT denotes the
T302 494-502 JJ denotes degraded
T303 503-508 NN denotes forms
T304 509-511 IN denotes of
T305 512-515 NN denotes CGN
T306 516-520 VB denotes have
T307 521-522 DT denotes a
T308 523-533 JJ denotes pronounced
T309 534-540 NN denotes effect
T310 541-543 IN denotes on
T311 544-553 NN denotes monocytes
T312 553-554 -COMMA- denotes ,
T313 555-569 JJ denotes characteristic
T314 570-572 IN denotes of
T315 573-575 DT denotes an
T316 576-588 JJ denotes inflammatory
T317 589-598 NN denotes phenotype
T776 614-625 NN denotes Carrageenan
T777 626-627 -LRB- denotes (
T778 627-630 NN denotes CGN
T779 630-631 -RRB- denotes )
T780 632-634 VB denotes is
T781 635-636 DT denotes a
T782 637-641 JJ denotes high
T783 642-651 JJ denotes molecular
T784 652-658 NN denotes weight
T785 659-668 VB denotes sulphated
T786 669-683 NN denotes polysaccharide
T787 684-685 -LRB- denotes (
T788 685-686 SYM denotes >
T789 686-689 CD denotes 200
T790 690-693 NN denotes kDa
T791 693-694 -RRB- denotes )
T792 695-702 VB denotes derived
T793 703-707 IN denotes from
T794 708-711 JJ denotes red
T795 712-717 NN denotes algae
T796 718-719 -LRB- denotes (
T797 719-731 NNP denotes Rhodophyceae
T798 731-732 -RRB- denotes )
T799 734-739 CD denotes Three
T800 740-744 JJ denotes main
T801 745-750 NN denotes forms
T802 751-753 IN denotes of
T803 754-757 NN denotes CGN
T804 758-762 VB denotes have
T805 763-767 VB denotes been
T806 768-778 VB denotes identified
T807 778-779 -COLON- denotes :
T808 780-785 NN denotes kappa
T809 785-786 -COMMA- denotes ,
T810 787-791 NN denotes iota
T811 791-792 -COMMA- denotes ,
T812 793-796 CC denotes and
T813 797-803 NN denotes lambda
T814 805-809 PRP denotes They
T815 810-816 VB denotes differ
T816 817-821 IN denotes from
T817 822-826 DT denotes each
T818 827-832 JJ denotes other
T819 833-835 IN denotes in
T820 836-846 NN denotes sulphation
T821 847-853 NN denotes degree
T822 854-857 CC denotes and
T823 858-868 NN denotes solubility
T824 869-870 -LRB- denotes [
T825 870-871 CD denotes 1
T826 871-872 -RRB- denotes ]
T827 872-873 -COMMA- denotes ,
T828 874-875 -LRB- denotes [
T829 875-876 CD denotes 2
T830 876-877 -RRB- denotes ]
T831 879-885 JJ denotes Native
T832 886-889 NN denotes CGN
T833 890-892 VB denotes is
T834 893-900 VB denotes thought
T835 901-903 TO denotes to
T836 904-906 VB denotes be
T837 907-915 JJ denotes harmless
T838 916-919 CC denotes and
T839 920-922 VB denotes is
T840 923-929 RB denotes widely
T841 930-934 VB denotes used
T842 935-937 IN denotes as
T843 938-939 DT denotes a
T844 940-944 NN denotes food
T845 945-953 JJ denotes additive
T846 954-956 TO denotes to
T847 957-964 VB denotes improve
T848 965-972 NN denotes texture
T849 974-976 PRP denotes It
T850 977-979 VB denotes is
T851 980-984 RB denotes also
T852 985-989 VB denotes used
T853 990-992 IN denotes in
T854 993-1002 NN denotes cosmetics
T855 1003-1006 CC denotes and
T856 1007-1022 NN denotes pharmaceuticals
T857 1024-1031 RB denotes However
T858 1031-1032 -COMMA- denotes ,
T859 1033-1037 NN denotes acid
T860 1038-1047 NN denotes treatment
T861 1048-1050 IN denotes at
T862 1051-1055 JJ denotes high
T863 1056-1067 NN denotes temperature
T864 1068-1069 -LRB- denotes (
T865 1069-1073 NN denotes 80°C
T866 1073-1074 -RRB- denotes )
T867 1075-1083 VB denotes triggers
T868 1084-1087 NN denotes CGN
T869 1088-1098 NN denotes hydrolysis
T870 1099-1101 TO denotes to
T871 1102-1107 VB denotes lower
T872 1108-1117 JJ denotes molecular
T873 1118-1124 NN denotes weight
T874 1125-1126 -LRB- denotes (
T875 1126-1127 SYM denotes <
T876 1127-1129 CD denotes 50
T877 1130-1133 NN denotes kDa
T878 1133-1134 -RRB- denotes )
T879 1135-1144 NN denotes compounds
T880 1145-1150 VB denotes known
T881 1151-1153 IN denotes as
T882 1154-1164 NN denotes poligeenan
T883 1165-1167 CC denotes or
T884 1168-1176 VB denotes degraded
T885 1177-1180 NN denotes CGN
T886 1181-1182 -LRB- denotes (
T887 1182-1186 NN denotes dCGN
T888 1186-1187 -RRB- denotes )
T889 1189-1194 DT denotes These
T890 1195-1200 NN denotes dCGNs
T891 1201-1207 VB denotes induce
T892 1208-1220 NN denotes inflammation
T893 1221-1224 CC denotes and
T894 1225-1229 VB denotes have
T895 1230-1234 VB denotes been
T896 1235-1241 RB denotes widely
T897 1242-1246 VB denotes used
T898 1247-1249 IN denotes as
T899 1250-1256 NN denotes models
T900 1257-1259 IN denotes of
T901 1260-1267 NN denotes colitis
T902 1268-1270 IN denotes in
T903 1271-1278 JJ denotes several
T904 1279-1286 NN denotes species
T905 1286-1287 -COMMA- denotes ,
T906 1288-1297 VB denotes including
T907 1298-1302 NN denotes rats
T908 1303-1304 -LRB- denotes [
T909 1304-1305 CD denotes 3
T910 1305-1306 -RRB- denotes ]
T911 1306-1307 -COMMA- denotes ,
T912 1308-1315 NN denotes rabbits
T913 1316-1317 -LRB- denotes [
T914 1317-1318 CD denotes 4
T915 1318-1319 -RRB- denotes ]
T916 1320-1323 CC denotes and
T917 1324-1330 NN denotes guinea
T918 1331-1335 NN denotes pigs
T919 1336-1337 -LRB- denotes [
T920 1337-1338 CD denotes 5
T921 1338-1339 -RRB- denotes ]
T922 1341-1344 DT denotes The
T923 1345-1349 NN denotes role
T924 1350-1352 IN denotes of
T925 1353-1357 NN denotes dCGN
T926 1358-1360 IN denotes as
T927 1361-1362 DT denotes a
T928 1363-1378 JJ denotes tumor-promoting
T929 1379-1385 NN denotes factor
T930 1386-1393 VB denotes remains
T931 1394-1407 JJ denotes controversial
T932 1408-1409 -LRB- denotes [
T933 1409-1410 CD denotes 4
T934 1410-1411 -RRB- denotes ]
T935 1411-1412 -COMMA- denotes ,
T936 1413-1414 -LRB- denotes [
T937 1414-1415 CD denotes 6
T938 1415-1416 -RRB- denotes ]
T939 1416-1417 -LRB- denotes –
T940 1417-1418 CD denotes [
T941 1418-1419 -RRB- denotes 8
T942 1422-1430 IN denotes Although
T943 1431-1434 DT denotes the
T944 1435-1441 JJ denotes native
T945 1442-1446 NN denotes form
T946 1447-1449 VB denotes is
T947 1450-1457 VB denotes thought
T948 1458-1460 TO denotes to
T949 1461-1463 VB denotes be
T950 1464-1472 JJ denotes harmless
T951 1473-1476 IN denotes for
T952 1477-1482 JJ denotes human
T953 1483-1494 NN denotes consumption
T954 1494-1495 -COMMA- denotes ,
T955 1496-1501 JJ denotes small
T956 1502-1509 NN denotes amounts
T957 1510-1512 IN denotes of
T958 1513-1517 NN denotes dCGN
T959 1518-1521 VB denotes are
T960 1522-1530 RB denotes probably
T961 1531-1539 VB denotes produced
T962 1540-1542 IN denotes by
T963 1543-1547 NN denotes acid
T964 1548-1558 NN denotes hydrolysis
T965 1559-1565 IN denotes during
T966 1566-1573 JJ denotes gastric
T967 1574-1583 NN denotes digestion
T968 1584-1585 -LRB- denotes [
T969 1585-1586 CD denotes 9
T970 1586-1587 -RRB- denotes ]
T971 1587-1588 -COMMA- denotes ,
T972 1589-1590 -LRB- denotes [
T973 1590-1592 CD denotes 10
T974 1592-1593 -RRB- denotes ]
T975 1594-1596 CC denotes or
T976 1597-1608 NN denotes interaction
T977 1609-1613 IN denotes with
T978 1614-1624 JJ denotes intestinal
T979 1625-1633 NN denotes bacteria
T980 1634-1635 -LRB- denotes [
T981 1635-1637 CD denotes 11
T982 1637-1638 -RRB- denotes ]
T983 1638-1639 -COMMA- denotes ,
T984 1640-1641 -LRB- denotes [
T985 1641-1643 CD denotes 12
T986 1643-1644 -RRB- denotes ]
T987 1646-1653 IN denotes Whereas
T988 1654-1657 DT denotes the
T989 1658-1665 NN denotes effects
T990 1666-1668 IN denotes of
T991 1669-1675 JJ denotes native
T992 1676-1679 CC denotes and
T993 1680-1684 NN denotes dCGN
T994 1685-1687 IN denotes on
T995 1688-1698 JJ denotes intestinal
T996 1699-1711 NN denotes inflammation
T997 1712-1716 VB denotes have
T998 1717-1721 VB denotes been
T999 1722-1733 RB denotes extensively
T1000 1734-1742 VB denotes analyzed
T1001 1743-1745 IN denotes in
T1002 1746-1752 NN denotes animal
T1003 1753-1759 NN denotes models
T1004 1759-1760 -COMMA- denotes ,
T1005 1761-1765 RB denotes only
T1006 1766-1769 JJ denotes few
T1007 1770-1777 NN denotes studies
T1008 1778-1782 VB denotes have
T1009 1783-1787 VB denotes been
T1010 1788-1797 VB denotes conducted
T1011 1798-1803 VB denotes using
T1012 1804-1809 JJ denotes human
T1013 1810-1814 NN denotes cell
T1014 1815-1820 NN denotes lines
T1015 1822-1828 JJ denotes Recent
T1016 1829-1836 NN denotes studies
T1017 1837-1841 VB denotes have
T1018 1842-1847 VB denotes shown
T1019 1848-1849 DT denotes a
T1020 1850-1854 NN denotes link
T1021 1855-1862 IN denotes between
T1022 1863-1871 NN denotes exposure
T1023 1872-1874 TO denotes to
T1024 1875-1881 JJ denotes native
T1025 1882-1886 NN denotes form
T1026 1887-1890 NN denotes CGN
T1027 1891-1894 CC denotes and
T1028 1895-1899 NN denotes IL-8
T1029 1900-1910 NN denotes production
T1030 1911-1913 IN denotes by
T1031 1914-1917 DT denotes the
T1032 1918-1923 JJ denotes human
T1033 1924-1934 JJ denotes intestinal
T1034 1935-1945 JJ denotes epithelial
T1035 1946-1950 NN denotes cell
T1036 1951-1955 NN denotes line
T1037 1955-1956 -COMMA- denotes ,
T1038 1957-1963 NN denotes NCM460
T1039 1963-1964 -COMMA- denotes ,
T1040 1965-1968 IN denotes via
T1041 1969-1976 JJ denotes Nuclear
T1042 1977-1986 NN denotes Factor-κB
T1043 1987-1988 -LRB- denotes (
T1044 1988-1993 NN denotes NF-κB
T1045 1993-1994 -RRB- denotes )
T1046 1995-2005 NN denotes activation
T1047 2006-2007 -LRB- denotes [
T1048 2007-2009 CD denotes 13
T1049 2009-2010 -RRB- denotes ]
T1050 2010-2011 -COMMA- denotes ,
T1051 2012-2013 -LRB- denotes [
T1052 2013-2015 CD denotes 14
T1053 2015-2016 -RRB- denotes ]
T1054 2018-2023 NN denotes NF-κB
T1055 2024-2026 VB denotes is
T1056 2027-2028 DT denotes a
T1057 2029-2042 NN denotes transcription
T1058 2043-2049 NN denotes factor
T1059 2050-2054 WDT denotes that
T1060 2055-2064 VB denotes regulates
T1061 2065-2068 DT denotes the
T1062 2069-2079 NN denotes expression
T1063 2080-2082 IN denotes of
T1064 2083-2088 NN denotes genes
T1065 2089-2099 VB denotes associated
T1066 2100-2104 IN denotes with
T1067 2105-2117 NN denotes inflammation
T1068 2118-2119 -LRB- denotes [
T1069 2119-2121 CD denotes 15
T1070 2121-2122 -RRB- denotes ]
T1071 2122-2123 -COMMA- denotes ,
T1072 2124-2125 -LRB- denotes [
T1073 2125-2127 CD denotes 16
T1074 2127-2128 -RRB- denotes ]
T1075 2130-2140 NN denotes Macrophage
T1076 2141-2153 NN denotes infiltration
T1077 2154-2157 CC denotes and
T1078 2158-2170 NN denotes accumulation
T1079 2171-2173 VB denotes is
T1080 2174-2175 DT denotes a
T1081 2176-2182 JJ denotes common
T1082 2183-2197 NN denotes characteristic
T1083 2198-2200 IN denotes of
T1084 2201-2211 JJ denotes intestinal
T1085 2212-2220 NN denotes diseases
T1086 2221-2222 -LRB- denotes [
T1087 2222-2224 CD denotes 17
T1088 2224-2225 -RRB- denotes ]
T1089 2227-2238 NN denotes Macrophages
T1090 2239-2248 VB denotes represent
T1091 2249-2251 CD denotes 10
T1092 2251-2252 NN denotes %
T1093 2253-2255 IN denotes of
T1094 2256-2261 JJ denotes total
T1095 2262-2268 NN denotes lamina
T1096 2269-2276 NN denotes propria
T1097 2277-2282 NN denotes cells
T1098 2282-2283 -COMMA- denotes ,
T1099 2284-2291 VB denotes secrete
T1100 2292-2293 DT denotes a
T1101 2294-2298 JJ denotes wide
T1102 2299-2304 NN denotes range
T1103 2305-2307 IN denotes of
T1104 2308-2320 RB denotes biologically
T1105 2321-2327 JJ denotes active
T1106 2328-2337 NN denotes compounds
T1107 2338-2341 CC denotes and
T1108 2342-2349 VB denotes express
T1109 2350-2363 NN denotes cell-adhesion
T1110 2364-2373 NN denotes molecules
T1111 2375-2378 DT denotes The
T1112 2379-2385 JJ denotes immune
T1113 2386-2390 NN denotes cell
T1114 2391-2399 NN denotes response
T1115 2400-2402 TO denotes to
T1116 2403-2405 DT denotes an
T1117 2406-2418 JJ denotes inflammatory
T1118 2419-2427 NN denotes stimulus
T1119 2428-2433 VB denotes seems
T1120 2434-2436 TO denotes to
T1121 2437-2439 VB denotes be
T1122 2440-2449 VB denotes amplified
T1123 2450-2452 CC denotes or
T1124 2453-2461 RB denotes directly
T1125 2462-2471 VB denotes generated
T1126 2472-2474 IN denotes by
T1127 2475-2480 NN denotes cells
T1128 2481-2488 VB denotes exposed
T1129 2489-2491 TO denotes to
T1130 2492-2501 JJ denotes sulphated
T1131 2502-2517 NN denotes polysaccharides
T1132 2518-2522 JJ denotes such
T1133 2523-2525 IN denotes as
T1134 2526-2538 NN denotes carrageenans
T1135 2540-2546 RB denotes Indeed
T1136 2546-2547 -COMMA- denotes ,
T1137 2548-2560 NN denotes inflammation
T1138 2561-2568 VB denotes induced
T1139 2569-2571 IN denotes by
T1140 2572-2576 NN denotes dCGN
T1141 2577-2580 VB denotes was
T1142 2581-2591 VB denotes associated
T1143 2592-2596 IN denotes with
T1144 2597-2608 NN denotes recruitment
T1145 2609-2611 IN denotes of
T1146 2612-2623 NN denotes macrophages
T1147 2624-2626 TO denotes to
T1148 2627-2639 NN denotes inflammation
T1149 2640-2645 NN denotes sites
T1150 2646-2647 -LRB- denotes [
T1151 2647-2649 CD denotes 18
T1152 2649-2650 -RRB- denotes ]
T1153 2650-2651 -COMMA- denotes ,
T1154 2652-2653 -LRB- denotes [
T1155 2653-2655 CD denotes 19
T1156 2655-2656 -RRB- denotes ]
T1157 2658-2662 RB denotes Also
T1158 2662-2663 -COMMA- denotes ,
T1159 2664-2676 NN denotes inflammation
T1160 2677-2684 VB denotes induced
T1161 2685-2687 IN denotes by
T1162 2688-2695 NNP denotes Dextran
T1163 2696-2704 NNP denotes Sulphate
T1164 2705-2711 NNP denotes Sodium
T1165 2712-2713 -LRB- denotes (
T1166 2713-2716 NN denotes DSS
T1167 2716-2717 -RRB- denotes )
T1168 2717-2718 -COMMA- denotes ,
T1169 2719-2726 DT denotes another
T1170 2727-2736 VB denotes sulphated
T1171 2737-2745 NN denotes compound
T1172 2745-2746 -COMMA- denotes ,
T1173 2747-2750 VB denotes was
T1174 2751-2759 RB denotes directly
T1175 2760-2770 VB denotes associated
T1176 2771-2775 IN denotes with
T1177 2776-2787 NN denotes macrophages
T1178 2788-2799 NN denotes recruitment
T1179 2800-2801 -LRB- denotes [
T1180 2801-2803 CD denotes 20
T1181 2803-2804 -RRB- denotes ]
T1182 2804-2805 -COMMA- denotes ,
T1183 2806-2811 IN denotes since
T1184 2812-2815 NN denotes DSS
T1185 2816-2821 RB denotes still
T1186 2822-2830 VB denotes provoked
T1187 2831-2843 NN denotes inflammation
T1188 2844-2849 IN denotes after
T1189 2850-2862 NN denotes T-lymphocyte
T1190 2863-2866 CC denotes and
T1191 2867-2869 NN denotes NK
T1192 2870-2874 NN denotes cell
T1193 2875-2884 NN denotes depletion
T1194 2885-2886 -LRB- denotes [
T1195 2886-2888 CD denotes 20
T1196 2888-2889 -RRB- denotes ]
T1197 2891-2899 IN denotes Although
T1198 2900-2912 NN denotes inflammation
T1199 2913-2916 MD denotes can
T1200 2917-2919 VB denotes be
T1201 2920-2927 VB denotes induced
T1202 2928-2930 IN denotes by
T1203 2931-2935 NN denotes dCGN
T1204 2935-2936 -COMMA- denotes ,
T1205 2937-2942 EX denotes there
T1206 2943-2946 VB denotes are
T1207 2947-2949 DT denotes no
T1208 2950-2954 NN denotes data
T1209 2955-2957 IN denotes on
T1210 2958-2963 JJ denotes human
T1211 2964-2972 NN denotes monocyte
T1212 2973-2982 NN denotes responses
T1213 2983-2985 TO denotes to
T1214 2986-2990 NN denotes dCGN
T1215 2991-2999 NN denotes exposure
T1216 3001-3010 RB denotes Therefore
T1217 3010-3011 -COMMA- denotes ,
T1218 3012-3014 TO denotes to
T1219 3015-3026 VB denotes investigate
T1220 3027-3030 DT denotes the
T1221 3031-3038 NN denotes effects
T1222 3039-3041 IN denotes of
T1223 3042-3046 NN denotes dCGN
T1224 3047-3049 IN denotes on
T1225 3050-3055 JJ denotes human
T1226 3056-3065 NN denotes monocytes
T1227 3065-3066 -COMMA- denotes ,
T1228 3067-3073 JJ denotes normal
T1229 3074-3084 JJ denotes Peripheral
T1230 3085-3090 NN denotes Blood
T1231 3091-3100 NN denotes Monocytes
T1232 3101-3102 -LRB- denotes (
T1233 3102-3105 NN denotes PBM
T1234 3105-3106 -RRB- denotes )
T1235 3107-3110 CC denotes and
T1236 3111-3118 JJ denotes tumoral
T1237 3119-3138 NN denotes monocyte/macrophage
T1238 3139-3144 NN denotes THP-1
T1239 3145-3150 NN denotes cells
T1240 3151-3155 VB denotes were
T1241 3156-3163 VB denotes exposed
T1242 3164-3166 TO denotes to
T1243 3167-3169 CD denotes 10
T1244 3170-3173 NN denotes kDa
T1245 3174-3177 CC denotes and
T1246 3178-3180 CD denotes 40
T1247 3181-3184 NN denotes kDa
T1248 3185-3189 NN denotes dCGN
T1249 3191-3193 PRP denotes We
T1250 3194-3199 VB denotes found
T1251 3200-3204 IN denotes that
T1252 3205-3209 NN denotes dCGN
T1253 3210-3219 VB denotes inhibited
T1254 3220-3225 NN denotes THP-1
T1255 3226-3230 NN denotes cell
T1256 3231-3244 NN denotes proliferation
T1257 3245-3247 FW denotes in
T1258 3248-3253 FW denotes vitro
T1259 3253-3254 -COMMA- denotes ,
T1260 3255-3264 VB denotes increased
T1261 3265-3271 NN denotes ICAM-1
T1262 3272-3282 NN denotes expression
T1263 3282-3283 -COMMA- denotes ,
T1264 3284-3294 VB denotes stimulated
T1265 3295-3311 JJ denotes ICAM-1-dependent
T1266 3312-3320 NN denotes monocyte
T1267 3321-3332 NN denotes aggregation
T1268 3332-3333 -COMMA- denotes ,
T1269 3334-3337 CC denotes and
T1270 3338-3348 VB denotes stimulated
T1271 3349-3354 NN denotes TNF-α
T1272 3355-3365 NN denotes expression
T1273 3366-3369 CC denotes and
T1274 3370-3379 NN denotes secretion
T1275 3381-3386 DT denotes These
T1276 3387-3396 NN denotes responses
T1277 3397-3401 VB denotes were
T1278 3402-3406 RB denotes more
T1279 3407-3417 JJ denotes pronounced
T1280 3418-3423 IN denotes after
T1281 3424-3426 CD denotes 40
T1282 3427-3430 NN denotes kDa
T1283 3431-3435 NN denotes dCGN
T1284 3436-3444 NN denotes exposure
T1285 3445-3448 CC denotes and
T1286 3449-3453 VB denotes were
T1287 3454-3460 VB denotes linked
T1288 3461-3463 TO denotes to
T1289 3464-3469 NN denotes NF-κB
T1290 3470-3480 NN denotes activation
T1291 3482-3484 IN denotes In
T1292 3485-3493 NN denotes addition
T1293 3493-3494 -COMMA- denotes ,
T1294 3495-3498 DT denotes the
T1295 3499-3501 CD denotes 40
T1296 3502-3505 NN denotes kDa
T1297 3506-3510 NN denotes dCGN
T1298 3510-3511 -COMMA- denotes ,
T1299 3512-3515 CC denotes but
T1300 3516-3519 RB denotes not
T1301 3520-3523 DT denotes the
T1302 3524-3526 CD denotes 10
T1303 3527-3530 NN denotes kDa
T1304 3531-3535 NN denotes dCGN
T1305 3536-3543 VB denotes induced
T1306 3544-3546 FW denotes in
T1307 3547-3551 FW denotes vivo
T1308 3552-3559 NN denotes colitis
T1309 3560-3562 IN denotes as
T1310 3563-3568 VB denotes shown
T1311 3569-3571 IN denotes by
T1312 3572-3575 DT denotes the
T1313 3576-3588 JJ denotes inflammatory
T1314 3589-3597 NN denotes response
T1315 3598-3600 IN denotes in
T1316 3601-3604 DT denotes the
T1317 3605-3608 NN denotes rat
T1318 3609-3614 NN denotes colon
T1319 3616-3621 DT denotes These
T1320 3622-3629 NN denotes results
T1321 3630-3637 VB denotes suggest
T1322 3638-3642 IN denotes that
T1323 3643-3646 DT denotes the
T1324 3647-3655 JJ denotes degraded
T1325 3656-3661 NN denotes forms
T1326 3662-3664 IN denotes of
T1327 3665-3668 NN denotes CGN
T1328 3669-3673 VB denotes have
T1329 3674-3676 DT denotes an
T1330 3677-3686 JJ denotes important
T1331 3687-3693 NN denotes effect
T1332 3694-3696 IN denotes on
T1333 3697-3706 NN denotes monocytes
T1334 3707-3716 VB denotes resulting
T1335 3717-3719 IN denotes in
T1336 3720-3722 DT denotes an
T1337 3723-3735 JJ denotes inflammatory
T1338 3736-3745 NN denotes phenotype
T2063 3771-3782 NN denotes Preparation
T2064 3783-3785 IN denotes of
T2065 3786-3794 NNP denotes Degraded
T2066 3795-3806 NNP denotes Carrageenan
T2067 3807-3810 CD denotes Two
T2068 3811-3823 NN denotes preparations
T2069 3824-3826 IN denotes of
T2070 3827-3835 VB denotes degraded
T2071 3836-3847 NN denotes carrageenan
T2072 3848-3852 IN denotes with
T2073 3853-3856 JJ denotes low
T2074 3856-3857 -COMMA- denotes ,
T2075 3858-3859 -LRB- denotes (
T2076 3859-3862 CD denotes ∼10
T2077 3863-3866 NN denotes kDa
T2078 3866-3867 -COLON- denotes ;
T2079 3868-3871 NN denotes C10
T2080 3871-3872 -RRB- denotes )
T2081 3872-3873 -COMMA- denotes ,
T2082 3874-3877 CC denotes and
T2083 3878-3884 NN denotes medium
T2084 3884-3885 -COMMA- denotes ,
T2085 3886-3887 -LRB- denotes (
T2086 3887-3890 CD denotes ∼40
T2087 3891-3894 NN denotes kDa
T2088 3894-3895 -COLON- denotes ;
T2089 3896-3899 NN denotes C40
T2090 3899-3900 -RRB- denotes )
T2091 3901-3910 JJ denotes molecular
T2092 3911-3917 NN denotes weight
T2093 3918-3922 VB denotes were
T2094 3923-3931 VB denotes prepared
T2095 3932-3936 IN denotes from
T2096 3937-3943 JJ denotes native
T2097 3944-3960 NN denotes iota-carrageenan
T2098 3961-3970 VB denotes extracted
T2099 3971-3975 IN denotes from
T2100 3976-3983 NNP denotes Euchema
T2101 3984-3992 NN denotes spinosum
T2102 3993-3994 -LRB- denotes (
T2103 3994-4004 RB denotes generously
T2104 4005-4013 VB denotes provided
T2105 4014-4016 IN denotes by
T2106 4017-4023 NNP denotes Sanofi
T2107 4024-4034 NNP denotes Biosystems
T2108 4035-4043 NNP denotes Industry
T2109 4043-4044 -COMMA- denotes ,
T2110 4045-4065 NNP denotes Boulogne-Billancourt
T2111 4065-4066 -COMMA- denotes ,
T2112 4067-4073 NNP denotes France
T2113 4073-4074 -RRB- denotes )
T2114 4076-4082 JJ denotes Native
T2115 4083-4094 NN denotes carrageenan
T2116 4095-4098 VB denotes was
T2117 4099-4108 VB denotes dissolved
T2118 4109-4111 IN denotes in
T2119 4112-4121 JJ denotes distilled
T2120 4122-4127 NN denotes water
T2121 4128-4129 -LRB- denotes (
T2122 4129-4130 CD denotes 5
T2123 4130-4131 NN denotes %
T2124 4132-4135 NN denotes w/v
T2125 4135-4136 -RRB- denotes )
T2126 4137-4142 IN denotes under
T2127 4143-4151 JJ denotes vigorous
T2128 4152-4160 VB denotes stirring
T2129 4161-4164 CC denotes and
T2130 4165-4171 VB denotes heated
T2131 4172-4174 TO denotes to
T2132 4175-4180 NNP denotes 60°C.
T2133 4181-4185 NNP denotes Then
T2134 4185-4186 -COMMA- denotes ,
T2135 4187-4190 DT denotes the
T2136 4191-4202 NN denotes carrageenan
T2137 4203-4211 NN denotes solution
T2138 4212-4215 VB denotes was
T2139 4216-4225 VB denotes submitted
T2140 4226-4228 TO denotes to
T2141 4229-4232 CD denotes two
T2142 4233-4242 JJ denotes different
T2143 4243-4253 NN denotes treatments
T2144 4254-4256 TO denotes to
T2145 4257-4263 VB denotes obtain
T2146 4264-4268 CC denotes both
T2147 4269-4272 JJ denotes low
T2148 4273-4276 CC denotes and
T2149 4277-4283 JJ denotes medium
T2150 4284-4293 JJ denotes molecular
T2151 4294-4300 NN denotes weight
T2152 4301-4310 NN denotes fractions
T2153 4312-4319 RB denotes Briefly
T2154 4319-4320 -COMMA- denotes ,
T2155 4321-4324 IN denotes for
T2156 4325-4328 DT denotes the
T2157 4329-4332 JJ denotes low
T2158 4333-4342 JJ denotes molecular
T2159 4343-4349 NN denotes weight
T2160 4350-4358 NN denotes fraction
T2161 4358-4359 -COMMA- denotes ,
T2162 4360-4371 NN denotes carrageenan
T2163 4372-4380 NN denotes solution
T2164 4381-4384 VB denotes was
T2165 4385-4395 VB denotes hydrolyzed
T2166 4396-4400 IN denotes with
T2167 4401-4404 CD denotes 0.3
T2168 4404-4405 NN denotes %
T2169 4406-4407 -LRB- denotes (
T2170 4407-4410 NN denotes v/v
T2171 4410-4411 -RRB- denotes )
T2172 4412-4424 VB denotes concentrated
T2173 4425-4434 JJ denotes sulphuric
T2174 4435-4439 NN denotes acid
T2175 4440-4443 IN denotes for
T2176 4444-4446 CD denotes 15
T2177 4447-4450 NN denotes min
T2178 4451-4453 IN denotes at
T2179 4454-4459 NNP denotes 80°C.
T2180 4460-4465 NNP denotes After
T2181 4466-4480 NN denotes neutralization
T2182 4481-4485 IN denotes with
T2183 4486-4490 NN denotes NaOH
T2184 4491-4493 NN denotes 4N
T2185 4493-4494 -COMMA- denotes ,
T2186 4495-4498 DT denotes the
T2187 4499-4507 NN denotes solution
T2188 4508-4511 VB denotes was
T2189 4512-4517 NN denotes ultra
T2190 4518-4526 VB denotes filtered
T2191 4527-4534 IN denotes through
T2192 4535-4536 DT denotes a
T2193 4537-4543 JJ denotes hollow
T2194 4544-4549 NN denotes fibre
T2195 4550-4559 NN denotes cartridge
T2196 4560-4564 IN denotes with
T2197 4565-4567 NN denotes MW
T2198 4568-4575 JJ denotes cut-off
T2199 4576-4577 CD denotes 5
T2200 4578-4581 NN denotes kDa
T2201 4581-4582 -COMMA- denotes ,
T2202 4583-4584 -LRB- denotes (
T2203 4584-4590 NNP denotes Amicon
T2204 4591-4594 NNP denotes Inc
T2205 4594-4595 -COMMA- denotes ,
T2206 4596-4603 NNP denotes Beverly
T2207 4603-4604 -COMMA- denotes ,
T2208 4605-4608 NNP denotes USA
T2209 4608-4609 -RRB- denotes )
T2210 4611-4614 IN denotes For
T2211 4615-4618 DT denotes the
T2212 4619-4625 NN denotes medium
T2213 4626-4635 JJ denotes molecular
T2214 4636-4642 NN denotes weight
T2215 4643-4651 NN denotes fraction
T2216 4651-4652 -COMMA- denotes ,
T2217 4653-4656 DT denotes the
T2218 4657-4668 NN denotes carrageenan
T2219 4669-4677 NN denotes solution
T2220 4678-4681 VB denotes was
T2221 4682-4692 VB denotes hydrolyzed
T2222 4693-4697 IN denotes with
T2223 4698-4701 CD denotes 0.3
T2224 4701-4702 NN denotes %
T2225 4703-4704 -LRB- denotes (
T2226 4704-4707 NN denotes v/v
T2227 4707-4708 -RRB- denotes )
T2228 4709-4721 VB denotes concentrated
T2229 4722-4731 JJ denotes sulphuric
T2230 4732-4736 NN denotes acid
T2231 4737-4740 IN denotes for
T2232 4741-4743 CD denotes 30
T2233 4744-4747 NN denotes min
T2234 4748-4750 IN denotes at
T2235 4751-4756 NNP denotes 60°C.
T2236 4757-4762 IN denotes After
T2237 4763-4777 NN denotes neutralization
T2238 4777-4778 -COMMA- denotes ,
T2239 4779-4782 DT denotes the
T2240 4783-4794 NN denotes supernatant
T2241 4795-4798 VB denotes was
T2242 4799-4804 AFX denotes ultra
T2243 4805-4813 VB denotes filtered
T2244 4814-4815 -LRB- denotes (
T2245 4815-4817 NN denotes MW
T2246 4818-4825 JJ denotes cut-off
T2247 4826-4829 CD denotes 100
T2248 4830-4833 NN denotes kDa
T2249 4833-4834 -RRB- denotes )
T2250 4836-4839 DT denotes The
T2251 4840-4848 NN denotes filtrate
T2252 4849-4852 VB denotes was
T2253 4853-4862 VB denotes submitted
T2254 4863-4865 TO denotes to
T2255 4866-4867 DT denotes a
T2256 4868-4874 JJ denotes second
T2257 4875-4880 NN denotes ultra
T2258 4881-4891 NN denotes filtration
T2259 4892-4893 -LRB- denotes (
T2260 4893-4895 NN denotes MW
T2261 4896-4903 NN denotes cut-off
T2262 4904-4905 CD denotes 5
T2263 4906-4909 NN denotes kDa
T2264 4909-4910 -RRB- denotes )
T2265 4912-4916 DT denotes Both
T2266 4917-4929 NN denotes preparations
T2267 4930-4932 IN denotes of
T2268 4933-4937 NN denotes dCGN
T2269 4938-4942 VB denotes were
T2270 4943-4955 VB denotes precipitated
T2271 4956-4960 IN denotes with
T2272 4961-4962 CD denotes 4
T2273 4963-4970 NN denotes volumes
T2274 4971-4973 IN denotes of
T2275 4974-4976 CD denotes 95
T2276 4976-4977 NN denotes %
T2277 4978-4985 NN denotes ethanol
T2278 4985-4986 -COMMA- denotes ,
T2279 4987-4992 VB denotes dried
T2280 4993-4995 IN denotes at
T2281 4996-5000 NN denotes room
T2282 5001-5012 NN denotes temperature
T2283 5013-5016 CC denotes and
T2284 5017-5023 NN denotes ground
T2285 5024-5026 TO denotes to
T2286 5027-5032 JJ denotes small
T2287 5033-5042 NN denotes particles
T2288 5043-5044 -LRB- denotes (
T2289 5044-5045 CD denotes 1
T2290 5046-5048 NN denotes mm
T2291 5049-5051 IN denotes in
T2292 5052-5060 NN denotes diameter
T2293 5060-5061 -RRB- denotes )
T2294 5063-5068 VB denotes Using
T2295 5069-5083 NN denotes gel-permeation
T2296 5084-5098 NN denotes chromatography
T2297 5099-5101 IN denotes in
T2298 5102-5113 NN denotes combination
T2299 5114-5118 IN denotes with
T2300 5119-5124 JJ denotes light
T2301 5125-5135 NN denotes scattering
T2302 5136-5148 NN denotes measurements
T2303 5149-5150 -LRB- denotes (
T2304 5150-5153 VB denotes see
T2305 5154-5160 NNP denotes Viebke
T2306 5161-5163 FW denotes et
T2307 5164-5167 FW denotes al.
T2308 5168-5169 -LRB- denotes [
T2309 5169-5171 CD denotes 21
T2310 5171-5172 -RRB- denotes ]
T2311 5172-5173 -RRB- denotes )
T2312 5173-5174 -COMMA- denotes ,
T2313 5175-5177 PRP denotes it
T2314 5178-5181 VB denotes was
T2315 5182-5191 VB denotes confirmed
T2316 5192-5196 IN denotes that
T2317 5197-5200 DT denotes the
T2318 5201-5204 JJ denotes low
T2319 5205-5213 NN denotes fraction
T2320 5214-5217 VB denotes had
T2321 5218-5220 DT denotes an
T2322 5221-5228 JJ denotes average
T2323 5229-5238 JJ denotes molecular
T2324 5239-5245 NN denotes weight
T2325 5246-5248 IN denotes of
T2326 5249-5251 CD denotes 10
T2327 5252-5255 NN denotes kDa
T2328 5255-5256 -COMMA- denotes ,
T2329 5257-5260 CC denotes and
T2330 5261-5264 DT denotes the
T2331 5265-5271 NN denotes medium
T2332 5272-5280 NN denotes fraction
T2333 5281-5283 IN denotes of
T2334 5284-5286 CD denotes 40
T2335 5287-5290 NN denotes kDa
T2336 5292-5295 DT denotes The
T2337 5296-5304 NN denotes sulphate
T2338 5305-5312 NN denotes content
T2339 5313-5315 IN denotes of
T2340 5316-5331 NN denotes polysaccharides
T2341 5332-5334 IN denotes in
T2342 5335-5339 DT denotes both
T2343 5340-5349 NN denotes fractions
T2344 5350-5353 VB denotes was
T2345 5354-5362 VB denotes measured
T2346 5363-5372 VB denotes following
T2347 5373-5376 DT denotes the
T2348 5377-5383 NN denotes method
T2349 5384-5386 IN denotes of
T2350 5387-5395 NNP denotes Quemener
T2351 5396-5398 FW denotes et
T2352 5399-5402 FW denotes al.
T2353 5403-5404 -LRB- denotes [
T2354 5404-5406 CD denotes 22
T2355 5406-5407 -RRB- denotes ]
T2356 5409-5416 RB denotes Finally
T2357 5416-5417 -COMMA- denotes ,
T2358 5418-5421 DT denotes the
T2359 5422-5429 NN denotes absence
T2360 5430-5432 IN denotes of
T2361 5433-5447 NN denotes polysaccharide
T2362 5448-5457 NN denotes structure
T2363 5458-5471 NN denotes modifications
T2364 5472-5474 IN denotes in
T2365 5475-5478 DT denotes the
T2366 5479-5482 CD denotes two
T2367 5483-5492 NN denotes fractions
T2368 5493-5496 VB denotes was
T2369 5497-5506 VB denotes confirmed
T2370 5507-5512 VB denotes using
T2371 5513-5519 NN denotes 2H-NMR
T2372 5520-5532 NN denotes spectroscopy
T2373 5534-5537 DT denotes The
T2374 5538-5545 NN denotes absence
T2375 5546-5548 IN denotes of
T2376 5549-5552 NN denotes LPS
T2377 5553-5566 NN denotes contamination
T2378 5567-5569 IN denotes in
T2379 5570-5573 DT denotes the
T2380 5574-5577 CD denotes two
T2381 5578-5587 NN denotes fractions
T2382 5588-5591 VB denotes was
T2383 5592-5601 VB denotes confirmed
T2384 5602-5607 VB denotes using
T2385 5608-5611 DT denotes the
T2386 5612-5621 NN denotes e-Toxate®
T2387 5622-5625 NN denotes kit
T2388 5626-5627 -LRB- denotes (
T2389 5627-5632 NNP denotes Sigma
T2390 5632-5633 -COMMA- denotes ,
T2391 5634-5636 NNP denotes St
T2392 5637-5644 NNP denotes Quentin
T2393 5645-5654 NNP denotes Fallavier
T2394 5654-5655 -COMMA- denotes ,
T2395 5656-5662 NNP denotes France
T2396 5662-5663 -RRB- denotes )
T2397 5665-5671 IN denotes Before
T2398 5672-5675 NN denotes use
T2399 5676-5678 IN denotes in
T2400 5679-5683 NN denotes cell
T2401 5684-5691 NN denotes culture
T2402 5691-5692 -COMMA- denotes ,
T2403 5693-5696 DT denotes the
T2404 5697-5700 CD denotes two
T2405 5701-5710 NN denotes fractions
T2406 5711-5715 VB denotes were
T2407 5716-5725 VB denotes dissolved
T2408 5726-5728 IN denotes in
T2409 5729-5737 JJ denotes complete
T2410 5738-5744 NN denotes medium
T2411 5745-5751 IN denotes during
T2412 5752-5754 CD denotes 30
T2413 5755-5758 NN denotes min
T2414 5759-5761 IN denotes at
T2415 5762-5766 NN denotes 56°C
T2818 5776-5777 -COMMA- denotes ,
T2819 5778-5787 NNP denotes Chemicals
T2820 5788-5791 CC denotes and
T2821 5792-5796 NNP denotes Diet
T2822 5797-5801 JJ denotes Male
T2823 5802-5808 NNP denotes Wistar
T2824 5809-5813 NN denotes rats
T2825 5814-5815 -LRB- denotes (
T2826 5815-5818 CD denotes 150
T2827 5819-5820 NN denotes g
T2828 5821-5828 JJ denotes average
T2829 5829-5835 NN denotes weight
T2830 5835-5836 -RRB- denotes )
T2831 5837-5841 VB denotes were
T2832 5842-5848 VB denotes housed
T2833 5849-5854 IN denotes under
T2834 5855-5863 JJ denotes standard
T2835 5864-5874 NN denotes conditions
T2836 5875-5878 CC denotes and
T2837 5879-5882 VB denotes fed
T2838 5883-5885 NN denotes ad
T2839 5886-5893 NN denotes libitum
T2840 5894-5898 IN denotes with
T2841 5899-5907 JJ denotes standard
T2842 5908-5914 NN denotes rodent
T2843 5915-5925 NN denotes laboratory
T2844 5926-5930 NN denotes chow
T2845 5932-5940 VB denotes Degraded
T2846 5941-5958 NN denotes iota-carrageenans
T2847 5959-5963 VB denotes were
T2848 5964-5976 VB denotes administered
T2849 5977-5979 IN denotes in
T2850 5980-5983 DT denotes the
T2851 5984-5992 NN denotes drinking
T2852 5993-5998 NN denotes water
T2853 5999-6000 -LRB- denotes (
T2854 6000-6001 CD denotes 5
T2855 6001-6002 NN denotes %
T2856 6003-6006 NN denotes w/v
T2857 6006-6007 -RRB- denotes )
T2858 6008-6011 IN denotes for
T2859 6012-6014 CD denotes 55
T2860 6015-6019 NN denotes days
T2861 6020-6022 TO denotes to
T2862 6023-6024 CD denotes 2
T2863 6025-6031 NN denotes groups
T2864 6032-6034 IN denotes of
T2865 6035-6038 CD denotes six
T2866 6039-6046 NN denotes animals
T2867 6047-6051 DT denotes each
T2868 6053-6056 DT denotes The
T2869 6057-6062 JJ denotes first
T2870 6063-6068 NN denotes group
T2871 6069-6077 VB denotes received
T2872 6078-6081 DT denotes the
T2873 6082-6085 JJ denotes low
T2874 6086-6095 JJ denotes molecular
T2875 6096-6102 NN denotes weight
T2876 6103-6114 NN denotes carrageenan
T2877 6115-6116 -LRB- denotes (
T2878 6116-6118 CD denotes 10
T2879 6119-6122 NN denotes kDa
T2880 6123-6127 NN denotes dCGN
T2881 6127-6128 -RRB- denotes )
T2882 6129-6132 CC denotes and
T2883 6133-6136 DT denotes the
T2884 6137-6143 NN denotes second
T2885 6144-6152 VB denotes received
T2886 6153-6156 DT denotes the
T2887 6157-6163 NN denotes medium
T2888 6164-6173 JJ denotes molecular
T2889 6174-6180 NN denotes weight
T2890 6181-6192 NN denotes carrageenan
T2891 6193-6194 -LRB- denotes (
T2892 6194-6196 CD denotes 40
T2893 6197-6200 NN denotes kDa
T2894 6201-6205 NN denotes dCGN
T2895 6205-6206 -RRB- denotes )
T2896 6208-6210 DT denotes An
T2897 6211-6221 JJ denotes additional
T2898 6222-6227 NN denotes group
T2899 6228-6230 IN denotes of
T2900 6231-6235 CD denotes four
T2901 6236-6240 NN denotes rats
T2902 6241-6245 VB denotes were
T2903 6246-6256 VB denotes maintained
T2904 6257-6259 IN denotes on
T2905 6260-6267 JJ denotes regular
T2906 6268-6271 NN denotes tap
T2907 6272-6277 NN denotes water
T2908 6278-6279 -LRB- denotes (
T2909 6279-6286 NN denotes control
T2910 6287-6292 NN denotes group
T2911 6292-6293 -RRB- denotes )
T2912 6295-6297 TO denotes To
T2913 6298-6306 VB denotes increase
T2914 6307-6319 NN denotes palatability
T2915 6320-6323 CD denotes 0.2
T2916 6323-6324 NN denotes %
T2917 6325-6332 NN denotes sucrose
T2918 6333-6336 VB denotes was
T2919 6337-6342 VB denotes added
T2920 6343-6345 TO denotes to
T2921 6346-6349 DT denotes the
T2922 6350-6358 NN denotes drinking
T2923 6359-6364 NN denotes water
T2924 6365-6367 IN denotes of
T2925 6368-6371 DT denotes all
T2926 6372-6378 NN denotes groups
T2927 6379-6380 -LRB- denotes (
T2928 6380-6383 NNP denotes Van
T2929 6384-6387 NNP denotes der
T2930 6388-6393 NNP denotes Waaji
T2931 6394-6396 FW denotes et
T2932 6397-6400 FW denotes al.
T2933 6400-6401 -COMMA- denotes ,
T2934 6402-6403 -LRB- denotes [
T2935 6403-6405 CD denotes 23
T2936 6405-6406 -RRB- denotes ]
T2937 6406-6407 -RRB- denotes )
T2938 6409-6414 JJ denotes Fresh
T2939 6415-6426 NN denotes carrageenan
T2940 6427-6436 NN denotes solutions
T2941 6437-6441 VB denotes were
T2942 6442-6450 VB denotes prepared
T2943 6451-6456 RB denotes daily
T3130 6459-6469 NN denotes Evaluation
T3131 6470-6472 IN denotes of
T3132 6473-6480 NN denotes Colitis
T3133 6481-6485 NNP denotes Body
T3134 6486-6492 NN denotes weight
T3135 6492-6493 -COMMA- denotes ,
T3136 6494-6500 NN denotes liquid
T3137 6501-6504 CC denotes and
T3138 6505-6509 NN denotes food
T3139 6510-6521 NN denotes consumption
T3140 6521-6522 -COMMA- denotes ,
T3141 6523-6531 NN denotes diarrhea
T3142 6532-6535 CC denotes and
T3143 6536-6542 JJ denotes rectal
T3144 6543-6551 NN denotes bleeding
T3145 6552-6553 -LRB- denotes (
T3146 6553-6561 VB denotes detected
T3147 6562-6564 IN denotes by
T3148 6565-6568 NN denotes eye
T3149 6569-6579 NN denotes inspection
T3150 6579-6580 -RRB- denotes )
T3151 6581-6585 VB denotes were
T3152 6586-6594 VB denotes recorded
T3153 6595-6605 IN denotes throughout
T3154 6606-6609 DT denotes the
T3155 6610-6617 NN denotes feeding
T3156 6618-6624 NN denotes period
T3157 6626-6631 IN denotes After
T3158 6632-6634 CD denotes 55
T3159 6635-6639 NN denotes days
T3160 6639-6640 -COMMA- denotes ,
T3161 6641-6648 NN denotes animals
T3162 6649-6653 VB denotes were
T3163 6654-6664 VB denotes sacrificed
T3164 6665-6667 IN denotes by
T3165 6668-6676 JJ denotes cervical
T3166 6677-6688 NN denotes dislocation
T3167 6690-6693 DT denotes The
T3168 6694-6700 NN denotes length
T3169 6701-6703 IN denotes of
T3170 6704-6707 DT denotes the
T3171 6708-6713 NN denotes colon
T3172 6714-6717 VB denotes was
T3173 6718-6726 VB denotes measured
T3174 6727-6729 IN denotes as
T3175 6730-6739 VB denotes described
T3176 6740-6742 IN denotes by
T3177 6743-6751 NNP denotes Okayashu
T3178 6752-6754 FW denotes et
T3179 6755-6758 FW denotes al.
T3180 6759-6760 -LRB- denotes [
T3181 6760-6762 CD denotes 24
T3182 6762-6763 -RRB- denotes ]
T3183 6765-6769 RB denotes Then
T3184 6769-6770 -COMMA- denotes ,
T3185 6771-6775 DT denotes each
T3186 6776-6781 NN denotes colon
T3187 6782-6785 VB denotes was
T3188 6786-6793 VB denotes ligated
T3189 6794-6796 IN denotes in
T3190 6797-6805 NN denotes sections
T3191 6806-6808 IN denotes of
T3192 6809-6810 CD denotes 2
T3193 6811-6813 NN denotes cm
T3194 6814-6817 CC denotes and
T3195 6818-6819 CD denotes 1
T3196 6820-6822 TO denotes to
T3197 6823-6824 CD denotes 2
T3198 6825-6827 NN denotes ml
T3199 6828-6830 IN denotes of
T3200 6831-6833 CD denotes 10
T3201 6833-6834 NN denotes %
T3202 6835-6843 NN denotes formalin
T3203 6844-6847 VB denotes was
T3204 6848-6855 VB denotes infused
T3205 6856-6860 IN denotes into
T3206 6861-6864 DT denotes the
T3207 6865-6875 JJ denotes intestinal
T3208 6876-6881 NN denotes lumen
T3209 6883-6886 DT denotes The
T3210 6887-6897 RB denotes moderately
T3211 6898-6907 VB denotes distended
T3212 6908-6915 NN denotes segment
T3213 6916-6919 VB denotes was
T3214 6920-6929 VB denotes sectioned
T3215 6930-6933 CC denotes and
T3216 6934-6939 VB denotes fixed
T3217 6940-6942 IN denotes in
T3218 6943-6945 CD denotes 10
T3219 6945-6946 NN denotes %
T3220 6947-6955 NN denotes formalin
T3221 6957-6960 DT denotes The
T3222 6961-6970 VB denotes following
T3223 6971-6974 NN denotes day
T3224 6974-6975 -COMMA- denotes ,
T3225 6976-6979 DT denotes the
T3226 6980-6990 JJ denotes intestinal
T3227 6991-6998 NN denotes content
T3228 6999-7002 VB denotes was
T3229 7003-7010 VB denotes removed
T3230 7011-7013 IN denotes by
T3231 7014-7023 NN denotes vortexing
T3232 7025-7028 DT denotes The
T3233 7029-7034 VB denotes fixed
T3234 7035-7042 NN denotes segment
T3235 7043-7046 VB denotes was
T3236 7047-7051 VB denotes kept
T3237 7052-7054 IN denotes in
T3238 7055-7057 CD denotes 10
T3239 7057-7058 NN denotes %
T3240 7059-7067 NN denotes formalin
T3241 7068-7070 IN denotes at
T3242 7071-7074 NN denotes 4°C
T3243 7075-7080 IN denotes until
T3244 7081-7084 DT denotes the
T3245 7085-7093 NN denotes paraffin
T3246 7094-7103 NN denotes embedding
T3247 7104-7113 NN denotes procedure
T3248 7115-7117 TO denotes To
T3249 7118-7126 VB denotes evaluate
T3250 7127-7130 DT denotes the
T3251 7131-7137 NN denotes degree
T3252 7138-7140 IN denotes of
T3253 7141-7153 NN denotes inflammation
T3254 7153-7154 -COMMA- denotes ,
T3255 7155-7159 DT denotes this
T3256 7160-7167 NN denotes segment
T3257 7168-7170 IN denotes of
T3258 7171-7176 NN denotes colon
T3259 7177-7180 VB denotes was
T3260 7181-7187 VB denotes opened
T3261 7188-7202 RB denotes longitudinally
T3262 7203-7206 CC denotes and
T3263 7207-7218 JJ denotes macroscopic
T3264 7219-7222 CC denotes and
T3265 7223-7235 JJ denotes histological
T3266 7236-7242 NN denotes scores
T3267 7243-7245 IN denotes of
T3268 7246-7258 NN denotes inflammation
T3269 7259-7263 VB denotes were
T3270 7264-7272 VB denotes recorded
T3271 7273-7275 IN denotes as
T3272 7276-7286 RB denotes previously
T3273 7287-7296 VB denotes described
T3274 7297-7298 -LRB- denotes [
T3275 7298-7300 CD denotes 25
T3276 7300-7301 -RRB- denotes ]
T3277 7301-7302 -COMMA- denotes ,
T3278 7303-7304 -LRB- denotes [
T3279 7304-7306 CD denotes 26
T3280 7306-7307 -RRB- denotes ]
T3281 7309-7312 DT denotes The
T3282 7313-7322 NN denotes toluidine
T3283 7323-7327 JJ denotes blue
T3284 7328-7336 NN denotes staining
T3285 7337-7340 VB denotes was
T3286 7341-7345 VB denotes used
T3287 7346-7349 IN denotes for
T3288 7350-7364 NN denotes identification
T3289 7365-7367 IN denotes of
T3290 7368-7377 VB denotes sulphated
T3291 7378-7393 NN denotes polysaccharides
T3292 7394-7396 IN denotes in
T3293 7397-7400 DT denotes the
T3294 7401-7411 JJ denotes intestinal
T3295 7412-7418 NN denotes mucosa
T3296 7420-7422 IN denotes On
T3297 7423-7426 DT denotes the
T3298 7427-7430 NN denotes day
T3299 7431-7433 IN denotes of
T3300 7434-7443 NN denotes sacrifice
T3301 7443-7444 -COMMA- denotes ,
T3302 7445-7446 DT denotes a
T3303 7447-7452 JJ denotes fresh
T3304 7453-7459 NN denotes sample
T3305 7460-7462 IN denotes of
T3306 7463-7467 DT denotes each
T3307 7468-7473 NN denotes colon
T3308 7474-7475 -LRB- denotes (
T3309 7475-7477 CD denotes 50
T3310 7478-7480 NN denotes mg
T3311 7480-7481 -RRB- denotes )
T3312 7482-7485 VB denotes was
T3313 7486-7495 VB denotes collected
T3314 7496-7499 IN denotes for
T3315 7500-7515 NN denotes myeloperoxidase
T3316 7516-7517 -LRB- denotes (
T3317 7517-7520 NN denotes MPO
T3318 7520-7521 -RRB- denotes )
T3319 7522-7527 NN denotes assay
T3320 7528-7537 VB denotes according
T3321 7538-7540 TO denotes to
T3322 7541-7548 NNP denotes Krawisz
T3323 7549-7551 FW denotes et
T3324 7552-7555 FW denotes al.
T3325 7555-7556 -COMMA- denotes ,
T3326 7557-7558 -LRB- denotes [
T3327 7558-7560 CD denotes 27
T3328 7560-7561 -RRB- denotes ]
T3329 7563-7566 DT denotes The
T3330 7567-7572 NN denotes level
T3331 7573-7575 IN denotes of
T3332 7576-7579 NN denotes MPO
T3333 7579-7580 -COMMA- denotes ,
T3334 7581-7587 RB denotes mainly
T3335 7588-7597 VB denotes expressed
T3336 7598-7600 IN denotes by
T3337 7601-7612 NN denotes neutrophils
T3338 7612-7613 -COMMA- denotes ,
T3339 7614-7623 VB denotes indicates
T3340 7624-7627 DT denotes the
T3341 7628-7632 NN denotes rate
T3342 7633-7635 IN denotes of
T3343 7636-7647 NN denotes recruitment
T3344 7648-7650 IN denotes of
T3345 7651-7662 NN denotes neutrophils
T3346 7663-7665 TO denotes to
T3347 7666-7669 DT denotes the
T3348 7670-7680 JJ denotes intestinal
T3349 7681-7687 NN denotes mucosa
T3350 7689-7692 CD denotes One
T3351 7693-7697 NN denotes unit
T3352 7698-7700 IN denotes of
T3353 7701-7704 NN denotes MPO
T3354 7705-7713 NN denotes activity
T3355 7714-7725 VB denotes corresponds
T3356 7726-7728 TO denotes to
T3357 7729-7732 DT denotes the
T3358 7733-7744 NN denotes degradation
T3359 7745-7747 IN denotes of
T3360 7748-7749 CD denotes 1
T3361 7750-7754 NN denotes µmol
T3362 7755-7757 IN denotes of
T3363 7758-7766 NN denotes peroxide
T3364 7767-7770 IN denotes per
T3365 7771-7777 NN denotes minute
T3366 7778-7780 IN denotes at
T3367 7781-7785 NN denotes 25°C
T3676 7788-7792 NNP denotes Cell
T3677 7793-7800 NNP denotes Culture
T3678 7801-7804 DT denotes All
T3679 7805-7811 NN denotes tissue
T3680 7812-7819 NN denotes culture
T3681 7820-7828 NN denotes reagents
T3682 7829-7833 VB denotes were
T3683 7834-7838 IN denotes from
T3684 7839-7849 NN denotes Invitrogen
T3685 7850-7851 -LRB- denotes (
T3686 7851-7856 NNP denotes Cergy
T3687 7857-7865 NNP denotes Pontoise
T3688 7865-7866 -COMMA- denotes ,
T3689 7867-7873 NNP denotes France
T3690 7873-7874 -RRB- denotes )
T3691 7876-7881 NN denotes THP-1
T3692 7882-7887 JJ denotes human
T3693 7888-7897 JJ denotes monocytic
T3694 7898-7903 NN denotes cells
T3695 7904-7908 VB denotes were
T3696 7909-7919 VB denotes maintained
T3697 7920-7922 IN denotes in
T3698 7923-7932 NN denotes RPMI-1640
T3699 7933-7945 VB denotes supplemented
T3700 7946-7950 IN denotes with
T3701 7951-7953 CD denotes 10
T3702 7953-7954 NN denotes %
T3703 7955-7958 NN denotes FCS
T3704 7958-7959 -COMMA- denotes ,
T3705 7960-7961 CD denotes 2
T3706 7962-7964 NN denotes mM
T3707 7965-7966 NN denotes L
T3708 7967-7977 NN denotes -glutamine
T3709 7977-7978 -COMMA- denotes ,
T3710 7979-7981 CD denotes 50
T3711 7982-7986 NN denotes U/ml
T3712 7987-7997 NN denotes penicillin
T3713 7998-8001 CC denotes and
T3714 8002-8004 CD denotes 50
T3715 8005-8010 NN denotes mg/ml
T3716 8011-8023 NN denotes streptomycin
T3717 8024-8026 IN denotes at
T3718 8027-8031 NN denotes 37°C
T3719 8032-8034 IN denotes in
T3720 8035-8036 DT denotes a
T3721 8037-8038 CD denotes 5
T3722 8038-8039 NN denotes %
T3723 8040-8043 NN denotes CO2
T3724 8044-8053 NN denotes incubator
T3725 8055-8060 JJ denotes Human
T3726 8061-8071 JJ denotes peripheral
T3727 8072-8077 NN denotes blood
T3728 8078-8089 JJ denotes mononuclear
T3729 8090-8095 NN denotes cells
T3730 8096-8100 VB denotes were
T3731 8101-8109 VB denotes obtained
T3732 8110-8114 IN denotes from
T3733 8115-8126 VB denotes heparinized
T3734 8127-8132 NN denotes blood
T3735 8133-8135 IN denotes by
T3736 8136-8150 JJ denotes Ficoll-Hypaque
T3737 8151-8158 NN denotes density
T3738 8159-8167 NN denotes gradient
T3739 8169-8178 NN denotes Monocytes
T3740 8179-8183 VB denotes were
T3741 8184-8188 RB denotes then
T3742 8189-8197 VB denotes isolated
T3743 8198-8200 IN denotes by
T3744 8201-8210 NN denotes adherence
T3745 8211-8213 TO denotes to
T3746 8214-8221 NN denotes culture
T3747 8222-8228 NN denotes flasks
T3748 8229-8231 IN denotes as
T3749 8232-8241 VB denotes described
T3750 8242-8243 -LRB- denotes [
T3751 8243-8245 CD denotes 28
T3752 8245-8246 -RRB- denotes ]
T3753 8248-8251 IN denotes For
T3754 8252-8256 NN denotes cell
T3755 8257-8268 NN denotes aggregation
T3756 8268-8269 -COMMA- denotes ,
T3757 8270-8279 NN denotes monocytes
T3758 8280-8284 VB denotes were
T3759 8285-8293 VB denotes cultured
T3760 8294-8296 IN denotes in
T3761 8297-8300 DT denotes the
T3762 8301-8309 NN denotes presence
T3763 8310-8312 CC denotes or
T3764 8313-8320 NN denotes absence
T3765 8321-8323 IN denotes of
T3766 8324-8327 NN denotes C10
T3767 8328-8330 CC denotes or
T3768 8331-8334 NN denotes C40
T3769 8335-8338 IN denotes for
T3770 8339-8341 CD denotes 72
T3771 8342-8343 NN denotes h
T3772 8345-8349 NN denotes Cell
T3773 8350-8358 NN denotes colonies
T3774 8359-8363 VB denotes were
T3775 8364-8373 VB denotes monitored
T3776 8374-8379 IN denotes under
T3777 8380-8382 DT denotes an
T3778 8383-8391 VB denotes inverted
T3779 8392-8397 NN denotes phase
T3780 8398-8406 NN denotes contrast
T3781 8407-8417 NN denotes microscope
T3782 8418-8425 VB denotes coupled
T3783 8426-8433 IN denotes through
T3784 8434-8435 DT denotes a
T3785 8436-8441 NN denotes video
T3786 8442-8448 NN denotes camera
T3787 8449-8451 TO denotes to
T3788 8452-8453 DT denotes a
T3789 8454-8462 NN denotes computer
T3790 8464-8466 IN denotes In
T3791 8467-8471 DT denotes some
T3792 8472-8477 NN denotes wells
T3793 8477-8478 -COMMA- denotes ,
T3794 8479-8491 VB denotes neutralizing
T3795 8492-8502 JJ denotes monoclonal
T3796 8503-8511 NN denotes antibody
T3797 8512-8514 TO denotes to
T3798 8515-8521 NN denotes ICAM-1
T3799 8522-8523 -LRB- denotes (
T3800 8523-8526 CD denotes 2.5
T3801 8527-8532 NN denotes µg/ml
T3802 8532-8533 -RRB- denotes )
T3803 8534-8535 -LRB- denotes (
T3804 8535-8539 NNP denotes Tebu
T3805 8539-8540 -COMMA- denotes ,
T3806 8541-8543 NNP denotes Le
T3807 8544-8550 NNP denotes Perray
T3808 8551-8553 NNP denotes en
T3809 8554-8562 NNP denotes Yvelines
T3810 8562-8563 -COMMA- denotes ,
T3811 8564-8570 NNP denotes France
T3812 8570-8571 -RRB- denotes )
T3813 8572-8575 VB denotes was
T3814 8576-8581 VB denotes added
T3986 8584-8588 NN denotes Cell
T3987 8589-8594 NN denotes Cycle
T3988 8595-8603 NN denotes Analysis
T3989 8604-8609 NN denotes THP-1
T3990 8610-8615 NN denotes cells
T3991 8616-8618 IN denotes in
T3992 8619-8630 JJ denotes exponential
T3993 8631-8637 NN denotes growth
T3994 8638-8643 NN denotes phase
T3995 8644-8648 VB denotes were
T3996 8649-8656 VB denotes exposed
T3997 8657-8659 TO denotes to
T3998 8660-8668 JJ denotes complete
T3999 8669-8675 NN denotes medium
T4000 8676-8678 IN denotes in
T4001 8679-8682 DT denotes the
T4002 8683-8691 NN denotes presence
T4003 8692-8694 CC denotes or
T4004 8695-8702 NN denotes absence
T4005 8703-8705 IN denotes of
T4006 8706-8718 NN denotes carrageenans
T4007 8719-8722 IN denotes for
T4008 8723-8725 CD denotes 24
T4009 8726-8727 NN denotes h
T4010 8728-8734 IN denotes before
T4011 8735-8740 VB denotes being
T4012 8741-8748 VB denotes stained
T4013 8749-8753 IN denotes with
T4014 8754-8763 NN denotes propidium
T4015 8764-8770 NN denotes iodide
T4016 8771-8776 VB denotes using
T4017 8777-8780 DT denotes the
T4018 8781-8789 JJ denotes DNA-Prep
T4019 8790-8797 NNP denotes Coulter
T4020 8798-8801 NN denotes kit
T4021 8802-8811 VB denotes according
T4022 8812-8814 TO denotes to
T4023 8815-8818 DT denotes the
T4024 8819-8831 NN denotes manufacturer
T4025 8831-8833 POS denotes 's
T4026 8834-8845 NN denotes instruction
T4027 8846-8847 -LRB- denotes (
T4028 8847-8862 NNP denotes Beckman-Coulter
T4029 8862-8863 -COMMA- denotes ,
T4030 8864-8874 NNP denotes Villepinte
T4031 8874-8875 -COMMA- denotes ,
T4032 8876-8882 NNP denotes France
T4033 8882-8883 -RRB- denotes )
T4034 8885-8889 NN denotes Cell
T4035 8890-8893 NN denotes DNA
T4036 8894-8901 NN denotes content
T4037 8902-8905 VB denotes was
T4038 8906-8910 RB denotes then
T4039 8911-8919 VB denotes analyzed
T4040 8920-8922 IN denotes by
T4041 8923-8927 NN denotes flow
T4042 8928-8937 NN denotes cytometry
T4043 8938-8943 VB denotes using
T4044 8944-8946 DT denotes an
T4045 8947-8952 NN denotes EPICS
T4046 8953-8956 NN denotes XL2
T4047 8957-8958 -LRB- denotes (
T4048 8958-8973 NN denotes Beckman-Coulter
T4049 8973-8974 -RRB- denotes )
T4050 8976-8979 NN denotes Raw
T4051 8980-8984 NN denotes data
T4052 8985-8988 IN denotes for
T4053 8989-8992 DT denotes the
T4054 8993-9005 NN denotes distribution
T4055 9006-9008 IN denotes of
T4056 9009-9012 NN denotes DNA
T4057 9013-9020 NN denotes content
T4058 9021-9023 IN denotes of
T4059 9024-9030 CD denotes 30,000
T4060 9031-9036 NN denotes cells
T4061 9037-9046 VB denotes retrieved
T4062 9047-9051 IN denotes from
T4063 9052-9055 DT denotes the
T4064 9056-9065 NN denotes cytometer
T4065 9066-9070 VB denotes were
T4066 9071-9080 VB denotes expressed
T4067 9081-9083 IN denotes as
T4068 9084-9087 DT denotes the
T4069 9088-9098 NN denotes percentage
T4070 9099-9101 IN denotes of
T4071 9102-9107 NN denotes G0/G1
T4072 9108-9115 IN denotes through
T4073 9116-9120 NN denotes G2/M
T4074 9121-9132 NN denotes populations
T4075 9134-9144 NNP denotes Multicycle
T4076 9145-9147 NN denotes AV
T4077 9148-9156 NN denotes software
T4078 9157-9158 -LRB- denotes (
T4079 9158-9165 NNP denotes Phoenix
T4080 9166-9170 NNP denotes Flow
T4081 9171-9178 NNP denotes Systems
T4082 9178-9179 -COMMA- denotes ,
T4083 9180-9183 NNP denotes San
T4084 9184-9189 NNP denotes Diego
T4085 9189-9190 -COMMA- denotes ,
T4086 9191-9193 NN denotes CA
T4087 9193-9194 -RRB- denotes )
T4088 9195-9198 VB denotes was
T4089 9199-9203 VB denotes used
T4090 9204-9206 TO denotes to
T4091 9207-9215 VB denotes generate
T4092 9216-9219 NN denotes DNA
T4093 9220-9227 NN denotes content
T4094 9228-9237 NN denotes frequency
T4095 9238-9248 NN denotes histograms
T4096 9249-9252 CC denotes and
T4097 9253-9263 VB denotes facilitate
T4098 9264-9268 NN denotes data
T4099 9269-9277 NN denotes analysis
T4242 9280-9284 NNP denotes Cell
T4243 9285-9292 NNP denotes Surface
T4244 9293-9300 NNP denotes Antigen
T4245 9301-9311 NN denotes Expression
T4246 9312-9320 NN denotes Analysis
T4247 9321-9331 JJ denotes Peripheral
T4248 9332-9337 NN denotes Blood
T4249 9338-9347 NN denotes Monocytes
T4250 9348-9350 CC denotes or
T4251 9351-9356 NN denotes THP-1
T4252 9357-9362 NN denotes cells
T4253 9363-9367 VB denotes were
T4254 9368-9375 VB denotes exposed
T4255 9376-9378 TO denotes to
T4256 9379-9387 JJ denotes complete
T4257 9388-9394 NN denotes medium
T4258 9395-9397 IN denotes in
T4259 9398-9401 DT denotes the
T4260 9402-9410 NN denotes presence
T4261 9411-9413 CC denotes or
T4262 9414-9421 NN denotes absence
T4263 9422-9424 IN denotes of
T4264 9425-9436 NN denotes carrageenan
T4265 9437-9440 IN denotes for
T4266 9441-9443 CD denotes 36
T4267 9444-9445 NN denotes h
T4268 9447-9452 IN denotes After
T4269 9453-9456 CD denotes two
T4270 9457-9463 NN denotes washes
T4271 9464-9466 IN denotes in
T4272 9467-9470 NNP denotes PBS
T4273 9471-9478 IN denotes without
T4274 9479-9483 NN denotes Ca2+
T4275 9484-9487 CC denotes and
T4276 9488-9492 NN denotes Mg2+
T4277 9492-9493 -COMMA- denotes ,
T4278 9494-9499 NN denotes cells
T4279 9500-9504 VB denotes were
T4280 9505-9514 VB denotes incubated
T4281 9515-9517 IN denotes in
T4282 9518-9521 NN denotes PBS
T4283 9522-9532 VB denotes containing
T4284 9533-9536 CD denotes 0.1
T4285 9536-9537 NN denotes %
T4286 9538-9545 NN denotes gelatin
T4287 9546-9549 CC denotes and
T4288 9550-9551 CD denotes 8
T4289 9551-9552 NN denotes %
T4290 9553-9555 NN denotes AB
T4291 9556-9561 JJ denotes human
T4292 9562-9567 NN denotes serum
T4293 9568-9570 TO denotes to
T4294 9571-9578 VB denotes prevent
T4295 9579-9586 NN denotes binding
T4296 9587-9589 TO denotes to
T4297 9590-9592 NN denotes Fc
T4298 9593-9602 NN denotes receptors
T4299 9604-9608 RB denotes Then
T4300 9608-9609 -COMMA- denotes ,
T4301 9610-9615 NN denotes 5×105
T4302 9616-9621 NN denotes cells
T4303 9622-9626 VB denotes were
T4304 9627-9636 VB denotes incubated
T4305 9637-9641 IN denotes with
T4306 9642-9649 JJ denotes primary
T4307 9650-9660 NN denotes antibodies
T4308 9661-9663 IN denotes at
T4309 9664-9667 NN denotes 4°C
T4310 9668-9671 IN denotes for
T4311 9672-9674 CD denotes 30
T4312 9675-9678 NN denotes min
T4313 9680-9683 CD denotes Two
T4314 9684-9689 JJ denotes other
T4315 9690-9696 NN denotes washes
T4316 9697-9699 IN denotes in
T4317 9700-9703 NNP denotes PBS
T4318 9704-9712 VB denotes preceded
T4319 9713-9723 NN denotes incubation
T4320 9724-9728 IN denotes with
T4321 9729-9744 JJ denotes FITC-conjugated
T4322 9745-9749 NN denotes goat
T4323 9750-9758 NN denotes antibody
T4324 9759-9769 JJ denotes anti-mouse
T4325 9770-9773 NN denotes IgG
T4326 9774-9781 VB denotes diluted
T4327 9782-9788 CD denotes 1/1000
T4328 9789-9791 IN denotes at
T4329 9792-9795 NN denotes 4°C
T4330 9796-9799 IN denotes for
T4331 9800-9802 CD denotes 30
T4332 9803-9806 NN denotes min
T4333 9807-9808 -LRB- denotes (
T4334 9808-9812 NN denotes Tebu
T4335 9812-9813 -RRB- denotes )
T4336 9815-9820 IN denotes After
T4337 9821-9824 CD denotes two
T4338 9825-9835 JJ denotes additional
T4339 9836-9842 NN denotes washes
T4340 9842-9843 -COMMA- denotes ,
T4341 9844-9852 NN denotes analysis
T4342 9853-9855 IN denotes of
T4343 9856-9863 VB denotes stained
T4344 9864-9869 NN denotes cells
T4345 9870-9873 VB denotes was
T4346 9874-9883 VB denotes performed
T4347 9884-9886 IN denotes on
T4348 9887-9889 DT denotes an
T4349 9890-9895 NN denotes EPICS
T4350 9896-9899 NN denotes XL2
T4351 9900-9901 -LRB- denotes (
T4352 9901-9916 NN denotes Beckman-Coulter
T4353 9916-9917 -RRB- denotes )
T4354 9919-9922 DT denotes The
T4355 9923-9927 NN denotes cell
T4356 9928-9938 NN denotes population
T4357 9939-9942 VB denotes was
T4358 9943-9948 VB denotes gated
T4359 9949-9958 VB denotes according
T4360 9959-9961 TO denotes to
T4361 9962-9965 PRP-DOLLAR- denotes its
T4362 9966-9973 JJ denotes forward
T4363 9974-9977 CC denotes and
T4364 9978-9988 JJ denotes wide-angle
T4365 9989-9994 NN denotes light
T4366 9995-10005 NN denotes scattering
T4367 10007-10011 NN denotes Data
T4368 10012-10016 VB denotes were
T4369 10017-10026 VB denotes expressed
T4370 10027-10029 IN denotes as
T4371 10030-10034 JJ denotes mean
T4372 10035-10043 JJ denotes relative
T4373 10044-10056 NN denotes fluorescence
T4374 10057-10066 NN denotes intensity
T4375 10067-10068 -LRB- denotes (
T4376 10068-10071 NN denotes MFI
T4377 10071-10072 -RRB- denotes )
T4378 10073-10075 IN denotes of
T4379 10076-10080 CD denotes 3000
T4380 10081-10086 NN denotes cells
T4587 10089-10092 NN denotes TNF
T4588 10093-10101 NNP denotes Activity
T4589 10102-10110 NNP denotes Bioassay
T4590 10111-10120 NN denotes Monocytes
T4591 10121-10123 CC denotes or
T4592 10124-10129 NN denotes THP-1
T4593 10130-10135 NN denotes cells
T4594 10136-10140 VB denotes were
T4595 10141-10149 VB denotes cultured
T4596 10150-10154 IN denotes with
T4597 10155-10157 CC denotes or
T4598 10158-10165 IN denotes without
T4599 10166-10175 JJ denotes different
T4600 10176-10190 NN denotes concentrations
T4601 10191-10193 IN denotes of
T4602 10194-10198 NN denotes CGNs
T4603 10199-10201 CC denotes or
T4604 10202-10205 NN denotes LPS
T4605 10206-10207 -LRB- denotes (
T4606 10207-10217 NN denotes Salmonella
T4607 10218-10225 NN denotes typhosa
T4608 10225-10226 -COMMA- denotes ,
T4609 10227-10232 NNP denotes Sigma
T4610 10232-10233 -RRB- denotes )
T4611 10234-10237 IN denotes for
T4612 10238-10240 CD denotes 24
T4613 10241-10242 NN denotes h
T4614 10243-10245 CC denotes or
T4615 10246-10249 DT denotes the
T4616 10250-10259 VB denotes indicated
T4617 10260-10264 NN denotes time
T4618 10266-10278 RB denotes Biologically
T4619 10279-10285 JJ denotes active
T4620 10286-10293 NN denotes TNF-α/β
T4621 10294-10296 IN denotes in
T4622 10297-10303 NN denotes tissue
T4623 10304-10311 NN denotes culture
T4624 10312-10323 NN denotes supernatant
T4625 10324-10327 VB denotes was
T4626 10328-10336 VB denotes measured
T4627 10337-10342 VB denotes using
T4628 10343-10346 DT denotes the
T4629 10347-10351 NN denotes WEHI
T4630 10352-10355 CD denotes 164
T4631 10356-10361 NN denotes clone
T4632 10362-10369 NN denotes 13-cell
T4633 10370-10377 NN denotes killing
T4634 10378-10383 NN denotes assay
T4635 10384-10385 -LRB- denotes [
T4636 10385-10387 CD denotes 29
T4637 10387-10388 -RRB- denotes ]
T4638 10390-10393 NN denotes TNF
T4639 10394-10408 NN denotes concentrations
T4640 10409-10412 VB denotes are
T4641 10413-10422 VB denotes expressed
T4642 10423-10425 IN denotes as
T4643 10426-10431 NN denotes pg/ml
T4750 10434-10440 NN denotes RT-PCR
T4751 10441-10449 NN denotes Analysis
T4752 10450-10455 JJ denotes Total
T4753 10456-10459 NN denotes RNA
T4754 10460-10464 IN denotes from
T4755 10465-10474 NN denotes monocytes
T4756 10475-10478 VB denotes was
T4757 10479-10487 VB denotes isolated
T4758 10488-10493 VB denotes using
T4759 10494-10500 NN denotes TRIzol
T4760 10501-10509 NN denotes Reagent™
T4761 10510-10511 -LRB- denotes (
T4762 10511-10521 NN denotes Invitrogen
T4763 10521-10522 -RRB- denotes )
T4764 10522-10526 NN denotes . cD
T4765 10526-10529 VB denotes NA
T4766 10529-10538 VB denotes was gener
T4767 10538-10540 IN denotes at
T4768 10540-10541 CD denotes e
T4769 10541-10543 NN denotes d
T4770 10543-10545 IN denotes on
T4771 10546-10551 JJ denotes 1 µg
T4772 10551-10554 NN denotes of
T4773 10554-10556 IN denotes to
T4774 10556-10557 DT denotes t
T4775 10557-10565 NN denotes al RNA i
T4776 10565-10571 NN denotes n a re
T4777 10571-10573 IN denotes ac
T4778 10573-10575 CD denotes ti
T4779 10575-10577 NN denotes on
T4780 10578-10579 -COMMA- denotes v
T4781 10579-10584 VB denotes olume
T4782 10585-10590 NN denotes of 20
T4783 10591-10598 JJ denotes µl, usi
T4784 10598-10611 NN denotes ng M-MLV reve
T4785 10611-10612 -LRB- denotes r
T4786 10612-10622 NN denotes se transcr
T4787 10622-10623 -RRB- denotes i
T4788 10643-10646 NN denotes PCR
T4789 10647-10650 VB denotes was
T4790 10651-10655 VB denotes done
T4791 10656-10658 IN denotes in
T4792 10659-10662 DT denotes the
T4793 10663-10669 JJ denotes linear
T4794 10670-10675 NN denotes range
T4795 10676-10678 IN denotes of
T4796 10679-10692 NN denotes amplification
T4797 10693-10694 -LRB- denotes (
T4798 10694-10704 VB denotes determined
T4799 10705-10708 IN denotes for
T4800 10709-10713 DT denotes each
T4801 10714-10720 NN denotes primer
T4802 10721-10730 NN denotes pair-cDNA
T4803 10731-10742 NN denotes combination
T4804 10742-10743 -RRB- denotes )
T4805 10745-10753 JJ denotes Standard
T4806 10754-10757 NN denotes PCR
T4807 10758-10767 NN denotes reactions
T4808 10768-10772 VB denotes were
T4809 10773-10782 VB denotes performed
T4810 10783-10787 IN denotes with
T4811 10788-10789 CD denotes 1
T4812 10790-10792 NN denotes µl
T4813 10793-10795 IN denotes of
T4814 10796-10799 DT denotes the
T4815 10800-10804 NN denotes cDNA
T4816 10805-10813 NN denotes solution
T4817 10813-10814 -COMMA- denotes ,
T4818 10815-10817 CD denotes 50
T4819 10818-10820 NN denotes µM
T4820 10821-10823 IN denotes of
T4821 10824-10828 DT denotes each
T4822 10829-10835 NN denotes primer
T4823 10836-10844 NN denotes solution
T4824 10844-10845 -COMMA- denotes ,
T4825 10846-10848 CD denotes 10
T4826 10849-10851 NN denotes mM
T4827 10852-10854 IN denotes of
T4828 10855-10859 DT denotes each
T4829 10860-10864 NN denotes dNTP
T4830 10864-10865 -COMMA- denotes ,
T4831 10866-10868 CD denotes 25
T4832 10869-10871 NN denotes mM
T4833 10872-10877 NN denotes MgCl2
T4834 10877-10878 -COMMA- denotes ,
T4835 10879-10882 NNP denotes 10X
T4836 10883-10891 NNP denotes Goldstar
T4837 10892-10895 NN denotes DNA
T4838 10896-10906 NN denotes polymerase
T4839 10907-10915 NN denotes reaction
T4840 10916-10922 NN denotes buffer
T4841 10922-10923 -COMMA- denotes ,
T4842 10924-10927 CC denotes and
T4843 10928-10931 CD denotes 0.5
T4844 10932-10937 NN denotes units
T4845 10938-10940 IN denotes of
T4846 10941-10949 NNP denotes Goldstar
T4847 10950-10953 NN denotes DNA
T4848 10954-10964 NN denotes polymerase
T4849 10965-10966 -LRB- denotes (
T4850 10966-10976 NNP denotes Eurogentec
T4851 10976-10977 -COMMA- denotes ,
T4852 10978-10985 NNP denotes Seraing
T4853 10985-10986 -COMMA- denotes ,
T4854 10987-10994 NNP denotes Belgium
T4855 10994-10995 -RRB- denotes )
T4856 10997-11002 JJ denotes First
T4857 11003-11006 NN denotes PCR
T4858 11007-11012 NN denotes cycle
T4859 11013-11022 VB denotes consisted
T4860 11023-11025 IN denotes of
T4861 11026-11027 CD denotes 1
T4862 11028-11031 NN denotes min
T4863 11032-11034 IN denotes at
T4864 11035-11039 NN denotes 92°C
T4865 11039-11040 -COMMA- denotes ,
T4866 11041-11042 CD denotes 1
T4867 11043-11046 NN denotes min
T4868 11047-11049 IN denotes at
T4869 11050-11054 NN denotes 58°C
T4870 11055-11058 CC denotes and
T4871 11059-11060 CD denotes 1
T4872 11061-11064 NN denotes min
T4873 11065-11067 IN denotes at
T4874 11068-11072 NN denotes 72°C
T4875 11072-11073 -COLON- denotes ;
T4876 11074-11078 RB denotes then
T4877 11079-11083 DT denotes each
T4878 11084-11087 NN denotes PCR
T4879 11088-11093 NN denotes cycle
T4880 11094-11103 VB denotes consisted
T4881 11104-11106 IN denotes of
T4882 11107-11109 CD denotes 40
T4883 11110-11113 NN denotes sec
T4884 11114-11116 IN denotes at
T4885 11117-11121 NN denotes 92°C
T4886 11121-11122 -COMMA- denotes ,
T4887 11123-11125 CD denotes 40
T4888 11126-11129 NN denotes sec
T4889 11130-11132 IN denotes at
T4890 11133-11137 NN denotes 58°C
T4891 11138-11141 CC denotes and
T4892 11142-11144 CD denotes 50
T4893 11145-11148 NN denotes sec
T4894 11149-11151 IN denotes at
T4895 11152-11157 NNP denotes 72°C.
T4896 11158-11162 NN denotes cDNA
T4897 11163-11166 IN denotes for
T4898 11167-11174 NN denotes β-actin
T4899 11175-11178 VB denotes was
T4900 11179-11188 VB denotes amplified
T4901 11189-11192 IN denotes for
T4902 11193-11195 CD denotes 28
T4903 11196-11202 NN denotes cycles
T4904 11203-11208 VB denotes using
T4905 11209-11212 DT denotes the
T4906 11213-11219 NN denotes oligos
T4907 11219-11220 -COLON- denotes :
T4908 11221-11226 NN denotes sense
T4909 11227-11252 NN denotes 5′-GGCATCGTGATGGACTCCG-3′
T4910 11253-11256 CC denotes and
T4911 11257-11266 JJ denotes antisense
T4912 11267-11292 JJ denotes 5′GCTGGAAGGTGGACAGCGA-3′.
T4913 11293-11297 NN denotes cDNA
T4914 11298-11301 IN denotes for
T4915 11302-11307 NN denotes TNF-α
T4916 11308-11311 VB denotes was
T4917 11312-11321 VB denotes amplified
T4918 11322-11325 IN denotes for
T4919 11326-11328 CD denotes 35
T4920 11329-11335 NN denotes cycles
T4921 11336-11341 VB denotes using
T4922 11342-11345 DT denotes the
T4923 11346-11352 NN denotes oligos
T4924 11352-11353 -COLON- denotes :
T4925 11354-11359 NN denotes sense
T4926 11360-11386 NN denotes 5′-AAGCCTGTAGCCCATGTTGT-3′
T4927 11387-11390 CC denotes and
T4928 11391-11400 JJ denotes antisense
T4929 11401-11428 JJ denotes 5′-CAGATAGATGGGCTCATACC-3′.
T4930 11429-11433 NN denotes cDNA
T4931 11434-11437 IN denotes for
T4932 11438-11444 NN denotes ICAM-1
T4933 11445-11448 VB denotes was
T4934 11449-11458 VB denotes amplified
T4935 11459-11462 IN denotes for
T4936 11463-11465 CD denotes 35
T4937 11466-11472 NN denotes cycles
T4938 11473-11478 VB denotes using
T4939 11479-11482 DT denotes the
T4940 11483-11489 NN denotes oligos
T4941 11490-11495 VB denotes sense
T4942 11496-11524 NN denotes 5′-GTAGCAGCCGCAGTCATAATGG-3′
T4943 11525-11528 CC denotes and
T4944 11529-11538 JJ denotes antisense
T4945 11539-11543 NN denotes 5′-A
T4946 11544-11568 NN denotes TGCTGTTGTATCTGACTGAGG-3′
T5239 11571-11576 NN denotes NF-kB
T5240 11577-11590 NN denotes Transcription
T5241 11591-11599 NNP denotes Reporter
T5242 11600-11604 NNP denotes Gene
T5243 11605-11610 NNP denotes Assay
T5244 11611-11614 DT denotes The
T5245 11615-11622 NN denotes plasmid
T5246 11623-11632 NN denotes 3XMHC-luc
T5247 11633-11634 -LRB- denotes (
T5248 11634-11635 DT denotes a
T5249 11636-11644 JJ denotes generous
T5250 11645-11649 NN denotes gift
T5251 11650-11654 IN denotes from
T5252 11655-11659 NNP denotes Drs.
T5253 11660-11662 NNP denotes J.
T5254 11663-11671 NNP denotes Westwick
T5255 11672-11675 CC denotes and
T5256 11676-11680 NNP denotes D.A.
T5257 11681-11688 NNP denotes Brenner
T5258 11688-11689 -COMMA- denotes ,
T5259 11690-11700 NNP denotes University
T5260 11701-11703 IN denotes of
T5261 11704-11709 NNP denotes North
T5262 11710-11718 NNP denotes Carolina
T5263 11718-11719 -COMMA- denotes ,
T5264 11720-11726 NNP denotes Chapel
T5265 11727-11731 NNP denotes Hill
T5266 11731-11732 -RRB- denotes )
T5267 11733-11741 VB denotes contains
T5268 11742-11747 CD denotes three
T5269 11748-11754 NN denotes copies
T5270 11755-11757 IN denotes of
T5271 11758-11774 JJ denotes NF-κB-responsive
T5272 11775-11782 NN denotes element
T5273 11783-11787 IN denotes from
T5274 11788-11791 DT denotes the
T5275 11792-11795 NN denotes MHC
T5276 11796-11801 NN denotes class
T5277 11802-11803 CD denotes I
T5278 11804-11809 NN denotes locus
T5279 11809-11810 -COMMA- denotes ,
T5280 11811-11817 VB denotes placed
T5281 11818-11826 RB denotes upstream
T5282 11827-11829 IN denotes of
T5283 11830-11833 DT denotes the
T5284 11834-11844 NN denotes luciferase
T5285 11845-11849 NN denotes gene
T5286 11851-11856 JJ denotes Human
T5287 11857-11866 JJ denotes monocytic
T5288 11867-11872 NN denotes THP-1
T5289 11873-11878 NN denotes cells
T5290 11879-11883 VB denotes were
T5291 11884-11895 RB denotes transiently
T5292 11896-11907 VB denotes transfected
T5293 11908-11910 IN denotes as
T5294 11911-11921 RB denotes previously
T5295 11922-11931 VB denotes described
T5296 11932-11933 -LRB- denotes [
T5297 11933-11935 CD denotes 30
T5298 11935-11936 -RRB- denotes ]
T5299 11936-11937 -COMMA- denotes ,
T5300 11938-11941 CC denotes and
T5301 11942-11946 RB denotes then
T5302 11947-11955 VB denotes cultured
T5303 11956-11959 IN denotes for
T5304 11960-11961 CD denotes 4
T5305 11962-11963 NN denotes h
T5306 11964-11969 RB denotes alone
T5307 11970-11972 CC denotes or
T5308 11973-11977 IN denotes with
T5309 11978-11988 VB denotes increasing
T5310 11989-12002 NN denotes concentration
T5311 12003-12005 IN denotes of
T5312 12006-12012 CC denotes either
T5313 12013-12016 NN denotes C10
T5314 12017-12019 CC denotes or
T5315 12020-12023 NN denotes C40
T5316 12025-12035 NN denotes Luciferase
T5317 12036-12044 NN denotes activity
T5318 12045-12048 VB denotes was
T5319 12049-12059 VB denotes determined
T5320 12060-12065 VB denotes using
T5321 12066-12067 DT denotes a
T5322 12068-12079 NN denotes luminometer
T5323 12080-12081 -LRB- denotes (
T5324 12081-12090 NN denotes Monolight
T5325 12091-12095 CD denotes 2010
T5326 12096-12107 NNP denotes Luminometer
T5327 12107-12108 -COMMA- denotes ,
T5328 12109-12112 NNP denotes Ann
T5329 12113-12118 NNP denotes Arbor
T5330 12118-12119 -COMMA- denotes ,
T5331 12120-12122 NNP denotes MI
T5332 12122-12123 -RRB- denotes )
T5492 12126-12133 NNP denotes Western
T5493 12134-12138 NNP denotes Blot
T5494 12139-12147 NN denotes Analysis
T5495 12148-12153 NN denotes THP-1
T5496 12154-12159 NN denotes cells
T5497 12160-12164 VB denotes were
T5498 12165-12175 VB denotes stimulated
T5499 12176-12179 IN denotes for
T5500 12180-12187 JJ denotes various
T5501 12188-12195 NN denotes lengths
T5502 12196-12198 IN denotes of
T5503 12199-12203 NN denotes time
T5504 12204-12208 IN denotes with
T5505 12209-12212 CD denotes 0.1
T5506 12213-12218 NN denotes mg/ml
T5507 12219-12222 NN denotes C10
T5508 12223-12225 CC denotes or
T5509 12226-12229 NN denotes C40
T5510 12229-12230 -COMMA- denotes ,
T5511 12231-12233 CC denotes or
T5512 12234-12236 CD denotes 10
T5513 12237-12242 NN denotes µg/ml
T5514 12243-12246 NN denotes LPS
T5515 12248-12253 NN denotes Cells
T5516 12254-12258 VB denotes were
T5517 12259-12263 RB denotes then
T5518 12264-12272 VB denotes pelleted
T5519 12272-12273 -COMMA- denotes ,
T5520 12274-12280 VB denotes washed
T5521 12281-12284 CC denotes and
T5522 12285-12296 VB denotes homogenised
T5523 12297-12299 IN denotes in
T5524 12300-12305 NN denotes lysis
T5525 12306-12312 NN denotes buffer
T5526 12313-12314 -LRB- denotes (
T5527 12314-12316 CD denotes 10
T5528 12317-12319 NN denotes mM
T5529 12320-12325 NNP denotes Hepes
T5530 12325-12326 -COMMA- denotes ,
T5531 12327-12329 NN denotes pH
T5532 12330-12333 CD denotes 7.9
T5533 12333-12334 -COMMA- denotes ,
T5534 12335-12338 CD denotes 150
T5535 12339-12341 NN denotes mM
T5536 12342-12346 NN denotes NaCl
T5537 12346-12347 -COMMA- denotes ,
T5538 12348-12349 CD denotes 1
T5539 12350-12352 NN denotes mM
T5540 12353-12357 NN denotes EDTA
T5541 12357-12358 -COMMA- denotes ,
T5542 12359-12362 CD denotes 0.6
T5543 12362-12363 NN denotes %
T5544 12364-12369 NN denotes NP-40
T5545 12369-12370 -COMMA- denotes ,
T5546 12371-12374 CC denotes and
T5547 12375-12378 CD denotes 0.5
T5548 12379-12381 NN denotes mM
T5549 12382-12386 NN denotes PMSF
T5550 12386-12387 -RRB- denotes )
T5551 12388-12390 IN denotes on
T5552 12391-12394 NN denotes ice
T5553 12396-12407 NN denotes Homogenates
T5554 12408-12412 VB denotes were
T5555 12413-12422 VB denotes sonicated
T5556 12422-12423 -COMMA- denotes ,
T5557 12424-12435 VB denotes centrifuged
T5558 12436-12438 IN denotes at
T5559 12439-12445 CD denotes 10,000
T5560 12446-12449 NN denotes rpm
T5561 12450-12452 TO denotes to
T5562 12453-12459 VB denotes remove
T5563 12460-12468 JJ denotes cellular
T5564 12469-12475 NN denotes debris
T5565 12475-12476 -COMMA- denotes ,
T5566 12477-12480 CC denotes and
T5567 12481-12492 NN denotes supernatant
T5568 12493-12502 VB denotes collected
T5569 12504-12511 NN denotes Protein
T5570 12512-12525 NN denotes concentration
T5571 12526-12529 VB denotes was
T5572 12530-12540 VB denotes determined
T5573 12541-12546 VB denotes using
T5574 12547-12550 DT denotes the
T5575 12551-12553 NN denotes DC
T5576 12554-12561 NN denotes Protein
T5577 12562-12567 NN denotes Assay
T5578 12568-12569 -LRB- denotes (
T5579 12569-12576 NN denotes Bio-Rad
T5580 12576-12577 -RRB- denotes )
T5581 12579-12587 NN denotes Proteins
T5582 12588-12590 IN denotes in
T5583 12591-12598 NN denotes samples
T5584 12599-12600 -LRB- denotes (
T5585 12600-12602 CD denotes 15
T5586 12603-12605 -COLON- denotes µg
T5587 12606-12611 JJ denotes total
T5588 12612-12620 NN denotes proteins
T5589 12620-12621 -RRB- denotes )
T5590 12622-12626 VB denotes were
T5591 12627-12635 VB denotes resolved
T5592 12636-12638 IN denotes in
T5593 12639-12640 DT denotes a
T5594 12641-12651 VB denotes denaturing
T5595 12652-12654 CD denotes 12
T5596 12654-12655 NN denotes %
T5597 12656-12670 NN denotes polyacrylamide
T5598 12671-12674 NN denotes gel
T5599 12675-12678 CC denotes and
T5600 12679-12690 VB denotes transferred
T5601 12691-12693 TO denotes to
T5602 12694-12695 DT denotes a
T5603 12696-12710 NN denotes nitrocellulose
T5604 12711-12719 NN denotes membrane
T5605 12721-12726 JJ denotes I-κBα
T5606 12727-12734 NN denotes protein
T5607 12735-12738 VB denotes was
T5608 12739-12747 VB denotes detected
T5609 12748-12753 VB denotes using
T5610 12754-12755 DT denotes a
T5611 12756-12762 NN denotes rabbit
T5612 12763-12773 JJ denotes polyclonal
T5613 12774-12782 NN denotes antibody
T5614 12783-12784 -LRB- denotes (
T5615 12784-12789 NNP denotes Santa
T5616 12790-12794 NNP denotes Cruz
T5617 12795-12808 NNP denotes Biotechnology
T5618 12808-12809 -COMMA- denotes ,
T5619 12810-12812 NN denotes CA
T5620 12812-12813 -RRB- denotes )
T5621 12814-12822 VB denotes followed
T5622 12823-12825 IN denotes by
T5623 12826-12827 DT denotes a
T5624 12828-12839 JJ denotes horseradish
T5625 12840-12858 JJ denotes peroxidase-coupled
T5626 12859-12863 NN denotes goat
T5627 12864-12874 JJ denotes polyclonal
T5628 12875-12883 NN denotes antibody
T5629 12884-12891 IN denotes against
T5630 12892-12898 NN denotes rabbit
T5631 12899-12901 NN denotes Ig
T5632 12902-12903 -LRB- denotes (
T5633 12903-12909 NNP denotes Caltag
T5634 12910-12922 NNP denotes Laboratories
T5635 12922-12923 -RRB- denotes )
T5636 12925-12932 RB denotes Finally
T5637 12932-12933 -COMMA- denotes ,
T5638 12934-12937 NN denotes IκB
T5639 12938-12943 NN denotes bands
T5640 12944-12948 VB denotes were
T5641 12949-12957 VB denotes revealed
T5642 12958-12963 VB denotes using
T5643 12964-12967 DT denotes the
T5644 12968-12972 NN denotes ECL™
T5645 12973-12982 NN denotes detection
T5646 12983-12989 NN denotes system
T5647 12990-12991 -LRB- denotes (
T5648 12991-12999 NNP denotes Amersham
T5649 13000-13009 NNP denotes Pharmacia
T5650 13010-13017 NNP denotes Biotech
T5651 13017-13018 -COMMA- denotes ,
T5652 13019-13022 NNP denotes Les
T5653 13023-13028 NNP denotes Ullis
T5654 13028-13029 -COMMA- denotes ,
T5655 13030-13036 NNP denotes France
T5656 13036-13037 -RRB- denotes )
T5657 13038-13047 VB denotes according
T5658 13048-13050 TO denotes to
T5659 13051-13054 DT denotes the
T5660 13055-13068 NN denotes manufacturers
T5661 13068-13069 POS denotes '
T5662 13070-13081 NN denotes instruction
T5663 13083-13091 NN denotes Antibody
T5664 13092-13094 TO denotes to
T5665 13095-13104 NN denotes α-Tubulin
T5666 13105-13106 -LRB- denotes (
T5667 13106-13111 NNP denotes Santa
T5668 13112-13116 NNP denotes Cruz
T5669 13116-13117 -RRB- denotes )
T5670 13118-13121 VB denotes was
T5671 13122-13125 NN denotes use
T5672 13126-13128 IN denotes as
T5673 13129-13136 VB denotes loading
T5674 13137-13144 NN denotes control
T5675 13146-13149 IN denotes For
T5676 13150-13157 JJ denotes nuclear
T5677 13158-13163 NN denotes NF-κB
T5678 13163-13164 -COMMA- denotes ,
T5679 13165-13170 NN denotes THP-1
T5680 13171-13176 NN denotes cells
T5681 13177-13181 VB denotes were
T5682 13182-13192 VB denotes stimulated
T5683 13193-13197 IN denotes with
T5684 13198-13199 CD denotes 1
T5685 13200-13205 NN denotes mg/ml
T5686 13206-13209 NN denotes C10
T5687 13210-13212 CC denotes or
T5688 13213-13216 NN denotes C40
T5689 13217-13220 IN denotes for
T5690 13221-13223 CD denotes 30
T5691 13224-13231 NN denotes minutes
T5692 13232-13234 IN denotes at
T5693 13235-13240 NNP denotes 37°C.
T5694 13241-13246 NNP denotes Cells
T5695 13247-13251 VB denotes were
T5696 13252-13256 RB denotes then
T5697 13257-13265 VB denotes pelleted
T5698 13266-13269 CC denotes and
T5699 13270-13276 NN denotes nuclei
T5700 13277-13286 VB denotes separated
T5701 13287-13289 IN denotes as
T5702 13290-13299 VB denotes described
T5703 13300-13301 -LRB- denotes [
T5704 13301-13303 CD denotes 31
T5705 13303-13304 -RRB- denotes ]
T5706 13306-13312 NN denotes Nuclei
T5707 13313-13317 VB denotes were
T5708 13318-13324 VB denotes washed
T5709 13325-13328 CC denotes and
T5710 13329-13340 VB denotes homogenized
T5711 13341-13349 RB denotes directly
T5712 13350-13352 IN denotes in
T5713 13353-13360 VB denotes loading
T5714 13361-13362 -LRB- denotes (
T5715 13362-13368 NNP denotes Laemli
T5716 13368-13369 -RRB- denotes )
T5717 13370-13376 NN denotes buffer
T5718 13377-13380 CC denotes and
T5719 13381-13387 VB denotes heated
T5720 13388-13391 IN denotes for
T5721 13392-13393 CD denotes 5
T5722 13394-13401 NN denotes minutes
T5723 13402-13404 IN denotes at
T5724 13405-13411 NNP denotes 100°C.
T5725 13412-13420 NNP denotes Proteins
T5726 13421-13423 IN denotes in
T5727 13424-13431 NN denotes samples
T5728 13432-13436 VB denotes were
T5729 13437-13445 VB denotes resolved
T5730 13446-13448 IN denotes in
T5731 13449-13450 DT denotes a
T5732 13451-13461 VB denotes denaturing
T5733 13462-13463 CD denotes 8
T5734 13463-13464 NN denotes %
T5735 13465-13479 NN denotes polyacrylamide
T5736 13480-13483 NN denotes gel
T5737 13484-13487 CC denotes and
T5738 13488-13499 VB denotes transferred
T5739 13500-13502 TO denotes to
T5740 13503-13504 DT denotes a
T5741 13505-13519 NN denotes polyvinylidine
T5742 13520-13528 NN denotes fluoride
T5743 13529-13530 -LRB- denotes (
T5744 13530-13534 NN denotes PVDF
T5745 13534-13535 -RRB- denotes )
T5746 13536-13544 NN denotes membrane
T5747 13545-13546 -LRB- denotes (
T5748 13546-13557 NN denotes Immobilon-P
T5749 13557-13558 -COLON- denotes ;
T5750 13559-13568 NNP denotes Millipore
T5751 13568-13569 -COMMA- denotes ,
T5752 13570-13577 NNP denotes Bedford
T5753 13577-13578 -COMMA- denotes ,
T5754 13579-13581 JJ denotes MA
T5755 13581-13582 -RRB- denotes )
T5756 13584-13593 NN denotes Membranes
T5757 13594-13598 VB denotes were
T5758 13599-13608 VB denotes incubated
T5759 13609-13611 IN denotes in
T5760 13612-13620 VB denotes blocking
T5761 13621-13627 NN denotes buffer
T5762 13628-13629 -LRB- denotes (
T5763 13629-13630 CD denotes 1
T5764 13630-13631 NN denotes %
T5765 13632-13635 NN denotes BSA
T5766 13635-13636 -COMMA- denotes ,
T5767 13637-13639 IN denotes in
T5768 13640-13643 NNP denotes PBS
T5769 13643-13644 -RRB- denotes )
T5770 13645-13648 IN denotes for
T5771 13649-13652 CD denotes two
T5772 13653-13658 NN denotes hours
T5773 13659-13661 IN denotes at
T5774 13662-13666 NN denotes room
T5775 13667-13678 NN denotes temperature
T5776 13680-13689 NN denotes Membranes
T5777 13690-13694 VB denotes were
T5778 13695-13707 RB denotes subsequently
T5779 13708-13714 VB denotes probed
T5780 13715-13719 IN denotes with
T5781 13720-13723 DT denotes the
T5782 13724-13737 JJ denotes corresponding
T5783 13738-13746 NN denotes antibody
T5784 13747-13749 IN denotes in
T5785 13750-13758 VB denotes blocking
T5786 13759-13765 NN denotes buffer
T5787 13765-13766 -COMMA- denotes ,
T5788 13767-13776 RB denotes overnight
T5789 13778-13784 NN denotes Rabbit
T5790 13785-13795 JJ denotes polyclonal
T5791 13796-13804 NN denotes antibody
T5792 13805-13815 JJ denotes anti-NF-κB
T5793 13816-13819 NN denotes p50
T5794 13820-13827 NN denotes subunit
T5795 13828-13829 -LRB- denotes (
T5796 13829-13830 -SHARP- denotes #
T5797 13831-13837 NN denotes sc-114
T5798 13837-13838 -RRB- denotes )
T5799 13839-13841 CC denotes or
T5800 13842-13852 JJ denotes anti-NF-κB
T5801 13853-13856 NN denotes p65
T5802 13857-13864 NN denotes subunit
T5803 13865-13866 -LRB- denotes (
T5804 13866-13867 -SHARP- denotes #
T5805 13868-13874 NN denotes sc-109
T5806 13874-13875 -RRB- denotes )
T5807 13876-13880 IN denotes from
T5808 13881-13886 NNP denotes Santa
T5809 13887-13891 NNP denotes Cruz
T5810 13892-13905 NNP denotes Biotechnology
T5811 13906-13910 VB denotes were
T5812 13911-13915 VB denotes used
T5813 13917-13926 NN denotes Membranes
T5814 13927-13931 VB denotes were
T5815 13932-13938 VB denotes washed
T5816 13939-13942 CD denotes six
T5817 13943-13948 NN denotes times
T5818 13949-13951 IN denotes in
T5819 13952-13955 NNP denotes PBS
T5820 13956-13960 IN denotes with
T5821 13961-13965 CD denotes 0.05
T5822 13965-13966 NN denotes %
T5823 13967-13972 CD denotes Tween
T5824 13973-13975 CD denotes 20
T5825 13975-13976 -COMMA- denotes ,
T5826 13977-13978 CD denotes 5
T5827 13979-13986 NN denotes minutes
T5828 13987-13991 DT denotes each
T5829 13992-13996 NN denotes time
T5830 13996-13997 -COMMA- denotes ,
T5831 13998-14001 CC denotes and
T5832 14002-14011 VB denotes incubated
T5833 14012-14016 IN denotes with
T5834 14017-14018 DT denotes a
T5835 14019-14025 CD denotes 1/3000
T5836 14026-14034 NN denotes dilution
T5837 14035-14037 IN denotes of
T5838 14038-14052 JJ denotes HRP-conjugated
T5839 14053-14054 NN denotes F
T5840 14054-14055 -LRB- denotes (
T5841 14055-14057 NN denotes ab
T5842 14057-14058 -DQE- denotes '
T5843 14058-14059 -RRB- denotes )
T5844 14059-14060 CD denotes 2
T5845 14061-14065 NN denotes goat
T5846 14066-14077 JJ denotes anti-rabbit
T5847 14078-14081 NN denotes IgG
T5848 14082-14084 IN denotes in
T5849 14085-14086 CD denotes 5
T5850 14086-14087 NN denotes %
T5851 14088-14094 JJ denotes nonfat
T5852 14095-14098 JJ denotes dry
T5853 14099-14103 NN denotes milk
T5854 14104-14107 CC denotes and
T5855 14108-14112 CD denotes 0.05
T5856 14112-14113 NN denotes %
T5857 14114-14119 CD denotes Tween
T5858 14120-14122 CD denotes 20
T5859 14123-14125 IN denotes in
T5860 14126-14129 NNP denotes PBS
T5861 14130-14133 IN denotes for
T5862 14134-14135 CD denotes 1
T5863 14136-14140 NN denotes hour
T5864 14141-14143 IN denotes at
T5865 14144-14148 NN denotes room
T5866 14149-14160 NN denotes temperature
T5867 14162-14167 IN denotes After
T5868 14168-14175 VB denotes washing
T5869 14176-14179 CD denotes six
T5870 14180-14184 JJ denotes more
T5871 14185-14190 NN denotes times
T5872 14191-14193 IN denotes in
T5873 14194-14197 NNP denotes PBS
T5874 14198-14202 IN denotes with
T5875 14203-14207 CD denotes 0.05
T5876 14207-14208 NN denotes %
T5877 14209-14214 CD denotes Tween
T5878 14215-14217 CD denotes 20
T5879 14217-14218 -COMMA- denotes ,
T5880 14219-14236 JJ denotes antibody-reactive
T5881 14237-14245 NN denotes proteins
T5882 14246-14250 VB denotes were
T5883 14251-14259 VB denotes detected
T5884 14260-14265 VB denotes using
T5885 14266-14267 DT denotes a
T5886 14268-14285 NN denotes chemiluminescence
T5887 14286-14295 NN denotes substrate
T5888 14296-14297 -LRB- denotes (
T5889 14297-14308 NNP denotes SuperSignal
T5890 14308-14309 -COLON- denotes ;
T5891 14310-14316 NNP denotes Pierce
T5892 14316-14317 -COMMA- denotes ,
T5893 14318-14326 NNP denotes Rockford
T5894 14326-14327 -COMMA- denotes ,
T5895 14328-14330 NN denotes IL
T5896 14330-14331 -RRB- denotes )
T5897 14332-14341 VB denotes according
T5898 14342-14344 TO denotes to
T5899 14345-14348 DT denotes the
T5900 14349-14361 NN denotes manufacturer
T5901 14361-14363 POS denotes 's
T5902 14364-14376 NN denotes instructions
T5903 14378-14380 TO denotes To
T5904 14381-14388 VB denotes confirm
T5905 14389-14393 IN denotes that
T5906 14394-14404 JJ denotes equivalent
T5907 14405-14412 NN denotes amounts
T5908 14413-14415 IN denotes of
T5909 14416-14423 NN denotes protein
T5910 14424-14428 VB denotes were
T5911 14429-14435 VB denotes loaded
T5912 14436-14438 IN denotes in
T5913 14439-14443 DT denotes each
T5914 14444-14448 NN denotes line
T5915 14448-14449 -COMMA- denotes ,
T5916 14450-14459 NN denotes membranes
T5917 14460-14464 VB denotes were
T5918 14465-14469 RB denotes also
T5919 14470-14477 JJ denotes Western
T5920 14478-14485 VB denotes blotted
T5921 14486-14489 IN denotes for
T5922 14490-14493 NN denotes ERK
T5923 14494-14496 IN denotes as
T5924 14497-14506 VB denotes described
T5925 14507-14508 -LRB- denotes [
T5926 14508-14510 CD denotes 32
T5927 14510-14511 -RRB- denotes ]
T6465 14514-14522 NN denotes Analysis
T6466 14523-14525 IN denotes of
T6467 14526-14531 NN denotes NF-κB
T6468 14532-14542 NN denotes Activation
T6469 14543-14545 IN denotes by
T6470 14546-14550 NN denotes Flow
T6471 14551-14560 NN denotes Cytometry
T6472 14561-14568 JJ denotes Nuclear
T6473 14569-14579 NN denotes activation
T6474 14580-14582 IN denotes of
T6475 14583-14588 NN denotes NF−κΒ
T6476 14589-14591 IN denotes by
T6477 14592-14596 NN denotes flow
T6478 14597-14606 NN denotes cytometry
T6479 14607-14610 VB denotes was
T6480 14611-14620 VB denotes performed
T6481 14621-14623 IN denotes as
T6482 14624-14633 VB denotes described
T6483 14634-14635 -LRB- denotes [
T6484 14635-14637 CD denotes 31
T6485 14637-14638 -RRB- denotes ]
T6520 14641-14652 JJ denotes Statistical
T6521 14653-14661 NN denotes Analysis
T6522 14662-14665 DT denotes The
T6523 14666-14673 NN denotes results
T6524 14674-14678 VB denotes were
T6525 14679-14688 VB denotes expressed
T6526 14689-14691 IN denotes as
T6527 14692-14695 DT denotes the
T6528 14696-14700 JJ denotes mean
T6529 14701-14706 NN denotes value
T6530 14707-14708 VB denotes ±
T6531 14709-14715 NNP denotes S.E.M.
T6532 14716-14718 IN denotes of
T6533 14719-14729 JJ denotes individual
T6534 14730-14741 NN denotes experiments
T6535 14743-14746 DT denotes The
T6536 14747-14758 JJ denotes statistical
T6537 14759-14771 NN denotes significance
T6538 14772-14774 IN denotes of
T6539 14775-14778 DT denotes the
T6540 14779-14790 NN denotes differences
T6541 14791-14798 IN denotes between
T6542 14799-14803 JJ denotes mean
T6543 14804-14810 NN denotes values
T6544 14811-14814 VB denotes was
T6545 14815-14823 VB denotes assessed
T6546 14824-14826 IN denotes by
T6547 14827-14830 DT denotes the
T6548 14831-14838 NN denotes Student
T6549 14838-14840 POS denotes 's
T6550 14841-14847 NN denotes t-test
T6551 14848-14851 CC denotes and
T6552 14852-14860 NN denotes analysis
T6553 14861-14863 IN denotes of
T6554 14864-14872 NN denotes variance
T6555 14873-14874 -LRB- denotes (
T6556 14874-14879 NN denotes ANOVA
T6557 14879-14880 -RRB- denotes )
T6623 14892-14900 VB denotes Degraded
T6624 14901-14904 NNP denotes CGN
T6625 14905-14911 NNP denotes Induce
T6626 14912-14919 NNP denotes Colonic
T6627 14920-14932 NNP denotes Inflammation
T6628 14933-14936 DT denotes All
T6629 14937-14941 NN denotes rats
T6630 14942-14951 VB denotes developed
T6631 14952-14960 NN denotes diarrhea
T6632 14961-14967 IN denotes during
T6633 14968-14976 VB denotes degraded
T6634 14977-14988 NN denotes carrageenan
T6635 14989-15003 NN denotes administration
T6636 15004-15007 CC denotes and
T6637 15008-15013 JJ denotes gross
T6638 15014-15022 NN denotes evidence
T6639 15023-15025 IN denotes of
T6640 15026-15031 NN denotes blood
T6641 15032-15035 VB denotes was
T6642 15036-15046 RB denotes frequently
T6643 15047-15055 VB denotes detected
T6644 15056-15058 IN denotes in
T6645 15059-15062 DT denotes the
T6646 15063-15069 NN denotes stools
T6647 15071-15076 NN denotes Colon
T6648 15077-15083 NN denotes length
T6649 15084-15096 RB denotes dramatically
T6650 15097-15106 VB denotes decreased
T6651 15107-15109 IN denotes in
T6652 15110-15113 DT denotes all
T6653 15114-15121 VB denotes treated
T6654 15122-15126 NN denotes rats
T6655 15127-15131 IN denotes with
T6656 15132-15133 DT denotes a
T6657 15134-15138 RB denotes more
T6658 15139-15149 JJ denotes pronounced
T6659 15150-15156 NN denotes effect
T6660 15157-15162 VB denotes being
T6661 15163-15171 VB denotes observed
T6662 15172-15174 IN denotes in
T6663 15175-15178 DT denotes the
T6664 15179-15181 CD denotes 40
T6665 15182-15185 NN denotes kDa
T6666 15186-15190 NN denotes dCGN
T6667 15191-15198 VB denotes treated
T6668 15199-15204 NN denotes group
T6669 15205-15206 -LRB- denotes (
T6670 15206-15210 NN denotes Fig.
T6671 15211-15213 NN denotes 1A
T6672 15213-15214 -RRB- denotes )
T6673 15216-15227 RB denotes Furthermore
T6674 15227-15228 -COMMA- denotes ,
T6675 15229-15238 VB denotes prolonged
T6676 15239-15247 NN denotes exposure
T6677 15248-15250 TO denotes to
T6678 15251-15253 CD denotes 40
T6679 15254-15257 NN denotes kDa
T6680 15258-15262 NN denotes dCGN
T6681 15263-15271 VB denotes resulted
T6682 15272-15274 IN denotes in
T6683 15275-15279 JJ denotes high
T6684 15280-15291 JJ denotes macroscopic
T6685 15292-15295 CC denotes and
T6686 15296-15308 JJ denotes histological
T6687 15309-15315 NN denotes scores
T6688 15316-15318 IN denotes of
T6689 15319-15331 NN denotes inflammation
T6690 15332-15333 -LRB- denotes (
T6691 15333-15337 NNP denotes Fig.
T6692 15338-15340 NN denotes 1B
T6693 15340-15341 -COMMA- denotes ,
T6694 15342-15343 NN denotes C
T6695 15343-15344 -RRB- denotes )
T6696 15346-15350 RB denotes Only
T6697 15351-15355 JJ denotes weak
T6698 15356-15371 NN denotes myeloperoxidase
T6699 15372-15380 NN denotes activity
T6700 15381-15384 VB denotes was
T6701 15385-15393 VB denotes detected
T6702 15394-15396 IN denotes in
T6703 15397-15401 CC denotes both
T6704 15402-15409 NN denotes control
T6705 15410-15413 CC denotes and
T6706 15414-15426 JJ denotes dCGN-treated
T6707 15427-15433 NN denotes groups
T6708 15434-15435 -LRB- denotes (
T6709 15435-15439 NNP denotes Fig.
T6710 15440-15442 NN denotes 1D
T6711 15442-15443 -RRB- denotes )
T6712 15443-15444 -COMMA- denotes ,
T6713 15445-15455 VB denotes indicating
T6714 15456-15460 IN denotes that
T6715 15461-15473 NN denotes granulocytes
T6716 15474-15477 VB denotes did
T6717 15478-15481 RB denotes not
T6718 15482-15486 VB denotes play
T6719 15487-15488 DT denotes a
T6720 15489-15494 JJ denotes major
T6721 15495-15499 NN denotes role
T6722 15500-15502 IN denotes in
T6723 15503-15506 DT denotes the
T6724 15507-15519 NN denotes inflammation
T6725 15520-15522 IN denotes at
T6726 15523-15527 DT denotes that
T6727 15528-15533 NN denotes stage
T6728 15535-15547 JJ denotes Histological
T6729 15548-15559 NN denotes examination
T6730 15560-15568 VB denotes revealed
T6731 15569-15576 JJ denotes various
T6732 15577-15584 NN denotes degrees
T6733 15585-15587 IN denotes of
T6734 15588-15595 JJ denotes mucosal
T6735 15596-15608 NN denotes inflammation
T6736 15610-15614 NN denotes Rats
T6737 15615-15622 VB denotes treated
T6738 15623-15627 IN denotes with
T6739 15628-15630 CD denotes 10
T6740 15631-15634 NN denotes kDa
T6741 15635-15639 NN denotes dCGN
T6742 15640-15646 VB denotes showed
T6743 15647-15652 NN denotes edema
T6744 15652-15653 -COMMA- denotes ,
T6745 15654-15664 NN denotes epithelium
T6746 15665-15672 NN denotes atrophy
T6747 15673-15676 CC denotes and
T6748 15677-15683 JJ denotes slight
T6749 15684-15694 NN denotes lymphocyte
T6750 15695-15707 NN denotes infiltration
T6751 15708-15709 -LRB- denotes (
T6752 15709-15713 NN denotes data
T6753 15714-15717 RB denotes not
T6754 15718-15723 VB denotes shown
T6755 15723-15724 -RRB- denotes )
T6756 15726-15731 DT denotes These
T6757 15732-15740 NN denotes symptoms
T6758 15741-15745 VB denotes were
T6759 15746-15753 RB denotes totally
T6760 15754-15760 JJ denotes absent
T6761 15761-15763 IN denotes in
T6762 15764-15767 DT denotes the
T6763 15768-15773 NN denotes colon
T6764 15774-15776 IN denotes of
T6765 15777-15784 NN denotes control
T6766 15785-15789 NN denotes rats
T6767 15790-15791 -LRB- denotes (
T6768 15791-15795 NNP denotes Fig.
T6769 15796-15798 NN denotes 1E
T6770 15798-15799 -RRB- denotes )
T6771 15801-15805 RB denotes More
T6772 15806-15812 JJ denotes severe
T6773 15813-15820 JJ denotes mucosal
T6774 15821-15829 NN denotes injuries
T6775 15830-15839 VB denotes including
T6776 15840-15850 NN denotes ulceration
T6777 15850-15851 -COMMA- denotes ,
T6778 15852-15864 JJ denotes hyperplastic
T6779 15865-15875 NN denotes epithelium
T6780 15875-15876 -COMMA- denotes ,
T6781 15877-15882 NN denotes crypt
T6782 15883-15893 NN denotes distortion
T6783 15894-15897 CC denotes and
T6784 15898-15899 DT denotes a
T6785 15900-15906 JJ denotes strong
T6786 15907-15917 NN denotes macrophage
T6787 15918-15930 NN denotes infiltration
T6788 15930-15931 -COMMA- denotes ,
T6789 15932-15936 VB denotes were
T6790 15937-15945 VB denotes observed
T6791 15946-15948 IN denotes in
T6792 15949-15952 DT denotes the
T6793 15953-15955 CD denotes 40
T6794 15956-15959 NN denotes kDa
T6795 15960-15972 JJ denotes dCGN-treated
T6796 15973-15977 NN denotes rats
T6797 15978-15979 -LRB- denotes (
T6798 15979-15983 NNP denotes Fig.
T6799 15984-15986 NN denotes 1F
T6800 15986-15987 -RRB- denotes )
T6801 15989-15991 DT denotes No
T6802 15992-16001 VB denotes sulphated
T6803 16002-16017 NN denotes polysaccharides
T6804 16018-16022 VB denotes were
T6805 16023-16031 VB denotes detected
T6806 16032-16034 IN denotes by
T6807 16035-16044 NN denotes toluidine
T6808 16045-16049 JJ denotes blue
T6809 16050-16058 NN denotes staining
T6810 16059-16061 IN denotes of
T6811 16062-16067 NN denotes colon
T6812 16068-16074 NN denotes mucosa
T6813 16075-16079 IN denotes from
T6814 16080-16084 NN denotes rats
T6815 16085-16092 VB denotes treated
T6816 16093-16097 IN denotes with
T6817 16098-16104 CC denotes either
T6818 16105-16108 DT denotes the
T6819 16109-16111 CD denotes 10
T6820 16112-16114 CC denotes or
T6821 16115-16117 CD denotes 40
T6822 16118-16121 NN denotes kDa
T6823 16122-16126 NN denotes dCGN
T6824 16127-16128 -LRB- denotes (
T6825 16128-16131 RB denotes not
T6826 16132-16137 VB denotes shown
T6827 16137-16138 -RRB- denotes )
T6828 16140-16148 IN denotes Although
T6829 16149-16151 PRP denotes we
T6830 16152-16155 MD denotes can
T6831 16155-16158 RB denotes not
T6832 16159-16166 VB denotes exclude
T6833 16167-16171 IN denotes that
T6834 16172-16176 NN denotes dCGN
T6835 16177-16180 NN denotes mat
T6836 16181-16184 RB denotes not
T6837 16185-16189 VB denotes have
T6838 16190-16198 VB denotes retained
T6839 16199-16201 IN denotes in
T6840 16202-16205 DT denotes the
T6841 16206-16213 NN denotes section
T6842 16214-16220 IN denotes during
T6843 16221-16224 DT denotes the
T6844 16225-16234 NN denotes histology
T6845 16235-16244 NN denotes procedure
T6846 16244-16245 -COMMA- denotes ,
T6847 16246-16250 DT denotes this
T6848 16251-16260 VB denotes indicates
T6849 16261-16265 IN denotes that
T6850 16266-16271 DT denotes these
T6851 16272-16280 NN denotes polymers
T6852 16281-16284 MD denotes may
T6853 16285-16288 RB denotes not
T6854 16289-16293 VB denotes have
T6855 16294-16298 VB denotes been
T6856 16299-16311 VB denotes phagocytosed
T7287 0-3 NN denotes CGN
T7288 4-16863 NNP denotes inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced-TNF-α
T15571 16357-16365 VB denotes Degraded
T15572 16366-16369 NN denotes CGN
T15573 16370-16377 VB denotes induced
T15574 16378-16383 NN denotes colon
T15575 16384-16396 NN denotes inflammation
T15576 16397-16399 IN denotes in
T15577 16400-16404 NN denotes rats
T15578 16406-16416 NN denotes Histograms
T15579 16417-16424 VB denotes showing
T15580 16425-16428 DT denotes the
T15581 16429-16435 NN denotes effect
T15582 16436-16438 IN denotes of
T15583 16439-16447 VB denotes degraded
T15584 16448-16451 NN denotes CGN
T15585 16452-16454 IN denotes on
T15586 16454-16455 -COLON- denotes :
T15587 16456-16461 NN denotes colon
T15588 16462-16468 NN denotes length
T15589 16469-16470 -LRB- denotes (
T15590 16470-16471 NN denotes A
T15591 16471-16472 -RRB- denotes )
T15592 16472-16473 -COLON- denotes ;
T15593 16474-16485 JJ denotes macroscopic
T15594 16486-16487 -LRB- denotes (
T15595 16487-16488 NN denotes B
T15596 16488-16489 -RRB- denotes )
T15597 16490-16493 CC denotes and
T15598 16494-16506 JJ denotes histological
T15599 16507-16508 -LRB- denotes (
T15600 16508-16509 NN denotes C
T15601 16509-16510 -RRB- denotes )
T15602 16511-16523 NN denotes inflammation
T15603 16524-16529 NN denotes score
T15604 16530-16532 IN denotes of
T15605 16533-16538 NN denotes colon
T15606 16538-16539 -COLON- denotes ;
T15607 16540-16555 NN denotes Myeloperoxidase
T15608 16556-16557 -LRB- denotes (
T15609 16557-16560 NN denotes MPO
T15610 16560-16561 -RRB- denotes )
T15611 16562-16570 NN denotes activity
T15612 16571-16572 -LRB- denotes (
T15613 16572-16573 NN denotes D
T15614 16573-16574 -RRB- denotes )
T15615 16576-16583 NN denotes Control
T15616 16584-16588 NN denotes rats
T15617 16589-16590 -LRB- denotes (
T15618 16590-16595 JJ denotes white
T15619 16596-16600 NN denotes bars
T15620 16600-16601 -RRB- denotes )
T15621 16601-16602 -COLON- denotes ;
T15622 16603-16605 CD denotes 10
T15623 16606-16609 NN denotes kDa
T15624 16610-16618 VB denotes degraded
T15625 16619-16630 JJ denotes CGN-treated
T15626 16631-16635 NN denotes rats
T15627 16636-16637 -LRB- denotes (
T15628 16637-16641 JJ denotes grey
T15629 16642-16646 NN denotes bars
T15630 16646-16647 -RRB- denotes )
T15631 16647-16648 -COLON- denotes ;
T15632 16649-16651 CD denotes 40
T15633 16652-16655 NN denotes kDa
T15634 16656-16664 VB denotes degraded
T15635 16665-16676 JJ denotes CGN-treated
T15636 16677-16681 NN denotes rats
T15637 16682-16683 -LRB- denotes (
T15638 16683-16688 JJ denotes black
T15639 16689-16693 NN denotes bars
T15640 16693-16694 -RRB- denotes )
T15641 16694-16695 SYM denotes .
T15642 16696-16697 NN denotes *
T15643 16698-16699 SYM denotes p
T15644 16699-16703 CD denotes <0.0
T15645 16703-16707 IN denotes 5 fr
T15646 16707-16715 NN denotes om contr
T15647 16715-16717 SYM denotes ol
T15648 16717-16718 NN denotes .
T15649 16719-16720 SYM denotes *
T15650 16720-16724 CD denotes * p<
T15651 16724-16728 IN denotes 0.01
T15652 16729-16736 NN denotes from co
T15653 16743-16755 JJ denotes Histological
T15654 16756-16764 NN denotes analysis
T15655 16765-16767 IN denotes of
T15656 16768-16773 NN denotes colon
T15657 16774-16778 IN denotes from
T15658 16779-16786 NN denotes control
T15659 16787-16791 NN denotes rats
T15660 16792-16793 -LRB- denotes (
T15661 16793-16794 NN denotes E
T15662 16794-16795 -RRB- denotes )
T15663 16795-16796 -COMMA- denotes ,
T15664 16797-16800 CC denotes and
T15665 16801-16805 IN denotes from
T15666 16806-16808 CD denotes 40
T15667 16809-16812 NN denotes kDa
T15668 16813-16825 JJ denotes dCGN-treated
T15669 16826-16830 NN denotes rats
T15670 16831-16832 -LRB- denotes (
T15671 16832-16833 NN denotes F
T15672 16833-16834 -RRB- denotes )
T2817 5769-5776 NN denotes Animals
R161 T221 T222 arg1Of CGN,inhibited
R162 T221 T229 arg1Of CGN,arresting
R163 T222 T227 arg1Of inhibited,vitro
R164 T222 T228 arg1Of inhibited,","
R165 T222 T229 modOf inhibited,arresting
R166 T225 T222 arg2Of proliferation,inhibited
R167 T225 T223 arg1Of proliferation,THP-1
R168 T225 T224 arg1Of proliferation,cell
R169 T227 T226 arg1Of vitro,in
R170 T231 T229 arg2Of cells,arresting
R171 T231 T230 arg1Of cells,the
R172 T231 T232 arg1Of cells,in
R173 T234 T232 arg2Of phase,in
R174 T234 T233 arg1Of phase,G1
R175 T236 T235 arg2Of addition,In
R176 T238 T239 arg1Of dCGN,increased
R177 T239 T235 arg1Of increased,In
R178 T239 T237 arg1Of increased,","
R179 T239 T242 arg1Of increased,in
R180 T239 T248 arg1Of increased,with
R181 T241 T239 arg2Of expression,increased
R182 T241 T240 arg1Of expression,ICAM-1
R183 T244 T245 arg1Of PBM,and
R184 T245 T242 arg2Of and,in
R185 T245 T243 arg1Of and,both
R186 T247 T245 arg2Of cells,and
R187 T247 T246 arg1Of cells,THP-1
R188 T251 T248 arg2Of effect,with
R189 T251 T249 arg1Of effect,a
R190 T251 T250 arg1Of effect,major
R191 T251 T252 arg2Of effect,seen
R192 T252 T253 arg1Of seen,after
R193 T257 T253 arg2Of exposure,after
R194 T257 T254 arg1Of exposure,40
R195 T257 T255 arg1Of exposure,kDa
R196 T257 T256 arg1Of exposure,dCGN
R197 T260 T261 arg1Of dCGN,stimulated
R198 T261 T258 arg1Of stimulated,Also
R199 T261 T259 arg1Of stimulated,","
R200 T263 T261 arg2Of aggregation,stimulated
R201 T263 T262 arg1Of aggregation,monocyte
R202 T263 T265 arg1Of aggregation,vitro
R203 T263 T266 arg1Of aggregation,that
R204 T263 T267 arg1Of aggregation,was
R205 T263 T268 arg2Of aggregation,prevented
R206 T265 T264 arg1Of vitro,in
R207 T268 T267 arg2Of prevented,was
R208 T270 T268 arg1Of incubation,prevented
R209 T270 T269 arg2Of incubation,by
R210 T270 T271 arg1Of incubation,with
R211 T273 T271 arg2Of antibody,with
R212 T273 T272 arg1Of antibody,anti-ICAM-1
R213 T276 T277 arg1Of dCGN,stimulated
R214 T277 T274 arg1Of stimulated,Finally
R215 T277 T275 arg1Of stimulated,","
R216 T277 T282 arg1Of stimulated,by
R217 T279 T280 arg1Of expression,and
R218 T280 T277 arg2Of and,stimulated
R219 T280 T278 arg1Of and,TNF-α
R220 T281 T280 arg2Of secretion,and
R221 T284 T285 arg1Of PBM,and
R222 T285 T282 arg2Of and,by
R223 T285 T283 arg1Of and,both
R224 T287 T285 arg2Of cells,and
R225 T287 T286 arg1Of cells,THP-1
R226 T290 T288 arg1Of effects,All
R227 T290 T289 arg1Of effects,these
R228 T290 T291 arg1Of effects,were
R229 T290 T292 arg2Of effects,linked
R230 T292 T291 arg2Of linked,were
R231 T292 T293 arg1Of linked,to
R232 T295 T293 arg2Of activation,to
R233 T295 T294 arg1Of activation,NF-κB
R234 T297 T296 arg1Of data,These
R235 T297 T299 arg1Of data,suggest
R236 T299 T298 arg1Of suggest,strongly
R237 T303 T301 arg1Of forms,the
R238 T303 T302 arg1Of forms,degraded
R239 T303 T304 arg1Of forms,of
R240 T303 T306 arg1Of forms,have
R241 T305 T304 arg2Of CGN,of
R242 T306 T299 arg2Of have,suggest
R243 T306 T300 arg1Of have,that
R244 T309 T306 arg2Of effect,have
R245 T309 T307 arg1Of effect,a
R246 T309 T308 arg1Of effect,pronounced
R247 T309 T310 arg1Of effect,on
R248 T309 T312 arg1Of effect,","
R249 T309 T313 arg1Of effect,characteristic
R250 T311 T310 arg2Of monocytes,on
R251 T313 T314 arg1Of characteristic,of
R252 T317 T314 arg2Of phenotype,of
R253 T317 T315 arg1Of phenotype,an
R254 T317 T316 arg1Of phenotype,inflammatory
R579 T776 T777 arg1Of Carrageenan,(
R580 T776 T780 arg1Of Carrageenan,is
R581 T778 T777 arg2Of CGN,(
R582 T779 T777 arg3Of ),(
R583 T784 T780 arg2Of weight,is
R584 T784 T781 arg1Of weight,a
R585 T784 T782 arg1Of weight,high
R586 T784 T783 arg1Of weight,molecular
R587 T784 T785 arg2Of weight,sulphated
R588 T786 T785 arg3Of polysaccharide,sulphated
R589 T786 T787 arg1Of polysaccharide,(
R590 T786 T792 arg2Of polysaccharide,derived
R591 T789 T788 arg1Of 200,>
R592 T790 T787 arg2Of kDa,(
R593 T790 T789 arg1Of kDa,200
R594 T791 T787 arg3Of ),(
R595 T792 T793 arg1Of derived,from
R596 T795 T793 arg2Of algae,from
R597 T795 T794 arg1Of algae,red
R598 T795 T796 arg1Of algae,(
R599 T797 T796 arg2Of Rhodophyceae,(
R600 T798 T796 arg3Of ),(
R601 T801 T799 arg1Of forms,Three
R602 T801 T800 arg1Of forms,main
R603 T801 T802 arg1Of forms,of
R604 T801 T804 arg1Of forms,have
R605 T801 T805 arg1Of forms,been
R606 T801 T806 arg2Of forms,identified
R607 T803 T802 arg2Of CGN,of
R608 T806 T804 arg2Of identified,have
R609 T806 T805 arg2Of identified,been
R610 T806 T807 arg1Of identified,:
R611 T808 T809 arg1Of kappa,","
R612 T809 T812 arg1Of ",",and
R613 T810 T809 arg2Of iota,","
R614 T812 T806 arg3Of and,identified
R615 T812 T811 arg1Of and,","
R616 T813 T812 arg2Of lambda,and
R617 T814 T815 arg1Of They,differ
R618 T815 T816 arg1Of differ,from
R619 T815 T819 arg1Of differ,in
R620 T815 T827 arg1Of differ,","
R621 T815 T828 arg1Of differ,[
R622 T818 T816 arg2Of other,from
R623 T818 T817 arg1Of other,each
R624 T821 T820 arg1Of degree,sulphation
R625 T821 T822 arg1Of degree,and
R626 T822 T819 arg2Of and,in
R627 T823 T822 arg2Of solubility,and
R628 T823 T824 arg1Of solubility,[
R629 T825 T824 arg2Of 1,[
R630 T826 T824 arg3Of ],[
R631 T829 T828 arg2Of 2,[
R632 T830 T828 arg3Of ],[
R633 T832 T831 arg1Of CGN,Native
R634 T832 T833 arg1Of CGN,is
R635 T832 T834 arg2Of CGN,thought
R636 T832 T836 arg1Of CGN,be
R637 T832 T837 arg1Of CGN,harmless
R638 T832 T839 arg1Of CGN,is
R639 T832 T841 arg2Of CGN,used
R640 T834 T833 arg2Of thought,is
R641 T834 T838 arg1Of thought,and
R642 T836 T834 arg3Of be,thought
R643 T836 T835 arg1Of be,to
R644 T837 T836 arg2Of harmless,be
R645 T841 T838 arg2Of used,and
R646 T841 T839 arg2Of used,is
R647 T841 T840 arg1Of used,widely
R648 T841 T842 arg1Of used,as
R649 T844 T842 arg2Of food,as
R650 T844 T843 arg1Of food,a
R651 T844 T845 arg1Of food,additive
R652 T847 T845 arg2Of improve,additive
R653 T847 T846 arg1Of improve,to
R654 T848 T847 arg2Of texture,improve
R655 T849 T850 arg1Of It,is
R656 T849 T852 arg2Of It,used
R657 T852 T850 arg2Of used,is
R658 T852 T851 arg1Of used,also
R659 T852 T853 arg1Of used,in
R660 T854 T855 arg1Of cosmetics,and
R661 T855 T853 arg2Of and,in
R662 T856 T855 arg2Of pharmaceuticals,and
R663 T860 T859 arg1Of treatment,acid
R664 T860 T861 arg1Of treatment,at
R665 T860 T867 arg1Of treatment,triggers
R666 T863 T861 arg2Of temperature,at
R667 T863 T862 arg1Of temperature,high
R668 T863 T864 arg1Of temperature,(
R669 T865 T864 arg2Of 80°C,(
R670 T866 T864 arg3Of ),(
R671 T867 T857 arg1Of triggers,However
R672 T867 T858 arg1Of triggers,","
R673 T867 T870 modOf triggers,to
R674 T869 T867 arg2Of hydrolysis,triggers
R675 T869 T868 arg1Of hydrolysis,CGN
R676 T871 T870 arg1Of lower,to
R677 T876 T875 arg1Of 50,<
R678 T877 T874 arg2Of kDa,(
R679 T877 T876 arg1Of kDa,50
R680 T878 T874 arg3Of ),(
R681 T879 T871 arg2Of compounds,lower
R682 T879 T872 arg1Of compounds,molecular
R683 T879 T873 arg1Of compounds,weight
R684 T879 T874 arg1Of compounds,(
R685 T879 T880 arg2Of compounds,known
R686 T880 T881 arg1Of known,as
R687 T882 T883 arg1Of poligeenan,or
R688 T883 T881 arg2Of or,as
R689 T885 T883 arg2Of CGN,or
R690 T885 T884 arg2Of CGN,degraded
R691 T885 T886 arg1Of CGN,(
R692 T887 T886 arg2Of dCGN,(
R693 T888 T886 arg3Of ),(
R694 T890 T889 arg1Of dCGNs,These
R695 T890 T891 arg1Of dCGNs,induce
R696 T890 T894 arg1Of dCGNs,have
R697 T890 T895 arg1Of dCGNs,been
R698 T890 T897 arg2Of dCGNs,used
R699 T891 T893 arg1Of induce,and
R700 T892 T891 arg2Of inflammation,induce
R701 T897 T893 arg2Of used,and
R702 T897 T894 arg2Of used,have
R703 T897 T895 arg2Of used,been
R704 T897 T896 arg1Of used,widely
R705 T897 T898 arg1Of used,as
R706 T899 T898 arg2Of models,as
R707 T899 T900 arg1Of models,of
R708 T901 T900 arg2Of colitis,of
R709 T901 T902 arg1Of colitis,in
R710 T904 T902 arg2Of species,in
R711 T904 T903 arg1Of species,several
R712 T904 T905 arg1Of species,","
R713 T904 T906 arg1Of species,including
R714 T907 T906 arg2Of rats,including
R715 T907 T908 arg1Of rats,[
R716 T907 T911 arg1Of rats,","
R717 T909 T908 arg2Of 3,[
R718 T910 T908 arg3Of ],[
R719 T912 T913 arg1Of rabbits,[
R720 T912 T916 arg1Of rabbits,and
R721 T914 T913 arg2Of 4,[
R722 T915 T913 arg3Of ],[
R723 T916 T911 arg2Of and,","
R724 T918 T916 arg2Of pigs,and
R725 T918 T917 arg1Of pigs,guinea
R726 T918 T919 arg1Of pigs,[
R727 T920 T919 arg2Of 5,[
R728 T921 T919 arg3Of ],[
R729 T923 T922 arg1Of role,The
R730 T923 T924 arg1Of role,of
R731 T923 T926 arg1Of role,as
R732 T923 T930 arg1Of role,remains
R733 T923 T931 arg1Of role,controversial
R734 T925 T924 arg2Of dCGN,of
R735 T929 T926 arg2Of factor,as
R736 T929 T927 arg1Of factor,a
R737 T929 T928 arg1Of factor,tumor-promoting
R738 T930 T932 arg1Of remains,[
R739 T930 T935 arg1Of remains,","
R740 T930 T936 arg1Of remains,[
R741 T930 T939 arg1Of remains,–
R742 T931 T930 arg2Of controversial,remains
R743 T933 T932 arg2Of 4,[
R744 T934 T932 arg3Of ],[
R745 T937 T936 arg2Of 6,[
R746 T938 T936 arg3Of ],[
R747 T940 T939 arg2Of [,–
R748 T941 T939 arg3Of 8,–
R749 T945 T943 arg1Of form,the
R750 T945 T944 arg1Of form,native
R751 T945 T946 arg1Of form,is
R752 T945 T947 arg2Of form,thought
R753 T945 T949 arg1Of form,be
R754 T945 T950 arg1Of form,harmless
R755 T947 T942 arg2Of thought,Although
R756 T947 T946 arg2Of thought,is
R757 T949 T947 arg3Of be,thought
R758 T949 T948 arg1Of be,to
R759 T950 T949 arg2Of harmless,be
R760 T950 T951 arg1Of harmless,for
R761 T953 T951 arg2Of consumption,for
R762 T953 T952 arg1Of consumption,human
R763 T956 T955 arg1Of amounts,small
R764 T956 T957 arg1Of amounts,of
R765 T956 T959 arg1Of amounts,are
R766 T956 T961 arg2Of amounts,produced
R767 T958 T957 arg2Of dCGN,of
R768 T961 T942 arg1Of produced,Although
R769 T961 T954 arg1Of produced,","
R770 T961 T959 arg2Of produced,are
R771 T961 T960 arg1Of produced,probably
R772 T961 T983 arg1Of produced,","
R773 T961 T984 arg1Of produced,[
R774 T964 T963 arg1Of hydrolysis,acid
R775 T964 T965 arg1Of hydrolysis,during
R776 T964 T972 arg1Of hydrolysis,[
R777 T964 T975 arg1Of hydrolysis,or
R778 T967 T965 arg2Of digestion,during
R779 T967 T966 arg1Of digestion,gastric
R780 T967 T968 arg1Of digestion,[
R781 T967 T971 arg1Of digestion,","
R782 T969 T968 arg2Of 9,[
R783 T970 T968 arg3Of ],[
R784 T973 T972 arg2Of 10,[
R785 T974 T972 arg3Of ],[
R786 T975 T961 arg1Of or,produced
R787 T975 T962 arg2Of or,by
R788 T976 T975 arg2Of interaction,or
R789 T976 T977 arg1Of interaction,with
R790 T979 T977 arg2Of bacteria,with
R791 T979 T978 arg1Of bacteria,intestinal
R792 T979 T980 arg1Of bacteria,[
R793 T981 T980 arg2Of 11,[
R794 T982 T980 arg3Of ],[
R795 T985 T984 arg2Of 12,[
R796 T986 T984 arg3Of ],[
R797 T989 T988 arg1Of effects,the
R798 T989 T990 arg1Of effects,of
R799 T989 T994 arg1Of effects,on
R800 T989 T997 arg1Of effects,have
R801 T989 T998 arg1Of effects,been
R802 T989 T1000 arg2Of effects,analyzed
R803 T991 T992 arg1Of native,and
R804 T993 T990 arg2Of dCGN,of
R805 T993 T991 arg1Of dCGN,native
R806 T996 T994 arg2Of inflammation,on
R807 T996 T995 arg1Of inflammation,intestinal
R808 T1000 T987 arg2Of analyzed,Whereas
R809 T1000 T997 arg2Of analyzed,have
R810 T1000 T998 arg2Of analyzed,been
R811 T1000 T999 arg1Of analyzed,extensively
R812 T1000 T1001 arg1Of analyzed,in
R813 T1003 T1001 arg2Of models,in
R814 T1003 T1002 arg1Of models,animal
R815 T1007 T1005 arg1Of studies,only
R816 T1007 T1006 arg1Of studies,few
R817 T1007 T1008 arg1Of studies,have
R818 T1007 T1009 arg1Of studies,been
R819 T1007 T1010 arg2Of studies,conducted
R820 T1010 T987 arg1Of conducted,Whereas
R821 T1010 T1004 arg1Of conducted,","
R822 T1010 T1008 arg2Of conducted,have
R823 T1010 T1009 arg2Of conducted,been
R824 T1010 T1011 modOf conducted,using
R825 T1014 T1011 arg2Of lines,using
R826 T1014 T1012 arg1Of lines,human
R827 T1014 T1013 arg1Of lines,cell
R828 T1016 T1015 arg1Of studies,Recent
R829 T1016 T1017 arg1Of studies,have
R830 T1016 T1018 arg1Of studies,shown
R831 T1018 T1017 arg2Of shown,have
R832 T1018 T1047 arg1Of shown,[
R833 T1018 T1050 arg1Of shown,","
R834 T1018 T1051 arg1Of shown,[
R835 T1020 T1018 arg2Of link,shown
R836 T1020 T1019 arg1Of link,a
R837 T1020 T1021 arg1Of link,between
R838 T1020 T1030 arg1Of link,by
R839 T1020 T1039 arg1Of link,","
R840 T1020 T1040 arg1Of link,via
R841 T1022 T1023 arg1Of exposure,to
R842 T1022 T1027 arg1Of exposure,and
R843 T1026 T1023 arg2Of CGN,to
R844 T1026 T1024 arg1Of CGN,native
R845 T1026 T1025 arg1Of CGN,form
R846 T1027 T1021 arg2Of and,between
R847 T1029 T1027 arg2Of production,and
R848 T1029 T1028 arg1Of production,IL-8
R849 T1036 T1030 arg2Of line,by
R850 T1036 T1031 arg1Of line,the
R851 T1036 T1032 arg1Of line,human
R852 T1036 T1033 arg1Of line,intestinal
R853 T1036 T1034 arg1Of line,epithelial
R854 T1036 T1035 arg1Of line,cell
R855 T1036 T1037 arg1Of line,","
R856 T1038 T1037 arg2Of NCM460,","
R857 T1044 T1043 arg2Of NF-κB,(
R858 T1045 T1043 arg3Of ),(
R859 T1046 T1040 arg2Of activation,via
R860 T1046 T1041 arg1Of activation,Nuclear
R861 T1046 T1042 arg1Of activation,Factor-κB
R862 T1046 T1043 arg1Of activation,(
R863 T1048 T1047 arg2Of 13,[
R864 T1049 T1047 arg3Of ],[
R865 T1052 T1051 arg2Of 14,[
R866 T1053 T1051 arg3Of ],[
R867 T1054 T1055 arg1Of NF-κB,is
R868 T1058 T1055 arg2Of factor,is
R869 T1058 T1056 arg1Of factor,a
R870 T1058 T1057 arg1Of factor,transcription
R871 T1058 T1059 arg1Of factor,that
R872 T1058 T1060 arg1Of factor,regulates
R873 T1060 T1071 arg1Of regulates,","
R874 T1060 T1072 arg1Of regulates,[
R875 T1062 T1060 arg2Of expression,regulates
R876 T1062 T1061 arg1Of expression,the
R877 T1062 T1063 arg1Of expression,of
R878 T1064 T1063 arg2Of genes,of
R879 T1064 T1065 arg2Of genes,associated
R880 T1065 T1066 arg1Of associated,with
R881 T1067 T1066 arg2Of inflammation,with
R882 T1067 T1068 arg1Of inflammation,[
R883 T1069 T1068 arg2Of 15,[
R884 T1070 T1068 arg3Of ],[
R885 T1073 T1072 arg2Of 16,[
R886 T1074 T1072 arg3Of ],[
R887 T1076 T1077 arg1Of infiltration,and
R888 T1077 T1075 arg1Of and,Macrophage
R889 T1077 T1079 arg1Of and,is
R890 T1078 T1077 arg2Of accumulation,and
R891 T1079 T1086 arg1Of is,[
R892 T1082 T1079 arg2Of characteristic,is
R893 T1082 T1080 arg1Of characteristic,a
R894 T1082 T1081 arg1Of characteristic,common
R895 T1082 T1083 arg1Of characteristic,of
R896 T1085 T1083 arg2Of diseases,of
R897 T1085 T1084 arg1Of diseases,intestinal
R898 T1087 T1086 arg2Of 17,[
R899 T1088 T1086 arg3Of ],[
R900 T1089 T1090 arg1Of Macrophages,represent
R901 T1089 T1099 arg1Of Macrophages,secrete
R902 T1089 T1108 arg1Of Macrophages,express
R903 T1090 T1098 arg1Of represent,","
R904 T1092 T1090 arg2Of %,represent
R905 T1092 T1091 arg1Of %,10
R906 T1092 T1093 arg1Of %,of
R907 T1097 T1093 arg2Of cells,of
R908 T1097 T1094 arg1Of cells,total
R909 T1097 T1095 arg1Of cells,lamina
R910 T1097 T1096 arg1Of cells,propria
R911 T1098 T1107 arg1Of ",",and
R912 T1099 T1098 arg2Of secrete,","
R913 T1102 T1099 arg2Of range,secrete
R914 T1102 T1100 arg1Of range,a
R915 T1102 T1101 arg1Of range,wide
R916 T1102 T1103 arg1Of range,of
R917 T1106 T1103 arg2Of compounds,of
R918 T1106 T1104 arg1Of compounds,biologically
R919 T1106 T1105 arg1Of compounds,active
R920 T1108 T1107 arg2Of express,and
R921 T1110 T1108 arg2Of molecules,express
R922 T1110 T1109 arg1Of molecules,cell-adhesion
R923 T1114 T1111 arg1Of response,The
R924 T1114 T1112 arg1Of response,immune
R925 T1114 T1113 arg1Of response,cell
R926 T1114 T1115 arg1Of response,to
R927 T1114 T1119 arg1Of response,seems
R928 T1114 T1121 arg1Of response,be
R929 T1114 T1122 arg2Of response,amplified
R930 T1114 T1125 arg2Of response,generated
R931 T1118 T1115 arg2Of stimulus,to
R932 T1118 T1116 arg1Of stimulus,an
R933 T1118 T1117 arg1Of stimulus,inflammatory
R934 T1122 T1123 arg1Of amplified,or
R935 T1123 T1119 arg2Of or,seems
R936 T1123 T1120 arg1Of or,to
R937 T1123 T1121 arg2Of or,be
R938 T1125 T1123 arg2Of generated,or
R939 T1125 T1124 arg1Of generated,directly
R940 T1125 T1126 arg1Of generated,by
R941 T1127 T1126 arg2Of cells,by
R942 T1127 T1128 arg2Of cells,exposed
R943 T1128 T1129 arg1Of exposed,to
R944 T1131 T1129 arg2Of polysaccharides,to
R945 T1131 T1130 arg1Of polysaccharides,sulphated
R946 T1131 T1133 arg1Of polysaccharides,as
R947 T1133 T1132 arg1Of as,such
R948 T1134 T1133 arg2Of carrageenans,as
R949 T1137 T1138 arg2Of inflammation,induced
R950 T1137 T1141 arg1Of inflammation,was
R951 T1137 T1142 arg2Of inflammation,associated
R952 T1140 T1138 arg1Of dCGN,induced
R953 T1140 T1139 arg2Of dCGN,by
R954 T1142 T1135 arg1Of associated,Indeed
R955 T1142 T1136 arg1Of associated,","
R956 T1142 T1141 arg2Of associated,was
R957 T1142 T1143 arg1Of associated,with
R958 T1142 T1153 arg1Of associated,","
R959 T1142 T1154 arg1Of associated,[
R960 T1144 T1143 arg2Of recruitment,with
R961 T1144 T1145 arg1Of recruitment,of
R962 T1144 T1147 arg1Of recruitment,to
R963 T1146 T1145 arg2Of macrophages,of
R964 T1149 T1147 arg2Of sites,to
R965 T1149 T1148 arg1Of sites,inflammation
R966 T1149 T1150 arg1Of sites,[
R967 T1151 T1150 arg2Of 18,[
R968 T1152 T1150 arg3Of ],[
R969 T1155 T1154 arg2Of 19,[
R970 T1156 T1154 arg3Of ],[
R971 T1159 T1160 arg2Of inflammation,induced
R972 T1159 T1173 arg1Of inflammation,was
R973 T1159 T1175 arg2Of inflammation,associated
R974 T1164 T1160 arg1Of Sodium,induced
R975 T1164 T1161 arg2Of Sodium,by
R976 T1164 T1162 arg1Of Sodium,Dextran
R977 T1164 T1163 arg1Of Sodium,Sulphate
R978 T1164 T1165 arg1Of Sodium,(
R979 T1164 T1168 arg1Of Sodium,","
R980 T1166 T1165 arg2Of DSS,(
R981 T1167 T1165 arg3Of ),(
R982 T1171 T1168 arg2Of compound,","
R983 T1171 T1169 arg1Of compound,another
R984 T1171 T1170 arg2Of compound,sulphated
R985 T1175 T1157 arg1Of associated,Also
R986 T1175 T1158 arg1Of associated,","
R987 T1175 T1172 arg1Of associated,","
R988 T1175 T1173 arg2Of associated,was
R989 T1175 T1174 arg1Of associated,directly
R990 T1175 T1176 arg1Of associated,with
R991 T1175 T1182 arg1Of associated,","
R992 T1175 T1183 arg1Of associated,since
R993 T1178 T1176 arg2Of recruitment,with
R994 T1178 T1177 arg1Of recruitment,macrophages
R995 T1178 T1179 arg1Of recruitment,[
R996 T1180 T1179 arg2Of 20,[
R997 T1181 T1179 arg3Of ],[
R998 T1184 T1186 arg1Of DSS,provoked
R999 T1186 T1183 arg2Of provoked,since
R1000 T1186 T1185 arg1Of provoked,still
R1001 T1186 T1188 arg1Of provoked,after
R1002 T1186 T1194 arg1Of provoked,[
R1003 T1187 T1186 arg2Of inflammation,provoked
R1004 T1189 T1190 arg1Of T-lymphocyte,and
R1005 T1191 T1190 arg2Of NK,and
R1006 T1193 T1188 arg2Of depletion,after
R1007 T1193 T1189 arg1Of depletion,T-lymphocyte
R1008 T1193 T1191 arg1Of depletion,NK
R1009 T1193 T1192 arg1Of depletion,cell
R1010 T1195 T1194 arg2Of 20,[
R1011 T1196 T1194 arg3Of ],[
R1012 T1198 T1199 arg1Of inflammation,can
R1013 T1198 T1200 arg1Of inflammation,be
R1014 T1198 T1201 arg2Of inflammation,induced
R1015 T1201 T1197 arg2Of induced,Although
R1016 T1201 T1199 arg2Of induced,can
R1017 T1201 T1200 arg2Of induced,be
R1018 T1203 T1201 arg1Of dCGN,induced
R1019 T1203 T1202 arg2Of dCGN,by
R1020 T1205 T1206 arg1Of there,are
R1021 T1206 T1197 arg1Of are,Although
R1022 T1206 T1204 arg1Of are,","
R1023 T1208 T1206 arg2Of data,are
R1024 T1208 T1207 arg1Of data,no
R1025 T1208 T1209 arg1Of data,on
R1026 T1212 T1209 arg2Of responses,on
R1027 T1212 T1210 arg1Of responses,human
R1028 T1212 T1211 arg1Of responses,monocyte
R1029 T1212 T1213 arg1Of responses,to
R1030 T1215 T1213 arg2Of exposure,to
R1031 T1215 T1214 arg1Of exposure,dCGN
R1032 T1219 T1218 arg1Of investigate,to
R1033 T1219 T1240 arg1Of investigate,were
R1034 T1219 T1241 arg2Of investigate,exposed
R1035 T1221 T1219 arg2Of effects,investigate
R1036 T1221 T1220 arg1Of effects,the
R1037 T1221 T1222 arg1Of effects,of
R1038 T1221 T1224 arg1Of effects,on
R1039 T1223 T1222 arg2Of dCGN,of
R1040 T1226 T1225 arg1Of monocytes,human
R1041 T1226 T1227 arg1Of monocytes,","
R1042 T1227 T1235 arg1Of ",",and
R1043 T1231 T1227 arg2Of Monocytes,","
R1044 T1231 T1228 arg1Of Monocytes,normal
R1045 T1231 T1229 arg1Of Monocytes,Peripheral
R1046 T1231 T1230 arg1Of Monocytes,Blood
R1047 T1231 T1232 arg1Of Monocytes,(
R1048 T1233 T1232 arg2Of PBM,(
R1049 T1234 T1232 arg3Of ),(
R1050 T1235 T1224 arg2Of and,on
R1051 T1239 T1235 arg2Of cells,and
R1052 T1239 T1236 arg1Of cells,tumoral
R1053 T1239 T1237 arg1Of cells,monocyte/macrophage
R1054 T1239 T1238 arg1Of cells,THP-1
R1055 T1241 T1216 arg1Of exposed,Therefore
R1056 T1241 T1217 arg1Of exposed,","
R1057 T1241 T1240 arg2Of exposed,were
R1058 T1241 T1242 arg1Of exposed,to
R1059 T1244 T1243 arg1Of kDa,10
R1060 T1244 T1245 arg1Of kDa,and
R1061 T1245 T1242 arg2Of and,to
R1062 T1248 T1245 arg2Of dCGN,and
R1063 T1248 T1246 arg1Of dCGN,40
R1064 T1248 T1247 arg1Of dCGN,kDa
R1065 T1249 T1250 arg1Of We,found
R1066 T1252 T1253 arg1Of dCGN,inhibited
R1067 T1253 T1258 arg1Of inhibited,vitro
R1068 T1256 T1253 arg2Of proliferation,inhibited
R1069 T1256 T1254 arg1Of proliferation,THP-1
R1070 T1256 T1255 arg1Of proliferation,cell
R1071 T1258 T1257 arg1Of vitro,in
R1072 T1262 T1260 arg2Of expression,increased
R1073 T1262 T1261 arg1Of expression,ICAM-1
R1074 T1262 T1264 arg1Of expression,stimulated
R1075 T1262 T1270 arg1Of expression,stimulated
R1076 T1264 T1269 arg1Of stimulated,and
R1077 T1267 T1264 arg2Of aggregation,stimulated
R1078 T1267 T1265 arg1Of aggregation,ICAM-1-dependent
R1079 T1267 T1266 arg1Of aggregation,monocyte
R1080 T1269 T1250 arg2Of and,found
R1081 T1269 T1251 arg1Of and,that
R1082 T1269 T1253 arg3Of and,inhibited
R1083 T1269 T1259 arg1Of and,","
R1084 T1269 T1263 arg1Of and,","
R1085 T1269 T1268 arg1Of and,","
R1086 T1270 T1269 arg2Of stimulated,and
R1087 T1272 T1273 arg1Of expression,and
R1088 T1273 T1270 arg2Of and,stimulated
R1089 T1273 T1271 arg1Of and,TNF-α
R1090 T1274 T1273 arg2Of secretion,and
R1091 T1276 T1275 arg1Of responses,These
R1092 T1276 T1277 arg1Of responses,were
R1093 T1276 T1279 arg1Of responses,pronounced
R1094 T1276 T1286 arg1Of responses,were
R1095 T1276 T1287 arg2Of responses,linked
R1096 T1277 T1280 arg1Of were,after
R1097 T1277 T1285 arg1Of were,and
R1098 T1279 T1277 arg2Of pronounced,were
R1099 T1279 T1278 arg1Of pronounced,more
R1100 T1284 T1280 arg2Of exposure,after
R1101 T1284 T1281 arg1Of exposure,40
R1102 T1284 T1282 arg1Of exposure,kDa
R1103 T1284 T1283 arg1Of exposure,dCGN
R1104 T1287 T1285 arg2Of linked,and
R1105 T1287 T1286 arg2Of linked,were
R1106 T1287 T1288 arg1Of linked,to
R1107 T1290 T1288 arg2Of activation,to
R1108 T1290 T1289 arg1Of activation,NF-κB
R1109 T1292 T1291 arg2Of addition,In
R1110 T1297 T1294 arg1Of dCGN,the
R1111 T1297 T1295 arg1Of dCGN,40
R1112 T1297 T1296 arg1Of dCGN,kDa
R1113 T1297 T1299 arg1Of dCGN,but
R1114 T1299 T1298 arg1Of but,","
R1115 T1299 T1300 arg1Of but,not
R1116 T1299 T1305 arg1Of but,induced
R1117 T1304 T1299 arg2Of dCGN,but
R1118 T1304 T1301 arg1Of dCGN,the
R1119 T1304 T1302 arg1Of dCGN,10
R1120 T1304 T1303 arg1Of dCGN,kDa
R1121 T1305 T1291 arg1Of induced,In
R1122 T1305 T1293 arg1Of induced,","
R1123 T1305 T1309 arg1Of induced,as
R1124 T1307 T1306 arg1Of vivo,in
R1125 T1308 T1305 arg2Of colitis,induced
R1126 T1308 T1307 arg1Of colitis,vivo
R1127 T1310 T1309 arg2Of shown,as
R1128 T1314 T1310 arg1Of response,shown
R1129 T1314 T1311 arg2Of response,by
R1130 T1314 T1312 arg1Of response,the
R1131 T1314 T1313 arg1Of response,inflammatory
R1132 T1314 T1315 arg1Of response,in
R1133 T1318 T1315 arg2Of colon,in
R1134 T1318 T1316 arg1Of colon,the
R1135 T1318 T1317 arg1Of colon,rat
R1136 T1320 T1319 arg1Of results,These
R1137 T1320 T1321 arg1Of results,suggest
R1138 T1325 T1323 arg1Of forms,the
R1139 T1325 T1324 arg1Of forms,degraded
R1140 T1325 T1326 arg1Of forms,of
R1141 T1325 T1328 arg1Of forms,have
R1142 T1327 T1326 arg2Of CGN,of
R1143 T1328 T1321 arg2Of have,suggest
R1144 T1328 T1322 arg1Of have,that
R1145 T1328 T1332 arg1Of have,on
R1146 T1331 T1328 arg2Of effect,have
R1147 T1331 T1329 arg1Of effect,an
R1148 T1331 T1330 arg1Of effect,important
R1149 T1333 T1332 arg2Of monocytes,on
R1150 T1333 T1334 arg1Of monocytes,resulting
R1151 T1334 T1335 arg1Of resulting,in
R1152 T1338 T1335 arg2Of phenotype,in
R1153 T1338 T1336 arg1Of phenotype,an
R1154 T1338 T1337 arg1Of phenotype,inflammatory
R1776 T2063 T2064 arg1Of Preparation,of
R1777 T2066 T2064 arg2Of Carrageenan,of
R1778 T2066 T2065 arg1Of Carrageenan,Degraded
R1779 T2068 T2067 arg1Of preparations,Two
R1780 T2068 T2069 arg1Of preparations,of
R1781 T2068 T2093 modOf preparations,were
R1784 T2071 T2069 arg2Of carrageenan,of
R1786 T2073 T2075 arg1Of low,(
R1787 T2073 T2082 arg1Of low,and
R1788 T2077 T2075 arg2Of kDa,(
R1789 T2077 T2076 arg1Of kDa,∼10
R1790 T2077 T2078 arg1Of kDa,;
R1791 T2079 T2078 arg2Of C10,;
R1792 T2080 T2075 arg3Of ),(
R1794 T2082 T2081 arg1Of and,","
R1795 T2083 T2082 arg2Of medium,and
R1796 T2087 T2085 arg2Of kDa,(
R1797 T2087 T2086 arg1Of kDa,∼40
R1798 T2087 T2088 arg1Of kDa,;
R1799 T2089 T2088 arg2Of C40,;
R1800 T2090 T2085 arg3Of ),(
R1801 T2092 T2085 arg1Of weight,(
R1802 T2092 T2091 arg1Of weight,molecular
R1803 T2092 T2093 arg1Of weight,were
R1804 T2092 T2094 arg2Of weight,prepared
R1805 T2094 T2084 arg1Of prepared,","
R1806 T2094 T2093 arg2Of prepared,were
R1807 T2094 T2095 arg1Of prepared,from
R1808 T2097 T2095 arg2Of iota-carrageenan,from
R1809 T2097 T2096 arg1Of iota-carrageenan,native
R1810 T2097 T2098 arg2Of iota-carrageenan,extracted
R1811 T2098 T2099 arg1Of extracted,from
R1812 T2101 T2099 arg2Of spinosum,from
R1813 T2101 T2100 arg1Of spinosum,Euchema
R1814 T2101 T2102 arg1Of spinosum,(
R1815 T2104 T2102 arg2Of provided,(
R1816 T2104 T2103 arg1Of provided,generously
R1817 T2108 T2104 arg1Of Industry,provided
R1818 T2108 T2105 arg2Of Industry,by
R1819 T2108 T2106 arg1Of Industry,Sanofi
R1820 T2108 T2107 arg1Of Industry,Biosystems
R1821 T2108 T2109 arg1Of Industry,","
R1822 T2110 T2109 arg2Of Boulogne-Billancourt,","
R1823 T2110 T2111 arg1Of Boulogne-Billancourt,","
R1824 T2112 T2111 arg2Of France,","
R1825 T2113 T2102 arg3Of ),(
R1826 T2115 T2114 arg1Of carrageenan,Native
R1827 T2115 T2116 arg1Of carrageenan,was
R1828 T2115 T2117 arg2Of carrageenan,dissolved
R1829 T2115 T2130 arg2Of carrageenan,heated
R1830 T2117 T2118 arg1Of dissolved,in
R1831 T2117 T2129 arg1Of dissolved,and
R1832 T2120 T2118 arg2Of water,in
R1833 T2120 T2119 arg1Of water,distilled
R1834 T2120 T2121 arg1Of water,(
R1835 T2120 T2126 arg1Of water,under
R1836 T2122 T2123 arg1Of 5,%
R1837 T2124 T2121 arg2Of w/v,(
R1838 T2124 T2122 arg1Of w/v,5
R1839 T2125 T2121 arg3Of ),(
R1840 T2128 T2126 arg2Of stirring,under
R1841 T2128 T2127 arg1Of stirring,vigorous
R1842 T2129 T2116 arg2Of and,was
R1843 T2130 T2129 arg2Of heated,and
R1844 T2130 T2131 arg1Of heated,to
R1845 T2133 T2131 arg2Of Then,to
R1846 T2133 T2132 arg1Of Then,60°C.
R1847 T2137 T2135 arg1Of solution,the
R1848 T2137 T2136 arg1Of solution,carrageenan
R1849 T2137 T2138 arg1Of solution,was
R1850 T2137 T2139 arg2Of solution,submitted
R1851 T2139 T2116 modOf submitted,was
R1852 T2139 T2134 arg1Of submitted,","
R1853 T2139 T2138 arg2Of submitted,was
R1854 T2139 T2140 arg1Of submitted,to
R1855 T2143 T2140 arg2Of treatments,to
R1856 T2143 T2141 arg1Of treatments,two
R1857 T2143 T2142 arg1Of treatments,different
R1858 T2145 T2139 arg3Of obtain,submitted
R1859 T2145 T2144 arg1Of obtain,to
R1860 T2147 T2148 arg1Of low,and
R1861 T2148 T2146 arg1Of and,both
R1862 T2149 T2148 arg2Of medium,and
R1863 T2152 T2145 arg2Of fractions,obtain
R1864 T2152 T2147 arg1Of fractions,low
R1865 T2152 T2149 arg1Of fractions,medium
R1866 T2152 T2150 arg1Of fractions,molecular
R1867 T2152 T2151 arg1Of fractions,weight
R1868 T2160 T2155 arg2Of fraction,for
R1869 T2160 T2156 arg1Of fraction,the
R1870 T2160 T2157 arg1Of fraction,low
R1871 T2160 T2158 arg1Of fraction,molecular
R1872 T2160 T2159 arg1Of fraction,weight
R1873 T2163 T2162 arg1Of solution,carrageenan
R1874 T2163 T2164 arg1Of solution,was
R1875 T2163 T2165 arg2Of solution,hydrolyzed
R1876 T2165 T2164 arg2Of hydrolyzed,was
R1877 T2165 T2166 arg1Of hydrolyzed,with
R1878 T2168 T2166 arg2Of %,with
R1879 T2168 T2167 arg1Of %,0.3
R1880 T2168 T2169 arg1Of %,(
R1881 T2168 T2172 arg2Of %,concentrated
R1882 T2170 T2169 arg2Of v/v,(
R1883 T2171 T2169 arg3Of ),(
R1884 T2174 T2172 arg1Of acid,concentrated
R1885 T2174 T2173 arg1Of acid,sulphuric
R1886 T2174 T2175 arg1Of acid,for
R1887 T2174 T2178 arg1Of acid,at
R1888 T2177 T2175 arg2Of min,for
R1889 T2177 T2176 arg1Of min,15
R1890 T2180 T2179 arg1Of After,80°C.
R1891 T2181 T2178 arg2Of neutralization,at
R1892 T2181 T2180 arg1Of neutralization,After
R1893 T2181 T2182 arg1Of neutralization,with
R1894 T2184 T2182 arg2Of 4N,with
R1895 T2184 T2183 arg1Of 4N,NaOH
R1896 T2187 T2186 arg1Of solution,the
R1897 T2187 T2188 arg1Of solution,was
R1898 T2188 T2153 arg1Of was,Briefly
R1899 T2188 T2154 arg1Of was,","
R1900 T2188 T2155 arg1Of was,for
R1901 T2188 T2161 arg1Of was,","
R1902 T2188 T2164 modOf was,was
R1903 T2188 T2185 arg1Of was,","
R1904 T2189 T2188 arg2Of ultra,was
R1905 T2189 T2190 arg2Of ultra,filtered
R1906 T2189 T2201 arg1Of ultra,","
R1907 T2189 T2202 arg1Of ultra,(
R1908 T2190 T2191 arg1Of filtered,through
R1909 T2195 T2191 arg2Of cartridge,through
R1910 T2195 T2192 arg1Of cartridge,a
R1911 T2195 T2193 arg1Of cartridge,hollow
R1912 T2195 T2194 arg1Of cartridge,fibre
R1913 T2195 T2196 arg1Of cartridge,with
R1914 T2200 T2196 arg2Of kDa,with
R1915 T2200 T2197 arg1Of kDa,MW
R1916 T2200 T2198 arg1Of kDa,cut-off
R1917 T2200 T2199 arg1Of kDa,5
R1918 T2204 T2202 arg2Of Inc,(
R1919 T2204 T2203 arg1Of Inc,Amicon
R1920 T2204 T2205 arg1Of Inc,","
R1921 T2206 T2205 arg2Of Beverly,","
R1922 T2206 T2207 arg1Of Beverly,","
R1923 T2208 T2207 arg2Of USA,","
R1924 T2209 T2202 arg3Of ),(
R1925 T2215 T2211 arg1Of fraction,the
R1926 T2215 T2212 arg1Of fraction,medium
R1927 T2215 T2213 arg1Of fraction,molecular
R1928 T2215 T2214 arg1Of fraction,weight
R1929 T2215 T2228 arg1Of fraction,concentrated
R1930 T2219 T2217 arg1Of solution,the
R1931 T2219 T2218 arg1Of solution,carrageenan
R1932 T2219 T2220 arg1Of solution,was
R1933 T2219 T2221 arg2Of solution,hydrolyzed
R1934 T2221 T2220 arg2Of hydrolyzed,was
R1935 T2221 T2222 arg1Of hydrolyzed,with
R1936 T2224 T2222 arg2Of %,with
R1937 T2224 T2223 arg1Of %,0.3
R1938 T2224 T2225 arg1Of %,(
R1939 T2226 T2225 arg2Of v/v,(
R1940 T2227 T2225 arg3Of ),(
R1941 T2228 T2210 arg1Of concentrated,For
R1942 T2228 T2216 arg1Of concentrated,","
R1943 T2228 T2220 modOf concentrated,was
R1944 T2228 T2244 arg1Of concentrated,(
R1945 T2230 T2228 arg2Of acid,concentrated
R1946 T2230 T2229 arg1Of acid,sulphuric
R1947 T2230 T2231 arg1Of acid,for
R1948 T2230 T2234 arg1Of acid,at
R1949 T2230 T2243 arg2Of acid,filtered
R1950 T2233 T2231 arg2Of min,for
R1951 T2233 T2232 arg1Of min,30
R1952 T2235 T2234 arg2Of 60°C.,at
R1953 T2235 T2236 arg1Of 60°C.,After
R1954 T2237 T2236 arg2Of neutralization,After
R1955 T2240 T2239 arg1Of supernatant,the
R1956 T2240 T2241 arg1Of supernatant,was
R1957 T2240 T2242 arg1Of supernatant,ultra
R1958 T2241 T2238 arg1Of was,","
R1959 T2242 T2241 arg2Of ultra,was
R1960 T2243 T2241 arg3Of filtered,was
R1961 T2245 T2244 arg2Of MW,(
R1962 T2245 T2246 arg1Of MW,cut-off
R1963 T2246 T2248 arg1Of cut-off,kDa
R1964 T2248 T2247 arg1Of kDa,100
R1965 T2249 T2244 arg3Of ),(
R1966 T2251 T2250 arg1Of filtrate,The
R1967 T2251 T2252 arg1Of filtrate,was
R1968 T2251 T2253 arg2Of filtrate,submitted
R1969 T2253 T2252 arg2Of submitted,was
R1970 T2253 T2254 arg1Of submitted,to
R1971 T2258 T2254 arg2Of filtration,to
R1972 T2258 T2255 arg1Of filtration,a
R1973 T2258 T2256 arg1Of filtration,second
R1974 T2258 T2257 arg1Of filtration,ultra
R1975 T2258 T2259 arg1Of filtration,(
R1976 T2263 T2259 arg2Of kDa,(
R1977 T2263 T2260 arg1Of kDa,MW
R1978 T2263 T2261 arg1Of kDa,cut-off
R1979 T2263 T2262 arg1Of kDa,5
R1980 T2264 T2259 arg3Of ),(
R1981 T2266 T2265 arg1Of preparations,Both
R1982 T2266 T2267 arg1Of preparations,of
R1983 T2266 T2269 arg1Of preparations,were
R1984 T2266 T2270 arg2Of preparations,precipitated
R1985 T2268 T2267 arg2Of dCGN,of
R1986 T2270 T2269 arg2Of precipitated,were
R1987 T2270 T2271 arg1Of precipitated,with
R1988 T2273 T2271 arg2Of volumes,with
R1989 T2273 T2272 arg1Of volumes,4
R1990 T2273 T2274 arg1Of volumes,of
R1991 T2273 T2279 arg2Of volumes,dried
R1992 T2275 T2276 arg1Of 95,%
R1993 T2277 T2274 arg2Of ethanol,of
R1994 T2277 T2275 arg1Of ethanol,95
R1995 T2279 T2278 arg1Of dried,","
R1996 T2279 T2280 arg1Of dried,at
R1997 T2282 T2281 arg1Of temperature,room
R1998 T2282 T2283 arg1Of temperature,and
R1999 T2283 T2280 arg2Of and,at
R2000 T2284 T2283 arg2Of ground,and
R2001 T2284 T2285 arg1Of ground,to
R2002 T2287 T2285 arg2Of particles,to
R2003 T2287 T2286 arg1Of particles,small
R2004 T2287 T2288 arg1Of particles,(
R2005 T2290 T2288 arg2Of mm,(
R2006 T2290 T2289 arg1Of mm,1
R2007 T2290 T2291 arg1Of mm,in
R2008 T2292 T2291 arg2Of diameter,in
R2009 T2293 T2288 arg3Of ),(
R2010 T2296 T2294 arg1Of chromatography,Using
R2011 T2296 T2295 arg1Of chromatography,gel-permeation
R2012 T2296 T2297 arg1Of chromatography,in
R2013 T2296 T2299 arg1Of chromatography,with
R2014 T2296 T2304 arg1Of chromatography,see
R2015 T2298 T2297 arg2Of combination,in
R2016 T2302 T2299 arg2Of measurements,with
R2017 T2302 T2300 arg1Of measurements,light
R2018 T2302 T2301 arg1Of measurements,scattering
R2019 T2304 T2303 arg1Of see,(
R2020 T2307 T2305 arg1Of al.,Viebke
R2021 T2307 T2306 arg1Of al.,et
R2022 T2307 T2308 arg1Of al.,[
R2023 T2307 T2311 arg1Of al.,)
R2024 T2307 T2312 arg1Of al.,","
R2025 T2307 T2314 arg1Of al.,was
R2026 T2307 T2315 arg2Of al.,confirmed
R2027 T2309 T2308 arg2Of 21,[
R2028 T2310 T2308 arg3Of ],[
R2029 T2313 T2312 arg2Of it,","
R2030 T2315 T2314 arg2Of confirmed,was
R2031 T2316 T2315 arg3Of that,confirmed
R2032 T2319 T2317 arg1Of fraction,the
R2033 T2319 T2318 arg1Of fraction,low
R2034 T2319 T2320 arg1Of fraction,had
R2035 T2320 T2304 arg2Of had,see
R2036 T2320 T2314 modOf had,was
R2037 T2324 T2321 arg1Of weight,an
R2038 T2324 T2322 arg1Of weight,average
R2039 T2324 T2323 arg1Of weight,molecular
R2040 T2324 T2325 arg1Of weight,of
R2041 T2324 T2329 arg1Of weight,and
R2042 T2327 T2325 arg2Of kDa,of
R2043 T2327 T2326 arg1Of kDa,10
R2044 T2329 T2320 arg2Of and,had
R2045 T2329 T2328 arg1Of and,","
R2046 T2332 T2329 arg2Of fraction,and
R2047 T2332 T2330 arg1Of fraction,the
R2048 T2332 T2331 arg1Of fraction,medium
R2049 T2332 T2333 arg1Of fraction,of
R2050 T2335 T2333 arg2Of kDa,of
R2051 T2335 T2334 arg1Of kDa,40
R2052 T2338 T2336 arg1Of content,The
R2053 T2338 T2337 arg1Of content,sulphate
R2054 T2338 T2339 arg1Of content,of
R2055 T2338 T2341 arg1Of content,in
R2056 T2338 T2344 arg1Of content,was
R2057 T2338 T2345 arg2Of content,measured
R2058 T2340 T2339 arg2Of polysaccharides,of
R2059 T2343 T2341 arg2Of fractions,in
R2060 T2343 T2342 arg1Of fractions,both
R2061 T2345 T2344 arg2Of measured,was
R2062 T2345 T2346 arg1Of measured,following
R2063 T2348 T2346 arg2Of method,following
R2064 T2348 T2347 arg1Of method,the
R2065 T2348 T2349 arg1Of method,of
R2066 T2352 T2349 arg2Of al.,of
R2067 T2352 T2350 arg1Of al.,Quemener
R2068 T2352 T2351 arg1Of al.,et
R2069 T2352 T2353 arg1Of al.,[
R2070 T2354 T2353 arg2Of 22,[
R2071 T2355 T2353 arg3Of ],[
R2072 T2359 T2358 arg1Of absence,the
R2073 T2359 T2360 arg1Of absence,of
R2074 T2359 T2368 arg1Of absence,was
R2075 T2359 T2369 arg2Of absence,confirmed
R2076 T2363 T2360 arg2Of modifications,of
R2077 T2363 T2361 arg1Of modifications,polysaccharide
R2078 T2363 T2362 arg1Of modifications,structure
R2079 T2363 T2364 arg1Of modifications,in
R2080 T2367 T2364 arg2Of fractions,in
R2081 T2367 T2365 arg1Of fractions,the
R2082 T2367 T2366 arg1Of fractions,two
R2083 T2369 T2356 arg1Of confirmed,Finally
R2084 T2369 T2357 arg1Of confirmed,","
R2085 T2369 T2368 arg2Of confirmed,was
R2086 T2370 T2369 arg3Of using,confirmed
R2087 T2372 T2370 arg2Of spectroscopy,using
R2088 T2372 T2371 arg1Of spectroscopy,2H-NMR
R2089 T2374 T2373 arg1Of absence,The
R2090 T2374 T2375 arg1Of absence,of
R2091 T2374 T2378 arg1Of absence,in
R2092 T2374 T2382 arg1Of absence,was
R2093 T2374 T2383 arg2Of absence,confirmed
R2094 T2377 T2375 arg2Of contamination,of
R2095 T2377 T2376 arg1Of contamination,LPS
R2096 T2381 T2378 arg2Of fractions,in
R2097 T2381 T2379 arg1Of fractions,the
R2098 T2381 T2380 arg1Of fractions,two
R2099 T2383 T2382 arg2Of confirmed,was
R2100 T2384 T2383 arg3Of using,confirmed
R2101 T2384 T2388 arg1Of using,(
R2102 T2387 T2384 arg2Of kit,using
R2103 T2387 T2385 arg1Of kit,the
R2104 T2387 T2386 arg1Of kit,e-Toxate®
R2105 T2389 T2388 arg2Of Sigma,(
R2106 T2389 T2390 arg1Of Sigma,","
R2107 T2393 T2390 arg2Of Fallavier,","
R2108 T2393 T2391 arg1Of Fallavier,St
R2109 T2393 T2392 arg1Of Fallavier,Quentin
R2110 T2393 T2394 arg1Of Fallavier,","
R2111 T2395 T2394 arg2Of France,","
R2112 T2396 T2388 arg3Of ),(
R2113 T2398 T2397 arg2Of use,Before
R2114 T2398 T2399 arg1Of use,in
R2115 T2401 T2399 arg2Of culture,in
R2116 T2401 T2400 arg1Of culture,cell
R2117 T2405 T2403 arg1Of fractions,the
R2118 T2405 T2404 arg1Of fractions,two
R2119 T2405 T2406 arg1Of fractions,were
R2120 T2405 T2407 arg2Of fractions,dissolved
R2121 T2407 T2397 arg1Of dissolved,Before
R2122 T2407 T2402 arg1Of dissolved,","
R2123 T2407 T2406 arg2Of dissolved,were
R2124 T2407 T2408 arg1Of dissolved,in
R2125 T2407 T2411 arg1Of dissolved,during
R2126 T2410 T2408 arg2Of medium,in
R2127 T2410 T2409 arg1Of medium,complete
R2128 T2413 T2411 arg2Of min,during
R2129 T2413 T2412 arg1Of min,30
R2130 T2413 T2414 arg1Of min,at
R2131 T2415 T2414 arg2Of 56°C,at
R2508 T2817 T2818 arg1Of Animals,","
R2509 T2818 T2820 arg1Of ",",and
R2510 T2819 T2818 arg2Of Chemicals,","
R2511 T2821 T2820 arg2Of Diet,and
R2512 T2824 T2822 arg1Of rats,Male
R2513 T2824 T2823 arg1Of rats,Wistar
R2514 T2824 T2825 arg1Of rats,(
R2515 T2824 T2831 arg1Of rats,were
R2516 T2824 T2832 arg2Of rats,housed
R2517 T2829 T2825 arg2Of weight,(
R2518 T2829 T2826 arg1Of weight,150
R2519 T2829 T2827 arg1Of weight,g
R2520 T2829 T2828 arg1Of weight,average
R2521 T2830 T2825 arg3Of ),(
R2522 T2832 T2831 arg2Of housed,were
R2523 T2832 T2833 arg1Of housed,under
R2524 T2835 T2834 arg1Of conditions,standard
R2525 T2835 T2836 arg1Of conditions,and
R2526 T2836 T2833 arg2Of and,under
R2527 T2836 T2840 arg1Of and,with
R2528 T2839 T2836 arg2Of libitum,and
R2529 T2839 T2837 arg2Of libitum,fed
R2530 T2839 T2838 arg1Of libitum,ad
R2531 T2844 T2840 arg2Of chow,with
R2532 T2844 T2841 arg1Of chow,standard
R2533 T2844 T2842 arg1Of chow,rodent
R2534 T2844 T2843 arg1Of chow,laboratory
R2535 T2846 T2845 arg2Of iota-carrageenans,Degraded
R2536 T2846 T2847 arg1Of iota-carrageenans,were
R2537 T2846 T2848 arg2Of iota-carrageenans,administered
R2538 T2848 T2847 arg2Of administered,were
R2539 T2848 T2849 arg1Of administered,in
R2540 T2848 T2858 arg1Of administered,for
R2541 T2852 T2849 arg2Of water,in
R2542 T2852 T2850 arg1Of water,the
R2543 T2852 T2851 arg1Of water,drinking
R2544 T2852 T2853 arg1Of water,(
R2545 T2854 T2855 arg1Of 5,%
R2546 T2856 T2853 arg2Of w/v,(
R2547 T2856 T2854 arg1Of w/v,5
R2548 T2857 T2853 arg3Of ),(
R2549 T2860 T2858 arg2Of days,for
R2550 T2860 T2859 arg1Of days,55
R2551 T2860 T2861 arg1Of days,to
R2552 T2863 T2861 arg2Of groups,to
R2553 T2863 T2862 arg1Of groups,2
R2554 T2863 T2864 arg1Of groups,of
R2555 T2866 T2864 arg2Of animals,of
R2556 T2866 T2865 arg1Of animals,six
R2557 T2866 T2867 arg1Of animals,each
R2558 T2870 T2868 arg1Of group,The
R2559 T2870 T2869 arg1Of group,first
R2560 T2870 T2871 arg1Of group,received
R2561 T2871 T2882 arg1Of received,and
R2562 T2876 T2871 arg2Of carrageenan,received
R2563 T2876 T2872 arg1Of carrageenan,the
R2564 T2876 T2873 arg1Of carrageenan,low
R2565 T2876 T2874 arg1Of carrageenan,molecular
R2566 T2876 T2875 arg1Of carrageenan,weight
R2567 T2876 T2877 arg1Of carrageenan,(
R2568 T2880 T2877 arg2Of dCGN,(
R2569 T2880 T2878 arg1Of dCGN,10
R2570 T2880 T2879 arg1Of dCGN,kDa
R2571 T2881 T2877 arg3Of ),(
R2572 T2884 T2883 arg1Of second,the
R2573 T2884 T2885 arg1Of second,received
R2574 T2885 T2882 arg2Of received,and
R2575 T2890 T2885 arg2Of carrageenan,received
R2576 T2890 T2886 arg1Of carrageenan,the
R2577 T2890 T2887 arg1Of carrageenan,medium
R2578 T2890 T2888 arg1Of carrageenan,molecular
R2579 T2890 T2889 arg1Of carrageenan,weight
R2580 T2890 T2891 arg1Of carrageenan,(
R2581 T2894 T2891 arg2Of dCGN,(
R2582 T2894 T2892 arg1Of dCGN,40
R2583 T2894 T2893 arg1Of dCGN,kDa
R2584 T2895 T2891 arg3Of ),(
R2585 T2898 T2896 arg1Of group,An
R2586 T2898 T2897 arg1Of group,additional
R2587 T2898 T2899 arg1Of group,of
R2588 T2898 T2902 arg1Of group,were
R2589 T2898 T2903 arg2Of group,maintained
R2590 T2901 T2899 arg2Of rats,of
R2591 T2901 T2900 arg1Of rats,four
R2592 T2903 T2902 arg2Of maintained,were
R2593 T2903 T2904 arg1Of maintained,on
R2594 T2907 T2904 arg2Of water,on
R2595 T2907 T2905 arg1Of water,regular
R2596 T2907 T2906 arg1Of water,tap
R2597 T2907 T2908 arg1Of water,(
R2598 T2910 T2908 arg2Of group,(
R2599 T2910 T2909 arg1Of group,control
R2600 T2911 T2908 arg3Of ),(
R2601 T2913 T2912 arg1Of increase,To
R2602 T2913 T2918 arg1Of increase,was
R2603 T2913 T2919 arg2Of increase,added
R2604 T2915 T2916 arg1Of 0.2,%
R2605 T2917 T2913 arg2Of sucrose,increase
R2606 T2917 T2914 arg1Of sucrose,palatability
R2607 T2917 T2915 arg1Of sucrose,0.2
R2608 T2919 T2918 arg2Of added,was
R2609 T2919 T2920 arg1Of added,to
R2610 T2919 T2927 arg1Of added,(
R2611 T2923 T2920 arg2Of water,to
R2612 T2923 T2921 arg1Of water,the
R2613 T2923 T2922 arg1Of water,drinking
R2614 T2923 T2924 arg1Of water,of
R2615 T2926 T2924 arg2Of groups,of
R2616 T2926 T2925 arg1Of groups,all
R2617 T2930 T2927 arg2Of Waaji,(
R2618 T2930 T2928 arg1Of Waaji,Van
R2619 T2930 T2929 arg1Of Waaji,der
R2620 T2930 T2931 arg1Of Waaji,et
R2621 T2931 T2932 arg1Of et,al.
R2622 T2931 T2933 arg1Of et,","
R2623 T2931 T2934 arg1Of et,[
R2624 T2935 T2934 arg2Of 23,[
R2625 T2936 T2934 arg3Of ],[
R2626 T2937 T2927 arg3Of ),(
R2627 T2940 T2938 arg1Of solutions,Fresh
R2628 T2940 T2939 arg1Of solutions,carrageenan
R2629 T2940 T2941 arg1Of solutions,were
R2630 T2940 T2942 arg2Of solutions,prepared
R2631 T2942 T2941 arg2Of prepared,were
R2632 T2942 T2943 arg1Of prepared,daily
R2772 T3130 T3131 arg1Of Evaluation,of
R2773 T3132 T3131 arg2Of Colitis,of
R2774 T3134 T3133 arg1Of weight,Body
R2775 T3134 T3135 arg1Of weight,","
R2776 T3135 T3140 arg1Of ",",","
R2777 T3136 T3137 arg1Of liquid,and
R2778 T3138 T3137 arg2Of food,and
R2779 T3139 T3135 arg2Of consumption,","
R2780 T3139 T3136 arg1Of consumption,liquid
R2781 T3139 T3138 arg1Of consumption,food
R2782 T3140 T3142 arg1Of ",",and
R2783 T3141 T3140 arg2Of diarrhea,","
R2784 T3142 T3151 arg1Of and,were
R2785 T3142 T3152 arg2Of and,recorded
R2786 T3144 T3142 arg2Of bleeding,and
R2787 T3144 T3143 arg1Of bleeding,rectal
R2788 T3144 T3145 arg1Of bleeding,(
R2789 T3146 T3145 arg2Of detected,(
R2790 T3149 T3146 arg1Of inspection,detected
R2791 T3149 T3147 arg2Of inspection,by
R2792 T3149 T3148 arg1Of inspection,eye
R2793 T3150 T3145 arg3Of ),(
R2794 T3152 T3151 arg2Of recorded,were
R2795 T3152 T3153 arg1Of recorded,throughout
R2796 T3156 T3153 arg2Of period,throughout
R2797 T3156 T3154 arg1Of period,the
R2798 T3156 T3155 arg1Of period,feeding
R2799 T3159 T3157 arg2Of days,After
R2800 T3159 T3158 arg1Of days,55
R2801 T3161 T3162 arg1Of animals,were
R2802 T3161 T3163 arg2Of animals,sacrificed
R2803 T3163 T3157 arg1Of sacrificed,After
R2804 T3163 T3160 arg1Of sacrificed,","
R2805 T3163 T3162 arg2Of sacrificed,were
R2806 T3166 T3163 arg1Of dislocation,sacrificed
R2807 T3166 T3164 arg2Of dislocation,by
R2808 T3166 T3165 arg1Of dislocation,cervical
R2809 T3168 T3167 arg1Of length,The
R2810 T3168 T3169 arg1Of length,of
R2811 T3168 T3172 arg1Of length,was
R2812 T3168 T3173 arg2Of length,measured
R2813 T3171 T3169 arg2Of colon,of
R2814 T3171 T3170 arg1Of colon,the
R2815 T3173 T3172 arg2Of measured,was
R2816 T3173 T3174 arg1Of measured,as
R2817 T3173 T3180 arg1Of measured,[
R2818 T3175 T3174 arg2Of described,as
R2819 T3177 T3175 arg1Of Okayashu,described
R2820 T3177 T3176 arg2Of Okayashu,by
R2821 T3177 T3179 arg1Of Okayashu,al.
R2822 T3179 T3178 arg1Of al.,et
R2823 T3181 T3180 arg2Of 24,[
R2824 T3182 T3180 arg3Of ],[
R2825 T3186 T3185 arg1Of colon,each
R2826 T3186 T3187 arg1Of colon,was
R2827 T3186 T3188 arg2Of colon,ligated
R2828 T3188 T3187 arg2Of ligated,was
R2829 T3188 T3189 arg1Of ligated,in
R2830 T3188 T3194 arg1Of ligated,and
R2831 T3190 T3189 arg2Of sections,in
R2832 T3190 T3191 arg1Of sections,of
R2833 T3193 T3191 arg2Of cm,of
R2834 T3193 T3192 arg1Of cm,2
R2835 T3194 T3183 arg1Of and,Then
R2836 T3194 T3184 arg1Of and,","
R2837 T3195 T3196 arg1Of 1,to
R2838 T3198 T3196 arg2Of ml,to
R2839 T3198 T3197 arg1Of ml,2
R2840 T3200 T3201 arg1Of 10,%
R2841 T3202 T3195 arg1Of formalin,1
R2842 T3202 T3199 arg1Of formalin,of
R2843 T3202 T3200 arg1Of formalin,10
R2844 T3202 T3203 arg1Of formalin,was
R2845 T3202 T3204 arg2Of formalin,infused
R2846 T3204 T3194 arg2Of infused,and
R2847 T3204 T3203 arg2Of infused,was
R2848 T3204 T3205 arg1Of infused,into
R2849 T3208 T3205 arg2Of lumen,into
R2850 T3208 T3206 arg1Of lumen,the
R2851 T3208 T3207 arg1Of lumen,intestinal
R2852 T3211 T3210 arg1Of distended,moderately
R2853 T3212 T3209 arg1Of segment,The
R2854 T3212 T3211 arg1Of segment,distended
R2855 T3212 T3213 arg1Of segment,was
R2856 T3212 T3214 arg2Of segment,sectioned
R2857 T3212 T3216 arg2Of segment,fixed
R2858 T3214 T3215 arg1Of sectioned,and
R2859 T3215 T3213 arg2Of and,was
R2860 T3215 T3217 arg1Of and,in
R2861 T3216 T3215 arg2Of fixed,and
R2862 T3218 T3219 arg1Of 10,%
R2863 T3220 T3217 arg2Of formalin,in
R2864 T3220 T3218 arg1Of formalin,10
R2865 T3223 T3221 arg1Of day,The
R2866 T3223 T3222 arg1Of day,following
R2867 T3227 T3225 arg1Of content,the
R2868 T3227 T3226 arg1Of content,intestinal
R2869 T3227 T3228 arg1Of content,was
R2870 T3227 T3229 arg2Of content,removed
R2871 T3229 T3223 arg1Of removed,day
R2872 T3229 T3224 arg1Of removed,","
R2873 T3229 T3228 arg2Of removed,was
R2874 T3231 T3229 arg1Of vortexing,removed
R2875 T3231 T3230 arg2Of vortexing,by
R2876 T3234 T3232 arg1Of segment,The
R2877 T3234 T3233 arg2Of segment,fixed
R2878 T3234 T3235 arg1Of segment,was
R2879 T3234 T3236 arg2Of segment,kept
R2880 T3236 T3235 arg2Of kept,was
R2881 T3236 T3237 arg1Of kept,in
R2882 T3236 T3241 arg1Of kept,at
R2883 T3236 T3243 arg1Of kept,until
R2884 T3238 T3239 arg1Of 10,%
R2885 T3240 T3237 arg2Of formalin,in
R2886 T3240 T3238 arg1Of formalin,10
R2887 T3242 T3241 arg2Of 4°C,at
R2888 T3247 T3243 arg2Of procedure,until
R2889 T3247 T3244 arg1Of procedure,the
R2890 T3247 T3245 arg1Of procedure,paraffin
R2891 T3247 T3246 arg1Of procedure,embedding
R2892 T3249 T3248 arg1Of evaluate,To
R2893 T3251 T3249 arg2Of degree,evaluate
R2894 T3251 T3250 arg1Of degree,the
R2895 T3251 T3252 arg1Of degree,of
R2896 T3253 T3252 arg2Of inflammation,of
R2897 T3256 T3255 arg1Of segment,this
R2898 T3256 T3257 arg1Of segment,of
R2899 T3256 T3259 arg1Of segment,was
R2900 T3256 T3260 arg2Of segment,opened
R2901 T3258 T3257 arg2Of colon,of
R2902 T3260 T3259 arg2Of opened,was
R2903 T3260 T3261 arg1Of opened,longitudinally
R2904 T3263 T3264 arg1Of macroscopic,and
R2905 T3265 T3264 arg2Of histological,and
R2906 T3266 T3262 arg1Of scores,and
R2907 T3266 T3263 arg1Of scores,macroscopic
R2908 T3266 T3265 arg1Of scores,histological
R2909 T3266 T3267 arg1Of scores,of
R2910 T3266 T3269 arg1Of scores,were
R2911 T3266 T3270 arg2Of scores,recorded
R2912 T3268 T3267 arg2Of inflammation,of
R2913 T3270 T3248 modOf recorded,To
R2914 T3270 T3254 arg1Of recorded,","
R2915 T3270 T3259 modOf recorded,was
R2916 T3270 T3269 arg2Of recorded,were
R2917 T3270 T3271 arg1Of recorded,as
R2918 T3270 T3277 arg1Of recorded,","
R2919 T3270 T3278 arg1Of recorded,[
R2920 T3273 T3272 arg1Of described,previously
R2921 T3275 T3271 arg2Of 25,as
R2922 T3275 T3273 arg1Of 25,described
R2923 T3275 T3274 arg2Of 25,[
R2924 T3276 T3274 arg3Of ],[
R2925 T3279 T3278 arg2Of 26,[
R2926 T3280 T3278 arg3Of ],[
R2927 T3284 T3281 arg1Of staining,The
R2928 T3284 T3282 arg1Of staining,toluidine
R2929 T3284 T3283 arg1Of staining,blue
R2930 T3284 T3285 arg1Of staining,was
R2931 T3284 T3286 arg2Of staining,used
R2932 T3286 T3285 arg2Of used,was
R2933 T3286 T3287 arg1Of used,for
R2934 T3288 T3287 arg2Of identification,for
R2935 T3288 T3289 arg1Of identification,of
R2936 T3288 T3292 arg1Of identification,in
R2937 T3291 T3289 arg2Of polysaccharides,of
R2938 T3291 T3290 arg2Of polysaccharides,sulphated
R2939 T3295 T3292 arg2Of mucosa,in
R2940 T3295 T3293 arg1Of mucosa,the
R2941 T3295 T3294 arg1Of mucosa,intestinal
R2942 T3298 T3296 arg2Of day,On
R2943 T3298 T3297 arg1Of day,the
R2944 T3298 T3299 arg1Of day,of
R2945 T3300 T3299 arg2Of sacrifice,of
R2946 T3304 T3302 arg1Of sample,a
R2947 T3304 T3303 arg1Of sample,fresh
R2948 T3304 T3305 arg1Of sample,of
R2949 T3304 T3312 arg1Of sample,was
R2950 T3304 T3313 arg2Of sample,collected
R2951 T3307 T3305 arg2Of colon,of
R2952 T3307 T3306 arg1Of colon,each
R2953 T3307 T3308 arg1Of colon,(
R2954 T3310 T3308 arg2Of mg,(
R2955 T3310 T3309 arg1Of mg,50
R2956 T3311 T3308 arg3Of ),(
R2957 T3313 T3296 arg1Of collected,On
R2958 T3313 T3301 arg1Of collected,","
R2959 T3313 T3312 arg2Of collected,was
R2960 T3313 T3314 arg1Of collected,for
R2961 T3313 T3320 arg1Of collected,according
R2962 T3313 T3325 arg1Of collected,","
R2963 T3313 T3326 arg1Of collected,[
R2964 T3317 T3316 arg2Of MPO,(
R2965 T3318 T3316 arg3Of ),(
R2966 T3319 T3314 arg2Of assay,for
R2967 T3319 T3315 arg1Of assay,myeloperoxidase
R2968 T3319 T3316 arg1Of assay,(
R2969 T3321 T3320 arg2Of to,according
R2970 T3322 T3321 arg2Of Krawisz,to
R2971 T3322 T3323 arg1Of Krawisz,et
R2972 T3322 T3324 arg1Of Krawisz,al.
R2973 T3327 T3326 arg2Of 27,[
R2974 T3328 T3326 arg3Of ],[
R2975 T3330 T3329 arg1Of level,The
R2976 T3330 T3331 arg1Of level,of
R2977 T3330 T3333 arg1Of level,","
R2978 T3330 T3335 arg2Of level,expressed
R2979 T3330 T3339 arg1Of level,indicates
R2980 T3332 T3331 arg2Of MPO,of
R2981 T3335 T3334 arg1Of expressed,mainly
R2982 T3337 T3335 arg1Of neutrophils,expressed
R2983 T3337 T3336 arg2Of neutrophils,by
R2984 T3339 T3338 arg1Of indicates,","
R2985 T3341 T3339 arg2Of rate,indicates
R2986 T3341 T3340 arg1Of rate,the
R2987 T3341 T3342 arg1Of rate,of
R2988 T3343 T3342 arg2Of recruitment,of
R2989 T3343 T3344 arg1Of recruitment,of
R2990 T3343 T3346 arg1Of recruitment,to
R2991 T3345 T3344 arg2Of neutrophils,of
R2992 T3349 T3346 arg2Of mucosa,to
R2993 T3349 T3347 arg1Of mucosa,the
R2994 T3349 T3348 arg1Of mucosa,intestinal
R2995 T3351 T3350 arg1Of unit,One
R2996 T3351 T3352 arg1Of unit,of
R2997 T3351 T3355 arg1Of unit,corresponds
R2998 T3354 T3352 arg2Of activity,of
R2999 T3354 T3353 arg1Of activity,MPO
R3000 T3355 T3356 arg1Of corresponds,to
R3001 T3358 T3356 arg2Of degradation,to
R3002 T3358 T3357 arg1Of degradation,the
R3003 T3358 T3359 arg1Of degradation,of
R3004 T3361 T3359 arg2Of µmol,of
R3005 T3361 T3360 arg1Of µmol,1
R3006 T3361 T3362 arg1Of µmol,of
R3007 T3361 T3364 arg1Of µmol,per
R3008 T3361 T3366 arg1Of µmol,at
R3009 T3363 T3362 arg2Of peroxide,of
R3010 T3365 T3364 arg2Of minute,per
R3011 T3367 T3366 arg2Of 25°C,at
R3275 T3677 T3676 arg1Of Culture,Cell
R3276 T3681 T3678 arg1Of reagents,All
R3277 T3681 T3679 arg1Of reagents,tissue
R3278 T3681 T3680 arg1Of reagents,culture
R3279 T3681 T3682 arg1Of reagents,were
R3280 T3681 T3683 arg1Of reagents,from
R3281 T3683 T3682 arg2Of from,were
R3282 T3684 T3683 arg2Of Invitrogen,from
R3283 T3684 T3685 arg1Of Invitrogen,(
R3284 T3687 T3685 arg2Of Pontoise,(
R3285 T3687 T3686 arg1Of Pontoise,Cergy
R3286 T3687 T3688 arg1Of Pontoise,","
R3287 T3689 T3688 arg2Of France,","
R3288 T3690 T3685 arg3Of ),(
R3289 T3694 T3691 arg1Of cells,THP-1
R3290 T3694 T3692 arg1Of cells,human
R3291 T3694 T3693 arg1Of cells,monocytic
R3292 T3694 T3695 arg1Of cells,were
R3293 T3694 T3696 arg2Of cells,maintained
R3294 T3696 T3695 arg2Of maintained,were
R3295 T3696 T3697 arg1Of maintained,in
R3296 T3698 T3697 arg2Of RPMI-1640,in
R3297 T3698 T3699 arg2Of RPMI-1640,supplemented
R3298 T3699 T3700 arg1Of supplemented,with
R3299 T3701 T3702 arg1Of 10,%
R3300 T3703 T3701 arg1Of FCS,10
R3301 T3703 T3704 arg1Of FCS,","
R3302 T3704 T3709 arg1Of ",",","
R3303 T3705 T3706 arg1Of 2,mM
R3304 T3708 T3704 arg2Of -glutamine,","
R3305 T3708 T3705 arg1Of -glutamine,2
R3306 T3708 T3707 arg1Of -glutamine,L
R3307 T3709 T3713 arg1Of ",",and
R3308 T3710 T3711 arg1Of 50,U/ml
R3309 T3712 T3709 arg2Of penicillin,","
R3310 T3712 T3710 arg1Of penicillin,50
R3311 T3713 T3700 arg2Of and,with
R3312 T3714 T3715 arg1Of 50,mg/ml
R3313 T3716 T3713 arg2Of streptomycin,and
R3314 T3716 T3714 arg1Of streptomycin,50
R3315 T3716 T3717 arg1Of streptomycin,at
R3316 T3716 T3719 arg1Of streptomycin,in
R3317 T3718 T3717 arg2Of 37°C,at
R3318 T3721 T3722 arg1Of 5,%
R3319 T3724 T3719 arg2Of incubator,in
R3320 T3724 T3720 arg1Of incubator,a
R3321 T3724 T3721 arg1Of incubator,5
R3322 T3724 T3723 arg1Of incubator,CO2
R3323 T3729 T3725 arg1Of cells,Human
R3324 T3729 T3726 arg1Of cells,peripheral
R3325 T3729 T3727 arg1Of cells,blood
R3326 T3729 T3728 arg1Of cells,mononuclear
R3327 T3729 T3730 arg1Of cells,were
R3328 T3729 T3731 arg2Of cells,obtained
R3329 T3731 T3730 arg2Of obtained,were
R3330 T3731 T3732 arg1Of obtained,from
R3331 T3734 T3732 arg2Of blood,from
R3332 T3734 T3733 arg2Of blood,heparinized
R3333 T3738 T3731 arg1Of gradient,obtained
R3334 T3738 T3735 arg2Of gradient,by
R3335 T3738 T3736 arg1Of gradient,Ficoll-Hypaque
R3336 T3738 T3737 arg1Of gradient,density
R3337 T3739 T3740 arg1Of Monocytes,were
R3338 T3739 T3742 arg2Of Monocytes,isolated
R3339 T3742 T3740 arg2Of isolated,were
R3340 T3742 T3741 arg1Of isolated,then
R3341 T3742 T3743 arg1Of isolated,by
R3342 T3742 T3745 arg1Of isolated,to
R3343 T3742 T3748 arg1Of isolated,as
R3344 T3744 T3743 arg2Of adherence,by
R3345 T3747 T3745 arg2Of flasks,to
R3346 T3747 T3746 arg1Of flasks,culture
R3347 T3749 T3748 arg2Of described,as
R3348 T3749 T3750 arg1Of described,[
R3349 T3751 T3750 arg2Of 28,[
R3350 T3752 T3750 arg3Of ],[
R3351 T3755 T3753 arg2Of aggregation,For
R3352 T3755 T3754 arg1Of aggregation,cell
R3353 T3757 T3758 arg1Of monocytes,were
R3354 T3757 T3759 arg2Of monocytes,cultured
R3355 T3759 T3753 arg1Of cultured,For
R3356 T3759 T3756 arg1Of cultured,","
R3357 T3759 T3758 arg2Of cultured,were
R3358 T3759 T3760 arg1Of cultured,in
R3359 T3762 T3763 arg1Of presence,or
R3360 T3763 T3760 arg2Of or,in
R3361 T3763 T3761 arg1Of or,the
R3362 T3763 T3765 arg1Of or,of
R3363 T3764 T3763 arg2Of absence,or
R3364 T3766 T3767 arg1Of C10,or
R3365 T3767 T3765 arg2Of or,of
R3366 T3767 T3769 arg1Of or,for
R3367 T3768 T3767 arg2Of C40,or
R3368 T3771 T3769 arg2Of h,for
R3369 T3771 T3770 arg1Of h,72
R3370 T3773 T3772 arg1Of colonies,Cell
R3371 T3773 T3774 arg1Of colonies,were
R3372 T3773 T3775 arg2Of colonies,monitored
R3373 T3775 T3774 arg2Of monitored,were
R3374 T3775 T3776 arg1Of monitored,under
R3375 T3781 T3776 arg2Of microscope,under
R3376 T3781 T3777 arg1Of microscope,an
R3377 T3781 T3778 arg2Of microscope,inverted
R3378 T3781 T3779 arg1Of microscope,phase
R3379 T3781 T3780 arg1Of microscope,contrast
R3380 T3781 T3782 arg2Of microscope,coupled
R3381 T3782 T3783 arg1Of coupled,through
R3382 T3786 T3783 arg2Of camera,through
R3383 T3786 T3784 arg1Of camera,a
R3384 T3786 T3785 arg1Of camera,video
R3385 T3786 T3787 arg1Of camera,to
R3386 T3789 T3787 arg2Of computer,to
R3387 T3789 T3788 arg1Of computer,a
R3388 T3792 T3790 arg2Of wells,In
R3389 T3792 T3791 arg1Of wells,some
R3390 T3794 T3813 arg1Of neutralizing,was
R3391 T3794 T3814 arg2Of neutralizing,added
R3392 T3796 T3794 arg2Of antibody,neutralizing
R3393 T3796 T3795 arg1Of antibody,monoclonal
R3394 T3796 T3797 arg1Of antibody,to
R3395 T3798 T3797 arg2Of ICAM-1,to
R3396 T3798 T3799 arg1Of ICAM-1,(
R3397 T3798 T3803 arg1Of ICAM-1,(
R3398 T3801 T3799 arg2Of µg/ml,(
R3399 T3801 T3800 arg1Of µg/ml,2.5
R3400 T3802 T3799 arg3Of ),(
R3401 T3804 T3803 arg2Of Tebu,(
R3402 T3804 T3805 arg1Of Tebu,","
R3403 T3804 T3810 arg1Of Tebu,","
R3404 T3809 T3805 arg2Of Yvelines,","
R3405 T3809 T3806 arg1Of Yvelines,Le
R3406 T3809 T3807 arg1Of Yvelines,Perray
R3407 T3809 T3808 arg1Of Yvelines,en
R3408 T3811 T3810 arg2Of France,","
R3409 T3812 T3803 arg3Of ),(
R3410 T3814 T3790 arg1Of added,In
R3411 T3814 T3793 arg1Of added,","
R3412 T3814 T3813 arg2Of added,was
R3563 T3988 T3986 arg1Of Analysis,Cell
R3564 T3988 T3987 arg1Of Analysis,Cycle
R3565 T3990 T3989 arg1Of cells,THP-1
R3566 T3990 T3991 arg1Of cells,in
R3567 T3990 T3995 arg1Of cells,were
R3568 T3990 T3996 arg2Of cells,exposed
R3569 T3990 T4011 arg1Of cells,being
R3570 T3990 T4012 arg2Of cells,stained
R3571 T3990 T4016 arg1Of cells,using
R3572 T3994 T3991 arg2Of phase,in
R3573 T3994 T3992 arg1Of phase,exponential
R3574 T3994 T3993 arg1Of phase,growth
R3575 T3996 T3995 arg2Of exposed,were
R3576 T3996 T3997 arg1Of exposed,to
R3577 T3996 T4000 arg1Of exposed,in
R3578 T3996 T4010 arg1Of exposed,before
R3579 T3999 T3997 arg2Of medium,to
R3580 T3999 T3998 arg1Of medium,complete
R3581 T4002 T4003 arg1Of presence,or
R3582 T4003 T4000 arg2Of or,in
R3583 T4003 T4001 arg1Of or,the
R3584 T4003 T4005 arg1Of or,of
R3585 T4003 T4007 arg1Of or,for
R3586 T4004 T4003 arg2Of absence,or
R3587 T4006 T4005 arg2Of carrageenans,of
R3588 T4009 T4007 arg2Of h,for
R3589 T4009 T4008 arg1Of h,24
R3590 T4012 T4010 arg2Of stained,before
R3591 T4012 T4011 arg2Of stained,being
R3592 T4012 T4013 arg1Of stained,with
R3593 T4012 T4016 modOf stained,using
R3594 T4015 T4013 arg2Of iodide,with
R3595 T4015 T4014 arg1Of iodide,propidium
R3596 T4016 T4021 arg1Of using,according
R3597 T4016 T4027 arg1Of using,(
R3598 T4020 T4016 arg2Of kit,using
R3599 T4020 T4017 arg1Of kit,the
R3600 T4020 T4018 arg1Of kit,DNA-Prep
R3601 T4020 T4019 arg1Of kit,Coulter
R3602 T4022 T4021 arg2Of to,according
R3603 T4024 T4023 arg1Of manufacturer,the
R3604 T4024 T4025 arg2Of manufacturer,'s
R3605 T4026 T4022 arg2Of instruction,to
R3606 T4026 T4025 arg1Of instruction,'s
R3607 T4028 T4029 arg1Of Beckman-Coulter,","
R3608 T4029 T4031 arg1Of ",",","
R3609 T4030 T4029 arg2Of Villepinte,","
R3610 T4031 T4027 arg2Of ",",(
R3611 T4032 T4031 arg2Of France,","
R3612 T4033 T4027 arg3Of ),(
R3613 T4036 T4034 arg1Of content,Cell
R3614 T4036 T4035 arg1Of content,DNA
R3615 T4036 T4037 arg1Of content,was
R3616 T4036 T4039 arg2Of content,analyzed
R3617 T4039 T4037 arg2Of analyzed,was
R3618 T4039 T4038 arg1Of analyzed,then
R3619 T4042 T4039 arg1Of cytometry,analyzed
R3620 T4042 T4040 arg2Of cytometry,by
R3621 T4042 T4041 arg1Of cytometry,flow
R3622 T4042 T4043 arg1Of cytometry,using
R3623 T4046 T4043 arg2Of XL2,using
R3624 T4046 T4044 arg1Of XL2,an
R3625 T4046 T4045 arg1Of XL2,EPICS
R3626 T4046 T4047 arg1Of XL2,(
R3627 T4048 T4047 arg2Of Beckman-Coulter,(
R3628 T4049 T4047 arg3Of ),(
R3629 T4051 T4050 arg1Of data,Raw
R3630 T4051 T4052 arg1Of data,for
R3631 T4051 T4065 arg1Of data,were
R3632 T4051 T4066 arg2Of data,expressed
R3633 T4054 T4052 arg2Of distribution,for
R3634 T4054 T4053 arg1Of distribution,the
R3635 T4054 T4055 arg1Of distribution,of
R3636 T4057 T4055 arg2Of content,of
R3637 T4057 T4056 arg1Of content,DNA
R3638 T4057 T4058 arg1Of content,of
R3639 T4060 T4058 arg2Of cells,of
R3640 T4060 T4059 arg1Of cells,"30,000"
R3641 T4060 T4061 arg2Of cells,retrieved
R3642 T4061 T4062 arg1Of retrieved,from
R3643 T4064 T4062 arg2Of cytometer,from
R3644 T4064 T4063 arg1Of cytometer,the
R3645 T4066 T4065 arg2Of expressed,were
R3646 T4066 T4067 arg1Of expressed,as
R3647 T4069 T4067 arg2Of percentage,as
R3648 T4069 T4068 arg1Of percentage,the
R3649 T4069 T4070 arg1Of percentage,of
R3650 T4069 T4072 arg1Of percentage,through
R3651 T4071 T4070 arg2Of G0/G1,of
R3652 T4074 T4072 arg2Of populations,through
R3653 T4074 T4073 arg1Of populations,G2/M
R3654 T4077 T4075 arg1Of software,Multicycle
R3655 T4077 T4076 arg1Of software,AV
R3656 T4077 T4078 arg1Of software,(
R3657 T4077 T4088 arg1Of software,was
R3658 T4077 T4089 arg2Of software,used
R3659 T4077 T4097 arg1Of software,facilitate
R3660 T4081 T4078 arg2Of Systems,(
R3661 T4081 T4079 arg1Of Systems,Phoenix
R3662 T4081 T4080 arg1Of Systems,Flow
R3663 T4081 T4082 arg1Of Systems,","
R3664 T4084 T4082 arg2Of Diego,","
R3665 T4084 T4083 arg1Of Diego,San
R3666 T4084 T4085 arg1Of Diego,","
R3667 T4086 T4085 arg2Of CA,","
R3668 T4087 T4078 arg3Of ),(
R3669 T4089 T4088 arg2Of used,was
R3670 T4089 T4090 modOf used,to
R3671 T4089 T4096 arg1Of used,and
R3672 T4091 T4090 arg1Of generate,to
R3673 T4095 T4091 arg2Of histograms,generate
R3674 T4095 T4092 arg1Of histograms,DNA
R3675 T4095 T4093 arg1Of histograms,content
R3676 T4095 T4094 arg1Of histograms,frequency
R3677 T4097 T4096 arg2Of facilitate,and
R3678 T4099 T4097 arg2Of analysis,facilitate
R3679 T4099 T4098 arg1Of analysis,data
R3798 T4244 T4242 arg1Of Antigen,Cell
R3799 T4244 T4243 arg1Of Antigen,Surface
R3800 T4246 T4244 arg1Of Analysis,Antigen
R3801 T4246 T4245 arg1Of Analysis,Expression
R3802 T4249 T4247 arg1Of Monocytes,Peripheral
R3803 T4249 T4248 arg1Of Monocytes,Blood
R3804 T4249 T4250 arg1Of Monocytes,or
R3805 T4250 T4253 arg1Of or,were
R3806 T4250 T4254 arg2Of or,exposed
R3807 T4252 T4250 arg2Of cells,or
R3808 T4252 T4251 arg1Of cells,THP-1
R3809 T4254 T4253 arg2Of exposed,were
R3810 T4254 T4255 arg1Of exposed,to
R3811 T4254 T4258 arg1Of exposed,in
R3812 T4257 T4255 arg2Of medium,to
R3813 T4257 T4256 arg1Of medium,complete
R3814 T4260 T4261 arg1Of presence,or
R3815 T4261 T4258 arg2Of or,in
R3816 T4261 T4259 arg1Of or,the
R3817 T4261 T4263 arg1Of or,of
R3818 T4261 T4265 arg1Of or,for
R3819 T4262 T4261 arg2Of absence,or
R3820 T4264 T4263 arg2Of carrageenan,of
R3821 T4267 T4265 arg2Of h,for
R3822 T4267 T4266 arg1Of h,36
R3823 T4270 T4268 arg2Of washes,After
R3824 T4270 T4269 arg1Of washes,two
R3825 T4270 T4271 arg1Of washes,in
R3826 T4270 T4273 arg1Of washes,without
R3827 T4272 T4271 arg2Of PBS,in
R3828 T4274 T4275 arg1Of Ca2+,and
R3829 T4275 T4273 arg2Of and,without
R3830 T4276 T4275 arg2Of Mg2+,and
R3831 T4278 T4279 arg1Of cells,were
R3832 T4278 T4280 arg2Of cells,incubated
R3833 T4280 T4268 arg1Of incubated,After
R3834 T4280 T4277 arg1Of incubated,","
R3835 T4280 T4279 arg2Of incubated,were
R3836 T4280 T4281 arg1Of incubated,in
R3837 T4280 T4293 modOf incubated,to
R3838 T4282 T4281 arg2Of PBS,in
R3839 T4282 T4283 arg1Of PBS,containing
R3840 T4284 T4285 arg1Of 0.1,%
R3841 T4286 T4284 arg1Of gelatin,0.1
R3842 T4286 T4287 arg1Of gelatin,and
R3843 T4287 T4283 arg2Of and,containing
R3844 T4288 T4289 arg1Of 8,%
R3845 T4292 T4287 arg2Of serum,and
R3846 T4292 T4288 arg1Of serum,8
R3847 T4292 T4290 arg1Of serum,AB
R3848 T4292 T4291 arg1Of serum,human
R3849 T4294 T4293 arg1Of prevent,to
R3850 T4295 T4294 arg2Of binding,prevent
R3851 T4295 T4296 arg1Of binding,to
R3852 T4298 T4296 arg2Of receptors,to
R3853 T4298 T4297 arg1Of receptors,Fc
R3854 T4302 T4301 arg1Of cells,5×105
R3855 T4302 T4303 arg1Of cells,were
R3856 T4302 T4304 arg2Of cells,incubated
R3857 T4304 T4299 arg1Of incubated,Then
R3858 T4304 T4300 arg1Of incubated,","
R3859 T4304 T4303 arg2Of incubated,were
R3860 T4304 T4305 arg1Of incubated,with
R3861 T4304 T4308 arg1Of incubated,at
R3862 T4304 T4310 arg1Of incubated,for
R3863 T4307 T4305 arg2Of antibodies,with
R3864 T4307 T4306 arg1Of antibodies,primary
R3865 T4309 T4308 arg2Of 4°C,at
R3866 T4312 T4310 arg2Of min,for
R3867 T4312 T4311 arg1Of min,30
R3868 T4315 T4313 arg1Of washes,Two
R3869 T4315 T4314 arg1Of washes,other
R3870 T4315 T4316 arg1Of washes,in
R3871 T4315 T4318 arg1Of washes,preceded
R3872 T4317 T4316 arg2Of PBS,in
R3873 T4319 T4318 arg2Of incubation,preceded
R3874 T4319 T4320 arg1Of incubation,with
R3875 T4325 T4320 arg2Of IgG,with
R3876 T4325 T4321 arg1Of IgG,FITC-conjugated
R3877 T4325 T4322 arg1Of IgG,goat
R3878 T4325 T4323 arg1Of IgG,antibody
R3879 T4325 T4324 arg1Of IgG,anti-mouse
R3880 T4325 T4326 arg2Of IgG,diluted
R3881 T4326 T4328 arg1Of diluted,at
R3882 T4326 T4330 arg1Of diluted,for
R3883 T4327 T4326 arg3Of 1/1000,diluted
R3884 T4329 T4328 arg2Of 4°C,at
R3885 T4332 T4330 arg2Of min,for
R3886 T4332 T4331 arg1Of min,30
R3887 T4332 T4333 arg1Of min,(
R3888 T4334 T4333 arg2Of Tebu,(
R3889 T4335 T4333 arg3Of ),(
R3890 T4339 T4336 arg2Of washes,After
R3891 T4339 T4337 arg1Of washes,two
R3892 T4339 T4338 arg1Of washes,additional
R3893 T4341 T4342 arg1Of analysis,of
R3894 T4341 T4345 arg1Of analysis,was
R3895 T4341 T4346 arg2Of analysis,performed
R3896 T4344 T4342 arg2Of cells,of
R3897 T4344 T4343 arg2Of cells,stained
R3898 T4346 T4336 arg1Of performed,After
R3899 T4346 T4340 arg1Of performed,","
R3900 T4346 T4345 arg2Of performed,was
R3901 T4346 T4347 arg1Of performed,on
R3902 T4350 T4347 arg2Of XL2,on
R3903 T4350 T4348 arg1Of XL2,an
R3904 T4350 T4349 arg1Of XL2,EPICS
R3905 T4350 T4351 arg1Of XL2,(
R3906 T4352 T4351 arg2Of Beckman-Coulter,(
R3907 T4353 T4351 arg3Of ),(
R3908 T4356 T4354 arg1Of population,The
R3909 T4356 T4355 arg1Of population,cell
R3910 T4356 T4357 arg1Of population,was
R3911 T4356 T4358 arg2Of population,gated
R3912 T4358 T4357 arg2Of gated,was
R3913 T4358 T4359 arg1Of gated,according
R3914 T4360 T4359 arg2Of to,according
R3915 T4362 T4363 arg1Of forward,and
R3916 T4364 T4363 arg2Of wide-angle,and
R3917 T4366 T4360 arg2Of scattering,to
R3918 T4366 T4361 arg1Of scattering,its
R3919 T4366 T4362 arg1Of scattering,forward
R3920 T4366 T4364 arg1Of scattering,wide-angle
R3921 T4366 T4365 arg1Of scattering,light
R3922 T4367 T4368 arg1Of Data,were
R3923 T4367 T4369 arg2Of Data,expressed
R3924 T4369 T4368 arg2Of expressed,were
R3925 T4369 T4370 arg1Of expressed,as
R3926 T4374 T4370 arg2Of intensity,as
R3927 T4374 T4371 arg1Of intensity,mean
R3928 T4374 T4372 arg1Of intensity,relative
R3929 T4374 T4373 arg1Of intensity,fluorescence
R3930 T4374 T4375 arg1Of intensity,(
R3931 T4374 T4378 arg1Of intensity,of
R3932 T4376 T4375 arg2Of MFI,(
R3933 T4377 T4375 arg3Of ),(
R3934 T4380 T4378 arg2Of cells,of
R3935 T4380 T4379 arg1Of cells,3000
R4092 T4589 T4587 arg1Of Bioassay,TNF
R4093 T4589 T4588 arg1Of Bioassay,Activity
R4094 T4590 T4591 arg1Of Monocytes,or
R4095 T4591 T4594 arg1Of or,were
R4096 T4591 T4595 arg2Of or,cultured
R4097 T4593 T4591 arg2Of cells,or
R4098 T4593 T4592 arg1Of cells,THP-1
R4099 T4595 T4594 arg2Of cultured,were
R4100 T4595 T4596 arg1Of cultured,with
R4101 T4595 T4598 arg1Of cultured,without
R4102 T4596 T4597 arg1Of with,or
R4103 T4598 T4597 arg2Of without,or
R4104 T4600 T4596 arg2Of concentrations,with
R4105 T4600 T4598 arg2Of concentrations,without
R4106 T4600 T4599 arg1Of concentrations,different
R4107 T4600 T4601 arg1Of concentrations,of
R4108 T4600 T4611 arg1Of concentrations,for
R4109 T4602 T4603 arg1Of CGNs,or
R4110 T4603 T4601 arg2Of or,of
R4111 T4603 T4605 arg1Of or,(
R4112 T4604 T4603 arg2Of LPS,or
R4113 T4607 T4605 arg2Of typhosa,(
R4114 T4607 T4606 arg1Of typhosa,Salmonella
R4115 T4607 T4608 arg1Of typhosa,","
R4116 T4609 T4608 arg2Of Sigma,","
R4117 T4610 T4605 arg3Of ),(
R4118 T4613 T4612 arg1Of h,24
R4119 T4613 T4614 arg1Of h,or
R4120 T4614 T4611 arg2Of or,for
R4121 T4617 T4614 arg2Of time,or
R4122 T4617 T4615 arg1Of time,the
R4123 T4617 T4616 arg2Of time,indicated
R4124 T4619 T4618 arg1Of active,Biologically
R4125 T4620 T4619 arg1Of TNF-α/β,active
R4126 T4620 T4621 arg1Of TNF-α/β,in
R4127 T4620 T4625 arg1Of TNF-α/β,was
R4128 T4620 T4626 arg2Of TNF-α/β,measured
R4129 T4624 T4621 arg2Of supernatant,in
R4130 T4624 T4622 arg1Of supernatant,tissue
R4131 T4624 T4623 arg1Of supernatant,culture
R4132 T4626 T4625 arg2Of measured,was
R4133 T4627 T4626 arg3Of using,measured
R4134 T4627 T4635 arg1Of using,[
R4135 T4634 T4627 arg2Of assay,using
R4136 T4634 T4628 arg1Of assay,the
R4137 T4634 T4629 arg1Of assay,WEHI
R4138 T4634 T4630 arg1Of assay,164
R4139 T4634 T4631 arg1Of assay,clone
R4140 T4634 T4632 arg1Of assay,13-cell
R4141 T4634 T4633 arg1Of assay,killing
R4142 T4636 T4635 arg2Of 29,[
R4143 T4637 T4635 arg3Of ],[
R4144 T4639 T4638 arg1Of concentrations,TNF
R4145 T4639 T4640 arg1Of concentrations,are
R4146 T4639 T4641 arg2Of concentrations,expressed
R4147 T4641 T4640 arg2Of expressed,are
R4148 T4641 T4642 arg1Of expressed,as
R4149 T4643 T4642 arg2Of pg/ml,as
R4213 T4751 T4750 arg1Of Analysis,RT-PCR
R4214 T4753 T4752 arg1Of RNA,Total
R4215 T4753 T4754 arg1Of RNA,from
R4216 T4753 T4756 arg1Of RNA,was
R4217 T4753 T4757 arg2Of RNA,isolated
R4218 T4755 T4754 arg2Of monocytes,from
R4219 T4757 T4756 arg2Of isolated,was
R4220 T4758 T4757 arg3Of using,isolated
R4221 T4760 T4758 arg2Of Reagent™,using
R4222 T4760 T4759 arg1Of Reagent™,TRIzol
R4223 T4760 T4761 arg1Of Reagent™,(
R4224 T4762 T4761 arg2Of Invitrogen,(
R4225 T4763 T4761 arg3Of ),(
R4226 T4764 T4765 arg1Of . cD,NA
R4227 T4764 T4766 arg2Of . cD,was gener
R4228 T4766 T4756 modOf was gener,was
R4229 T4766 T4765 arg2Of was gener,NA
R4230 T4766 T4767 arg1Of was gener,at
R4231 T4766 T4780 arg1Of was gener,v
R4232 T4769 T4767 arg2Of d ,at
R4233 T4769 T4768 arg1Of d ,e
R4234 T4769 T4770 arg1Of d ,on
R4235 T4769 T4773 arg1Of d ,to
R4236 T4772 T4770 arg2Of of ,on
R4237 T4772 T4771 arg1Of of ,1 µg
R4238 T4776 T4773 arg2Of n a re,to
R4239 T4776 T4774 arg1Of n a re,t
R4240 T4776 T4775 arg1Of n a re,al RNA i
R4241 T4776 T4777 arg1Of n a re,ac
R4242 T4779 T4777 arg2Of on,ac
R4243 T4779 T4778 arg1Of on,ti
R4244 T4781 T4766 arg3Of olume,was gener
R4245 T4784 T4781 arg2Of ng M-MLV reve,olume
R4246 T4784 T4782 arg1Of ng M-MLV reve,of 20
R4247 T4784 T4783 arg1Of ng M-MLV reve,"µl, usi"
R4248 T4784 T4785 arg1Of ng M-MLV reve,r
R4249 T4786 T4785 arg2Of se transcr,r
R4250 T4787 T4785 arg3Of i,r
R4251 T4788 T4789 arg1Of PCR,was
R4252 T4788 T4790 arg2Of PCR,done
R4253 T4790 T4789 arg2Of done,was
R4254 T4790 T4791 arg1Of done,in
R4255 T4794 T4791 arg2Of range,in
R4256 T4794 T4792 arg1Of range,the
R4257 T4794 T4793 arg1Of range,linear
R4258 T4794 T4795 arg1Of range,of
R4259 T4796 T4795 arg2Of amplification,of
R4260 T4796 T4797 arg1Of amplification,(
R4261 T4798 T4797 arg2Of determined,(
R4262 T4798 T4799 arg1Of determined,for
R4263 T4803 T4799 arg2Of combination,for
R4264 T4803 T4800 arg1Of combination,each
R4265 T4803 T4801 arg1Of combination,primer
R4266 T4803 T4802 arg1Of combination,pair-cDNA
R4267 T4804 T4797 arg3Of ),(
R4268 T4807 T4805 arg1Of reactions,Standard
R4269 T4807 T4806 arg1Of reactions,PCR
R4270 T4807 T4808 arg1Of reactions,were
R4271 T4807 T4809 arg2Of reactions,performed
R4272 T4809 T4808 arg2Of performed,were
R4273 T4809 T4810 arg1Of performed,with
R4274 T4809 T4849 arg1Of performed,(
R4275 T4812 T4811 arg1Of µl,1
R4276 T4812 T4813 arg1Of µl,of
R4277 T4812 T4817 arg1Of µl,","
R4278 T4816 T4813 arg2Of solution,of
R4279 T4816 T4814 arg1Of solution,the
R4280 T4816 T4815 arg1Of solution,cDNA
R4281 T4817 T4824 arg1Of ",",","
R4282 T4819 T4817 arg2Of µM,","
R4283 T4819 T4818 arg1Of µM,50
R4284 T4819 T4820 arg1Of µM,of
R4285 T4823 T4820 arg2Of solution,of
R4286 T4823 T4821 arg1Of solution,each
R4287 T4823 T4822 arg1Of solution,primer
R4288 T4824 T4830 arg1Of ",",","
R4289 T4826 T4824 arg2Of mM,","
R4290 T4826 T4825 arg1Of mM,10
R4291 T4826 T4827 arg1Of mM,of
R4292 T4829 T4827 arg2Of dNTP,of
R4293 T4829 T4828 arg1Of dNTP,each
R4294 T4830 T4834 arg1Of ",",","
R4295 T4830 T4842 arg1Of ",",and
R4296 T4831 T4832 arg1Of 25,mM
R4297 T4833 T4830 arg2Of MgCl2,","
R4298 T4833 T4831 arg1Of MgCl2,25
R4299 T4836 T4835 arg1Of Goldstar,10X
R4300 T4840 T4834 arg2Of buffer,","
R4301 T4840 T4836 arg1Of buffer,Goldstar
R4302 T4840 T4837 arg1Of buffer,DNA
R4303 T4840 T4838 arg1Of buffer,polymerase
R4304 T4840 T4839 arg1Of buffer,reaction
R4305 T4842 T4810 arg2Of and,with
R4306 T4842 T4841 arg1Of and,","
R4307 T4844 T4842 arg2Of units,and
R4308 T4844 T4843 arg1Of units,0.5
R4309 T4844 T4845 arg1Of units,of
R4310 T4848 T4845 arg2Of polymerase,of
R4311 T4848 T4846 arg1Of polymerase,Goldstar
R4312 T4848 T4847 arg1Of polymerase,DNA
R4313 T4850 T4851 arg1Of Eurogentec,","
R4314 T4851 T4849 arg2Of ",",(
R4315 T4851 T4853 arg1Of ",",","
R4316 T4852 T4851 arg2Of Seraing,","
R4317 T4854 T4853 arg2Of Belgium,","
R4318 T4855 T4849 arg3Of ),(
R4319 T4858 T4856 arg1Of cycle,First
R4320 T4858 T4857 arg1Of cycle,PCR
R4321 T4858 T4859 arg1Of cycle,consisted
R4322 T4859 T4860 arg1Of consisted,of
R4323 T4859 T4863 arg1Of consisted,at
R4324 T4859 T4875 arg1Of consisted,;
R4325 T4862 T4860 arg2Of min,of
R4326 T4862 T4861 arg1Of min,1
R4327 T4864 T4865 arg1Of 92°C,","
R4328 T4865 T4870 arg1Of ",",and
R4329 T4867 T4865 arg2Of min,","
R4330 T4867 T4866 arg1Of min,1
R4331 T4867 T4868 arg1Of min,at
R4332 T4869 T4868 arg2Of 58°C,at
R4333 T4870 T4863 arg2Of and,at
R4334 T4870 T4873 arg1Of and,at
R4335 T4872 T4870 arg2Of min,and
R4336 T4872 T4871 arg1Of min,1
R4337 T4874 T4873 arg2Of 72°C,at
R4338 T4879 T4877 arg1Of cycle,each
R4339 T4879 T4878 arg1Of cycle,PCR
R4340 T4879 T4880 arg1Of cycle,consisted
R4341 T4880 T4875 arg2Of consisted,;
R4342 T4880 T4876 arg1Of consisted,then
R4343 T4880 T4881 arg1Of consisted,of
R4344 T4880 T4884 arg1Of consisted,at
R4345 T4883 T4881 arg2Of sec,of
R4346 T4883 T4882 arg1Of sec,40
R4347 T4885 T4884 arg2Of 92°C,at
R4348 T4888 T4887 arg1Of sec,40
R4349 T4888 T4889 arg1Of sec,at
R4350 T4888 T4894 arg1Of sec,at
R4351 T4888 T4899 arg1Of sec,was
R4352 T4888 T4900 arg2Of sec,amplified
R4353 T4888 T4941 arg1Of sec,sense
R4354 T4890 T4891 arg1Of 58°C,and
R4355 T4891 T4889 arg2Of and,at
R4356 T4893 T4891 arg2Of sec,and
R4357 T4893 T4892 arg1Of sec,50
R4358 T4896 T4894 arg2Of cDNA,at
R4359 T4896 T4895 arg1Of cDNA,72°C.
R4360 T4896 T4897 arg1Of cDNA,for
R4361 T4898 T4897 arg2Of β-actin,for
R4362 T4900 T4899 arg2Of amplified,was
R4363 T4900 T4901 arg1Of amplified,for
R4364 T4900 T4907 arg1Of amplified,:
R4365 T4903 T4901 arg2Of cycles,for
R4366 T4903 T4902 arg1Of cycles,28
R4367 T4903 T4904 arg1Of cycles,using
R4368 T4906 T4904 arg2Of oligos,using
R4369 T4906 T4905 arg1Of oligos,the
R4370 T4907 T4875 arg3Of :,;
R4371 T4907 T4886 arg1Of :,","
R4372 T4909 T4908 arg1Of 5′-GGCATCGTGATGGACTCCG-3′,sense
R4373 T4909 T4910 arg1Of 5′-GGCATCGTGATGGACTCCG-3′,and
R4374 T4910 T4914 arg1Of and,for
R4375 T4910 T4916 arg1Of and,was
R4376 T4910 T4917 arg2Of and,amplified
R4377 T4913 T4910 arg2Of cDNA,and
R4378 T4913 T4911 arg1Of cDNA,antisense
R4379 T4913 T4912 arg1Of cDNA,5′GCTGGAAGGTGGACAGCGA-3′.
R4380 T4915 T4914 arg2Of TNF-α,for
R4381 T4917 T4916 arg2Of amplified,was
R4382 T4917 T4918 arg1Of amplified,for
R4383 T4917 T4924 arg1Of amplified,:
R4384 T4920 T4918 arg2Of cycles,for
R4385 T4920 T4919 arg1Of cycles,35
R4386 T4920 T4921 arg1Of cycles,using
R4387 T4923 T4921 arg2Of oligos,using
R4388 T4923 T4922 arg1Of oligos,the
R4389 T4926 T4925 arg1Of 5′-AAGCCTGTAGCCCATGTTGT-3′,sense
R4390 T4926 T4927 arg1Of 5′-AAGCCTGTAGCCCATGTTGT-3′,and
R4391 T4927 T4931 arg1Of and,for
R4392 T4927 T4933 arg1Of and,was
R4393 T4927 T4934 arg2Of and,amplified
R4394 T4930 T4927 arg2Of cDNA,and
R4395 T4930 T4928 arg1Of cDNA,antisense
R4396 T4930 T4929 arg1Of cDNA,5′-CAGATAGATGGGCTCATACC-3′.
R4397 T4932 T4931 arg2Of ICAM-1,for
R4398 T4934 T4924 arg2Of amplified,:
R4399 T4934 T4933 arg2Of amplified,was
R4400 T4934 T4935 arg1Of amplified,for
R4401 T4937 T4935 arg2Of cycles,for
R4402 T4937 T4936 arg1Of cycles,35
R4403 T4937 T4938 arg1Of cycles,using
R4404 T4940 T4938 arg2Of oligos,using
R4405 T4940 T4939 arg1Of oligos,the
R4406 T4941 T4907 arg2Of sense,:
R4407 T4941 T4916 modOf sense,was
R4408 T4942 T4943 arg1Of 5′-GTAGCAGCCGCAGTCATAATGG-3′,and
R4409 T4944 T4943 arg2Of antisense,and
R4410 T4946 T4941 arg2Of TGCTGTTGTATCTGACTGAGG-3′,sense
R4411 T4946 T4942 arg1Of TGCTGTTGTATCTGACTGAGG-3′,5′-GTAGCAGCCGCAGTCATAATGG-3′
R4412 T4946 T4944 arg1Of TGCTGTTGTATCTGACTGAGG-3′,antisense
R4413 T4946 T4945 arg1Of TGCTGTTGTATCTGACTGAGG-3′,5′-A
R4654 T5243 T5239 arg1Of Assay,NF-kB
R4655 T5243 T5240 arg1Of Assay,Transcription
R4656 T5243 T5241 arg1Of Assay,Reporter
R4657 T5243 T5242 arg1Of Assay,Gene
R4658 T5246 T5244 arg1Of 3XMHC-luc,The
R4659 T5246 T5245 arg1Of 3XMHC-luc,plasmid
R4660 T5246 T5247 arg1Of 3XMHC-luc,(
R4661 T5246 T5267 arg1Of 3XMHC-luc,contains
R4662 T5250 T5248 arg1Of gift,a
R4663 T5250 T5249 arg1Of gift,generous
R4664 T5250 T5251 arg1Of gift,from
R4665 T5250 T5255 arg1Of gift,and
R4666 T5254 T5251 arg2Of Westwick,from
R4667 T5254 T5252 arg1Of Westwick,Drs.
R4668 T5254 T5253 arg1Of Westwick,J.
R4669 T5255 T5247 arg2Of and,(
R4670 T5257 T5255 arg2Of Brenner,and
R4671 T5257 T5256 arg1Of Brenner,D.A.
R4672 T5257 T5258 arg1Of Brenner,","
R4673 T5257 T5263 arg1Of Brenner,","
R4674 T5259 T5258 arg2Of University,","
R4675 T5259 T5260 arg1Of University,of
R4676 T5262 T5260 arg2Of Carolina,of
R4677 T5262 T5261 arg1Of Carolina,North
R4678 T5265 T5263 arg2Of Hill,","
R4679 T5265 T5264 arg1Of Hill,Chapel
R4680 T5266 T5247 arg3Of ),(
R4681 T5269 T5267 arg2Of copies,contains
R4682 T5269 T5268 arg1Of copies,three
R4683 T5269 T5270 arg1Of copies,of
R4684 T5269 T5273 arg1Of copies,from
R4685 T5269 T5279 arg1Of copies,","
R4686 T5269 T5280 arg2Of copies,placed
R4687 T5272 T5270 arg2Of element,of
R4688 T5272 T5271 arg1Of element,NF-κB-responsive
R4689 T5278 T5273 arg2Of locus,from
R4690 T5278 T5274 arg1Of locus,the
R4691 T5278 T5275 arg1Of locus,MHC
R4692 T5278 T5276 arg1Of locus,class
R4693 T5278 T5277 arg1Of locus,I
R4694 T5280 T5281 arg1Of placed,upstream
R4695 T5281 T5282 arg1Of upstream,of
R4696 T5285 T5282 arg2Of gene,of
R4697 T5285 T5283 arg1Of gene,the
R4698 T5285 T5284 arg1Of gene,luciferase
R4699 T5289 T5286 arg1Of cells,Human
R4700 T5289 T5287 arg1Of cells,monocytic
R4701 T5289 T5288 arg1Of cells,THP-1
R4702 T5289 T5290 arg1Of cells,were
R4703 T5289 T5292 arg2Of cells,transfected
R4704 T5289 T5302 arg2Of cells,cultured
R4705 T5292 T5293 arg1Of transfected,as
R4706 T5292 T5300 arg1Of transfected,and
R4707 T5295 T5294 arg1Of described,previously
R4708 T5297 T5293 arg2Of 30,as
R4709 T5297 T5295 arg1Of 30,described
R4710 T5297 T5296 arg2Of 30,[
R4711 T5298 T5296 arg3Of ],[
R4712 T5300 T5290 arg2Of and,were
R4713 T5300 T5291 arg1Of and,transiently
R4714 T5300 T5299 arg1Of and,","
R4715 T5302 T5300 arg2Of cultured,and
R4716 T5302 T5301 arg1Of cultured,then
R4717 T5302 T5303 arg1Of cultured,for
R4718 T5305 T5303 arg2Of h,for
R4719 T5305 T5304 arg1Of h,4
R4720 T5305 T5306 arg1Of h,alone
R4721 T5305 T5308 arg1Of h,with
R4722 T5306 T5307 arg1Of alone,or
R4723 T5308 T5307 arg2Of with,or
R4724 T5310 T5308 arg2Of concentration,with
R4725 T5310 T5309 arg1Of concentration,increasing
R4726 T5310 T5311 arg1Of concentration,of
R4727 T5313 T5314 arg1Of C10,or
R4728 T5314 T5311 arg2Of or,of
R4729 T5314 T5312 arg1Of or,either
R4730 T5315 T5314 arg2Of C40,or
R4731 T5317 T5316 arg1Of activity,Luciferase
R4732 T5317 T5318 arg1Of activity,was
R4733 T5317 T5319 arg2Of activity,determined
R4734 T5319 T5318 arg2Of determined,was
R4735 T5320 T5319 arg3Of using,determined
R4736 T5322 T5320 arg2Of luminometer,using
R4737 T5322 T5321 arg1Of luminometer,a
R4738 T5322 T5323 arg1Of luminometer,(
R4739 T5326 T5323 arg2Of Luminometer,(
R4740 T5326 T5324 arg1Of Luminometer,Monolight
R4741 T5326 T5325 arg1Of Luminometer,2010
R4742 T5326 T5327 arg1Of Luminometer,","
R4743 T5329 T5327 arg2Of Arbor,","
R4744 T5329 T5328 arg1Of Arbor,Ann
R4745 T5329 T5330 arg1Of Arbor,","
R4746 T5331 T5330 arg2Of MI,","
R4747 T5332 T5323 arg3Of ),(
R4845 T5493 T5492 arg1Of Blot,Western
R4846 T5494 T5493 arg1Of Analysis,Blot
R4847 T5496 T5495 arg1Of cells,THP-1
R4848 T5496 T5497 arg1Of cells,were
R4849 T5496 T5498 arg2Of cells,stimulated
R4850 T5498 T5497 arg2Of stimulated,were
R4851 T5498 T5499 arg1Of stimulated,for
R4852 T5501 T5499 arg2Of lengths,for
R4853 T5501 T5500 arg1Of lengths,various
R4854 T5501 T5502 arg1Of lengths,of
R4855 T5503 T5502 arg2Of time,of
R4856 T5503 T5504 arg1Of time,with
R4857 T5505 T5506 arg1Of 0.1,mg/ml
R4858 T5507 T5505 arg1Of C10,0.1
R4859 T5507 T5508 arg1Of C10,or
R4860 T5508 T5511 arg1Of or,or
R4861 T5509 T5508 arg2Of C40,or
R4862 T5511 T5504 arg2Of or,with
R4863 T5511 T5510 arg1Of or,","
R4864 T5514 T5511 arg2Of LPS,or
R4865 T5514 T5512 arg1Of LPS,10
R4866 T5514 T5513 arg1Of LPS,µg/ml
R4867 T5515 T5516 arg1Of Cells,were
R4868 T5515 T5518 arg2Of Cells,pelleted
R4869 T5515 T5520 arg2Of Cells,washed
R4870 T5515 T5522 arg2Of Cells,homogenised
R4871 T5518 T5519 arg1Of pelleted,","
R4872 T5519 T5521 arg1Of ",",and
R4873 T5520 T5519 arg2Of washed,","
R4874 T5521 T5516 arg2Of and,were
R4875 T5521 T5517 arg1Of and,then
R4876 T5522 T5521 arg2Of homogenised,and
R4877 T5522 T5523 arg1Of homogenised,in
R4878 T5525 T5523 arg2Of buffer,in
R4879 T5525 T5524 arg1Of buffer,lysis
R4880 T5525 T5526 arg1Of buffer,(
R4881 T5525 T5551 arg1Of buffer,on
R4882 T5529 T5526 arg2Of Hepes,(
R4883 T5529 T5527 arg1Of Hepes,10
R4884 T5529 T5528 arg1Of Hepes,mM
R4885 T5529 T5530 arg1Of Hepes,","
R4886 T5531 T5532 arg1Of pH,7.9
R4887 T5531 T5533 arg1Of pH,","
R4888 T5533 T5537 arg1Of ",",","
R4889 T5534 T5535 arg1Of 150,mM
R4890 T5536 T5533 arg2Of NaCl,","
R4891 T5536 T5534 arg1Of NaCl,150
R4892 T5537 T5541 arg1Of ",",","
R4893 T5538 T5539 arg1Of 1,mM
R4894 T5540 T5537 arg2Of EDTA,","
R4895 T5540 T5538 arg1Of EDTA,1
R4896 T5541 T5546 arg1Of ",",and
R4897 T5542 T5543 arg1Of 0.6,%
R4898 T5544 T5541 arg2Of NP-40,","
R4899 T5544 T5542 arg1Of NP-40,0.6
R4900 T5546 T5530 arg2Of and,","
R4901 T5546 T5545 arg1Of and,","
R4902 T5547 T5548 arg1Of 0.5,mM
R4903 T5549 T5546 arg2Of PMSF,and
R4904 T5549 T5547 arg1Of PMSF,0.5
R4905 T5550 T5526 arg3Of ),(
R4906 T5552 T5551 arg2Of ice,on
R4907 T5553 T5554 arg1Of Homogenates,were
R4908 T5553 T5555 arg2Of Homogenates,sonicated
R4909 T5553 T5557 arg2Of Homogenates,centrifuged
R4910 T5555 T5556 arg1Of sonicated,","
R4911 T5556 T5554 arg2Of ",",were
R4912 T5556 T5561 modOf ",",to
R4913 T5557 T5556 arg2Of centrifuged,","
R4914 T5557 T5558 arg1Of centrifuged,at
R4915 T5560 T5558 arg2Of rpm,at
R4916 T5560 T5559 arg1Of rpm,"10,000"
R4917 T5562 T5561 arg1Of remove,to
R4918 T5564 T5563 arg1Of debris,cellular
R4919 T5564 T5566 arg1Of debris,and
R4920 T5566 T5562 arg2Of and,remove
R4921 T5566 T5565 arg1Of and,","
R4922 T5566 T5568 arg2Of and,collected
R4923 T5567 T5566 arg2Of supernatant,and
R4924 T5570 T5569 arg1Of concentration,Protein
R4925 T5570 T5571 arg1Of concentration,was
R4926 T5570 T5572 arg2Of concentration,determined
R4927 T5572 T5571 arg2Of determined,was
R4928 T5573 T5572 arg3Of using,determined
R4929 T5577 T5573 arg2Of Assay,using
R4930 T5577 T5574 arg1Of Assay,the
R4931 T5577 T5575 arg1Of Assay,DC
R4932 T5577 T5576 arg1Of Assay,Protein
R4933 T5577 T5578 arg1Of Assay,(
R4934 T5579 T5578 arg2Of Bio-Rad,(
R4935 T5580 T5578 arg3Of ),(
R4936 T5581 T5582 arg1Of Proteins,in
R4937 T5581 T5590 arg1Of Proteins,were
R4938 T5581 T5591 arg2Of Proteins,resolved
R4939 T5581 T5600 arg2Of Proteins,transferred
R4940 T5583 T5582 arg2Of samples,in
R4941 T5583 T5584 arg1Of samples,(
R4942 T5588 T5584 arg2Of proteins,(
R4943 T5588 T5585 arg1Of proteins,15
R4944 T5588 T5586 arg1Of proteins,µg
R4945 T5588 T5587 arg1Of proteins,total
R4946 T5589 T5584 arg3Of ),(
R4947 T5591 T5592 arg1Of resolved,in
R4948 T5591 T5599 arg1Of resolved,and
R4949 T5595 T5596 arg1Of 12,%
R4950 T5598 T5592 arg2Of gel,in
R4951 T5598 T5593 arg1Of gel,a
R4952 T5598 T5594 arg1Of gel,denaturing
R4953 T5598 T5595 arg1Of gel,12
R4954 T5598 T5597 arg1Of gel,polyacrylamide
R4955 T5599 T5590 arg2Of and,were
R4956 T5600 T5599 arg2Of transferred,and
R4957 T5600 T5601 arg1Of transferred,to
R4958 T5604 T5601 arg2Of membrane,to
R4959 T5604 T5602 arg1Of membrane,a
R4960 T5604 T5603 arg1Of membrane,nitrocellulose
R4961 T5606 T5605 arg1Of protein,I-κBα
R4962 T5606 T5607 arg1Of protein,was
R4963 T5606 T5608 arg2Of protein,detected
R4964 T5606 T5609 arg1Of protein,using
R4965 T5608 T5607 arg2Of detected,was
R4966 T5609 T5608 arg3Of using,detected
R4967 T5609 T5621 modOf using,followed
R4968 T5613 T5609 arg2Of antibody,using
R4969 T5613 T5610 arg1Of antibody,a
R4970 T5613 T5611 arg1Of antibody,rabbit
R4971 T5613 T5612 arg1Of antibody,polyclonal
R4972 T5613 T5614 arg1Of antibody,(
R4973 T5617 T5614 arg2Of Biotechnology,(
R4974 T5617 T5615 arg1Of Biotechnology,Santa
R4975 T5617 T5616 arg1Of Biotechnology,Cruz
R4976 T5617 T5618 arg1Of Biotechnology,","
R4977 T5619 T5618 arg2Of CA,","
R4978 T5620 T5614 arg3Of ),(
R4979 T5628 T5621 arg1Of antibody,followed
R4980 T5628 T5622 arg2Of antibody,by
R4981 T5628 T5623 arg1Of antibody,a
R4982 T5628 T5624 arg1Of antibody,horseradish
R4983 T5628 T5625 arg1Of antibody,peroxidase-coupled
R4984 T5628 T5626 arg1Of antibody,goat
R4985 T5628 T5627 arg1Of antibody,polyclonal
R4986 T5628 T5629 arg1Of antibody,against
R4987 T5631 T5629 arg2Of Ig,against
R4988 T5631 T5630 arg1Of Ig,rabbit
R4989 T5631 T5632 arg1Of Ig,(
R4990 T5634 T5632 arg2Of Laboratories,(
R4991 T5634 T5633 arg1Of Laboratories,Caltag
R4992 T5635 T5632 arg3Of ),(
R4993 T5639 T5638 arg1Of bands,IκB
R4994 T5639 T5640 arg1Of bands,were
R4995 T5639 T5641 arg2Of bands,revealed
R4996 T5641 T5636 arg1Of revealed,Finally
R4997 T5641 T5637 arg1Of revealed,","
R4998 T5641 T5640 arg2Of revealed,were
R4999 T5641 T5642 modOf revealed,using
R5000 T5641 T5647 arg1Of revealed,(
R5001 T5641 T5657 arg1Of revealed,according
R5002 T5646 T5642 arg2Of system,using
R5003 T5646 T5643 arg1Of system,the
R5004 T5646 T5644 arg1Of system,ECL™
R5005 T5646 T5645 arg1Of system,detection
R5006 T5650 T5647 arg2Of Biotech,(
R5007 T5650 T5648 arg1Of Biotech,Amersham
R5008 T5650 T5649 arg1Of Biotech,Pharmacia
R5009 T5650 T5651 arg1Of Biotech,","
R5010 T5653 T5651 arg2Of Ullis,","
R5011 T5653 T5652 arg1Of Ullis,Les
R5012 T5653 T5654 arg1Of Ullis,","
R5013 T5655 T5654 arg2Of France,","
R5014 T5656 T5647 arg3Of ),(
R5015 T5658 T5657 arg2Of to,according
R5016 T5660 T5659 arg1Of manufacturers,the
R5017 T5660 T5661 arg2Of manufacturers,'
R5018 T5662 T5658 arg2Of instruction,to
R5019 T5662 T5661 arg1Of instruction,'
R5020 T5663 T5664 arg1Of Antibody,to
R5021 T5663 T5670 arg1Of Antibody,was
R5022 T5665 T5664 arg2Of α-Tubulin,to
R5023 T5665 T5666 arg1Of α-Tubulin,(
R5024 T5668 T5666 arg2Of Cruz,(
R5025 T5668 T5667 arg1Of Cruz,Santa
R5026 T5669 T5666 arg3Of ),(
R5027 T5670 T5672 arg1Of was,as
R5028 T5671 T5670 arg2Of use,was
R5029 T5673 T5672 arg2Of loading,as
R5030 T5674 T5673 arg2Of control,loading
R5031 T5677 T5675 arg2Of NF-κB,For
R5032 T5677 T5676 arg1Of NF-κB,nuclear
R5033 T5680 T5679 arg1Of cells,THP-1
R5034 T5680 T5681 arg1Of cells,were
R5035 T5680 T5682 arg2Of cells,stimulated
R5036 T5682 T5681 arg2Of stimulated,were
R5037 T5682 T5683 arg1Of stimulated,with
R5038 T5686 T5687 arg1Of C10,or
R5039 T5687 T5683 arg2Of or,with
R5040 T5687 T5684 arg1Of or,1
R5041 T5687 T5685 arg1Of or,mg/ml
R5042 T5687 T5689 arg1Of or,for
R5043 T5688 T5687 arg2Of C40,or
R5044 T5691 T5690 arg1Of minutes,30
R5045 T5691 T5692 arg1Of minutes,at
R5046 T5691 T5695 arg1Of minutes,were
R5047 T5691 T5697 arg2Of minutes,pelleted
R5048 T5694 T5692 arg2Of Cells,at
R5049 T5694 T5693 arg1Of Cells,37°C.
R5050 T5697 T5675 arg1Of pelleted,For
R5051 T5697 T5678 arg1Of pelleted,","
R5052 T5697 T5681 modOf pelleted,were
R5053 T5697 T5695 arg2Of pelleted,were
R5054 T5697 T5696 arg1Of pelleted,then
R5055 T5697 T5698 arg1Of pelleted,and
R5056 T5699 T5697 arg3Of nuclei,pelleted
R5057 T5699 T5700 arg2Of nuclei,separated
R5058 T5700 T5701 arg1Of separated,as
R5059 T5702 T5701 arg2Of described,as
R5060 T5702 T5703 arg1Of described,[
R5061 T5704 T5703 arg2Of 31,[
R5062 T5705 T5703 arg3Of ],[
R5063 T5706 T5707 arg1Of Nuclei,were
R5064 T5706 T5708 arg2Of Nuclei,washed
R5065 T5706 T5710 arg2Of Nuclei,homogenized
R5066 T5706 T5719 arg2Of Nuclei,heated
R5067 T5708 T5709 arg1Of washed,and
R5068 T5709 T5707 arg2Of and,were
R5069 T5709 T5726 arg1Of and,in
R5070 T5710 T5711 arg1Of homogenized,directly
R5071 T5710 T5712 arg1Of homogenized,in
R5072 T5710 T5718 arg1Of homogenized,and
R5073 T5715 T5714 arg2Of Laemli,(
R5074 T5716 T5714 arg3Of ),(
R5075 T5717 T5712 arg2Of buffer,in
R5076 T5717 T5713 arg1Of buffer,loading
R5077 T5717 T5714 arg1Of buffer,(
R5078 T5718 T5709 arg2Of and,and
R5079 T5719 T5718 arg2Of heated,and
R5080 T5719 T5720 arg1Of heated,for
R5081 T5722 T5720 arg2Of minutes,for
R5082 T5722 T5721 arg1Of minutes,5
R5083 T5722 T5723 arg1Of minutes,at
R5084 T5725 T5723 arg2Of Proteins,at
R5085 T5725 T5724 arg1Of Proteins,100°C.
R5086 T5727 T5728 arg1Of samples,were
R5087 T5727 T5729 arg2Of samples,resolved
R5088 T5727 T5738 arg2Of samples,transferred
R5089 T5729 T5730 arg1Of resolved,in
R5090 T5729 T5737 arg1Of resolved,and
R5091 T5733 T5734 arg1Of 8,%
R5092 T5736 T5730 arg2Of gel,in
R5093 T5736 T5731 arg1Of gel,a
R5094 T5736 T5732 arg1Of gel,denaturing
R5095 T5736 T5733 arg1Of gel,8
R5096 T5736 T5735 arg1Of gel,polyacrylamide
R5097 T5737 T5726 arg2Of and,in
R5098 T5737 T5728 arg2Of and,were
R5099 T5738 T5737 arg2Of transferred,and
R5100 T5738 T5739 arg1Of transferred,to
R5101 T5744 T5743 arg2Of PVDF,(
R5102 T5745 T5743 arg3Of ),(
R5103 T5746 T5739 arg2Of membrane,to
R5104 T5746 T5740 arg1Of membrane,a
R5105 T5746 T5741 arg1Of membrane,polyvinylidine
R5106 T5746 T5742 arg1Of membrane,fluoride
R5107 T5746 T5743 arg1Of membrane,(
R5108 T5746 T5747 arg1Of membrane,(
R5109 T5748 T5747 arg2Of Immobilon-P,(
R5110 T5748 T5749 arg1Of Immobilon-P,;
R5111 T5748 T5753 arg1Of Immobilon-P,","
R5112 T5748 T5754 arg1Of Immobilon-P,MA
R5113 T5750 T5751 arg1Of Millipore,","
R5114 T5751 T5749 arg2Of ",",;
R5115 T5752 T5751 arg2Of Bedford,","
R5116 T5755 T5747 arg3Of ),(
R5117 T5756 T5757 arg1Of Membranes,were
R5118 T5756 T5758 arg2Of Membranes,incubated
R5119 T5758 T5757 arg2Of incubated,were
R5120 T5758 T5759 arg1Of incubated,in
R5121 T5760 T5759 arg2Of blocking,in
R5122 T5761 T5760 arg2Of buffer,blocking
R5123 T5761 T5762 arg1Of buffer,(
R5124 T5761 T5770 arg1Of buffer,for
R5125 T5763 T5764 arg1Of 1,%
R5126 T5765 T5762 arg2Of BSA,(
R5127 T5765 T5763 arg1Of BSA,1
R5128 T5765 T5766 arg1Of BSA,","
R5129 T5765 T5767 arg1Of BSA,in
R5130 T5768 T5767 arg2Of PBS,in
R5131 T5769 T5762 arg3Of ),(
R5132 T5772 T5770 arg2Of hours,for
R5133 T5772 T5771 arg1Of hours,two
R5134 T5772 T5773 arg1Of hours,at
R5135 T5775 T5773 arg2Of temperature,at
R5136 T5775 T5774 arg1Of temperature,room
R5137 T5776 T5777 arg1Of Membranes,were
R5138 T5776 T5779 arg2Of Membranes,probed
R5139 T5779 T5777 arg2Of probed,were
R5140 T5779 T5778 arg1Of probed,subsequently
R5141 T5779 T5780 arg1Of probed,with
R5142 T5779 T5787 arg1Of probed,","
R5143 T5779 T5788 arg1Of probed,overnight
R5144 T5783 T5780 arg2Of antibody,with
R5145 T5783 T5781 arg1Of antibody,the
R5146 T5783 T5782 arg1Of antibody,corresponding
R5147 T5783 T5784 arg1Of antibody,in
R5148 T5785 T5784 arg2Of blocking,in
R5149 T5786 T5785 arg2Of buffer,blocking
R5150 T5794 T5789 arg1Of subunit,Rabbit
R5151 T5794 T5790 arg1Of subunit,polyclonal
R5152 T5794 T5791 arg1Of subunit,antibody
R5153 T5794 T5792 arg1Of subunit,anti-NF-κB
R5154 T5794 T5793 arg1Of subunit,p50
R5155 T5794 T5795 arg1Of subunit,(
R5156 T5794 T5799 arg1Of subunit,or
R5157 T5797 T5795 arg2Of sc-114,(
R5158 T5797 T5796 arg1Of sc-114,#
R5159 T5798 T5795 arg3Of ),(
R5160 T5799 T5807 arg1Of or,from
R5161 T5799 T5811 arg1Of or,were
R5162 T5799 T5812 arg2Of or,used
R5163 T5802 T5799 arg2Of subunit,or
R5164 T5802 T5800 arg1Of subunit,anti-NF-κB
R5165 T5802 T5801 arg1Of subunit,p65
R5166 T5802 T5803 arg1Of subunit,(
R5167 T5805 T5803 arg2Of sc-109,(
R5168 T5805 T5804 arg1Of sc-109,#
R5169 T5806 T5803 arg3Of ),(
R5170 T5810 T5807 arg2Of Biotechnology,from
R5171 T5810 T5808 arg1Of Biotechnology,Santa
R5172 T5810 T5809 arg1Of Biotechnology,Cruz
R5173 T5812 T5811 arg2Of used,were
R5174 T5813 T5814 arg1Of Membranes,were
R5175 T5813 T5815 arg2Of Membranes,washed
R5176 T5813 T5832 arg2Of Membranes,incubated
R5177 T5815 T5817 arg1Of washed,times
R5178 T5815 T5818 arg1Of washed,in
R5179 T5815 T5820 arg1Of washed,with
R5180 T5815 T5824 arg1Of washed,20
R5181 T5815 T5827 arg1Of washed,minutes
R5182 T5815 T5829 arg1Of washed,time
R5183 T5815 T5831 arg1Of washed,and
R5184 T5817 T5816 arg1Of times,six
R5185 T5819 T5818 arg2Of PBS,in
R5186 T5822 T5820 arg2Of %,with
R5187 T5822 T5821 arg1Of %,0.05
R5188 T5824 T5823 arg1Of 20,Tween
R5189 T5824 T5825 arg1Of 20,","
R5190 T5827 T5825 arg2Of minutes,","
R5191 T5827 T5826 arg1Of minutes,5
R5192 T5829 T5828 arg1Of time,each
R5193 T5831 T5814 arg2Of and,were
R5194 T5831 T5830 arg1Of and,","
R5195 T5832 T5831 arg2Of incubated,and
R5196 T5832 T5833 arg1Of incubated,with
R5197 T5832 T5861 arg1Of incubated,for
R5198 T5833 T5859 arg1Of with,in
R5199 T5836 T5834 arg1Of dilution,a
R5200 T5836 T5835 arg1Of dilution,1/3000
R5201 T5836 T5837 arg1Of dilution,of
R5202 T5839 T5837 arg2Of F,of
R5203 T5839 T5838 arg1Of F,HRP-conjugated
R5204 T5841 T5840 arg2Of ab,(
R5205 T5841 T5842 arg1Of ab,'
R5206 T5843 T5840 arg3Of ),(
R5207 T5847 T5833 arg2Of IgG,with
R5208 T5847 T5836 arg1Of IgG,dilution
R5209 T5847 T5840 arg1Of IgG,(
R5210 T5847 T5844 arg1Of IgG,2
R5211 T5847 T5845 arg1Of IgG,goat
R5212 T5847 T5846 arg1Of IgG,anti-rabbit
R5213 T5847 T5848 arg1Of IgG,in
R5214 T5849 T5850 arg1Of 5,%
R5215 T5853 T5849 arg1Of milk,5
R5216 T5853 T5851 arg1Of milk,nonfat
R5217 T5853 T5852 arg1Of milk,dry
R5218 T5853 T5854 arg1Of milk,and
R5219 T5854 T5848 arg2Of and,in
R5220 T5856 T5855 arg1Of %,0.05
R5221 T5858 T5854 arg2Of 20,and
R5222 T5858 T5856 arg1Of 20,%
R5223 T5858 T5857 arg1Of 20,Tween
R5224 T5860 T5859 arg2Of PBS,in
R5225 T5863 T5861 arg2Of hour,for
R5226 T5863 T5862 arg1Of hour,1
R5227 T5863 T5864 arg1Of hour,at
R5228 T5866 T5864 arg2Of temperature,at
R5229 T5866 T5865 arg1Of temperature,room
R5230 T5868 T5867 arg2Of washing,After
R5231 T5868 T5872 arg1Of washing,in
R5232 T5868 T5874 arg1Of washing,with
R5233 T5870 T5869 arg1Of more,six
R5234 T5871 T5868 arg2Of times,washing
R5235 T5871 T5870 arg1Of times,more
R5236 T5873 T5872 arg2Of PBS,in
R5237 T5876 T5875 arg1Of %,0.05
R5238 T5878 T5874 arg2Of 20,with
R5239 T5878 T5876 arg1Of 20,%
R5240 T5878 T5877 arg1Of 20,Tween
R5241 T5881 T5868 arg1Of proteins,washing
R5242 T5881 T5880 arg1Of proteins,antibody-reactive
R5243 T5881 T5882 arg1Of proteins,were
R5244 T5881 T5883 arg2Of proteins,detected
R5245 T5883 T5867 arg1Of detected,After
R5246 T5883 T5879 arg1Of detected,","
R5247 T5883 T5882 arg2Of detected,were
R5248 T5884 T5883 arg3Of using,detected
R5249 T5884 T5888 arg1Of using,(
R5250 T5884 T5897 arg1Of using,according
R5251 T5887 T5884 arg2Of substrate,using
R5252 T5887 T5885 arg1Of substrate,a
R5253 T5887 T5886 arg1Of substrate,chemiluminescence
R5254 T5889 T5890 arg1Of SuperSignal,;
R5255 T5890 T5888 arg2Of ;,(
R5256 T5890 T5894 arg1Of ;,","
R5257 T5891 T5892 arg1Of Pierce,","
R5258 T5892 T5890 arg2Of ",",;
R5259 T5893 T5892 arg2Of Rockford,","
R5260 T5895 T5894 arg2Of IL,","
R5261 T5896 T5888 arg3Of ),(
R5262 T5898 T5897 arg2Of to,according
R5263 T5900 T5899 arg1Of manufacturer,the
R5264 T5900 T5901 arg2Of manufacturer,'s
R5265 T5902 T5898 arg2Of instructions,to
R5266 T5902 T5901 arg1Of instructions,'s
R5267 T5904 T5903 arg1Of confirm,To
R5268 T5907 T5906 arg1Of amounts,equivalent
R5269 T5907 T5908 arg1Of amounts,of
R5270 T5907 T5910 arg1Of amounts,were
R5271 T5907 T5911 arg2Of amounts,loaded
R5272 T5909 T5908 arg2Of protein,of
R5273 T5911 T5904 arg2Of loaded,confirm
R5274 T5911 T5905 arg1Of loaded,that
R5275 T5911 T5910 arg2Of loaded,were
R5276 T5911 T5912 arg1Of loaded,in
R5277 T5914 T5912 arg2Of line,in
R5278 T5914 T5913 arg1Of line,each
R5279 T5916 T5904 arg1Of membranes,confirm
R5280 T5916 T5917 arg1Of membranes,were
R5281 T5916 T5920 arg1Of membranes,blotted
R5282 T5917 T5903 modOf were,To
R5283 T5917 T5915 arg1Of were,","
R5284 T5917 T5918 arg1Of were,also
R5285 T5917 T5923 arg1Of were,as
R5286 T5920 T5917 arg2Of blotted,were
R5287 T5920 T5919 arg1Of blotted,Western
R5288 T5920 T5921 arg1Of blotted,for
R5289 T5922 T5921 arg2Of ERK,for
R5290 T5924 T5923 arg2Of described,as
R5291 T5924 T5925 arg1Of described,[
R5292 T5926 T5925 arg2Of 32,[
R5293 T5927 T5925 arg3Of ],[
R5753 T6465 T6466 arg1Of Analysis,of
R5754 T6465 T6469 arg1Of Analysis,by
R5755 T6468 T6466 arg2Of Activation,of
R5756 T6468 T6467 arg1Of Activation,NF-κB
R5757 T6471 T6469 arg2Of Cytometry,by
R5758 T6471 T6470 arg1Of Cytometry,Flow
R5759 T6473 T6472 arg1Of activation,Nuclear
R5760 T6473 T6474 arg1Of activation,of
R5761 T6473 T6476 arg1Of activation,by
R5762 T6473 T6479 arg1Of activation,was
R5763 T6473 T6480 arg2Of activation,performed
R5764 T6475 T6474 arg2Of NF−κΒ,of
R5765 T6478 T6476 arg2Of cytometry,by
R5766 T6478 T6477 arg1Of cytometry,flow
R5767 T6480 T6479 arg2Of performed,was
R5768 T6480 T6481 arg1Of performed,as
R5769 T6482 T6481 arg2Of described,as
R5770 T6482 T6483 arg1Of described,[
R5771 T6484 T6483 arg2Of 31,[
R5772 T6485 T6483 arg3Of ],[
R5800 T6521 T6520 arg1Of Analysis,Statistical
R5801 T6523 T6522 arg1Of results,The
R5802 T6523 T6524 arg1Of results,were
R5803 T6523 T6525 arg2Of results,expressed
R5804 T6525 T6524 arg2Of expressed,were
R5805 T6525 T6526 arg1Of expressed,as
R5806 T6529 T6527 arg1Of value,the
R5807 T6529 T6528 arg1Of value,mean
R5808 T6529 T6530 arg1Of value,±
R5809 T6530 T6526 arg2Of ±,as
R5810 T6531 T6530 arg2Of S.E.M.,±
R5811 T6531 T6532 arg1Of S.E.M.,of
R5812 T6534 T6532 arg2Of experiments,of
R5813 T6534 T6533 arg1Of experiments,individual
R5814 T6537 T6535 arg1Of significance,The
R5815 T6537 T6536 arg1Of significance,statistical
R5816 T6537 T6538 arg1Of significance,of
R5817 T6537 T6544 arg1Of significance,was
R5818 T6537 T6545 arg2Of significance,assessed
R5819 T6540 T6538 arg2Of differences,of
R5820 T6540 T6539 arg1Of differences,the
R5821 T6540 T6541 arg1Of differences,between
R5822 T6543 T6541 arg2Of values,between
R5823 T6543 T6542 arg1Of values,mean
R5824 T6545 T6544 arg2Of assessed,was
R5825 T6550 T6551 arg1Of t-test,and
R5826 T6551 T6545 arg1Of and,assessed
R5827 T6551 T6546 arg2Of and,by
R5828 T6551 T6547 arg1Of and,the
R5829 T6551 T6548 arg1Of and,Student
R5830 T6551 T6549 arg1Of and,'s
R5831 T6551 T6553 arg1Of and,of
R5832 T6552 T6551 arg2Of analysis,and
R5833 T6554 T6553 arg2Of variance,of
R5834 T6554 T6555 arg1Of variance,(
R5835 T6556 T6555 arg2Of ANOVA,(
R5836 T6557 T6555 arg3Of ),(
R5877 T6627 T6623 arg2Of Inflammation,Degraded
R5878 T6627 T6624 arg1Of Inflammation,CGN
R5879 T6627 T6625 arg1Of Inflammation,Induce
R5880 T6627 T6626 arg1Of Inflammation,Colonic
R5881 T6629 T6628 arg1Of rats,All
R5882 T6629 T6630 arg1Of rats,developed
R5883 T6630 T6632 arg1Of developed,during
R5884 T6630 T6636 arg1Of developed,and
R5885 T6631 T6630 arg2Of diarrhea,developed
R5886 T6635 T6632 arg2Of administration,during
R5887 T6635 T6633 arg2Of administration,degraded
R5888 T6635 T6634 arg1Of administration,carrageenan
R5889 T6638 T6637 arg1Of evidence,gross
R5890 T6638 T6639 arg1Of evidence,of
R5891 T6638 T6641 arg1Of evidence,was
R5892 T6638 T6643 arg2Of evidence,detected
R5893 T6640 T6639 arg2Of blood,of
R5894 T6643 T6636 arg2Of detected,and
R5895 T6643 T6641 arg2Of detected,was
R5896 T6643 T6642 arg1Of detected,frequently
R5897 T6643 T6644 arg1Of detected,in
R5898 T6646 T6644 arg2Of stools,in
R5899 T6646 T6645 arg1Of stools,the
R5900 T6648 T6647 arg1Of length,Colon
R5901 T6648 T6650 arg2Of length,decreased
R5902 T6650 T6649 arg1Of decreased,dramatically
R5903 T6650 T6651 arg1Of decreased,in
R5904 T6650 T6655 arg1Of decreased,with
R5905 T6654 T6651 arg2Of rats,in
R5906 T6654 T6652 arg1Of rats,all
R5907 T6654 T6653 arg2Of rats,treated
R5908 T6658 T6657 arg1Of pronounced,more
R5909 T6659 T6655 arg2Of effect,with
R5910 T6659 T6656 arg1Of effect,a
R5911 T6659 T6658 arg1Of effect,pronounced
R5912 T6659 T6660 arg1Of effect,being
R5913 T6659 T6661 arg2Of effect,observed
R5914 T6661 T6660 arg2Of observed,being
R5915 T6661 T6662 arg1Of observed,in
R5916 T6667 T6666 arg1Of treated,dCGN
R5917 T6668 T6662 arg2Of group,in
R5918 T6668 T6663 arg1Of group,the
R5919 T6668 T6664 arg1Of group,40
R5920 T6668 T6665 arg1Of group,kDa
R5921 T6668 T6667 arg1Of group,treated
R5922 T6668 T6669 arg1Of group,(
R5923 T6671 T6669 arg2Of 1A,(
R5924 T6671 T6670 arg1Of 1A,Fig.
R5925 T6672 T6669 arg3Of ),(
R5926 T6676 T6675 arg2Of exposure,prolonged
R5927 T6676 T6677 arg1Of exposure,to
R5928 T6676 T6681 arg1Of exposure,resulted
R5929 T6680 T6677 arg2Of dCGN,to
R5930 T6680 T6678 arg1Of dCGN,40
R5931 T6680 T6679 arg1Of dCGN,kDa
R5932 T6681 T6673 arg1Of resulted,Furthermore
R5933 T6681 T6674 arg1Of resulted,","
R5934 T6681 T6682 arg1Of resulted,in
R5935 T6681 T6690 arg1Of resulted,(
R5936 T6684 T6685 arg1Of macroscopic,and
R5937 T6686 T6685 arg2Of histological,and
R5938 T6687 T6682 arg2Of scores,in
R5939 T6687 T6683 arg1Of scores,high
R5940 T6687 T6684 arg1Of scores,macroscopic
R5941 T6687 T6686 arg1Of scores,histological
R5942 T6687 T6688 arg1Of scores,of
R5943 T6689 T6688 arg2Of inflammation,of
R5944 T6692 T6690 arg2Of 1B,(
R5945 T6692 T6691 arg1Of 1B,Fig.
R5946 T6692 T6693 arg1Of 1B,","
R5947 T6694 T6693 arg2Of C,","
R5948 T6695 T6690 arg3Of ),(
R5949 T6699 T6696 arg1Of activity,Only
R5950 T6699 T6697 arg1Of activity,weak
R5951 T6699 T6698 arg1Of activity,myeloperoxidase
R5952 T6699 T6700 arg1Of activity,was
R5953 T6699 T6701 arg2Of activity,detected
R5954 T6701 T6700 arg2Of detected,was
R5955 T6701 T6702 arg1Of detected,in
R5956 T6701 T6712 arg1Of detected,","
R5957 T6701 T6713 modOf detected,indicating
R5958 T6704 T6705 arg1Of control,and
R5959 T6706 T6705 arg2Of dCGN-treated,and
R5960 T6707 T6702 arg2Of groups,in
R5961 T6707 T6703 arg1Of groups,both
R5962 T6707 T6704 arg1Of groups,control
R5963 T6707 T6706 arg1Of groups,dCGN-treated
R5964 T6707 T6708 arg1Of groups,(
R5965 T6710 T6708 arg2Of 1D,(
R5966 T6710 T6709 arg1Of 1D,Fig.
R5967 T6711 T6708 arg3Of ),(
R5968 T6715 T6716 arg1Of granulocytes,did
R5969 T6715 T6718 arg1Of granulocytes,play
R5970 T6718 T6713 arg2Of play,indicating
R5971 T6718 T6714 arg1Of play,that
R5972 T6718 T6716 arg2Of play,did
R5973 T6718 T6717 arg1Of play,not
R5974 T6718 T6722 arg1Of play,in
R5975 T6718 T6725 arg1Of play,at
R5976 T6721 T6718 arg2Of role,play
R5977 T6721 T6719 arg1Of role,a
R5978 T6721 T6720 arg1Of role,major
R5979 T6724 T6722 arg2Of inflammation,in
R5980 T6724 T6723 arg1Of inflammation,the
R5981 T6727 T6725 arg2Of stage,at
R5982 T6727 T6726 arg1Of stage,that
R5983 T6729 T6728 arg1Of examination,Histological
R5984 T6729 T6730 arg1Of examination,revealed
R5985 T6732 T6730 arg2Of degrees,revealed
R5986 T6732 T6731 arg1Of degrees,various
R5987 T6732 T6733 arg1Of degrees,of
R5988 T6735 T6733 arg2Of inflammation,of
R5989 T6735 T6734 arg1Of inflammation,mucosal
R5990 T6736 T6737 arg2Of Rats,treated
R5991 T6736 T6742 arg1Of Rats,showed
R5992 T6737 T6738 arg1Of treated,with
R5993 T6741 T6738 arg2Of dCGN,with
R5994 T6741 T6739 arg1Of dCGN,10
R5995 T6741 T6740 arg1Of dCGN,kDa
R5996 T6743 T6744 arg1Of edema,","
R5997 T6744 T6747 arg1Of ",",and
R5998 T6746 T6744 arg2Of atrophy,","
R5999 T6746 T6745 arg1Of atrophy,epithelium
R6000 T6747 T6742 arg2Of and,showed
R6001 T6750 T6747 arg2Of infiltration,and
R6002 T6750 T6748 arg1Of infiltration,slight
R6003 T6750 T6749 arg1Of infiltration,lymphocyte
R6004 T6750 T6751 arg1Of infiltration,(
R6005 T6752 T6751 arg2Of data,(
R6006 T6752 T6754 arg2Of data,shown
R6007 T6754 T6753 arg1Of shown,not
R6008 T6755 T6751 arg3Of ),(
R6009 T6757 T6756 arg1Of symptoms,These
R6010 T6757 T6758 arg1Of symptoms,were
R6011 T6757 T6760 arg1Of symptoms,absent
R6012 T6760 T6758 arg2Of absent,were
R6013 T6760 T6759 arg1Of absent,totally
R6014 T6760 T6761 arg1Of absent,in
R6015 T6763 T6761 arg2Of colon,in
R6016 T6763 T6762 arg1Of colon,the
R6017 T6763 T6764 arg1Of colon,of
R6018 T6766 T6764 arg2Of rats,of
R6019 T6766 T6765 arg1Of rats,control
R6020 T6766 T6767 arg1Of rats,(
R6021 T6769 T6767 arg2Of 1E,(
R6022 T6769 T6768 arg1Of 1E,Fig.
R6023 T6770 T6767 arg3Of ),(
R6024 T6772 T6771 arg1Of severe,More
R6025 T6774 T6772 arg1Of injuries,severe
R6026 T6774 T6773 arg1Of injuries,mucosal
R6027 T6774 T6775 arg1Of injuries,including
R6028 T6774 T6789 arg1Of injuries,were
R6029 T6774 T6790 arg2Of injuries,observed
R6030 T6776 T6777 arg1Of ulceration,","
R6031 T6777 T6780 arg1Of ",",","
R6032 T6779 T6777 arg2Of epithelium,","
R6033 T6779 T6778 arg1Of epithelium,hyperplastic
R6034 T6780 T6783 arg1Of ",",and
R6035 T6782 T6780 arg2Of distortion,","
R6036 T6782 T6781 arg1Of distortion,crypt
R6037 T6783 T6775 arg2Of and,including
R6038 T6787 T6783 arg2Of infiltration,and
R6039 T6787 T6784 arg1Of infiltration,a
R6040 T6787 T6785 arg1Of infiltration,strong
R6041 T6787 T6786 arg1Of infiltration,macrophage
R6042 T6790 T6788 arg1Of observed,","
R6043 T6790 T6789 arg2Of observed,were
R6044 T6790 T6791 arg1Of observed,in
R6045 T6796 T6791 arg2Of rats,in
R6046 T6796 T6792 arg1Of rats,the
R6047 T6796 T6793 arg1Of rats,40
R6048 T6796 T6794 arg1Of rats,kDa
R6049 T6796 T6795 arg1Of rats,dCGN-treated
R6050 T6796 T6797 arg1Of rats,(
R6051 T6799 T6797 arg2Of 1F,(
R6052 T6799 T6798 arg1Of 1F,Fig.
R6053 T6800 T6797 arg3Of ),(
R6054 T6803 T6801 arg1Of polysaccharides,No
R6055 T6803 T6802 arg2Of polysaccharides,sulphated
R6056 T6803 T6804 arg1Of polysaccharides,were
R6057 T6803 T6805 arg2Of polysaccharides,detected
R6058 T6805 T6804 arg2Of detected,were
R6059 T6809 T6805 arg1Of staining,detected
R6060 T6809 T6806 arg2Of staining,by
R6061 T6809 T6807 arg1Of staining,toluidine
R6062 T6809 T6808 arg1Of staining,blue
R6063 T6809 T6810 arg1Of staining,of
R6064 T6812 T6810 arg2Of mucosa,of
R6065 T6812 T6811 arg1Of mucosa,colon
R6066 T6812 T6813 arg1Of mucosa,from
R6067 T6814 T6813 arg2Of rats,from
R6068 T6814 T6815 arg2Of rats,treated
R6069 T6815 T6816 arg1Of treated,with
R6070 T6819 T6820 arg1Of 10,or
R6071 T6821 T6820 arg2Of 40,or
R6072 T6823 T6816 arg2Of dCGN,with
R6073 T6823 T6817 arg1Of dCGN,either
R6074 T6823 T6818 arg1Of dCGN,the
R6075 T6823 T6819 arg1Of dCGN,10
R6076 T6823 T6821 arg1Of dCGN,40
R6077 T6823 T6822 arg1Of dCGN,kDa
R6078 T6823 T6824 arg1Of dCGN,(
R6079 T6826 T6824 arg2Of shown,(
R6080 T6826 T6825 arg1Of shown,not
R6081 T6827 T6824 arg3Of ),(
R6082 T6829 T6830 arg1Of we,can
R6083 T6829 T6832 arg1Of we,exclude
R6084 T6832 T6828 arg2Of exclude,Although
R6085 T6832 T6830 arg2Of exclude,can
R6086 T6832 T6831 arg1Of exclude,not
R6087 T6833 T6832 arg2Of that,exclude
R6088 T6835 T6834 arg1Of mat,dCGN
R6089 T6835 T6837 arg1Of mat,have
R6090 T6835 T6838 arg1Of mat,retained
R6091 T6838 T6833 arg1Of retained,that
R6092 T6838 T6836 arg1Of retained,not
R6093 T6838 T6837 arg2Of retained,have
R6094 T6838 T6839 arg1Of retained,in
R6095 T6838 T6842 arg1Of retained,during
R6096 T6841 T6839 arg2Of section,in
R6097 T6841 T6840 arg1Of section,the
R6098 T6845 T6842 arg2Of procedure,during
R6099 T6845 T6843 arg1Of procedure,the
R6100 T6845 T6844 arg1Of procedure,histology
R6101 T6847 T6848 arg1Of this,indicates
R6102 T6848 T6828 arg1Of indicates,Although
R6103 T6848 T6846 arg1Of indicates,","
R6104 T6851 T6850 arg1Of polymers,these
R6105 T6851 T6852 arg1Of polymers,may
R6106 T6851 T6854 arg1Of polymers,have
R6107 T6851 T6855 arg1Of polymers,been
R6108 T6851 T6856 arg2Of polymers,phagocytosed
R6109 T6856 T6848 arg2Of phagocytosed,indicates
R6110 T6856 T6849 arg1Of phagocytosed,that
R6111 T6856 T6852 arg2Of phagocytosed,may
R6112 T6856 T6853 arg1Of phagocytosed,not
R6113 T6856 T6854 arg2Of phagocytosed,have
R6114 T6856 T6855 arg2Of phagocytosed,been
R13350 T15572 T15571 arg2Of CGN,Degraded
R13351 T15572 T15573 arg1Of CGN,induced
R13352 T15575 T15573 arg2Of inflammation,induced
R13353 T15575 T15574 arg1Of inflammation,colon
R13354 T15575 T15576 arg1Of inflammation,in
R13355 T15577 T15576 arg2Of rats,in
R13356 T15578 T15579 arg1Of Histograms,showing
R13357 T15578 T15592 arg1Of Histograms,;
R13358 T15581 T15579 arg2Of effect,showing
R13359 T15581 T15580 arg1Of effect,the
R13360 T15581 T15582 arg1Of effect,of
R13361 T15581 T15585 arg1Of effect,on
R13362 T15584 T15582 arg2Of CGN,of
R13363 T15584 T15583 arg2Of CGN,degraded
R13364 T15585 T15586 arg1Of on,:
R13365 T15588 T15585 arg2Of length,on
R13366 T15588 T15587 arg1Of length,colon
R13367 T15588 T15589 arg1Of length,(
R13368 T15590 T15589 arg2Of A,(
R13369 T15591 T15589 arg3Of ),(
R13370 T15593 T15594 arg1Of macroscopic,(
R13371 T15593 T15597 arg1Of macroscopic,and
R13372 T15595 T15594 arg2Of B,(
R13373 T15596 T15594 arg3Of ),(
R13374 T15597 T15592 arg2Of and,;
R13375 T15600 T15599 arg2Of C,(
R13376 T15601 T15599 arg3Of ),(
R13377 T15603 T15598 arg1Of score,histological
R13378 T15603 T15599 arg1Of score,(
R13379 T15603 T15602 arg1Of score,inflammation
R13380 T15603 T15604 arg1Of score,of
R13381 T15603 T15606 arg1Of score,;
R13382 T15605 T15604 arg2Of colon,of
R13383 T15606 T15597 arg2Of ;,and
R13384 T15609 T15608 arg2Of MPO,(
R13385 T15610 T15608 arg3Of ),(
R13386 T15611 T15606 arg2Of activity,;
R13387 T15611 T15607 arg1Of activity,Myeloperoxidase
R13388 T15611 T15608 arg1Of activity,(
R13389 T15611 T15612 arg1Of activity,(
R13390 T15613 T15612 arg2Of D,(
R13391 T15614 T15612 arg3Of ),(
R13392 T15616 T15615 arg1Of rats,Control
R13393 T15616 T15617 arg1Of rats,(
R13394 T15616 T15621 arg1Of rats,;
R13395 T15619 T15617 arg2Of bars,(
R13396 T15619 T15618 arg1Of bars,white
R13397 T15620 T15617 arg3Of ),(
R13398 T15621 T15624 arg1Of ;,degraded
R13399 T15623 T15621 arg2Of kDa,;
R13400 T15623 T15622 arg1Of kDa,10
R13401 T15624 T15631 arg1Of degraded,;
R13402 T15626 T15624 arg2Of rats,degraded
R13403 T15626 T15625 arg1Of rats,CGN-treated
R13404 T15626 T15627 arg1Of rats,(
R13405 T15629 T15627 arg2Of bars,(
R13406 T15629 T15628 arg1Of bars,grey
R13407 T15630 T15627 arg3Of ),(
R13408 T15633 T15632 arg1Of kDa,40
R13409 T15633 T15634 arg1Of kDa,degraded
R13410 T15634 T15631 arg2Of degraded,;
R13411 T15636 T15634 arg2Of rats,degraded
R13412 T15636 T15635 arg1Of rats,CGN-treated
R13413 T15636 T15637 arg1Of rats,(
R13414 T15639 T15637 arg2Of bars,(
R13415 T15639 T15638 arg1Of bars,black
R13416 T15640 T15637 arg3Of ),(
R13417 T15642 T15634 arg3Of *,degraded
R13418 T15642 T15641 arg1Of *,.
R13419 T15642 T15643 arg1Of *,p
R13420 T15642 T15649 arg1Of *,*
R13421 T15644 T15645 arg1Of <0.0,5 fr
R13422 T15648 T15645 arg2Of .,5 fr
R13423 T15648 T15646 arg1Of .,om contr
R13424 T15648 T15647 arg1Of .,ol
R13425 T15649 T15644 arg1Of *,<0.0
R13426 T15649 T15650 arg1Of *,* p<
R13427 T15649 T15651 arg1Of *,0.01
R13428 T15652 T15651 arg2Of from co,0.01
R13429 T15654 T15653 arg1Of analysis,Histological
R13430 T15654 T15655 arg1Of analysis,of
R13431 T15654 T15657 arg1Of analysis,from
R13432 T15654 T15665 arg1Of analysis,from
R13433 T15656 T15655 arg2Of colon,of
R13434 T15657 T15664 arg1Of from,and
R13435 T15659 T15657 arg2Of rats,from
R13436 T15659 T15658 arg1Of rats,control
R13437 T15659 T15660 arg1Of rats,(
R13438 T15661 T15660 arg2Of E,(
R13439 T15662 T15660 arg3Of ),(
R13440 T15664 T15663 arg1Of and,","
R13441 T15665 T15664 arg2Of from,and
R13442 T15669 T15665 arg2Of rats,from
R13443 T15669 T15666 arg1Of rats,40
R13444 T15669 T15667 arg1Of rats,kDa
R13445 T15669 T15668 arg1Of rats,dCGN-treated
R13446 T15669 T15670 arg1Of rats,(
R13447 T15671 T15670 arg2Of F,(
R13448 T15672 T15670 arg3Of ),(
R1782 T2071 T2070 arg2Of carrageenan,degraded
R1783 T2071 T2072 arg1Of carrageenan,with
R1785 T2073 T2074 arg1Of low,","
R1793 T2082 T2072 arg2Of and,with

bionlp-st-ge-2016-spacy-parsed

Id Subject Object Predicate Lexical cue
T468 0-3 NNP denotes CGN
T469 4-13 VBD denotes inhibited
T470 14-19 CD denotes THP-1
T471 20-24 NN denotes cell
T472 25-38 NN denotes proliferation
T473 39-41 IN denotes in
T474 42-47 NN denotes vitro
T475 47-48 , denotes ,
T476 49-58 VBG denotes arresting
T477 59-62 DT denotes the
T478 63-68 NNS denotes cells
T479 69-71 IN denotes in
T480 72-74 CD denotes G1
T481 75-80 NN denotes phase
T482 80-81 . denotes .
T483 82-84 IN denotes In
T484 85-93 NN denotes addition
T485 93-94 , denotes ,
T486 95-99 NNP denotes dCGN
T487 100-109 VBD denotes increased
T488 110-116 NNP denotes ICAM-1
T489 117-127 NN denotes expression
T490 128-130 IN denotes in
T491 131-135 DT denotes both
T492 136-139 NNP denotes PBM
T493 140-143 CC denotes and
T494 144-149 NNP denotes THP-1
T495 150-155 NNS denotes cells
T496 156-160 IN denotes with
T497 161-162 DT denotes a
T498 163-168 JJ denotes major
T499 169-175 NN denotes effect
T500 176-180 VBN denotes seen
T501 181-186 IN denotes after
T502 187-189 CD denotes 40
T503 190-193 NNP denotes kDa
T504 194-198 NNP denotes dCGN
T505 199-207 NN denotes exposure
T506 207-208 . denotes .
T507 209-213 RB denotes Also
T508 213-214 , denotes ,
T509 215-219 NNP denotes dCGN
T510 220-230 VBD denotes stimulated
T511 231-239 NN denotes monocyte
T512 240-251 NN denotes aggregation
T513 252-254 IN denotes in
T514 255-260 NN denotes vitro
T515 261-265 WDT denotes that
T516 266-269 VBD denotes was
T517 270-279 VBN denotes prevented
T518 280-282 IN denotes by
T519 283-293 NN denotes incubation
T520 294-298 IN denotes with
T521 299-310 JJ denotes anti-ICAM-1
T522 311-319 NN denotes antibody
T523 319-320 . denotes .
T524 321-328 RB denotes Finally
T525 328-329 , denotes ,
T526 330-334 NNP denotes dCGN
T527 335-345 VBD denotes stimulated
T528 346-351 JJ denotes TNF-α
T529 352-362 NN denotes expression
T530 363-366 CC denotes and
T531 367-376 NN denotes secretion
T532 377-379 IN denotes by
T533 380-384 DT denotes both
T534 385-388 NNP denotes PBM
T535 389-392 CC denotes and
T536 393-398 NNP denotes THP-1
T537 399-404 NNS denotes cells
T538 404-405 . denotes .
T539 406-409 PDT denotes All
T540 410-415 DT denotes these
T541 416-423 NNS denotes effects
T542 424-428 VBD denotes were
T543 429-435 VBN denotes linked
T544 436-438 TO denotes to
T545 439-444 NNP denotes NF-κB
T546 445-455 NN denotes activation
T547 455-456 . denotes .
T548 457-462 DT denotes These
T549 463-467 NNS denotes data
T550 468-476 RB denotes strongly
T551 477-484 VBP denotes suggest
T552 485-489 IN denotes that
T553 490-493 DT denotes the
T554 494-502 JJ denotes degraded
T555 503-508 NNS denotes forms
T556 509-511 IN denotes of
T557 512-515 NNP denotes CGN
T558 516-520 VBP denotes have
T559 521-522 DT denotes a
T560 523-533 JJ denotes pronounced
T561 534-540 NN denotes effect
T562 541-543 IN denotes on
T563 544-553 NNS denotes monocytes
T564 553-554 , denotes ,
T565 555-569 NN denotes characteristic
T566 570-572 IN denotes of
T567 573-575 DT denotes an
T568 576-588 JJ denotes inflammatory
T569 589-598 NN denotes phenotype
T570 598-599 . denotes .
T1344 614-625 NNP denotes Carrageenan
T1345 626-627 -LRB- denotes (
T1346 627-630 NNP denotes CGN
T1347 630-631 -RRB- denotes )
T1348 632-634 VBZ denotes is
T1349 635-636 DT denotes a
T1350 637-641 JJ denotes high
T1351 642-651 JJ denotes molecular
T1352 652-658 NN denotes weight
T1353 659-668 VBN denotes sulphated
T1354 669-683 NN denotes polysaccharide
T1355 684-685 -LRB- denotes (
T1356 685-686 FW denotes >
T1357 686-689 CD denotes 200
T1358 690-693 NNP denotes kDa
T1359 693-694 -RRB- denotes )
T1360 695-702 VBN denotes derived
T1361 703-707 IN denotes from
T1362 708-711 JJ denotes red
T1363 712-717 NN denotes algae
T1364 718-719 -LRB- denotes (
T1365 719-731 NNP denotes Rhodophyceae
T1366 731-732 -RRB- denotes )
T1367 732-733 . denotes .
T1368 734-739 CD denotes Three
T1369 740-744 JJ denotes main
T1370 745-750 NNS denotes forms
T1371 751-753 IN denotes of
T1372 754-757 NNP denotes CGN
T1373 758-762 VBP denotes have
T1374 763-767 VBN denotes been
T1375 768-778 VBN denotes identified
T1376 778-779 : denotes :
T1377 780-785 NN denotes kappa
T1378 785-786 , denotes ,
T1379 787-791 NN denotes iota
T1380 791-792 , denotes ,
T1381 793-796 CC denotes and
T1382 797-803 NN denotes lambda
T1383 803-804 . denotes .
T1384 805-809 PRP denotes They
T1385 810-816 VBP denotes differ
T1386 817-821 IN denotes from
T1387 822-826 DT denotes each
T1388 827-832 JJ denotes other
T1389 833-835 IN denotes in
T1390 836-846 NN denotes sulphation
T1391 847-853 NN denotes degree
T1392 854-857 CC denotes and
T1393 858-868 NN denotes solubility
T1394 869-870 NN denotes [
T1395 870-871 CD denotes 1
T1396 871-872 NNP denotes ]
T1397 872-873 , denotes ,
T1398 874-875 NNP denotes [
T1399 875-876 CD denotes 2
T1400 876-877 NNP denotes ]
T1401 877-878 . denotes .
T1402 879-885 JJ denotes Native
T1403 886-889 NNP denotes CGN
T1404 890-892 VBZ denotes is
T1405 893-900 VBN denotes thought
T1406 901-903 TO denotes to
T1407 904-906 VB denotes be
T1408 907-915 JJ denotes harmless
T1409 916-919 CC denotes and
T1410 920-922 VBZ denotes is
T1411 923-929 RB denotes widely
T1412 930-934 VBN denotes used
T1413 935-937 IN denotes as
T1414 938-939 DT denotes a
T1415 940-944 NN denotes food
T1416 945-953 JJ denotes additive
T1417 954-956 TO denotes to
T1418 957-964 VB denotes improve
T1419 965-972 NN denotes texture
T1420 972-973 . denotes .
T1421 974-976 PRP denotes It
T1422 977-979 VBZ denotes is
T1423 980-984 RB denotes also
T1424 985-989 VBN denotes used
T1425 990-992 IN denotes in
T1426 993-1002 NNS denotes cosmetics
T1427 1003-1006 CC denotes and
T1428 1007-1022 NNS denotes pharmaceuticals
T1429 1022-1023 . denotes .
T1430 1024-1031 RB denotes However
T1431 1031-1032 , denotes ,
T1432 1033-1037 NN denotes acid
T1433 1038-1047 NN denotes treatment
T1434 1048-1050 IN denotes at
T1435 1051-1055 JJ denotes high
T1436 1056-1067 NN denotes temperature
T1437 1068-1069 -LRB- denotes (
T1438 1069-1071 CD denotes 80
T1439 1071-1072 CD denotes °
T1440 1072-1073 NNP denotes C
T1441 1073-1074 -RRB- denotes )
T1442 1075-1083 VBZ denotes triggers
T1443 1084-1087 NNP denotes CGN
T1444 1088-1098 NN denotes hydrolysis
T1445 1099-1101 TO denotes to
T1446 1102-1107 JJR denotes lower
T1447 1108-1117 JJ denotes molecular
T1448 1118-1124 NN denotes weight
T1449 1125-1126 -LRB- denotes (
T1450 1126-1127 FW denotes <
T1451 1127-1129 CD denotes 50
T1452 1130-1133 NNP denotes kDa
T1453 1133-1134 -RRB- denotes )
T1454 1135-1144 VBZ denotes compounds
T1455 1145-1150 VBN denotes known
T1456 1151-1153 IN denotes as
T1457 1154-1164 NN denotes poligeenan
T1458 1165-1167 CC denotes or
T1459 1168-1176 JJ denotes degraded
T1460 1177-1180 NNP denotes CGN
T1461 1181-1182 -LRB- denotes (
T1462 1182-1186 NNP denotes dCGN
T1463 1186-1187 -RRB- denotes )
T1464 1187-1188 . denotes .
T1465 1189-1194 DT denotes These
T1466 1195-1200 NNS denotes dCGNs
T1467 1201-1207 VBP denotes induce
T1468 1208-1220 NN denotes inflammation
T1469 1221-1224 CC denotes and
T1470 1225-1229 VBP denotes have
T1471 1230-1234 VBN denotes been
T1472 1235-1241 RB denotes widely
T1473 1242-1246 VBN denotes used
T1474 1247-1249 IN denotes as
T1475 1250-1256 NNS denotes models
T1476 1257-1259 IN denotes of
T1477 1260-1267 NN denotes colitis
T1478 1268-1270 IN denotes in
T1479 1271-1278 JJ denotes several
T1480 1279-1286 NNS denotes species
T1481 1286-1287 , denotes ,
T1482 1288-1297 VBG denotes including
T1483 1298-1302 NNS denotes rats
T1484 1303-1304 JJ denotes [
T1485 1304-1305 CD denotes 3
T1486 1305-1306 NNP denotes ]
T1487 1306-1307 , denotes ,
T1488 1308-1315 VBZ denotes rabbits
T1489 1316-1317 NNP denotes [
T1490 1317-1318 CD denotes 4
T1491 1318-1319 NNP denotes ]
T1492 1320-1323 CC denotes and
T1493 1324-1330 NN denotes guinea
T1494 1331-1335 NNS denotes pigs
T1495 1336-1337 VBP denotes [
T1496 1337-1338 CD denotes 5
T1497 1338-1339 NNP denotes ]
T1498 1339-1340 . denotes .
T1499 1341-1344 DT denotes The
T1500 1345-1349 NN denotes role
T1501 1350-1352 IN denotes of
T1502 1353-1357 NNP denotes dCGN
T1503 1358-1360 IN denotes as
T1504 1361-1362 DT denotes a
T1505 1363-1378 JJ denotes tumor-promoting
T1506 1379-1385 NN denotes factor
T1507 1386-1393 VBZ denotes remains
T1508 1394-1407 JJ denotes controversial
T1509 1408-1409 NN denotes [
T1510 1409-1410 CD denotes 4
T1511 1410-1411 NNP denotes ]
T1512 1411-1412 , denotes ,
T1513 1413-1414 NNP denotes [
T1514 1414-1415 CD denotes 6
T1515 1415-1416 NNP denotes ]
T1516 1417-1418 NN denotes [
T1517 1418-1419 CD denotes 8
T1518 1419-1420 NNP denotes ]
T1519 1420-1421 . denotes .
T1520 1422-1430 IN denotes Although
T1521 1431-1434 DT denotes the
T1522 1435-1441 JJ denotes native
T1523 1442-1446 NN denotes form
T1524 1447-1449 VBZ denotes is
T1525 1450-1457 VBN denotes thought
T1526 1458-1460 TO denotes to
T1527 1461-1463 VB denotes be
T1528 1464-1472 JJ denotes harmless
T1529 1473-1476 IN denotes for
T1530 1477-1482 JJ denotes human
T1531 1483-1494 NN denotes consumption
T1532 1494-1495 , denotes ,
T1533 1496-1501 JJ denotes small
T1534 1502-1509 NNS denotes amounts
T1535 1510-1512 IN denotes of
T1536 1513-1517 NNP denotes dCGN
T1537 1518-1521 VBP denotes are
T1538 1522-1530 RB denotes probably
T1539 1531-1539 VBN denotes produced
T1540 1540-1542 IN denotes by
T1541 1543-1547 NN denotes acid
T1542 1548-1558 NN denotes hydrolysis
T1543 1559-1565 IN denotes during
T1544 1566-1573 JJ denotes gastric
T1545 1574-1583 NN denotes digestion
T1546 1584-1585 NN denotes [
T1547 1585-1586 CD denotes 9
T1548 1586-1587 NNP denotes ]
T1549 1587-1588 , denotes ,
T1550 1589-1590 NNP denotes [
T1551 1590-1592 CD denotes 10
T1552 1592-1593 NNP denotes ]
T1553 1594-1596 CC denotes or
T1554 1597-1608 NN denotes interaction
T1555 1609-1613 IN denotes with
T1556 1614-1624 JJ denotes intestinal
T1557 1625-1633 NNS denotes bacteria
T1558 1634-1635 NNP denotes [
T1559 1635-1637 CD denotes 11
T1560 1637-1638 NNP denotes ]
T1561 1638-1639 , denotes ,
T1562 1640-1641 NNP denotes [
T1563 1641-1643 CD denotes 12
T1564 1643-1644 NNP denotes ]
T1565 1644-1645 . denotes .
T1566 1646-1653 IN denotes Whereas
T1567 1654-1657 DT denotes the
T1568 1658-1665 NNS denotes effects
T1569 1666-1668 IN denotes of
T1570 1669-1675 JJ denotes native
T1571 1676-1679 CC denotes and
T1572 1680-1684 NNP denotes dCGN
T1573 1685-1687 IN denotes on
T1574 1688-1698 JJ denotes intestinal
T1575 1699-1711 NN denotes inflammation
T1576 1712-1716 VBP denotes have
T1577 1717-1721 VBN denotes been
T1578 1722-1733 RB denotes extensively
T1579 1734-1742 VBN denotes analyzed
T1580 1743-1745 IN denotes in
T1581 1746-1752 NN denotes animal
T1582 1753-1759 NNS denotes models
T1583 1759-1760 , denotes ,
T1584 1761-1765 RB denotes only
T1585 1766-1769 JJ denotes few
T1586 1770-1777 NNS denotes studies
T1587 1778-1782 VBP denotes have
T1588 1783-1787 VBN denotes been
T1589 1788-1797 VBN denotes conducted
T1590 1798-1803 VBG denotes using
T1591 1804-1809 JJ denotes human
T1592 1810-1814 NN denotes cell
T1593 1815-1820 NNS denotes lines
T1594 1820-1821 . denotes .
T1595 1822-1828 JJ denotes Recent
T1596 1829-1836 NNS denotes studies
T1597 1837-1841 VBP denotes have
T1598 1842-1847 VBN denotes shown
T1599 1848-1849 DT denotes a
T1600 1850-1854 NN denotes link
T1601 1855-1862 IN denotes between
T1602 1863-1871 NN denotes exposure
T1603 1872-1874 TO denotes to
T1604 1875-1881 JJ denotes native
T1605 1882-1886 NN denotes form
T1606 1887-1890 NNP denotes CGN
T1607 1891-1894 CC denotes and
T1608 1895-1899 JJ denotes IL-8
T1609 1900-1910 NN denotes production
T1610 1911-1913 IN denotes by
T1611 1914-1917 DT denotes the
T1612 1918-1923 JJ denotes human
T1613 1924-1934 JJ denotes intestinal
T1614 1935-1945 JJ denotes epithelial
T1615 1946-1950 NN denotes cell
T1616 1951-1955 NN denotes line
T1617 1955-1956 , denotes ,
T1618 1957-1963 NNP denotes NCM460
T1619 1963-1964 , denotes ,
T1620 1965-1968 IN denotes via
T1621 1969-1976 NNP denotes Nuclear
T1622 1977-1986 NNP denotes Factor-κB
T1623 1987-1988 -LRB- denotes (
T1624 1988-1993 NNP denotes NF-κB
T1625 1993-1994 -RRB- denotes )
T1626 1995-2005 NN denotes activation
T1627 2006-2007 NNP denotes [
T1628 2007-2009 CD denotes 13
T1629 2009-2010 NNP denotes ]
T1630 2010-2011 , denotes ,
T1631 2012-2013 NNP denotes [
T1632 2013-2015 CD denotes 14
T1633 2015-2016 NNP denotes ]
T1634 2016-2017 . denotes .
T1635 2018-2023 NN denotes NF-κB
T1636 2024-2026 VBZ denotes is
T1637 2027-2028 DT denotes a
T1638 2029-2042 NN denotes transcription
T1639 2043-2049 NN denotes factor
T1640 2050-2054 WDT denotes that
T1641 2055-2064 VBZ denotes regulates
T1642 2065-2068 DT denotes the
T1643 2069-2079 NN denotes expression
T1644 2080-2082 IN denotes of
T1645 2083-2088 NNS denotes genes
T1646 2089-2099 VBN denotes associated
T1647 2100-2104 IN denotes with
T1648 2105-2117 NN denotes inflammation
T1649 2118-2119 NNP denotes [
T1650 2119-2121 CD denotes 15
T1651 2121-2122 NNP denotes ]
T1652 2122-2123 , denotes ,
T1653 2124-2125 NNP denotes [
T1654 2125-2127 CD denotes 16
T1655 2127-2128 NNP denotes ]
T1656 2128-2129 . denotes .
T1657 2130-2140 NNP denotes Macrophage
T1658 2141-2153 NN denotes infiltration
T1659 2154-2157 CC denotes and
T1660 2158-2170 NN denotes accumulation
T1661 2171-2173 VBZ denotes is
T1662 2174-2175 DT denotes a
T1663 2176-2182 JJ denotes common
T1664 2183-2197 NN denotes characteristic
T1665 2198-2200 IN denotes of
T1666 2201-2211 JJ denotes intestinal
T1667 2212-2220 NNS denotes diseases
T1668 2221-2222 NNP denotes [
T1669 2222-2224 CD denotes 17
T1670 2224-2225 NNP denotes ]
T1671 2225-2226 . denotes .
T1672 2227-2238 NNS denotes Macrophages
T1673 2239-2248 VBP denotes represent
T1674 2249-2251 CD denotes 10
T1675 2251-2252 NN denotes %
T1676 2253-2255 IN denotes of
T1677 2256-2261 JJ denotes total
T1678 2262-2268 NN denotes lamina
T1679 2269-2276 NN denotes propria
T1680 2277-2282 NNS denotes cells
T1681 2282-2283 , denotes ,
T1682 2284-2291 VBP denotes secrete
T1683 2292-2293 DT denotes a
T1684 2294-2298 JJ denotes wide
T1685 2299-2304 NN denotes range
T1686 2305-2307 IN denotes of
T1687 2308-2320 RB denotes biologically
T1688 2321-2327 JJ denotes active
T1689 2328-2337 NNS denotes compounds
T1690 2338-2341 CC denotes and
T1691 2342-2349 VB denotes express
T1692 2350-2363 NN denotes cell-adhesion
T1693 2364-2373 NNS denotes molecules
T1694 2373-2374 . denotes .
T1695 2375-2378 DT denotes The
T1696 2379-2385 JJ denotes immune
T1697 2386-2390 NN denotes cell
T1698 2391-2399 NN denotes response
T1699 2400-2402 TO denotes to
T1700 2403-2405 DT denotes an
T1701 2406-2418 JJ denotes inflammatory
T1702 2419-2427 NN denotes stimulus
T1703 2428-2433 VBZ denotes seems
T1704 2434-2436 TO denotes to
T1705 2437-2439 VB denotes be
T1706 2440-2449 VBN denotes amplified
T1707 2450-2452 CC denotes or
T1708 2453-2461 RB denotes directly
T1709 2462-2471 VBN denotes generated
T1710 2472-2474 IN denotes by
T1711 2475-2480 NNS denotes cells
T1712 2481-2488 VBN denotes exposed
T1713 2489-2491 TO denotes to
T1714 2492-2501 VBN denotes sulphated
T1715 2502-2517 NNS denotes polysaccharides
T1716 2518-2522 JJ denotes such
T1717 2523-2525 IN denotes as
T1718 2526-2538 NNS denotes carrageenans
T1719 2538-2539 . denotes .
T1720 2540-2546 RB denotes Indeed
T1721 2546-2547 , denotes ,
T1722 2548-2560 NN denotes inflammation
T1723 2561-2568 VBN denotes induced
T1724 2569-2571 IN denotes by
T1725 2572-2576 NNP denotes dCGN
T1726 2577-2580 VBD denotes was
T1727 2581-2591 VBN denotes associated
T1728 2592-2596 IN denotes with
T1729 2597-2608 NN denotes recruitment
T1730 2609-2611 IN denotes of
T1731 2612-2623 NNS denotes macrophages
T1732 2624-2626 TO denotes to
T1733 2627-2639 NN denotes inflammation
T1734 2640-2645 NNS denotes sites
T1735 2646-2647 NNP denotes [
T1736 2647-2649 CD denotes 18
T1737 2649-2650 NNP denotes ]
T1738 2650-2651 , denotes ,
T1739 2652-2653 NNP denotes [
T1740 2653-2655 CD denotes 19
T1741 2655-2656 NNP denotes ]
T1742 2656-2657 . denotes .
T1743 2658-2662 RB denotes Also
T1744 2662-2663 , denotes ,
T1745 2664-2676 NN denotes inflammation
T1746 2677-2684 VBN denotes induced
T1747 2685-2687 IN denotes by
T1748 2688-2695 NNP denotes Dextran
T1749 2696-2704 NNP denotes Sulphate
T1750 2705-2711 NNP denotes Sodium
T1751 2712-2713 -LRB- denotes (
T1752 2713-2716 NNP denotes DSS
T1753 2716-2717 -RRB- denotes )
T1754 2717-2718 , denotes ,
T1755 2719-2726 DT denotes another
T1756 2727-2736 VBN denotes sulphated
T1757 2737-2745 NN denotes compound
T1758 2745-2746 , denotes ,
T1759 2747-2750 VBD denotes was
T1760 2751-2759 RB denotes directly
T1761 2760-2770 VBN denotes associated
T1762 2771-2775 IN denotes with
T1763 2776-2787 NNS denotes macrophages
T1764 2788-2799 NN denotes recruitment
T1765 2800-2801 NNP denotes [
T1766 2801-2803 CD denotes 20
T1767 2803-2804 NNP denotes ]
T1768 2804-2805 , denotes ,
T1769 2806-2811 IN denotes since
T1770 2812-2815 NNP denotes DSS
T1771 2816-2821 RB denotes still
T1772 2822-2830 VBD denotes provoked
T1773 2831-2843 NN denotes inflammation
T1774 2844-2849 IN denotes after
T1775 2850-2862 NN denotes T-lymphocyte
T1776 2863-2866 CC denotes and
T1777 2867-2869 NNP denotes NK
T1778 2870-2874 NN denotes cell
T1779 2875-2884 NN denotes depletion
T1780 2885-2886 NNP denotes [
T1781 2886-2888 CD denotes 20
T1782 2888-2889 NNP denotes ]
T1783 2889-2890 . denotes .
T1784 2891-2899 IN denotes Although
T1785 2900-2912 NN denotes inflammation
T1786 2913-2916 MD denotes can
T1787 2917-2919 VB denotes be
T1788 2920-2927 VBN denotes induced
T1789 2928-2930 IN denotes by
T1790 2931-2935 NNP denotes dCGN
T1791 2935-2936 , denotes ,
T1792 2937-2942 EX denotes there
T1793 2943-2946 VBP denotes are
T1794 2947-2949 DT denotes no
T1795 2950-2954 NNS denotes data
T1796 2955-2957 IN denotes on
T1797 2958-2963 JJ denotes human
T1798 2964-2972 NN denotes monocyte
T1799 2973-2982 NNS denotes responses
T1800 2983-2985 TO denotes to
T1801 2986-2990 NNP denotes dCGN
T1802 2991-2999 NN denotes exposure
T1803 2999-3000 . denotes .
T1804 3001-3010 RB denotes Therefore
T1805 3010-3011 , denotes ,
T1806 3012-3014 TO denotes to
T1807 3015-3026 VB denotes investigate
T1808 3027-3030 DT denotes the
T1809 3031-3038 NNS denotes effects
T1810 3039-3041 IN denotes of
T1811 3042-3046 NNP denotes dCGN
T1812 3047-3049 IN denotes on
T1813 3050-3055 JJ denotes human
T1814 3056-3065 NNS denotes monocytes
T1815 3065-3066 , denotes ,
T1816 3067-3073 JJ denotes normal
T1817 3074-3084 NNP denotes Peripheral
T1818 3085-3090 NNP denotes Blood
T1819 3091-3100 NNP denotes Monocytes
T1820 3101-3102 -LRB- denotes (
T1821 3102-3105 NNP denotes PBM
T1822 3105-3106 -RRB- denotes )
T1823 3107-3110 CC denotes and
T1824 3111-3118 JJ denotes tumoral
T1825 3119-3138 NN denotes monocyte/macrophage
T1826 3139-3144 CD denotes THP-1
T1827 3145-3150 NNS denotes cells
T1828 3151-3155 VBD denotes were
T1829 3156-3163 VBN denotes exposed
T1830 3164-3166 TO denotes to
T1831 3167-3169 CD denotes 10
T1832 3170-3173 NN denotes kDa
T1833 3174-3177 CC denotes and
T1834 3178-3180 CD denotes 40
T1835 3181-3184 NNP denotes kDa
T1836 3185-3189 NNP denotes dCGN
T1837 3189-3190 . denotes .
T1838 3191-3193 PRP denotes We
T1839 3194-3199 VBD denotes found
T1840 3200-3204 IN denotes that
T1841 3205-3209 NNP denotes dCGN
T1842 3210-3219 VBD denotes inhibited
T1843 3220-3225 CD denotes THP-1
T1844 3226-3230 NN denotes cell
T1845 3231-3244 NN denotes proliferation
T1846 3245-3247 IN denotes in
T1847 3248-3253 NN denotes vitro
T1848 3253-3254 , denotes ,
T1849 3255-3264 VBD denotes increased
T1850 3265-3271 NNP denotes ICAM-1
T1851 3272-3282 NN denotes expression
T1852 3282-3283 , denotes ,
T1853 3284-3294 VBN denotes stimulated
T1854 3295-3311 JJ denotes ICAM-1-dependent
T1855 3312-3320 NN denotes monocyte
T1856 3321-3332 NN denotes aggregation
T1857 3332-3333 , denotes ,
T1858 3334-3337 CC denotes and
T1859 3338-3348 VBD denotes stimulated
T1860 3349-3354 NNP denotes TNF-α
T1861 3355-3365 NN denotes expression
T1862 3366-3369 CC denotes and
T1863 3370-3379 NN denotes secretion
T1864 3379-3380 . denotes .
T1865 3381-3386 DT denotes These
T1866 3387-3396 NNS denotes responses
T1867 3397-3401 VBD denotes were
T1868 3402-3406 RBR denotes more
T1869 3407-3417 JJ denotes pronounced
T1870 3418-3423 IN denotes after
T1871 3424-3426 CD denotes 40
T1872 3427-3430 NNP denotes kDa
T1873 3431-3435 NNP denotes dCGN
T1874 3436-3444 NN denotes exposure
T1875 3445-3448 CC denotes and
T1876 3449-3453 VBD denotes were
T1877 3454-3460 VBN denotes linked
T1878 3461-3463 TO denotes to
T1879 3464-3469 NNP denotes NF-κB
T1880 3470-3480 NN denotes activation
T1881 3480-3481 . denotes .
T1882 3482-3484 IN denotes In
T1883 3485-3493 NN denotes addition
T1884 3493-3494 , denotes ,
T1885 3495-3498 DT denotes the
T1886 3499-3501 CD denotes 40
T1887 3502-3505 NNP denotes kDa
T1888 3506-3510 NNP denotes dCGN
T1889 3510-3511 , denotes ,
T1890 3512-3515 CC denotes but
T1891 3516-3519 RB denotes not
T1892 3520-3523 DT denotes the
T1893 3524-3526 CD denotes 10
T1894 3527-3530 NNP denotes kDa
T1895 3531-3535 NNP denotes dCGN
T1896 3536-3543 VBD denotes induced
T1897 3544-3546 IN denotes in
T1898 3547-3551 NN denotes vivo
T1899 3552-3559 NNS denotes colitis
T1900 3560-3562 IN denotes as
T1901 3563-3568 VBN denotes shown
T1902 3569-3571 IN denotes by
T1903 3572-3575 DT denotes the
T1904 3576-3588 JJ denotes inflammatory
T1905 3589-3597 NN denotes response
T1906 3598-3600 IN denotes in
T1907 3601-3604 DT denotes the
T1908 3605-3608 NN denotes rat
T1909 3609-3614 NN denotes colon
T1910 3614-3615 . denotes .
T1911 3616-3621 DT denotes These
T1912 3622-3629 NNS denotes results
T1913 3630-3637 VBP denotes suggest
T1914 3638-3642 IN denotes that
T1915 3643-3646 DT denotes the
T1916 3647-3655 JJ denotes degraded
T1917 3656-3661 NNS denotes forms
T1918 3662-3664 IN denotes of
T1919 3665-3668 NNP denotes CGN
T1920 3669-3673 VBP denotes have
T1921 3674-3676 DT denotes an
T1922 3677-3686 JJ denotes important
T1923 3687-3693 NN denotes effect
T1924 3694-3696 IN denotes on
T1925 3697-3706 NNS denotes monocytes
T1926 3707-3716 VBG denotes resulting
T1927 3717-3719 IN denotes in
T1928 3720-3722 DT denotes an
T1929 3723-3735 JJ denotes inflammatory
T1930 3736-3745 NN denotes phenotype
T1931 3745-3746 . denotes .
T2416 3771-3782 NNP denotes Preparation
T2417 3783-3785 IN denotes of
T2418 3786-3794 NNP denotes Degraded
T2419 3795-3806 NNP denotes Carrageenan
T2420 3807-3810 CD denotes Two
T2421 3811-3823 NNS denotes preparations
T2422 3824-3826 IN denotes of
T2423 3827-3835 JJ denotes degraded
T2424 3836-3847 NN denotes carrageenan
T2425 3848-3852 IN denotes with
T2426 3853-3856 JJ denotes low
T2427 3856-3857 , denotes ,
T2428 3858-3859 -LRB- denotes (
T2429 3859-3860 FW denotes ∼
T2430 3860-3862 CD denotes 10
T2431 3863-3866 NN denotes kDa
T2432 3866-3867 : denotes ;
T2433 3868-3871 NNP denotes C10
T2434 3871-3872 -RRB- denotes )
T2435 3872-3873 , denotes ,
T2436 3874-3877 CC denotes and
T2437 3878-3884 NN denotes medium
T2438 3884-3885 , denotes ,
T2439 3886-3887 -LRB- denotes (
T2440 3887-3888 FW denotes ∼
T2441 3888-3890 CD denotes 40
T2442 3891-3894 NN denotes kDa
T2443 3894-3895 : denotes ;
T2444 3896-3899 NNP denotes C40
T2445 3899-3900 -RRB- denotes )
T2446 3901-3910 JJ denotes molecular
T2447 3911-3917 NN denotes weight
T2448 3918-3922 VBD denotes were
T2449 3923-3931 VBN denotes prepared
T2450 3932-3936 IN denotes from
T2451 3937-3943 JJ denotes native
T2452 3944-3960 JJ denotes iota-carrageenan
T2453 3961-3970 VBN denotes extracted
T2454 3971-3975 IN denotes from
T2455 3976-3983 NNP denotes Euchema
T2456 3984-3992 NN denotes spinosum
T2457 3993-3994 -LRB- denotes (
T2458 3994-4004 RB denotes generously
T2459 4005-4013 VBN denotes provided
T2460 4014-4016 IN denotes by
T2461 4017-4023 NNP denotes Sanofi
T2462 4024-4034 NNPS denotes Biosystems
T2463 4035-4043 NNP denotes Industry
T2464 4043-4044 , denotes ,
T2465 4045-4065 NNP denotes Boulogne-Billancourt
T2466 4065-4066 , denotes ,
T2467 4067-4073 NNP denotes France
T2468 4073-4074 -RRB- denotes )
T2469 4074-4075 . denotes .
T2470 4076-4082 JJ denotes Native
T2471 4083-4094 NN denotes carrageenan
T2472 4095-4098 VBD denotes was
T2473 4099-4108 VBN denotes dissolved
T2474 4109-4111 IN denotes in
T2475 4112-4121 JJ denotes distilled
T2476 4122-4127 NN denotes water
T2477 4128-4129 -LRB- denotes (
T2478 4129-4130 CD denotes 5
T2479 4130-4131 NN denotes %
T2480 4132-4135 NN denotes w/v
T2481 4135-4136 -RRB- denotes )
T2482 4137-4142 IN denotes under
T2483 4143-4151 JJ denotes vigorous
T2484 4152-4160 VBG denotes stirring
T2485 4161-4164 CC denotes and
T2486 4165-4171 VBN denotes heated
T2487 4172-4174 TO denotes to
T2488 4175-4177 CD denotes 60
T2489 4177-4178 CD denotes °
T2490 4178-4180 NNP denotes C.
T2491 4181-4185 RB denotes Then
T2492 4185-4186 , denotes ,
T2493 4187-4190 DT denotes the
T2494 4191-4202 NN denotes carrageenan
T2495 4203-4211 NN denotes solution
T2496 4212-4215 VBD denotes was
T2497 4216-4225 VBN denotes submitted
T2498 4226-4228 TO denotes to
T2499 4229-4232 CD denotes two
T2500 4233-4242 JJ denotes different
T2501 4243-4253 NNS denotes treatments
T2502 4254-4256 TO denotes to
T2503 4257-4263 VB denotes obtain
T2504 4264-4268 DT denotes both
T2505 4269-4272 JJ denotes low
T2506 4273-4276 CC denotes and
T2507 4277-4283 JJ denotes medium
T2508 4284-4293 JJ denotes molecular
T2509 4294-4300 NN denotes weight
T2510 4301-4310 NNS denotes fractions
T2511 4310-4311 . denotes .
T2512 4312-4319 RB denotes Briefly
T2513 4319-4320 , denotes ,
T2514 4321-4324 IN denotes for
T2515 4325-4328 DT denotes the
T2516 4329-4332 JJ denotes low
T2517 4333-4342 JJ denotes molecular
T2518 4343-4349 NN denotes weight
T2519 4350-4358 NN denotes fraction
T2520 4358-4359 , denotes ,
T2521 4360-4371 NN denotes carrageenan
T2522 4372-4380 NN denotes solution
T2523 4381-4384 VBD denotes was
T2524 4385-4395 VBN denotes hydrolyzed
T2525 4396-4400 IN denotes with
T2526 4401-4404 CD denotes 0.3
T2527 4404-4405 NN denotes %
T2528 4406-4407 -LRB- denotes (
T2529 4407-4410 FW denotes v/v
T2530 4410-4411 -RRB- denotes )
T2531 4412-4424 VBD denotes concentrated
T2532 4425-4434 JJ denotes sulphuric
T2533 4435-4439 NN denotes acid
T2534 4440-4443 IN denotes for
T2535 4444-4446 CD denotes 15
T2536 4447-4450 NN denotes min
T2537 4451-4453 IN denotes at
T2538 4454-4456 CD denotes 80
T2539 4456-4457 CD denotes °
T2540 4457-4458 NNP denotes C
T2541 4458-4459 . denotes .
T2542 4460-4465 IN denotes After
T2543 4466-4480 NN denotes neutralization
T2544 4481-4485 IN denotes with
T2545 4486-4490 NNP denotes NaOH
T2546 4491-4493 NNP denotes 4N
T2547 4493-4494 , denotes ,
T2548 4495-4498 DT denotes the
T2549 4499-4507 NN denotes solution
T2550 4508-4511 VBD denotes was
T2551 4512-4517 JJ denotes ultra
T2552 4518-4526 VBN denotes filtered
T2553 4527-4534 IN denotes through
T2554 4535-4536 DT denotes a
T2555 4537-4543 JJ denotes hollow
T2556 4544-4549 NN denotes fibre
T2557 4550-4559 NN denotes cartridge
T2558 4560-4564 IN denotes with
T2559 4565-4567 NNP denotes MW
T2560 4568-4575 NN denotes cut-off
T2561 4576-4577 CD denotes 5
T2562 4578-4581 NNP denotes kDa
T2563 4581-4582 , denotes ,
T2564 4583-4584 -LRB- denotes (
T2565 4584-4590 NNP denotes Amicon
T2566 4591-4594 NNP denotes Inc
T2567 4594-4595 , denotes ,
T2568 4596-4603 NNP denotes Beverly
T2569 4603-4604 , denotes ,
T2570 4605-4608 NNP denotes USA
T2571 4608-4609 -RRB- denotes )
T2572 4609-4610 . denotes .
T2573 4611-4614 IN denotes For
T2574 4615-4618 DT denotes the
T2575 4619-4625 NN denotes medium
T2576 4626-4635 JJ denotes molecular
T2577 4636-4642 NN denotes weight
T2578 4643-4651 NN denotes fraction
T2579 4651-4652 , denotes ,
T2580 4653-4656 DT denotes the
T2581 4657-4668 NN denotes carrageenan
T2582 4669-4677 NN denotes solution
T2583 4678-4681 VBD denotes was
T2584 4682-4692 VBN denotes hydrolyzed
T2585 4693-4697 IN denotes with
T2586 4698-4701 CD denotes 0.3
T2587 4701-4702 NN denotes %
T2588 4703-4704 -LRB- denotes (
T2589 4704-4707 FW denotes v/v
T2590 4707-4708 -RRB- denotes )
T2591 4709-4721 VBD denotes concentrated
T2592 4722-4731 JJ denotes sulphuric
T2593 4732-4736 NN denotes acid
T2594 4737-4740 IN denotes for
T2595 4741-4743 CD denotes 30
T2596 4744-4747 NN denotes min
T2597 4748-4750 IN denotes at
T2598 4751-4753 CD denotes 60
T2599 4753-4754 CD denotes °
T2600 4754-4755 NNP denotes C
T2601 4755-4756 . denotes .
T2602 4757-4762 IN denotes After
T2603 4763-4777 NN denotes neutralization
T2604 4777-4778 , denotes ,
T2605 4779-4782 DT denotes the
T2606 4783-4794 NN denotes supernatant
T2607 4795-4798 VBD denotes was
T2608 4799-4804 JJ denotes ultra
T2609 4805-4813 VBN denotes filtered
T2610 4814-4815 -LRB- denotes (
T2611 4815-4817 NNP denotes MW
T2612 4818-4825 NN denotes cut-off
T2613 4826-4829 CD denotes 100
T2614 4830-4833 NNP denotes kDa
T2615 4833-4834 -RRB- denotes )
T2616 4834-4835 . denotes .
T2617 4836-4839 DT denotes The
T2618 4840-4848 NN denotes filtrate
T2619 4849-4852 VBD denotes was
T2620 4853-4862 VBN denotes submitted
T2621 4863-4865 TO denotes to
T2622 4866-4867 DT denotes a
T2623 4868-4874 JJ denotes second
T2624 4875-4880 NN denotes ultra
T2625 4881-4891 NN denotes filtration
T2626 4892-4893 -LRB- denotes (
T2627 4893-4895 NNP denotes MW
T2628 4896-4903 NN denotes cut-off
T2629 4904-4905 CD denotes 5
T2630 4906-4909 NNP denotes kDa
T2631 4909-4910 -RRB- denotes )
T2632 4910-4911 . denotes .
T2633 4912-4916 DT denotes Both
T2634 4917-4929 NNS denotes preparations
T2635 4930-4932 IN denotes of
T2636 4933-4937 NNP denotes dCGN
T2637 4938-4942 VBD denotes were
T2638 4943-4955 VBN denotes precipitated
T2639 4956-4960 IN denotes with
T2640 4961-4962 CD denotes 4
T2641 4963-4970 NNS denotes volumes
T2642 4971-4973 IN denotes of
T2643 4974-4976 CD denotes 95
T2644 4976-4977 NN denotes %
T2645 4978-4985 NN denotes ethanol
T2646 4985-4986 , denotes ,
T2647 4987-4992 VBD denotes dried
T2648 4993-4995 IN denotes at
T2649 4996-5000 NN denotes room
T2650 5001-5012 NN denotes temperature
T2651 5013-5016 CC denotes and
T2652 5017-5023 NN denotes ground
T2653 5024-5026 TO denotes to
T2654 5027-5032 JJ denotes small
T2655 5033-5042 NNS denotes particles
T2656 5043-5044 -LRB- denotes (
T2657 5044-5045 CD denotes 1
T2658 5046-5048 NN denotes mm
T2659 5049-5051 IN denotes in
T2660 5052-5060 NN denotes diameter
T2661 5060-5061 -RRB- denotes )
T2662 5061-5062 . denotes .
T2663 5063-5068 VBG denotes Using
T2664 5069-5083 NN denotes gel-permeation
T2665 5084-5098 NN denotes chromatography
T2666 5099-5101 IN denotes in
T2667 5102-5113 NN denotes combination
T2668 5114-5118 IN denotes with
T2669 5119-5124 JJ denotes light
T2670 5125-5135 VBG denotes scattering
T2671 5136-5148 NNS denotes measurements
T2672 5149-5150 -LRB- denotes (
T2673 5150-5153 VB denotes see
T2674 5154-5160 NNP denotes Viebke
T2675 5161-5163 FW denotes et
T2676 5164-5167 FW denotes al.
T2677 5168-5169 NNP denotes [
T2678 5169-5171 CD denotes 21
T2679 5171-5172 NNP denotes ]
T2680 5172-5173 -RRB- denotes )
T2681 5173-5174 , denotes ,
T2682 5175-5177 PRP denotes it
T2683 5178-5181 VBD denotes was
T2684 5182-5191 VBN denotes confirmed
T2685 5192-5196 IN denotes that
T2686 5197-5200 DT denotes the
T2687 5201-5204 JJ denotes low
T2688 5205-5213 NN denotes fraction
T2689 5214-5217 VBD denotes had
T2690 5218-5220 DT denotes an
T2691 5221-5228 JJ denotes average
T2692 5229-5238 JJ denotes molecular
T2693 5239-5245 NN denotes weight
T2694 5246-5248 IN denotes of
T2695 5249-5251 CD denotes 10
T2696 5252-5255 NN denotes kDa
T2697 5255-5256 , denotes ,
T2698 5257-5260 CC denotes and
T2699 5261-5264 DT denotes the
T2700 5265-5271 NN denotes medium
T2701 5272-5280 NN denotes fraction
T2702 5281-5283 IN denotes of
T2703 5284-5286 CD denotes 40
T2704 5287-5290 NN denotes kDa
T2705 5290-5291 . denotes .
T2706 5292-5295 DT denotes The
T2707 5296-5304 NN denotes sulphate
T2708 5305-5312 NN denotes content
T2709 5313-5315 IN denotes of
T2710 5316-5331 NNS denotes polysaccharides
T2711 5332-5334 IN denotes in
T2712 5335-5339 DT denotes both
T2713 5340-5349 NNS denotes fractions
T2714 5350-5353 VBD denotes was
T2715 5354-5362 VBN denotes measured
T2716 5363-5372 VBG denotes following
T2717 5373-5376 DT denotes the
T2718 5377-5383 NN denotes method
T2719 5384-5386 IN denotes of
T2720 5387-5395 NNP denotes Quemener
T2721 5396-5398 FW denotes et
T2722 5399-5402 FW denotes al.
T2723 5403-5404 NNP denotes [
T2724 5404-5406 CD denotes 22
T2725 5406-5407 NNP denotes ]
T2726 5407-5408 . denotes .
T2727 5409-5416 RB denotes Finally
T2728 5416-5417 , denotes ,
T2729 5418-5421 DT denotes the
T2730 5422-5429 NN denotes absence
T2731 5430-5432 IN denotes of
T2732 5433-5447 NN denotes polysaccharide
T2733 5448-5457 NN denotes structure
T2734 5458-5471 NNS denotes modifications
T2735 5472-5474 IN denotes in
T2736 5475-5478 DT denotes the
T2737 5479-5482 CD denotes two
T2738 5483-5492 NNS denotes fractions
T2739 5493-5496 VBD denotes was
T2740 5497-5506 VBN denotes confirmed
T2741 5507-5512 VBG denotes using
T2742 5513-5519 JJ denotes 2H-NMR
T2743 5520-5532 NN denotes spectroscopy
T2744 5532-5533 . denotes .
T2745 5534-5537 DT denotes The
T2746 5538-5545 NN denotes absence
T2747 5546-5548 IN denotes of
T2748 5549-5552 NNP denotes LPS
T2749 5553-5566 NN denotes contamination
T2750 5567-5569 IN denotes in
T2751 5570-5573 DT denotes the
T2752 5574-5577 CD denotes two
T2753 5578-5587 NNS denotes fractions
T2754 5588-5591 VBD denotes was
T2755 5592-5601 VBN denotes confirmed
T2756 5602-5607 VBG denotes using
T2757 5608-5611 DT denotes the
T2758 5612-5620 JJ denotes e-Toxate
T2759 5620-5621 NN denotes ®
T2760 5622-5625 NN denotes kit
T2761 5626-5627 -LRB- denotes (
T2762 5627-5632 NNP denotes Sigma
T2763 5632-5633 , denotes ,
T2764 5634-5636 NNP denotes St
T2765 5637-5644 NNP denotes Quentin
T2766 5645-5654 NNP denotes Fallavier
T2767 5654-5655 , denotes ,
T2768 5656-5662 NNP denotes France
T2769 5662-5663 -RRB- denotes )
T2770 5663-5664 . denotes .
T2771 5665-5671 IN denotes Before
T2772 5672-5675 NN denotes use
T2773 5676-5678 IN denotes in
T2774 5679-5683 NN denotes cell
T2775 5684-5691 NN denotes culture
T2776 5691-5692 , denotes ,
T2777 5693-5696 DT denotes the
T2778 5697-5700 CD denotes two
T2779 5701-5710 NNS denotes fractions
T2780 5711-5715 VBD denotes were
T2781 5716-5725 VBN denotes dissolved
T2782 5726-5728 IN denotes in
T2783 5729-5737 JJ denotes complete
T2784 5738-5744 NN denotes medium
T2785 5745-5751 IN denotes during
T2786 5752-5754 CD denotes 30
T2787 5755-5758 NN denotes min
T2788 5759-5761 IN denotes at
T2789 5762-5764 CD denotes 56
T2790 5764-5765 CD denotes °
T2791 5765-5767 NNP denotes C.
T2945 5769-5776 NNS denotes Animals
T2946 5776-5777 , denotes ,
T2947 5778-5787 NNPS denotes Chemicals
T2948 5788-5791 CC denotes and
T2949 5792-5796 NNP denotes Diet
T2950 5797-5801 NNP denotes Male
T2951 5802-5808 NNP denotes Wistar
T2952 5809-5813 NNS denotes rats
T2953 5814-5815 -LRB- denotes (
T2954 5815-5818 CD denotes 150
T2955 5819-5820 JJ denotes g
T2956 5821-5828 JJ denotes average
T2957 5829-5835 NN denotes weight
T2958 5835-5836 -RRB- denotes )
T2959 5837-5841 VBD denotes were
T2960 5842-5848 VBN denotes housed
T2961 5849-5854 IN denotes under
T2962 5855-5863 JJ denotes standard
T2963 5864-5874 NNS denotes conditions
T2964 5875-5878 CC denotes and
T2965 5879-5882 VBN denotes fed
T2966 5883-5885 NN denotes ad
T2967 5886-5893 NN denotes libitum
T2968 5894-5898 IN denotes with
T2969 5899-5907 JJ denotes standard
T2970 5908-5914 NN denotes rodent
T2971 5915-5925 NN denotes laboratory
T2972 5926-5930 NN denotes chow
T2973 5930-5931 . denotes .
T2974 5932-5940 JJ denotes Degraded
T2975 5941-5958 NNS denotes iota-carrageenans
T2976 5959-5963 VBD denotes were
T2977 5964-5976 VBN denotes administered
T2978 5977-5979 IN denotes in
T2979 5980-5983 DT denotes the
T2980 5984-5992 NN denotes drinking
T2981 5993-5998 NN denotes water
T2982 5999-6000 -LRB- denotes (
T2983 6000-6001 CD denotes 5
T2984 6001-6002 NN denotes %
T2985 6003-6006 NN denotes w/v
T2986 6006-6007 -RRB- denotes )
T2987 6008-6011 IN denotes for
T2988 6012-6014 CD denotes 55
T2989 6015-6019 NNS denotes days
T2990 6020-6022 TO denotes to
T2991 6023-6024 CD denotes 2
T2992 6025-6031 NNS denotes groups
T2993 6032-6034 IN denotes of
T2994 6035-6038 CD denotes six
T2995 6039-6046 NNS denotes animals
T2996 6047-6051 DT denotes each
T2997 6051-6052 . denotes .
T2998 6053-6056 DT denotes The
T2999 6057-6062 JJ denotes first
T3000 6063-6068 NN denotes group
T3001 6069-6077 VBD denotes received
T3002 6078-6081 DT denotes the
T3003 6082-6085 JJ denotes low
T3004 6086-6095 JJ denotes molecular
T3005 6096-6102 NN denotes weight
T3006 6103-6114 NN denotes carrageenan
T3007 6115-6116 -LRB- denotes (
T3008 6116-6118 CD denotes 10
T3009 6119-6122 NNP denotes kDa
T3010 6123-6127 NNP denotes dCGN
T3011 6127-6128 -RRB- denotes )
T3012 6129-6132 CC denotes and
T3013 6133-6136 DT denotes the
T3014 6137-6143 JJ denotes second
T3015 6144-6152 VBD denotes received
T3016 6153-6156 DT denotes the
T3017 6157-6163 NN denotes medium
T3018 6164-6173 JJ denotes molecular
T3019 6174-6180 NN denotes weight
T3020 6181-6192 NN denotes carrageenan
T3021 6193-6194 -LRB- denotes (
T3022 6194-6196 CD denotes 40
T3023 6197-6200 NNP denotes kDa
T3024 6201-6205 NNP denotes dCGN
T3025 6205-6206 -RRB- denotes )
T3026 6206-6207 . denotes .
T3027 6208-6210 DT denotes An
T3028 6211-6221 JJ denotes additional
T3029 6222-6227 NN denotes group
T3030 6228-6230 IN denotes of
T3031 6231-6235 CD denotes four
T3032 6236-6240 NNS denotes rats
T3033 6241-6245 VBD denotes were
T3034 6246-6256 VBN denotes maintained
T3035 6257-6259 IN denotes on
T3036 6260-6267 JJ denotes regular
T3037 6268-6271 VBP denotes tap
T3038 6272-6277 NN denotes water
T3039 6278-6279 -LRB- denotes (
T3040 6279-6286 NN denotes control
T3041 6287-6292 NN denotes group
T3042 6292-6293 -RRB- denotes )
T3043 6293-6294 . denotes .
T3044 6295-6297 TO denotes To
T3045 6298-6306 VB denotes increase
T3046 6307-6319 NN denotes palatability
T3047 6320-6323 CD denotes 0.2
T3048 6323-6324 NN denotes %
T3049 6325-6332 NN denotes sucrose
T3050 6333-6336 VBD denotes was
T3051 6337-6342 VBN denotes added
T3052 6343-6345 TO denotes to
T3053 6346-6349 DT denotes the
T3054 6350-6358 NN denotes drinking
T3055 6359-6364 NN denotes water
T3056 6365-6367 IN denotes of
T3057 6368-6371 DT denotes all
T3058 6372-6378 NNS denotes groups
T3059 6379-6380 -LRB- denotes (
T3060 6380-6383 NNP denotes Van
T3061 6384-6387 NN denotes der
T3062 6388-6393 NNP denotes Waaji
T3063 6394-6396 FW denotes et
T3064 6397-6400 FW denotes al.
T3065 6400-6401 , denotes ,
T3066 6402-6403 NNP denotes [
T3067 6403-6405 CD denotes 23
T3068 6405-6406 NNP denotes ]
T3069 6406-6407 -RRB- denotes )
T3070 6407-6408 . denotes .
T3071 6409-6414 JJ denotes Fresh
T3072 6415-6426 NN denotes carrageenan
T3073 6427-6436 NNS denotes solutions
T3074 6437-6441 VBD denotes were
T3075 6442-6450 JJ denotes prepared
T3076 6451-6456 JJ denotes daily
T3077 6456-6457 . denotes .
T3368 6459-6469 NN denotes Evaluation
T3369 6470-6472 IN denotes of
T3370 6473-6480 NNP denotes Colitis
T3371 6481-6485 NNP denotes Body
T3372 6486-6492 NN denotes weight
T3373 6492-6493 , denotes ,
T3374 6494-6500 NN denotes liquid
T3375 6501-6504 CC denotes and
T3376 6505-6509 NN denotes food
T3377 6510-6521 NN denotes consumption
T3378 6521-6522 , denotes ,
T3379 6523-6531 NN denotes diarrhea
T3380 6532-6535 CC denotes and
T3381 6536-6542 JJ denotes rectal
T3382 6543-6551 NN denotes bleeding
T3383 6552-6553 -LRB- denotes (
T3384 6553-6561 VBN denotes detected
T3385 6562-6564 IN denotes by
T3386 6565-6568 NN denotes eye
T3387 6569-6579 NN denotes inspection
T3388 6579-6580 -RRB- denotes )
T3389 6581-6585 VBD denotes were
T3390 6586-6594 VBN denotes recorded
T3391 6595-6605 IN denotes throughout
T3392 6606-6609 DT denotes the
T3393 6610-6617 NN denotes feeding
T3394 6618-6624 NN denotes period
T3395 6624-6625 . denotes .
T3396 6626-6631 IN denotes After
T3397 6632-6634 CD denotes 55
T3398 6635-6639 NNS denotes days
T3399 6639-6640 , denotes ,
T3400 6641-6648 NNS denotes animals
T3401 6649-6653 VBD denotes were
T3402 6654-6664 VBN denotes sacrificed
T3403 6665-6667 IN denotes by
T3404 6668-6676 JJ denotes cervical
T3405 6677-6688 NN denotes dislocation
T3406 6688-6689 . denotes .
T3407 6690-6693 DT denotes The
T3408 6694-6700 NN denotes length
T3409 6701-6703 IN denotes of
T3410 6704-6707 DT denotes the
T3411 6708-6713 NN denotes colon
T3412 6714-6717 VBD denotes was
T3413 6718-6726 VBN denotes measured
T3414 6727-6729 IN denotes as
T3415 6730-6739 VBN denotes described
T3416 6740-6742 IN denotes by
T3417 6743-6751 NNP denotes Okayashu
T3418 6752-6754 FW denotes et
T3419 6755-6758 FW denotes al.
T3420 6759-6760 NNP denotes [
T3421 6760-6762 CD denotes 24
T3422 6762-6763 NNP denotes ]
T3423 6763-6764 . denotes .
T3424 6765-6769 RB denotes Then
T3425 6769-6770 , denotes ,
T3426 6771-6775 DT denotes each
T3427 6776-6781 NN denotes colon
T3428 6782-6785 VBD denotes was
T3429 6786-6793 VBN denotes ligated
T3430 6794-6796 IN denotes in
T3431 6797-6805 NNS denotes sections
T3432 6806-6808 IN denotes of
T3433 6809-6810 CD denotes 2
T3434 6811-6813 NN denotes cm
T3435 6814-6817 CC denotes and
T3436 6818-6819 CD denotes 1
T3437 6820-6822 TO denotes to
T3438 6823-6824 CD denotes 2
T3439 6825-6827 NN denotes ml
T3440 6828-6830 IN denotes of
T3441 6831-6833 CD denotes 10
T3442 6833-6834 NN denotes %
T3443 6835-6843 NN denotes formalin
T3444 6844-6847 VBD denotes was
T3445 6848-6855 VBN denotes infused
T3446 6856-6860 IN denotes into
T3447 6861-6864 DT denotes the
T3448 6865-6875 JJ denotes intestinal
T3449 6876-6881 NNS denotes lumen
T3450 6881-6882 . denotes .
T3451 6883-6886 DT denotes The
T3452 6887-6897 RB denotes moderately
T3453 6898-6907 JJ denotes distended
T3454 6908-6915 NN denotes segment
T3455 6916-6919 VBD denotes was
T3456 6920-6929 VBN denotes sectioned
T3457 6930-6933 CC denotes and
T3458 6934-6939 VBN denotes fixed
T3459 6940-6942 IN denotes in
T3460 6943-6945 CD denotes 10
T3461 6945-6946 NN denotes %
T3462 6947-6955 NN denotes formalin
T3463 6955-6956 . denotes .
T3464 6957-6960 DT denotes The
T3465 6961-6970 JJ denotes following
T3466 6971-6974 NN denotes day
T3467 6974-6975 , denotes ,
T3468 6976-6979 DT denotes the
T3469 6980-6990 JJ denotes intestinal
T3470 6991-6998 NN denotes content
T3471 6999-7002 VBD denotes was
T3472 7003-7010 VBN denotes removed
T3473 7011-7013 IN denotes by
T3474 7014-7023 VBG denotes vortexing
T3475 7023-7024 . denotes .
T3476 7025-7028 DT denotes The
T3477 7029-7034 VBN denotes fixed
T3478 7035-7042 NN denotes segment
T3479 7043-7046 VBD denotes was
T3480 7047-7051 VBN denotes kept
T3481 7052-7054 IN denotes in
T3482 7055-7057 CD denotes 10
T3483 7057-7058 NN denotes %
T3484 7059-7067 NN denotes formalin
T3485 7068-7070 IN denotes at
T3486 7071-7072 CD denotes 4
T3487 7072-7073 CD denotes °
T3488 7073-7074 NNP denotes C
T3489 7075-7080 IN denotes until
T3490 7081-7084 DT denotes the
T3491 7085-7093 NN denotes paraffin
T3492 7094-7103 NN denotes embedding
T3493 7104-7113 NN denotes procedure
T3494 7113-7114 . denotes .
T3495 7115-7117 TO denotes To
T3496 7118-7126 VB denotes evaluate
T3497 7127-7130 DT denotes the
T3498 7131-7137 NN denotes degree
T3499 7138-7140 IN denotes of
T3500 7141-7153 NN denotes inflammation
T3501 7153-7154 , denotes ,
T3502 7155-7159 DT denotes this
T3503 7160-7167 NN denotes segment
T3504 7168-7170 IN denotes of
T3505 7171-7176 NN denotes colon
T3506 7177-7180 VBD denotes was
T3507 7181-7187 VBN denotes opened
T3508 7188-7202 RB denotes longitudinally
T3509 7203-7206 CC denotes and
T3510 7207-7218 JJ denotes macroscopic
T3511 7219-7222 CC denotes and
T3512 7223-7235 JJ denotes histological
T3513 7236-7242 NNS denotes scores
T3514 7243-7245 IN denotes of
T3515 7246-7258 NN denotes inflammation
T3516 7259-7263 VBD denotes were
T3517 7264-7272 VBN denotes recorded
T3518 7273-7275 IN denotes as
T3519 7276-7286 RB denotes previously
T3520 7287-7296 VBN denotes described
T3521 7297-7298 NNP denotes [
T3522 7298-7300 CD denotes 25
T3523 7300-7301 NNP denotes ]
T3524 7301-7302 , denotes ,
T3525 7303-7304 NNP denotes [
T3526 7304-7306 CD denotes 26
T3527 7306-7307 NNP denotes ]
T3528 7307-7308 . denotes .
T3529 7309-7312 DT denotes The
T3530 7313-7322 NN denotes toluidine
T3531 7323-7327 JJ denotes blue
T3532 7328-7336 NN denotes staining
T3533 7337-7340 VBD denotes was
T3534 7341-7345 VBN denotes used
T3535 7346-7349 IN denotes for
T3536 7350-7364 NN denotes identification
T3537 7365-7367 IN denotes of
T3538 7368-7377 VBN denotes sulphated
T3539 7378-7393 NNS denotes polysaccharides
T3540 7394-7396 IN denotes in
T3541 7397-7400 DT denotes the
T3542 7401-7411 JJ denotes intestinal
T3543 7412-7418 NN denotes mucosa
T3544 7418-7419 . denotes .
T3545 7420-7422 IN denotes On
T3546 7423-7426 DT denotes the
T3547 7427-7430 NN denotes day
T3548 7431-7433 IN denotes of
T3549 7434-7443 NN denotes sacrifice
T3550 7443-7444 , denotes ,
T3551 7445-7446 DT denotes a
T3552 7447-7452 JJ denotes fresh
T3553 7453-7459 NN denotes sample
T3554 7460-7462 IN denotes of
T3555 7463-7467 DT denotes each
T3556 7468-7473 NN denotes colon
T3557 7474-7475 -LRB- denotes (
T3558 7475-7477 CD denotes 50
T3559 7478-7480 NN denotes mg
T3560 7480-7481 -RRB- denotes )
T3561 7482-7485 VBD denotes was
T3562 7486-7495 VBN denotes collected
T3563 7496-7499 IN denotes for
T3564 7500-7515 NN denotes myeloperoxidase
T3565 7516-7517 -LRB- denotes (
T3566 7517-7520 NNP denotes MPO
T3567 7520-7521 -RRB- denotes )
T3568 7522-7527 JJ denotes assay
T3569 7528-7537 VBG denotes according
T3570 7538-7540 TO denotes to
T3571 7541-7548 NNP denotes Krawisz
T3572 7549-7551 FW denotes et
T3573 7552-7555 FW denotes al.
T3574 7555-7556 , denotes ,
T3575 7557-7558 NNP denotes [
T3576 7558-7560 CD denotes 27
T3577 7560-7561 NNP denotes ]
T3578 7561-7562 . denotes .
T3579 7563-7566 DT denotes The
T3580 7567-7572 NN denotes level
T3581 7573-7575 IN denotes of
T3582 7576-7579 NNP denotes MPO
T3583 7579-7580 , denotes ,
T3584 7581-7587 RB denotes mainly
T3585 7588-7597 VBN denotes expressed
T3586 7598-7600 IN denotes by
T3587 7601-7612 NNS denotes neutrophils
T3588 7612-7613 , denotes ,
T3589 7614-7623 VBZ denotes indicates
T3590 7624-7627 DT denotes the
T3591 7628-7632 NN denotes rate
T3592 7633-7635 IN denotes of
T3593 7636-7647 NN denotes recruitment
T3594 7648-7650 IN denotes of
T3595 7651-7662 NNS denotes neutrophils
T3596 7663-7665 TO denotes to
T3597 7666-7669 DT denotes the
T3598 7670-7680 JJ denotes intestinal
T3599 7681-7687 NN denotes mucosa
T3600 7687-7688 . denotes .
T3601 7689-7692 CD denotes One
T3602 7693-7697 NN denotes unit
T3603 7698-7700 IN denotes of
T3604 7701-7704 NNP denotes MPO
T3605 7705-7713 NN denotes activity
T3606 7714-7725 VBZ denotes corresponds
T3607 7726-7728 TO denotes to
T3608 7729-7732 DT denotes the
T3609 7733-7744 NN denotes degradation
T3610 7745-7747 IN denotes of
T3611 7748-7749 CD denotes 1
T3612 7750-7754 NN denotes µmol
T3613 7755-7757 IN denotes of
T3614 7758-7766 NN denotes peroxide
T3615 7767-7770 IN denotes per
T3616 7771-7777 NN denotes minute
T3617 7778-7780 IN denotes at
T3618 7781-7783 CD denotes 25
T3619 7783-7784 CD denotes °
T3620 7784-7786 NNP denotes C.
T3816 7788-7792 NNP denotes Cell
T3817 7793-7800 NNP denotes Culture
T3818 7801-7804 NNP denotes All
T3819 7805-7811 NN denotes tissue
T3820 7812-7819 NN denotes culture
T3821 7820-7828 NNS denotes reagents
T3822 7829-7833 VBD denotes were
T3823 7834-7838 IN denotes from
T3824 7839-7849 NNP denotes Invitrogen
T3825 7850-7851 -LRB- denotes (
T3826 7851-7856 NNP denotes Cergy
T3827 7857-7865 NNP denotes Pontoise
T3828 7865-7866 , denotes ,
T3829 7867-7873 NNP denotes France
T3830 7873-7874 -RRB- denotes )
T3831 7874-7875 . denotes .
T3832 7876-7881 CD denotes THP-1
T3833 7882-7887 JJ denotes human
T3834 7888-7897 JJ denotes monocytic
T3835 7898-7903 NNS denotes cells
T3836 7904-7908 VBD denotes were
T3837 7909-7919 VBN denotes maintained
T3838 7920-7922 IN denotes in
T3839 7923-7932 NN denotes RPMI-1640
T3840 7933-7945 VBN denotes supplemented
T3841 7946-7950 IN denotes with
T3842 7951-7953 CD denotes 10
T3843 7953-7954 NN denotes %
T3844 7955-7958 NNP denotes FCS
T3845 7958-7959 , denotes ,
T3846 7960-7961 CD denotes 2
T3847 7962-7964 NNP denotes mM
T3848 7965-7966 NNP denotes L
T3849 7967-7968 : denotes -
T3850 7968-7977 NN denotes glutamine
T3851 7977-7978 , denotes ,
T3852 7979-7981 CD denotes 50
T3853 7982-7986 NN denotes U/ml
T3854 7987-7997 NN denotes penicillin
T3855 7998-8001 CC denotes and
T3856 8002-8004 CD denotes 50
T3857 8005-8010 JJ denotes mg/ml
T3858 8011-8023 NN denotes streptomycin
T3859 8024-8026 IN denotes at
T3860 8027-8029 CD denotes 37
T3861 8029-8030 CD denotes °
T3862 8030-8031 NNP denotes C
T3863 8032-8034 IN denotes in
T3864 8035-8036 DT denotes a
T3865 8037-8038 CD denotes 5
T3866 8038-8039 NN denotes %
T3867 8040-8043 CD denotes CO2
T3868 8044-8053 NN denotes incubator
T3869 8053-8054 . denotes .
T3870 8055-8060 JJ denotes Human
T3871 8061-8071 JJ denotes peripheral
T3872 8072-8077 NN denotes blood
T3873 8078-8089 NN denotes mononuclear
T3874 8090-8095 NNS denotes cells
T3875 8096-8100 VBD denotes were
T3876 8101-8109 VBN denotes obtained
T3877 8110-8114 IN denotes from
T3878 8115-8126 VBN denotes heparinized
T3879 8127-8132 NN denotes blood
T3880 8133-8135 IN denotes by
T3881 8136-8150 JJ denotes Ficoll-Hypaque
T3882 8151-8158 NN denotes density
T3883 8159-8167 NN denotes gradient
T3884 8167-8168 . denotes .
T3885 8169-8178 NNS denotes Monocytes
T3886 8179-8183 VBD denotes were
T3887 8184-8188 RB denotes then
T3888 8189-8197 VBN denotes isolated
T3889 8198-8200 IN denotes by
T3890 8201-8210 NN denotes adherence
T3891 8211-8213 TO denotes to
T3892 8214-8221 NN denotes culture
T3893 8222-8228 NNS denotes flasks
T3894 8229-8231 IN denotes as
T3895 8232-8241 VBN denotes described
T3896 8242-8243 NNP denotes [
T3897 8243-8245 CD denotes 28
T3898 8245-8246 NNP denotes ]
T3899 8246-8247 . denotes .
T3900 8248-8251 IN denotes For
T3901 8252-8256 NN denotes cell
T3902 8257-8268 NN denotes aggregation
T3903 8268-8269 , denotes ,
T3904 8270-8279 NNS denotes monocytes
T3905 8280-8284 VBD denotes were
T3906 8285-8293 VBN denotes cultured
T3907 8294-8296 IN denotes in
T3908 8297-8300 DT denotes the
T3909 8301-8309 NN denotes presence
T3910 8310-8312 CC denotes or
T3911 8313-8320 NN denotes absence
T3912 8321-8323 IN denotes of
T3913 8324-8327 CD denotes C10
T3914 8328-8330 CC denotes or
T3915 8331-8334 CD denotes C40
T3916 8335-8338 IN denotes for
T3917 8339-8341 CD denotes 72
T3918 8342-8344 NN denotes h.
T3919 8345-8349 NNP denotes Cell
T3920 8350-8358 NNS denotes colonies
T3921 8359-8363 VBD denotes were
T3922 8364-8373 VBN denotes monitored
T3923 8374-8379 IN denotes under
T3924 8380-8382 DT denotes an
T3925 8383-8391 JJ denotes inverted
T3926 8392-8397 NN denotes phase
T3927 8398-8406 NN denotes contrast
T3928 8407-8417 NN denotes microscope
T3929 8418-8425 VBN denotes coupled
T3930 8426-8433 IN denotes through
T3931 8434-8435 DT denotes a
T3932 8436-8441 NN denotes video
T3933 8442-8448 NN denotes camera
T3934 8449-8451 TO denotes to
T3935 8452-8453 DT denotes a
T3936 8454-8462 NN denotes computer
T3937 8462-8463 . denotes .
T3938 8464-8466 IN denotes In
T3939 8467-8471 DT denotes some
T3940 8472-8477 NNS denotes wells
T3941 8477-8478 , denotes ,
T3942 8479-8491 VBG denotes neutralizing
T3943 8492-8502 JJ denotes monoclonal
T3944 8503-8511 NN denotes antibody
T3945 8512-8514 TO denotes to
T3946 8515-8521 NNP denotes ICAM-1
T3947 8522-8523 -LRB- denotes (
T3948 8523-8526 CD denotes 2.5
T3949 8527-8529 NN denotes µg
T3950 8529-8530 NN denotes /
T3951 8530-8532 NN denotes ml
T3952 8532-8533 -RRB- denotes )
T3953 8534-8535 -LRB- denotes (
T3954 8535-8539 NNP denotes Tebu
T3955 8539-8540 , denotes ,
T3956 8541-8543 NNP denotes Le
T3957 8544-8550 NNP denotes Perray
T3958 8551-8553 IN denotes en
T3959 8554-8562 NNP denotes Yvelines
T3960 8562-8563 , denotes ,
T3961 8564-8570 NNP denotes France
T3962 8570-8571 -RRB- denotes )
T3963 8572-8575 VBD denotes was
T3964 8576-8581 VBN denotes added
T3965 8581-8582 . denotes .
T4100 8584-8588 NNP denotes Cell
T4101 8589-8594 NNP denotes Cycle
T4102 8595-8603 NNP denotes Analysis
T4103 8604-8609 NNP denotes THP-1
T4104 8610-8615 NNS denotes cells
T4105 8616-8618 IN denotes in
T4106 8619-8630 JJ denotes exponential
T4107 8631-8637 NN denotes growth
T4108 8638-8643 NN denotes phase
T4109 8644-8648 VBD denotes were
T4110 8649-8656 VBN denotes exposed
T4111 8657-8659 TO denotes to
T4112 8660-8668 VB denotes complete
T4113 8669-8675 NN denotes medium
T4114 8676-8678 IN denotes in
T4115 8679-8682 DT denotes the
T4116 8683-8691 NN denotes presence
T4117 8692-8694 CC denotes or
T4118 8695-8702 NN denotes absence
T4119 8703-8705 IN denotes of
T4120 8706-8718 NNS denotes carrageenans
T4121 8719-8722 IN denotes for
T4122 8723-8725 CD denotes 24
T4123 8726-8727 NN denotes h
T4124 8728-8734 IN denotes before
T4125 8735-8740 VBG denotes being
T4126 8741-8748 VBN denotes stained
T4127 8749-8753 IN denotes with
T4128 8754-8763 NN denotes propidium
T4129 8764-8770 NN denotes iodide
T4130 8771-8776 VBG denotes using
T4131 8777-8780 DT denotes the
T4132 8781-8789 NNP denotes DNA-Prep
T4133 8790-8797 NNP denotes Coulter
T4134 8798-8801 NN denotes kit
T4135 8802-8811 VBG denotes according
T4136 8812-8814 TO denotes to
T4137 8815-8818 DT denotes the
T4138 8819-8831 NN denotes manufacturer
T4139 8831-8833 POS denotes 's
T4140 8834-8845 NN denotes instruction
T4141 8846-8847 -LRB- denotes (
T4142 8847-8862 NNP denotes Beckman-Coulter
T4143 8862-8863 , denotes ,
T4144 8864-8874 NNP denotes Villepinte
T4145 8874-8875 , denotes ,
T4146 8876-8882 NNP denotes France
T4147 8882-8883 -RRB- denotes )
T4148 8883-8884 . denotes .
T4149 8885-8889 NNP denotes Cell
T4150 8890-8893 NNP denotes DNA
T4151 8894-8901 NN denotes content
T4152 8902-8905 VBD denotes was
T4153 8906-8910 RB denotes then
T4154 8911-8919 VBN denotes analyzed
T4155 8920-8922 IN denotes by
T4156 8923-8927 NN denotes flow
T4157 8928-8937 NN denotes cytometry
T4158 8938-8943 VBG denotes using
T4159 8944-8946 DT denotes an
T4160 8947-8952 NNP denotes EPICS
T4161 8953-8956 NNP denotes XL2
T4162 8957-8958 -LRB- denotes (
T4163 8958-8973 NNP denotes Beckman-Coulter
T4164 8973-8974 -RRB- denotes )
T4165 8974-8975 . denotes .
T4166 8976-8979 JJ denotes Raw
T4167 8980-8984 NNS denotes data
T4168 8985-8988 IN denotes for
T4169 8989-8992 DT denotes the
T4170 8993-9005 NN denotes distribution
T4171 9006-9008 IN denotes of
T4172 9009-9012 NNP denotes DNA
T4173 9013-9020 NN denotes content
T4174 9021-9023 IN denotes of
T4175 9024-9030 CD denotes 30,000
T4176 9031-9036 NNS denotes cells
T4177 9037-9046 VBN denotes retrieved
T4178 9047-9051 IN denotes from
T4179 9052-9055 DT denotes the
T4180 9056-9065 NN denotes cytometer
T4181 9066-9070 VBD denotes were
T4182 9071-9080 VBN denotes expressed
T4183 9081-9083 IN denotes as
T4184 9084-9087 DT denotes the
T4185 9088-9098 NN denotes percentage
T4186 9099-9101 IN denotes of
T4187 9102-9107 CD denotes G0/G1
T4188 9108-9115 IN denotes through
T4189 9116-9120 NNP denotes G2/M
T4190 9121-9132 NNS denotes populations
T4191 9132-9133 . denotes .
T4192 9134-9144 NNP denotes Multicycle
T4193 9145-9147 NNP denotes AV
T4194 9148-9156 NN denotes software
T4195 9157-9158 -LRB- denotes (
T4196 9158-9165 NNP denotes Phoenix
T4197 9166-9170 NNP denotes Flow
T4198 9171-9178 NNPS denotes Systems
T4199 9178-9179 , denotes ,
T4200 9180-9183 NNP denotes San
T4201 9184-9189 NNP denotes Diego
T4202 9189-9190 , denotes ,
T4203 9191-9193 NNP denotes CA
T4204 9193-9194 -RRB- denotes )
T4205 9195-9198 VBD denotes was
T4206 9199-9203 VBN denotes used
T4207 9204-9206 TO denotes to
T4208 9207-9215 VB denotes generate
T4209 9216-9219 NNP denotes DNA
T4210 9220-9227 JJ denotes content
T4211 9228-9237 NN denotes frequency
T4212 9238-9248 NNS denotes histograms
T4213 9249-9252 CC denotes and
T4214 9253-9263 VB denotes facilitate
T4215 9264-9268 NNS denotes data
T4216 9269-9277 NN denotes analysis
T4217 9277-9278 . denotes .
T4382 9280-9284 NNP denotes Cell
T4383 9285-9292 NNP denotes Surface
T4384 9293-9300 NNP denotes Antigen
T4385 9301-9311 NNP denotes Expression
T4386 9312-9320 NNP denotes Analysis
T4387 9321-9331 NNP denotes Peripheral
T4388 9332-9337 NNP denotes Blood
T4389 9338-9347 NNPS denotes Monocytes
T4390 9348-9350 CC denotes or
T4391 9351-9356 CD denotes THP-1
T4392 9357-9362 NNS denotes cells
T4393 9363-9367 VBD denotes were
T4394 9368-9375 VBN denotes exposed
T4395 9376-9378 TO denotes to
T4396 9379-9387 VB denotes complete
T4397 9388-9394 NN denotes medium
T4398 9395-9397 IN denotes in
T4399 9398-9401 DT denotes the
T4400 9402-9410 NN denotes presence
T4401 9411-9413 CC denotes or
T4402 9414-9421 NN denotes absence
T4403 9422-9424 IN denotes of
T4404 9425-9436 NN denotes carrageenan
T4405 9437-9440 IN denotes for
T4406 9441-9443 CD denotes 36
T4407 9444-9445 NN denotes h
T4408 9445-9446 . denotes .
T4409 9447-9452 IN denotes After
T4410 9453-9456 CD denotes two
T4411 9457-9463 NNS denotes washes
T4412 9464-9466 IN denotes in
T4413 9467-9470 NNP denotes PBS
T4414 9471-9478 IN denotes without
T4415 9479-9482 NNP denotes Ca2
T4416 9482-9483 NNP denotes +
T4417 9484-9487 CC denotes and
T4418 9488-9491 NNP denotes Mg2
T4419 9491-9492 NNP denotes +
T4420 9492-9493 , denotes ,
T4421 9494-9499 NNS denotes cells
T4422 9500-9504 VBD denotes were
T4423 9505-9514 VBN denotes incubated
T4424 9515-9517 IN denotes in
T4425 9518-9521 NNP denotes PBS
T4426 9522-9532 VBG denotes containing
T4427 9533-9536 CD denotes 0.1
T4428 9536-9537 NN denotes %
T4429 9538-9545 NN denotes gelatin
T4430 9546-9549 CC denotes and
T4431 9550-9551 CD denotes 8
T4432 9551-9552 NN denotes %
T4433 9553-9555 NNP denotes AB
T4434 9556-9561 JJ denotes human
T4435 9562-9567 NN denotes serum
T4436 9568-9570 TO denotes to
T4437 9571-9578 VB denotes prevent
T4438 9579-9586 JJ denotes binding
T4439 9587-9589 TO denotes to
T4440 9590-9592 VB denotes Fc
T4441 9593-9602 NNS denotes receptors
T4442 9602-9603 . denotes .
T4443 9604-9608 RB denotes Then
T4444 9608-9609 , denotes ,
T4445 9610-9611 CD denotes 5
T4446 9611-9612 NN denotes ×
T4447 9612-9615 CD denotes 105
T4448 9616-9621 NNS denotes cells
T4449 9622-9626 VBD denotes were
T4450 9627-9636 VBN denotes incubated
T4451 9637-9641 IN denotes with
T4452 9642-9649 JJ denotes primary
T4453 9650-9660 NNS denotes antibodies
T4454 9661-9663 IN denotes at
T4455 9664-9665 CD denotes 4
T4456 9665-9666 CD denotes °
T4457 9666-9667 NNP denotes C
T4458 9668-9671 IN denotes for
T4459 9672-9674 CD denotes 30
T4460 9675-9678 NN denotes min
T4461 9678-9679 . denotes .
T4462 9680-9683 CD denotes Two
T4463 9684-9689 JJ denotes other
T4464 9690-9696 NNS denotes washes
T4465 9697-9699 IN denotes in
T4466 9700-9703 NNP denotes PBS
T4467 9704-9712 VBD denotes preceded
T4468 9713-9723 NN denotes incubation
T4469 9724-9728 IN denotes with
T4470 9729-9744 JJ denotes FITC-conjugated
T4471 9745-9749 NN denotes goat
T4472 9750-9758 NN denotes antibody
T4473 9759-9769 JJ denotes anti-mouse
T4474 9770-9773 NNP denotes IgG
T4475 9774-9781 VBD denotes diluted
T4476 9782-9788 CD denotes 1/1000
T4477 9789-9791 IN denotes at
T4478 9792-9793 CD denotes 4
T4479 9793-9794 CD denotes °
T4480 9794-9795 NNP denotes C
T4481 9796-9799 IN denotes for
T4482 9800-9802 CD denotes 30
T4483 9803-9806 NN denotes min
T4484 9807-9808 -LRB- denotes (
T4485 9808-9812 NNP denotes Tebu
T4486 9812-9813 -RRB- denotes )
T4487 9813-9814 . denotes .
T4488 9815-9820 IN denotes After
T4489 9821-9824 CD denotes two
T4490 9825-9835 JJ denotes additional
T4491 9836-9842 NNS denotes washes
T4492 9842-9843 , denotes ,
T4493 9844-9852 NN denotes analysis
T4494 9853-9855 IN denotes of
T4495 9856-9863 JJ denotes stained
T4496 9864-9869 NNS denotes cells
T4497 9870-9873 VBD denotes was
T4498 9874-9883 VBN denotes performed
T4499 9884-9886 IN denotes on
T4500 9887-9889 DT denotes an
T4501 9890-9895 NNP denotes EPICS
T4502 9896-9899 NNP denotes XL2
T4503 9900-9901 -LRB- denotes (
T4504 9901-9916 NNP denotes Beckman-Coulter
T4505 9916-9917 -RRB- denotes )
T4506 9917-9918 . denotes .
T4507 9919-9922 DT denotes The
T4508 9923-9927 NN denotes cell
T4509 9928-9938 NN denotes population
T4510 9939-9942 VBD denotes was
T4511 9943-9948 VBN denotes gated
T4512 9949-9958 VBG denotes according
T4513 9959-9961 TO denotes to
T4514 9962-9965 PRP$ denotes its
T4515 9966-9973 JJ denotes forward
T4516 9974-9977 CC denotes and
T4517 9978-9988 JJ denotes wide-angle
T4518 9989-9994 JJ denotes light
T4519 9995-10005 NN denotes scattering
T4520 10005-10006 . denotes .
T4521 10007-10011 NNS denotes Data
T4522 10012-10016 VBD denotes were
T4523 10017-10026 VBN denotes expressed
T4524 10027-10029 IN denotes as
T4525 10030-10034 JJ denotes mean
T4526 10035-10043 JJ denotes relative
T4527 10044-10056 NN denotes fluorescence
T4528 10057-10066 NN denotes intensity
T4529 10067-10068 -LRB- denotes (
T4530 10068-10071 NNP denotes MFI
T4531 10071-10072 -RRB- denotes )
T4532 10073-10075 IN denotes of
T4533 10076-10080 CD denotes 3000
T4534 10081-10086 NNS denotes cells
T4535 10086-10087 . denotes .
T4648 10089-10092 NNP denotes TNF
T4649 10093-10101 NNP denotes Activity
T4650 10102-10110 NNP denotes Bioassay
T4651 10111-10120 NNPS denotes Monocytes
T4652 10121-10123 CC denotes or
T4653 10124-10129 CD denotes THP-1
T4654 10130-10135 NNS denotes cells
T4655 10136-10140 VBD denotes were
T4656 10141-10149 VBN denotes cultured
T4657 10150-10154 IN denotes with
T4658 10155-10157 CC denotes or
T4659 10158-10165 IN denotes without
T4660 10166-10175 JJ denotes different
T4661 10176-10190 NNS denotes concentrations
T4662 10191-10193 IN denotes of
T4663 10194-10198 NNS denotes CGNs
T4664 10199-10201 CC denotes or
T4665 10202-10205 NNP denotes LPS
T4666 10206-10207 -LRB- denotes (
T4667 10207-10217 NNP denotes Salmonella
T4668 10218-10225 NN denotes typhosa
T4669 10225-10226 , denotes ,
T4670 10227-10232 NNP denotes Sigma
T4671 10232-10233 -RRB- denotes )
T4672 10234-10237 IN denotes for
T4673 10238-10240 CD denotes 24
T4674 10241-10242 NN denotes h
T4675 10243-10245 CC denotes or
T4676 10246-10249 DT denotes the
T4677 10250-10259 JJ denotes indicated
T4678 10260-10264 NN denotes time
T4679 10264-10265 . denotes .
T4680 10266-10278 RB denotes Biologically
T4681 10279-10285 JJ denotes active
T4682 10286-10291 NNP denotes TNF-α
T4683 10291-10292 NN denotes /
T4684 10292-10293 NN denotes β
T4685 10294-10296 IN denotes in
T4686 10297-10303 NN denotes tissue
T4687 10304-10311 NN denotes culture
T4688 10312-10323 NN denotes supernatant
T4689 10324-10327 VBD denotes was
T4690 10328-10336 VBN denotes measured
T4691 10337-10342 VBG denotes using
T4692 10343-10346 DT denotes the
T4693 10347-10351 NNP denotes WEHI
T4694 10352-10355 CD denotes 164
T4695 10356-10361 NN denotes clone
T4696 10362-10369 JJ denotes 13-cell
T4697 10370-10377 VBG denotes killing
T4698 10378-10383 NN denotes assay
T4699 10384-10385 NNP denotes [
T4700 10385-10387 CD denotes 29
T4701 10387-10388 NNP denotes ]
T4702 10388-10389 . denotes .
T4703 10390-10393 NNP denotes TNF
T4704 10394-10408 NNS denotes concentrations
T4705 10409-10412 VBP denotes are
T4706 10413-10422 VBN denotes expressed
T4707 10423-10425 IN denotes as
T4708 10426-10431 NN denotes pg/ml
T4709 10431-10432 . denotes .
T4951 10434-10440 NNP denotes RT-PCR
T4952 10441-10449 NNP denotes Analysis
T4953 10450-10455 NNP denotes Total
T4954 10456-10459 NNP denotes RNA
T4955 10460-10464 IN denotes from
T4956 10465-10474 NNS denotes monocytes
T4957 10475-10478 VBD denotes was
T4958 10479-10487 VBN denotes isolated
T4959 10488-10493 VBG denotes using
T4960 10494-10500 NNP denotes TRIzol
T4961 10501-10508 NNP denotes Reagent
T4962 10508-10509 NNP denotes ™
T4963 10510-10511 -LRB- denotes (
T4964 10511-10521 NNP denotes Invitrogen
T4965 10521-10522 -RRB- denotes )
T4966 10522-10523 . denotes .
T4967 10524-10528 NNP denotes cDNA
T4968 10529-10532 VBD denotes was
T4969 10533-10542 VBN denotes generated
T4970 10543-10545 IN denotes on
T4971 10546-10547 CD denotes 1
T4972 10548-10550 NN denotes µg
T4973 10551-10553 IN denotes of
T4974 10554-10559 JJ denotes total
T4975 10560-10563 NNP denotes RNA
T4976 10564-10566 IN denotes in
T4977 10567-10568 DT denotes a
T4978 10569-10577 NN denotes reaction
T4979 10578-10584 NN denotes volume
T4980 10585-10587 IN denotes of
T4981 10588-10590 CD denotes 20
T4982 10591-10593 NN denotes µl
T4983 10593-10594 , denotes ,
T4984 10595-10600 VBG denotes using
T4985 10601-10606 NNP denotes M-MLV
T4986 10607-10614 VB denotes reverse
T4987 10615-10628 NN denotes transcriptase
T4988 10629-10630 -LRB- denotes (
T4989 10630-10640 NNP denotes Invitrogen
T4990 10640-10641 -RRB- denotes )
T4991 10641-10642 . denotes .
T4992 10643-10646 NNP denotes PCR
T4993 10647-10650 VBD denotes was
T4994 10651-10655 VBN denotes done
T4995 10656-10658 IN denotes in
T4996 10659-10662 DT denotes the
T4997 10663-10669 JJ denotes linear
T4998 10670-10675 NN denotes range
T4999 10676-10678 IN denotes of
T5000 10679-10692 NN denotes amplification
T5001 10693-10694 -LRB- denotes (
T5002 10694-10704 VBN denotes determined
T5003 10705-10708 IN denotes for
T5004 10709-10713 DT denotes each
T5005 10714-10720 NN denotes primer
T5006 10721-10730 NN denotes pair-cDNA
T5007 10731-10742 NN denotes combination
T5008 10742-10743 -RRB- denotes )
T5009 10743-10744 . denotes .
T5010 10745-10753 NNP denotes Standard
T5011 10754-10757 NNP denotes PCR
T5012 10758-10767 NNS denotes reactions
T5013 10768-10772 VBD denotes were
T5014 10773-10782 VBN denotes performed
T5015 10783-10787 IN denotes with
T5016 10788-10789 CD denotes 1
T5017 10790-10792 NN denotes µl
T5018 10793-10795 IN denotes of
T5019 10796-10799 DT denotes the
T5020 10800-10804 NNP denotes cDNA
T5021 10805-10813 NN denotes solution
T5022 10813-10814 , denotes ,
T5023 10815-10817 CD denotes 50
T5024 10818-10820 NN denotes µM
T5025 10821-10823 IN denotes of
T5026 10824-10828 DT denotes each
T5027 10829-10835 NN denotes primer
T5028 10836-10844 NN denotes solution
T5029 10844-10845 , denotes ,
T5030 10846-10848 CD denotes 10
T5031 10849-10851 NN denotes mM
T5032 10852-10854 IN denotes of
T5033 10855-10859 DT denotes each
T5034 10860-10864 NNP denotes dNTP
T5035 10864-10865 , denotes ,
T5036 10866-10868 CD denotes 25
T5037 10869-10871 NNP denotes mM
T5038 10872-10877 NNP denotes MgCl2
T5039 10877-10878 , denotes ,
T5040 10879-10882 NNP denotes 10X
T5041 10883-10891 NNP denotes Goldstar
T5042 10892-10895 NNP denotes DNA
T5043 10896-10906 NN denotes polymerase
T5044 10907-10915 NN denotes reaction
T5045 10916-10922 NN denotes buffer
T5046 10922-10923 , denotes ,
T5047 10924-10927 CC denotes and
T5048 10928-10931 CD denotes 0.5
T5049 10932-10937 NNS denotes units
T5050 10938-10940 IN denotes of
T5051 10941-10949 NNP denotes Goldstar
T5052 10950-10953 NNP denotes DNA
T5053 10954-10964 NN denotes polymerase
T5054 10965-10966 -LRB- denotes (
T5055 10966-10976 NNP denotes Eurogentec
T5056 10976-10977 , denotes ,
T5057 10978-10985 NNP denotes Seraing
T5058 10985-10986 , denotes ,
T5059 10987-10994 NNP denotes Belgium
T5060 10994-10995 -RRB- denotes )
T5061 10995-10996 . denotes .
T5062 10997-11002 NNP denotes First
T5063 11003-11006 NNP denotes PCR
T5064 11007-11012 NN denotes cycle
T5065 11013-11022 VBD denotes consisted
T5066 11023-11025 IN denotes of
T5067 11026-11027 CD denotes 1
T5068 11028-11031 NN denotes min
T5069 11032-11034 IN denotes at
T5070 11035-11037 CD denotes 92
T5071 11037-11038 CD denotes °
T5072 11038-11039 NNP denotes C
T5073 11039-11040 , denotes ,
T5074 11041-11042 CD denotes 1
T5075 11043-11046 NN denotes min
T5076 11047-11049 IN denotes at
T5077 11050-11052 CD denotes 58
T5078 11052-11053 CD denotes °
T5079 11053-11054 NNP denotes C
T5080 11055-11058 CC denotes and
T5081 11059-11060 CD denotes 1
T5082 11061-11064 NN denotes min
T5083 11065-11067 IN denotes at
T5084 11068-11070 CD denotes 72
T5085 11070-11071 CD denotes °
T5086 11071-11072 NNP denotes C
T5087 11072-11073 : denotes ;
T5088 11074-11078 RB denotes then
T5089 11079-11083 DT denotes each
T5090 11084-11087 NNP denotes PCR
T5091 11088-11093 NN denotes cycle
T5092 11094-11103 VBD denotes consisted
T5093 11104-11106 IN denotes of
T5094 11107-11109 CD denotes 40
T5095 11110-11113 NN denotes sec
T5096 11114-11116 IN denotes at
T5097 11117-11119 CD denotes 92
T5098 11119-11120 CD denotes °
T5099 11120-11121 NNP denotes C
T5100 11121-11122 , denotes ,
T5101 11123-11125 CD denotes 40
T5102 11126-11129 NN denotes sec
T5103 11130-11132 IN denotes at
T5104 11133-11135 CD denotes 58
T5105 11135-11136 CD denotes °
T5106 11136-11137 NNP denotes C
T5107 11138-11141 CC denotes and
T5108 11142-11144 CD denotes 50
T5109 11145-11148 NN denotes sec
T5110 11149-11151 IN denotes at
T5111 11152-11154 CD denotes 72
T5112 11154-11155 CD denotes °
T5113 11155-11157 NNP denotes C.
T5114 11158-11162 NNP denotes cDNA
T5115 11163-11166 IN denotes for
T5116 11167-11174 NN denotes β-actin
T5117 11175-11178 VBD denotes was
T5118 11179-11188 VBN denotes amplified
T5119 11189-11192 IN denotes for
T5120 11193-11195 CD denotes 28
T5121 11196-11202 NNS denotes cycles
T5122 11203-11208 VBG denotes using
T5123 11209-11212 DT denotes the
T5124 11213-11219 NNS denotes oligos
T5125 11219-11220 : denotes :
T5126 11221-11226 NN denotes sense
T5127 11227-11228 CD denotes 5
T5128 11228-11229 SYM denotes ′
T5129 11229-11230 : denotes -
T5130 11230-11251 NN denotes GGCATCGTGATGGACTCCG-3
T5131 11251-11252 NN denotes ′
T5132 11253-11256 CC denotes and
T5133 11257-11266 JJ denotes antisense
T5134 11267-11268 CD denotes 5
T5135 11268-11269 CD denotes ′
T5136 11269-11290 NNP denotes GCTGGAAGGTGGACAGCGA-3
T5137 11290-11291 NN denotes ′
T5138 11291-11292 . denotes .
T5139 11293-11297 NNP denotes cDNA
T5140 11298-11301 IN denotes for
T5141 11302-11307 NNP denotes TNF-α
T5142 11308-11311 VBD denotes was
T5143 11312-11321 VBN denotes amplified
T5144 11322-11325 IN denotes for
T5145 11326-11328 CD denotes 35
T5146 11329-11335 NNS denotes cycles
T5147 11336-11341 VBG denotes using
T5148 11342-11345 DT denotes the
T5149 11346-11352 NNS denotes oligos
T5150 11352-11353 : denotes :
T5151 11354-11359 NN denotes sense
T5152 11360-11361 CD denotes 5
T5153 11361-11362 SYM denotes ′
T5154 11362-11363 : denotes -
T5155 11363-11385 NN denotes AAGCCTGTAGCCCATGTTGT-3
T5156 11385-11386 NN denotes ′
T5157 11387-11390 CC denotes and
T5158 11391-11400 JJ denotes antisense
T5159 11401-11402 CD denotes 5
T5160 11402-11403 SYM denotes ′
T5161 11403-11404 : denotes -
T5162 11404-11426 NN denotes CAGATAGATGGGCTCATACC-3
T5163 11426-11427 NN denotes ′
T5164 11427-11428 . denotes .
T5165 11429-11433 NNP denotes cDNA
T5166 11434-11437 IN denotes for
T5167 11438-11444 NNP denotes ICAM-1
T5168 11445-11448 VBD denotes was
T5169 11449-11458 VBN denotes amplified
T5170 11459-11462 IN denotes for
T5171 11463-11465 CD denotes 35
T5172 11466-11472 NNS denotes cycles
T5173 11473-11478 VBG denotes using
T5174 11479-11482 DT denotes the
T5175 11483-11489 NNS denotes oligos
T5176 11490-11495 VBP denotes sense
T5177 11496-11497 CD denotes 5
T5178 11497-11498 SYM denotes ′
T5179 11498-11499 : denotes -
T5180 11499-11523 NN denotes GTAGCAGCCGCAGTCATAATGG-3
T5181 11523-11524 NN denotes ′
T5182 11525-11528 CC denotes and
T5183 11529-11538 JJ denotes antisense
T5184 11539-11540 CD denotes 5
T5185 11540-11541 SYM denotes ′
T5186 11541-11542 : denotes -
T5187 11542-11543 DT denotes A
T5188 11544-11567 JJ denotes TGCTGTTGTATCTGACTGAGG-3
T5189 11567-11568 NN denotes ′
T5190 11568-11569 . denotes .
T5335 11571-11576 NNP denotes NF-kB
T5336 11577-11590 NNP denotes Transcription
T5337 11591-11599 NNP denotes Reporter
T5338 11600-11604 NNP denotes Gene
T5339 11605-11610 NNP denotes Assay
T5340 11611-11614 NNP denotes The
T5341 11615-11622 VBD denotes plasmid
T5342 11623-11632 NNP denotes 3XMHC-luc
T5343 11633-11634 -LRB- denotes (
T5344 11634-11635 DT denotes a
T5345 11636-11644 JJ denotes generous
T5346 11645-11649 NN denotes gift
T5347 11650-11654 IN denotes from
T5348 11655-11659 NNP denotes Drs.
T5349 11660-11662 NNP denotes J.
T5350 11663-11671 NNP denotes Westwick
T5351 11672-11675 CC denotes and
T5352 11676-11680 NNP denotes D.A.
T5353 11681-11688 NNP denotes Brenner
T5354 11688-11689 , denotes ,
T5355 11690-11700 NNP denotes University
T5356 11701-11703 IN denotes of
T5357 11704-11709 NNP denotes North
T5358 11710-11718 NNP denotes Carolina
T5359 11718-11719 , denotes ,
T5360 11720-11726 NNP denotes Chapel
T5361 11727-11731 NNP denotes Hill
T5362 11731-11732 -RRB- denotes )
T5363 11733-11741 VBZ denotes contains
T5364 11742-11747 CD denotes three
T5365 11748-11754 NNS denotes copies
T5366 11755-11757 IN denotes of
T5367 11758-11774 JJ denotes NF-κB-responsive
T5368 11775-11782 NN denotes element
T5369 11783-11787 IN denotes from
T5370 11788-11791 DT denotes the
T5371 11792-11795 NNP denotes MHC
T5372 11796-11801 NN denotes class
T5373 11802-11803 PRP denotes I
T5374 11804-11809 VBZ denotes locus
T5375 11809-11810 , denotes ,
T5376 11811-11817 VBN denotes placed
T5377 11818-11826 RB denotes upstream
T5378 11827-11829 IN denotes of
T5379 11830-11833 DT denotes the
T5380 11834-11844 NN denotes luciferase
T5381 11845-11849 NN denotes gene
T5382 11849-11850 . denotes .
T5383 11851-11856 JJ denotes Human
T5384 11857-11866 JJ denotes monocytic
T5385 11867-11872 NN denotes THP-1
T5386 11873-11878 NNS denotes cells
T5387 11879-11883 VBD denotes were
T5388 11884-11895 RB denotes transiently
T5389 11896-11907 VBN denotes transfected
T5390 11908-11910 IN denotes as
T5391 11911-11921 RB denotes previously
T5392 11922-11931 VBN denotes described
T5393 11932-11933 NNP denotes [
T5394 11933-11935 CD denotes 30
T5395 11935-11936 NNP denotes ]
T5396 11936-11937 , denotes ,
T5397 11938-11941 CC denotes and
T5398 11942-11946 RB denotes then
T5399 11947-11955 VBD denotes cultured
T5400 11956-11959 IN denotes for
T5401 11960-11961 CD denotes 4
T5402 11962-11963 NN denotes h
T5403 11964-11969 RB denotes alone
T5404 11970-11972 CC denotes or
T5405 11973-11977 IN denotes with
T5406 11978-11988 VBG denotes increasing
T5407 11989-12002 NN denotes concentration
T5408 12003-12005 IN denotes of
T5409 12006-12012 DT denotes either
T5410 12013-12016 NNP denotes C10
T5411 12017-12019 CC denotes or
T5412 12020-12023 NNP denotes C40
T5413 12023-12024 . denotes .
T5414 12025-12035 JJ denotes Luciferase
T5415 12036-12044 NN denotes activity
T5416 12045-12048 VBD denotes was
T5417 12049-12059 VBN denotes determined
T5418 12060-12065 VBG denotes using
T5419 12066-12067 DT denotes a
T5420 12068-12079 NN denotes luminometer
T5421 12080-12081 -LRB- denotes (
T5422 12081-12090 NNP denotes Monolight
T5423 12091-12095 CD denotes 2010
T5424 12096-12107 NNP denotes Luminometer
T5425 12107-12108 , denotes ,
T5426 12109-12112 NNP denotes Ann
T5427 12113-12118 NNP denotes Arbor
T5428 12118-12119 , denotes ,
T5429 12120-12122 NNP denotes MI
T5430 12122-12123 -RRB- denotes )
T5431 12123-12124 . denotes .
T5933 12126-12133 JJ denotes Western
T5934 12134-12138 NNP denotes Blot
T5935 12139-12147 NNP denotes Analysis
T5936 12148-12153 NNP denotes THP-1
T5937 12154-12159 NNS denotes cells
T5938 12160-12164 VBD denotes were
T5939 12165-12175 VBN denotes stimulated
T5940 12176-12179 IN denotes for
T5941 12180-12187 JJ denotes various
T5942 12188-12195 NNS denotes lengths
T5943 12196-12198 IN denotes of
T5944 12199-12203 NN denotes time
T5945 12204-12208 IN denotes with
T5946 12209-12212 CD denotes 0.1
T5947 12213-12218 NN denotes mg/ml
T5948 12219-12222 CD denotes C10
T5949 12223-12225 CC denotes or
T5950 12226-12229 CD denotes C40
T5951 12229-12230 , denotes ,
T5952 12231-12233 CC denotes or
T5953 12234-12236 CD denotes 10
T5954 12237-12239 NN denotes µg
T5955 12239-12240 NN denotes /
T5956 12240-12242 NN denotes ml
T5957 12243-12246 NNP denotes LPS
T5958 12246-12247 . denotes .
T5959 12248-12253 NNS denotes Cells
T5960 12254-12258 VBD denotes were
T5961 12259-12263 RB denotes then
T5962 12264-12272 VBN denotes pelleted
T5963 12272-12273 , denotes ,
T5964 12274-12280 VBN denotes washed
T5965 12281-12284 CC denotes and
T5966 12285-12296 VBN denotes homogenised
T5967 12297-12299 IN denotes in
T5968 12300-12305 NN denotes lysis
T5969 12306-12312 NN denotes buffer
T5970 12313-12314 -LRB- denotes (
T5971 12314-12316 CD denotes 10
T5972 12317-12319 NNP denotes mM
T5973 12320-12325 NNP denotes Hepes
T5974 12325-12326 , denotes ,
T5975 12327-12329 NNP denotes pH
T5976 12330-12333 CD denotes 7.9
T5977 12333-12334 , denotes ,
T5978 12335-12338 CD denotes 150
T5979 12339-12341 NNP denotes mM
T5980 12342-12346 NNP denotes NaCl
T5981 12346-12347 , denotes ,
T5982 12348-12349 CD denotes 1
T5983 12350-12352 NNP denotes mM
T5984 12353-12357 NNP denotes EDTA
T5985 12357-12358 , denotes ,
T5986 12359-12362 CD denotes 0.6
T5987 12362-12363 NN denotes %
T5988 12364-12369 NN denotes NP-40
T5989 12369-12370 , denotes ,
T5990 12371-12374 CC denotes and
T5991 12375-12378 CD denotes 0.5
T5992 12379-12381 NNP denotes mM
T5993 12382-12386 NNP denotes PMSF
T5994 12386-12387 -RRB- denotes )
T5995 12388-12390 IN denotes on
T5996 12391-12394 NN denotes ice
T5997 12394-12395 . denotes .
T5998 12396-12407 NNS denotes Homogenates
T5999 12408-12412 VBD denotes were
T6000 12413-12422 VBN denotes sonicated
T6001 12422-12423 , denotes ,
T6002 12424-12435 VBN denotes centrifuged
T6003 12436-12438 IN denotes at
T6004 12439-12445 CD denotes 10,000
T6005 12446-12449 NN denotes rpm
T6006 12450-12452 TO denotes to
T6007 12453-12459 VB denotes remove
T6008 12460-12468 JJ denotes cellular
T6009 12469-12475 NN denotes debris
T6010 12475-12476 , denotes ,
T6011 12477-12480 CC denotes and
T6012 12481-12492 JJ denotes supernatant
T6013 12493-12502 VBN denotes collected
T6014 12502-12503 . denotes .
T6015 12504-12511 NN denotes Protein
T6016 12512-12525 NN denotes concentration
T6017 12526-12529 VBD denotes was
T6018 12530-12540 VBN denotes determined
T6019 12541-12546 VBG denotes using
T6020 12547-12550 DT denotes the
T6021 12551-12553 NNP denotes DC
T6022 12554-12561 NNP denotes Protein
T6023 12562-12567 NNP denotes Assay
T6024 12568-12569 -LRB- denotes (
T6025 12569-12576 NNP denotes Bio-Rad
T6026 12576-12577 -RRB- denotes )
T6027 12577-12578 . denotes .
T6028 12579-12587 NNPS denotes Proteins
T6029 12588-12590 IN denotes in
T6030 12591-12598 NNS denotes samples
T6031 12599-12600 -LRB- denotes (
T6032 12600-12602 CD denotes 15
T6033 12603-12605 NN denotes µg
T6034 12606-12611 JJ denotes total
T6035 12612-12620 NNS denotes proteins
T6036 12620-12621 -RRB- denotes )
T6037 12622-12626 VBD denotes were
T6038 12627-12635 VBN denotes resolved
T6039 12636-12638 IN denotes in
T6040 12639-12640 DT denotes a
T6041 12641-12651 VBG denotes denaturing
T6042 12652-12654 CD denotes 12
T6043 12654-12655 NN denotes %
T6044 12656-12670 NN denotes polyacrylamide
T6045 12671-12674 NN denotes gel
T6046 12675-12678 CC denotes and
T6047 12679-12690 VBN denotes transferred
T6048 12691-12693 TO denotes to
T6049 12694-12695 DT denotes a
T6050 12696-12710 JJ denotes nitrocellulose
T6051 12711-12719 NN denotes membrane
T6052 12719-12720 . denotes .
T6053 12721-12726 JJ denotes I-κBα
T6054 12727-12734 NN denotes protein
T6055 12735-12738 VBD denotes was
T6056 12739-12747 VBN denotes detected
T6057 12748-12753 VBG denotes using
T6058 12754-12755 DT denotes a
T6059 12756-12762 NN denotes rabbit
T6060 12763-12773 NN denotes polyclonal
T6061 12774-12782 NN denotes antibody
T6062 12783-12784 -LRB- denotes (
T6063 12784-12789 NNP denotes Santa
T6064 12790-12794 NNP denotes Cruz
T6065 12795-12808 NNP denotes Biotechnology
T6066 12808-12809 , denotes ,
T6067 12810-12812 NNP denotes CA
T6068 12812-12813 -RRB- denotes )
T6069 12814-12822 VBN denotes followed
T6070 12823-12825 IN denotes by
T6071 12826-12827 DT denotes a
T6072 12828-12839 JJ denotes horseradish
T6073 12840-12858 JJ denotes peroxidase-coupled
T6074 12859-12863 NN denotes goat
T6075 12864-12874 NN denotes polyclonal
T6076 12875-12883 NN denotes antibody
T6077 12884-12891 IN denotes against
T6078 12892-12898 NN denotes rabbit
T6079 12899-12901 NNP denotes Ig
T6080 12902-12903 -LRB- denotes (
T6081 12903-12909 NNP denotes Caltag
T6082 12910-12922 NNPS denotes Laboratories
T6083 12922-12923 -RRB- denotes )
T6084 12923-12924 . denotes .
T6085 12925-12932 RB denotes Finally
T6086 12932-12933 , denotes ,
T6087 12934-12937 NNP denotes IκB
T6088 12938-12943 NNS denotes bands
T6089 12944-12948 VBD denotes were
T6090 12949-12957 VBN denotes revealed
T6091 12958-12963 VBG denotes using
T6092 12964-12967 DT denotes the
T6093 12968-12971 NNP denotes ECL
T6094 12971-12972 NN denotes ™
T6095 12973-12982 NN denotes detection
T6096 12983-12989 NN denotes system
T6097 12990-12991 -LRB- denotes (
T6098 12991-12999 NNP denotes Amersham
T6099 13000-13009 NNP denotes Pharmacia
T6100 13010-13017 NNP denotes Biotech
T6101 13017-13018 , denotes ,
T6102 13019-13022 NNP denotes Les
T6103 13023-13028 NNP denotes Ullis
T6104 13028-13029 , denotes ,
T6105 13030-13036 NNP denotes France
T6106 13036-13037 -RRB- denotes )
T6107 13038-13047 VBG denotes according
T6108 13048-13050 TO denotes to
T6109 13051-13054 DT denotes the
T6110 13055-13068 NNS denotes manufacturers
T6111 13068-13069 POS denotes '
T6112 13070-13081 NN denotes instruction
T6113 13081-13082 . denotes .
T6114 13083-13091 NN denotes Antibody
T6115 13092-13094 TO denotes to
T6116 13095-13104 NNP denotes α-Tubulin
T6117 13105-13106 -LRB- denotes (
T6118 13106-13111 NNP denotes Santa
T6119 13112-13116 NNP denotes Cruz
T6120 13116-13117 -RRB- denotes )
T6121 13118-13121 VBD denotes was
T6122 13122-13125 NN denotes use
T6123 13126-13128 IN denotes as
T6124 13129-13136 VBG denotes loading
T6125 13137-13144 NN denotes control
T6126 13144-13145 . denotes .
T6127 13146-13149 IN denotes For
T6128 13150-13157 JJ denotes nuclear
T6129 13158-13163 NNP denotes NF-κB
T6130 13163-13164 , denotes ,
T6131 13165-13170 NNP denotes THP-1
T6132 13171-13176 NNS denotes cells
T6133 13177-13181 VBD denotes were
T6134 13182-13192 VBN denotes stimulated
T6135 13193-13197 IN denotes with
T6136 13198-13199 CD denotes 1
T6137 13200-13205 NN denotes mg/ml
T6138 13206-13209 CD denotes C10
T6139 13210-13212 CC denotes or
T6140 13213-13216 CD denotes C40
T6141 13217-13220 IN denotes for
T6142 13221-13223 CD denotes 30
T6143 13224-13231 NNS denotes minutes
T6144 13232-13234 IN denotes at
T6145 13235-13237 CD denotes 37
T6146 13237-13238 NN denotes °
T6147 13238-13240 NNP denotes C.
T6148 13241-13246 NNP denotes Cells
T6149 13247-13251 VBD denotes were
T6150 13252-13256 RB denotes then
T6151 13257-13265 VBN denotes pelleted
T6152 13266-13269 CC denotes and
T6153 13270-13276 JJ denotes nuclei
T6154 13277-13286 VBN denotes separated
T6155 13287-13289 IN denotes as
T6156 13290-13299 VBN denotes described
T6157 13300-13301 NNP denotes [
T6158 13301-13303 CD denotes 31
T6159 13303-13304 NNP denotes ]
T6160 13304-13305 . denotes .
T6161 13306-13312 NNP denotes Nuclei
T6162 13313-13317 VBD denotes were
T6163 13318-13324 VBN denotes washed
T6164 13325-13328 CC denotes and
T6165 13329-13340 VBN denotes homogenized
T6166 13341-13349 RB denotes directly
T6167 13350-13352 IN denotes in
T6168 13353-13360 VBG denotes loading
T6169 13361-13362 -LRB- denotes (
T6170 13362-13368 NNP denotes Laemli
T6171 13368-13369 -RRB- denotes )
T6172 13370-13376 NN denotes buffer
T6173 13377-13380 CC denotes and
T6174 13381-13387 VBN denotes heated
T6175 13388-13391 IN denotes for
T6176 13392-13393 CD denotes 5
T6177 13394-13401 NNS denotes minutes
T6178 13402-13404 IN denotes at
T6179 13405-13408 CD denotes 100
T6180 13408-13409 CD denotes °
T6181 13409-13411 NNP denotes C.
T6182 13412-13420 NNPS denotes Proteins
T6183 13421-13423 IN denotes in
T6184 13424-13431 NNS denotes samples
T6185 13432-13436 VBD denotes were
T6186 13437-13445 VBN denotes resolved
T6187 13446-13448 IN denotes in
T6188 13449-13450 DT denotes a
T6189 13451-13461 NN denotes denaturing
T6190 13462-13463 CD denotes 8
T6191 13463-13464 NN denotes %
T6192 13465-13479 NN denotes polyacrylamide
T6193 13480-13483 NN denotes gel
T6194 13484-13487 CC denotes and
T6195 13488-13499 VBN denotes transferred
T6196 13500-13502 TO denotes to
T6197 13503-13504 DT denotes a
T6198 13505-13519 NN denotes polyvinylidine
T6199 13520-13528 NN denotes fluoride
T6200 13529-13530 -LRB- denotes (
T6201 13530-13534 NNP denotes PVDF
T6202 13534-13535 -RRB- denotes )
T6203 13536-13544 NN denotes membrane
T6204 13545-13546 -LRB- denotes (
T6205 13546-13557 JJ denotes Immobilon-P
T6206 13557-13558 : denotes ;
T6207 13559-13568 NNP denotes Millipore
T6208 13568-13569 , denotes ,
T6209 13570-13577 NNP denotes Bedford
T6210 13577-13578 , denotes ,
T6211 13579-13581 NNP denotes MA
T6212 13581-13582 -RRB- denotes )
T6213 13582-13583 . denotes .
T6214 13584-13593 NNS denotes Membranes
T6215 13594-13598 VBD denotes were
T6216 13599-13608 VBN denotes incubated
T6217 13609-13611 IN denotes in
T6218 13612-13620 VBG denotes blocking
T6219 13621-13627 NN denotes buffer
T6220 13628-13629 -LRB- denotes (
T6221 13629-13630 CD denotes 1
T6222 13630-13631 NN denotes %
T6223 13632-13635 NNP denotes BSA
T6224 13635-13636 , denotes ,
T6225 13637-13639 IN denotes in
T6226 13640-13643 NNP denotes PBS
T6227 13643-13644 -RRB- denotes )
T6228 13645-13648 IN denotes for
T6229 13649-13652 CD denotes two
T6230 13653-13658 NNS denotes hours
T6231 13659-13661 IN denotes at
T6232 13662-13666 NN denotes room
T6233 13667-13678 NN denotes temperature
T6234 13678-13679 . denotes .
T6235 13680-13689 NNS denotes Membranes
T6236 13690-13694 VBD denotes were
T6237 13695-13707 RB denotes subsequently
T6238 13708-13714 VBN denotes probed
T6239 13715-13719 IN denotes with
T6240 13720-13723 DT denotes the
T6241 13724-13737 JJ denotes corresponding
T6242 13738-13746 NN denotes antibody
T6243 13747-13749 IN denotes in
T6244 13750-13758 VBG denotes blocking
T6245 13759-13765 NN denotes buffer
T6246 13765-13766 , denotes ,
T6247 13767-13776 JJ denotes overnight
T6248 13776-13777 . denotes .
T6249 13778-13784 NN denotes Rabbit
T6250 13785-13795 NN denotes polyclonal
T6251 13796-13804 NN denotes antibody
T6252 13805-13815 JJ denotes anti-NF-κB
T6253 13816-13819 JJ denotes p50
T6254 13820-13827 NN denotes subunit
T6255 13828-13829 -LRB- denotes (
T6256 13829-13830 # denotes #
T6257 13831-13837 CD denotes sc-114
T6258 13837-13838 -RRB- denotes )
T6259 13839-13841 CC denotes or
T6260 13842-13852 JJ denotes anti-NF-κB
T6261 13853-13856 NNS denotes p65
T6262 13857-13864 VBN denotes subunit
T6263 13865-13866 -LRB- denotes (
T6264 13866-13867 # denotes #
T6265 13868-13874 CD denotes sc-109
T6266 13874-13875 -RRB- denotes )
T6267 13876-13880 IN denotes from
T6268 13881-13886 NNP denotes Santa
T6269 13887-13891 NNP denotes Cruz
T6270 13892-13905 NNP denotes Biotechnology
T6271 13906-13910 VBD denotes were
T6272 13911-13915 VBN denotes used
T6273 13915-13916 . denotes .
T6274 13917-13926 NNS denotes Membranes
T6275 13927-13931 VBD denotes were
T6276 13932-13938 VBN denotes washed
T6277 13939-13942 CD denotes six
T6278 13943-13948 NNS denotes times
T6279 13949-13951 IN denotes in
T6280 13952-13955 NNP denotes PBS
T6281 13956-13960 IN denotes with
T6282 13961-13965 CD denotes 0.05
T6283 13965-13966 NN denotes %
T6284 13967-13972 CD denotes Tween
T6285 13973-13975 CD denotes 20
T6286 13975-13976 , denotes ,
T6287 13977-13978 CD denotes 5
T6288 13979-13986 NNS denotes minutes
T6289 13987-13991 DT denotes each
T6290 13992-13996 NN denotes time
T6291 13996-13997 , denotes ,
T6292 13998-14001 CC denotes and
T6293 14002-14011 VBN denotes incubated
T6294 14012-14016 IN denotes with
T6295 14017-14018 DT denotes a
T6296 14019-14025 CD denotes 1/3000
T6297 14026-14034 NN denotes dilution
T6298 14035-14037 IN denotes of
T6299 14038-14052 JJ denotes HRP-conjugated
T6300 14053-14054 NN denotes F
T6301 14054-14055 -LRB- denotes (
T6302 14055-14057 NN denotes ab
T6303 14057-14058 '' denotes '
T6304 14058-14059 -RRB- denotes )
T6305 14059-14060 CD denotes 2
T6306 14061-14065 NN denotes goat
T6307 14066-14077 JJ denotes anti-rabbit
T6308 14078-14081 NNP denotes IgG
T6309 14082-14084 IN denotes in
T6310 14085-14086 CD denotes 5
T6311 14086-14087 NN denotes %
T6312 14088-14094 JJ denotes nonfat
T6313 14095-14098 JJ denotes dry
T6314 14099-14103 NN denotes milk
T6315 14104-14107 CC denotes and
T6316 14108-14112 CD denotes 0.05
T6317 14112-14113 NN denotes %
T6318 14114-14119 CD denotes Tween
T6319 14120-14122 CD denotes 20
T6320 14123-14125 IN denotes in
T6321 14126-14129 NNP denotes PBS
T6322 14130-14133 IN denotes for
T6323 14134-14135 CD denotes 1
T6324 14136-14140 NN denotes hour
T6325 14141-14143 IN denotes at
T6326 14144-14148 NN denotes room
T6327 14149-14160 NN denotes temperature
T6328 14160-14161 . denotes .
T6329 14162-14167 IN denotes After
T6330 14168-14175 VBG denotes washing
T6331 14176-14179 CD denotes six
T6332 14180-14184 JJR denotes more
T6333 14185-14190 NNS denotes times
T6334 14191-14193 IN denotes in
T6335 14194-14197 NNP denotes PBS
T6336 14198-14202 IN denotes with
T6337 14203-14207 CD denotes 0.05
T6338 14207-14208 NN denotes %
T6339 14209-14214 CD denotes Tween
T6340 14215-14217 CD denotes 20
T6341 14217-14218 , denotes ,
T6342 14219-14236 JJ denotes antibody-reactive
T6343 14237-14245 NNS denotes proteins
T6344 14246-14250 VBD denotes were
T6345 14251-14259 VBN denotes detected
T6346 14260-14265 VBG denotes using
T6347 14266-14267 DT denotes a
T6348 14268-14285 NN denotes chemiluminescence
T6349 14286-14295 NN denotes substrate
T6350 14296-14297 -LRB- denotes (
T6351 14297-14308 NNP denotes SuperSignal
T6352 14308-14309 : denotes ;
T6353 14310-14316 NNP denotes Pierce
T6354 14316-14317 , denotes ,
T6355 14318-14326 NNP denotes Rockford
T6356 14326-14327 , denotes ,
T6357 14328-14330 NNP denotes IL
T6358 14330-14331 -RRB- denotes )
T6359 14332-14341 VBG denotes according
T6360 14342-14344 TO denotes to
T6361 14345-14348 DT denotes the
T6362 14349-14361 NN denotes manufacturer
T6363 14361-14363 POS denotes 's
T6364 14364-14376 NNS denotes instructions
T6365 14376-14377 . denotes .
T6366 14378-14380 TO denotes To
T6367 14381-14388 VB denotes confirm
T6368 14389-14393 DT denotes that
T6369 14394-14404 JJ denotes equivalent
T6370 14405-14412 NNS denotes amounts
T6371 14413-14415 IN denotes of
T6372 14416-14423 NN denotes protein
T6373 14424-14428 VBD denotes were
T6374 14429-14435 VBN denotes loaded
T6375 14436-14438 IN denotes in
T6376 14439-14443 DT denotes each
T6377 14444-14448 NN denotes line
T6378 14448-14449 , denotes ,
T6379 14450-14459 NNS denotes membranes
T6380 14460-14464 VBD denotes were
T6381 14465-14469 RB denotes also
T6382 14470-14477 JJ denotes Western
T6383 14478-14485 VBN denotes blotted
T6384 14486-14489 IN denotes for
T6385 14490-14493 NNP denotes ERK
T6386 14494-14496 IN denotes as
T6387 14497-14506 VBN denotes described
T6388 14507-14508 NNP denotes [
T6389 14508-14510 CD denotes 32
T6390 14510-14511 NNP denotes ]
T6391 14511-14512 . denotes .
T6486 14514-14522 NN denotes Analysis
T6487 14523-14525 IN denotes of
T6488 14526-14531 NNP denotes NF-κB
T6489 14532-14542 NNP denotes Activation
T6490 14543-14545 IN denotes by
T6491 14546-14550 NNP denotes Flow
T6492 14551-14560 NNP denotes Cytometry
T6493 14561-14568 NNP denotes Nuclear
T6494 14569-14579 NN denotes activation
T6495 14580-14582 IN denotes of
T6496 14583-14585 NNP denotes NF
T6497 14585-14586 NNP denotes −
T6498 14586-14588 NNP denotes κΒ
T6499 14589-14591 IN denotes by
T6500 14592-14596 NN denotes flow
T6501 14597-14606 NN denotes cytometry
T6502 14607-14610 VBD denotes was
T6503 14611-14620 VBN denotes performed
T6504 14621-14623 IN denotes as
T6505 14624-14633 VBN denotes described
T6506 14634-14635 NNP denotes [
T6507 14635-14637 CD denotes 31
T6508 14637-14638 NNP denotes ]
T6509 14638-14639 . denotes .
T6558 14641-14652 NNP denotes Statistical
T6559 14653-14661 NNP denotes Analysis
T6560 14662-14665 NNP denotes The
T6561 14666-14673 NNS denotes results
T6562 14674-14678 VBD denotes were
T6563 14679-14688 VBN denotes expressed
T6564 14689-14691 IN denotes as
T6565 14692-14695 DT denotes the
T6566 14696-14700 JJ denotes mean
T6567 14701-14706 NN denotes value
T6568 14707-14708 NN denotes ±
T6569 14709-14715 NNP denotes S.E.M.
T6570 14716-14718 IN denotes of
T6571 14719-14729 JJ denotes individual
T6572 14730-14741 NNS denotes experiments
T6573 14741-14742 . denotes .
T6574 14743-14746 DT denotes The
T6575 14747-14758 JJ denotes statistical
T6576 14759-14771 NN denotes significance
T6577 14772-14774 IN denotes of
T6578 14775-14778 DT denotes the
T6579 14779-14790 NNS denotes differences
T6580 14791-14798 IN denotes between
T6581 14799-14803 JJ denotes mean
T6582 14804-14810 NNS denotes values
T6583 14811-14814 VBD denotes was
T6584 14815-14823 VBN denotes assessed
T6585 14824-14826 IN denotes by
T6586 14827-14830 DT denotes the
T6587 14831-14838 NN denotes Student
T6588 14838-14840 POS denotes 's
T6589 14841-14847 JJS denotes t-test
T6590 14848-14851 CC denotes and
T6591 14852-14860 NN denotes analysis
T6592 14861-14863 IN denotes of
T6593 14864-14872 NN denotes variance
T6594 14873-14874 -LRB- denotes (
T6595 14874-14879 NNP denotes ANOVA
T6596 14879-14880 -RRB- denotes )
T6597 14880-14881 . denotes .
T6857 14892-14900 JJ denotes Degraded
T6858 14901-14904 NNP denotes CGN
T6859 14905-14911 NNP denotes Induce
T6860 14912-14919 NNP denotes Colonic
T6861 14920-14932 NNP denotes Inflammation
T6862 14933-14936 NNP denotes All
T6863 14937-14941 NNS denotes rats
T6864 14942-14951 VBD denotes developed
T6865 14952-14960 NN denotes diarrhea
T6866 14961-14967 IN denotes during
T6867 14968-14976 JJ denotes degraded
T6868 14977-14988 NN denotes carrageenan
T6869 14989-15003 NN denotes administration
T6870 15004-15007 CC denotes and
T6871 15008-15013 JJ denotes gross
T6872 15014-15022 NN denotes evidence
T6873 15023-15025 IN denotes of
T6874 15026-15031 NN denotes blood
T6875 15032-15035 VBD denotes was
T6876 15036-15046 RB denotes frequently
T6877 15047-15055 VBN denotes detected
T6878 15056-15058 IN denotes in
T6879 15059-15062 DT denotes the
T6880 15063-15069 NNS denotes stools
T6881 15069-15070 . denotes .
T6882 15071-15076 NN denotes Colon
T6883 15077-15083 NN denotes length
T6884 15084-15096 RB denotes dramatically
T6885 15097-15106 VBD denotes decreased
T6886 15107-15109 IN denotes in
T6887 15110-15113 DT denotes all
T6888 15114-15121 VBN denotes treated
T6889 15122-15126 NNS denotes rats
T6890 15127-15131 IN denotes with
T6891 15132-15133 DT denotes a
T6892 15134-15138 RBR denotes more
T6893 15139-15149 JJ denotes pronounced
T6894 15150-15156 NN denotes effect
T6895 15157-15162 VBG denotes being
T6896 15163-15171 VBN denotes observed
T6897 15172-15174 IN denotes in
T6898 15175-15178 DT denotes the
T6899 15179-15181 CD denotes 40
T6900 15182-15185 NNP denotes kDa
T6901 15186-15190 NNP denotes dCGN
T6902 15191-15198 VBD denotes treated
T6903 15199-15204 NN denotes group
T6904 15205-15206 -LRB- denotes (
T6905 15206-15210 NNP denotes Fig.
T6906 15211-15213 NNP denotes 1A
T6907 15213-15214 -RRB- denotes )
T6908 15214-15215 . denotes .
T6909 15216-15227 RB denotes Furthermore
T6910 15227-15228 , denotes ,
T6911 15229-15238 VBN denotes prolonged
T6912 15239-15247 NN denotes exposure
T6913 15248-15250 TO denotes to
T6914 15251-15253 CD denotes 40
T6915 15254-15257 NNP denotes kDa
T6916 15258-15262 NNP denotes dCGN
T6917 15263-15271 VBD denotes resulted
T6918 15272-15274 IN denotes in
T6919 15275-15279 JJ denotes high
T6920 15280-15291 JJ denotes macroscopic
T6921 15292-15295 CC denotes and
T6922 15296-15308 JJ denotes histological
T6923 15309-15315 NNS denotes scores
T6924 15316-15318 IN denotes of
T6925 15319-15331 NN denotes inflammation
T6926 15332-15333 -LRB- denotes (
T6927 15333-15337 NNP denotes Fig.
T6928 15338-15340 NNP denotes 1B
T6929 15340-15341 , denotes ,
T6930 15342-15343 NNP denotes C
T6931 15343-15344 -RRB- denotes )
T6932 15344-15345 . denotes .
T6933 15346-15350 RB denotes Only
T6934 15351-15355 JJ denotes weak
T6935 15356-15371 NN denotes myeloperoxidase
T6936 15372-15380 NN denotes activity
T6937 15381-15384 VBD denotes was
T6938 15385-15393 VBN denotes detected
T6939 15394-15396 IN denotes in
T6940 15397-15401 DT denotes both
T6941 15402-15409 NN denotes control
T6942 15410-15413 CC denotes and
T6943 15414-15426 JJ denotes dCGN-treated
T6944 15427-15433 NNS denotes groups
T6945 15434-15435 -LRB- denotes (
T6946 15435-15439 NNP denotes Fig.
T6947 15440-15442 NNP denotes 1D
T6948 15442-15443 -RRB- denotes )
T6949 15443-15444 , denotes ,
T6950 15445-15455 VBG denotes indicating
T6951 15456-15460 IN denotes that
T6952 15461-15473 NNS denotes granulocytes
T6953 15474-15477 VBD denotes did
T6954 15478-15481 RB denotes not
T6955 15482-15486 VB denotes play
T6956 15487-15488 DT denotes a
T6957 15489-15494 JJ denotes major
T6958 15495-15499 NN denotes role
T6959 15500-15502 IN denotes in
T6960 15503-15506 DT denotes the
T6961 15507-15519 NN denotes inflammation
T6962 15520-15522 IN denotes at
T6963 15523-15527 DT denotes that
T6964 15528-15533 NN denotes stage
T6965 15533-15534 . denotes .
T6966 15535-15547 JJ denotes Histological
T6967 15548-15559 NN denotes examination
T6968 15560-15568 VBD denotes revealed
T6969 15569-15576 JJ denotes various
T6970 15577-15584 NNS denotes degrees
T6971 15585-15587 IN denotes of
T6972 15588-15595 JJ denotes mucosal
T6973 15596-15608 NN denotes inflammation
T6974 15608-15609 . denotes .
T6975 15610-15614 NNS denotes Rats
T6976 15615-15622 VBN denotes treated
T6977 15623-15627 IN denotes with
T6978 15628-15630 CD denotes 10
T6979 15631-15634 NNP denotes kDa
T6980 15635-15639 NNP denotes dCGN
T6981 15640-15646 VBD denotes showed
T6982 15647-15652 NN denotes edema
T6983 15652-15653 , denotes ,
T6984 15654-15664 NN denotes epithelium
T6985 15665-15672 NN denotes atrophy
T6986 15673-15676 CC denotes and
T6987 15677-15683 JJ denotes slight
T6988 15684-15694 NN denotes lymphocyte
T6989 15695-15707 NN denotes infiltration
T6990 15708-15709 -LRB- denotes (
T6991 15709-15713 NNS denotes data
T6992 15714-15717 RB denotes not
T6993 15718-15723 VBN denotes shown
T6994 15723-15724 -RRB- denotes )
T6995 15724-15725 . denotes .
T6996 15726-15731 DT denotes These
T6997 15732-15740 NNS denotes symptoms
T6998 15741-15745 VBD denotes were
T6999 15746-15753 RB denotes totally
T7000 15754-15760 JJ denotes absent
T7001 15761-15763 IN denotes in
T7002 15764-15767 DT denotes the
T7003 15768-15773 NN denotes colon
T7004 15774-15776 IN denotes of
T7005 15777-15784 NN denotes control
T7006 15785-15789 NNS denotes rats
T7007 15790-15791 -LRB- denotes (
T7008 15791-15795 NNP denotes Fig.
T7009 15796-15798 NNP denotes 1E
T7010 15798-15799 -RRB- denotes )
T7011 15799-15800 . denotes .
T7012 15801-15805 RBR denotes More
T7013 15806-15812 JJ denotes severe
T7014 15813-15820 JJ denotes mucosal
T7015 15821-15829 NNS denotes injuries
T7016 15830-15839 VBG denotes including
T7017 15840-15850 NN denotes ulceration
T7018 15850-15851 , denotes ,
T7019 15852-15864 JJ denotes hyperplastic
T7020 15865-15875 NN denotes epithelium
T7021 15875-15876 , denotes ,
T7022 15877-15882 JJ denotes crypt
T7023 15883-15893 NN denotes distortion
T7024 15894-15897 CC denotes and
T7025 15898-15899 DT denotes a
T7026 15900-15906 JJ denotes strong
T7027 15907-15917 NN denotes macrophage
T7028 15918-15930 NN denotes infiltration
T7029 15930-15931 , denotes ,
T7030 15932-15936 VBD denotes were
T7031 15937-15945 VBN denotes observed
T7032 15946-15948 IN denotes in
T7033 15949-15952 DT denotes the
T7034 15953-15955 CD denotes 40
T7035 15956-15959 NNP denotes kDa
T7036 15960-15972 NNP denotes dCGN-treated
T7037 15973-15977 NNS denotes rats
T7038 15978-15979 -LRB- denotes (
T7039 15979-15983 NNP denotes Fig.
T7040 15984-15986 NNP denotes 1F
T7041 15986-15987 -RRB- denotes )
T7042 15987-15988 . denotes .
T7043 15989-15991 DT denotes No
T7044 15992-16001 VBN denotes sulphated
T7045 16002-16017 NNS denotes polysaccharides
T7046 16018-16022 VBD denotes were
T7047 16023-16031 VBN denotes detected
T7048 16032-16034 IN denotes by
T7049 16035-16044 NN denotes toluidine
T7050 16045-16049 JJ denotes blue
T7051 16050-16058 NN denotes staining
T7052 16059-16061 IN denotes of
T7053 16062-16067 NN denotes colon
T7054 16068-16074 NN denotes mucosa
T7055 16075-16079 IN denotes from
T7056 16080-16084 NNS denotes rats
T7057 16085-16092 VBN denotes treated
T7058 16093-16097 IN denotes with
T7059 16098-16104 CC denotes either
T7060 16105-16108 DT denotes the
T7061 16109-16111 CD denotes 10
T7062 16112-16114 CC denotes or
T7063 16115-16117 CD denotes 40
T7064 16118-16121 NNP denotes kDa
T7065 16122-16126 NNP denotes dCGN
T7066 16127-16128 -LRB- denotes (
T7067 16128-16131 RB denotes not
T7068 16132-16137 VBN denotes shown
T7069 16137-16138 -RRB- denotes )
T7070 16138-16139 . denotes .
T7071 16140-16148 IN denotes Although
T7072 16149-16151 PRP denotes we
T7073 16152-16155 MD denotes can
T7074 16155-16158 RB denotes not
T7075 16159-16166 VB denotes exclude
T7076 16167-16171 DT denotes that
T7077 16172-16176 NNP denotes dCGN
T7078 16177-16180 NN denotes mat
T7079 16181-16184 RB denotes not
T7080 16185-16189 VB denotes have
T7081 16190-16198 VBN denotes retained
T7082 16199-16201 IN denotes in
T7083 16202-16205 DT denotes the
T7084 16206-16213 NN denotes section
T7085 16214-16220 IN denotes during
T7086 16221-16224 DT denotes the
T7087 16225-16234 NN denotes histology
T7088 16235-16244 NN denotes procedure
T7089 16244-16245 , denotes ,
T7090 16246-16250 DT denotes this
T7091 16251-16260 VBZ denotes indicates
T7092 16261-16265 IN denotes that
T7093 16266-16271 DT denotes these
T7094 16272-16280 NNS denotes polymers
T7095 16281-16284 MD denotes may
T7096 16285-16288 RB denotes not
T7097 16289-16293 VB denotes have
T7098 16294-16298 VBN denotes been
T7099 16299-16311 VBN denotes phagocytosed
T7100 16311-16312 . denotes .
T7658 0-3 NNP denotes CGN
T7659 4-16863 NNP denotes inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced-TNF-α
T15673 16357-16365 JJ denotes Degraded
T15674 16366-16369 NNP denotes CGN
T15675 16370-16377 VBD denotes induced
T15676 16378-16383 NN denotes colon
T15677 16384-16396 NN denotes inflammation
T15678 16397-16399 IN denotes in
T15679 16400-16404 NNS denotes rats
T15680 16404-16405 . denotes .
T15681 16406-16416 NNP denotes Histograms
T15682 16417-16424 VBG denotes showing
T15683 16425-16428 DT denotes the
T15684 16429-16435 NN denotes effect
T15685 16436-16438 IN denotes of
T15686 16439-16447 JJ denotes degraded
T15687 16448-16451 NNP denotes CGN
T15688 16452-16454 IN denotes on
T15689 16454-16455 : denotes :
T15690 16456-16461 NN denotes colon
T15691 16462-16468 NN denotes length
T15692 16469-16470 -LRB- denotes (
T15693 16470-16471 NNP denotes A
T15694 16471-16472 -RRB- denotes )
T15695 16472-16473 : denotes ;
T15696 16474-16485 JJ denotes macroscopic
T15697 16486-16487 -LRB- denotes (
T15698 16487-16488 NNP denotes B
T15699 16488-16489 -RRB- denotes )
T15700 16490-16493 CC denotes and
T15701 16494-16506 JJ denotes histological
T15702 16507-16508 -LRB- denotes (
T15703 16508-16509 NNP denotes C
T15704 16509-16510 -RRB- denotes )
T15705 16511-16523 NN denotes inflammation
T15706 16524-16529 NN denotes score
T15707 16530-16532 IN denotes of
T15708 16533-16538 NN denotes colon
T15709 16538-16539 : denotes ;
T15710 16540-16555 NNP denotes Myeloperoxidase
T15711 16556-16557 -LRB- denotes (
T15712 16557-16560 NNP denotes MPO
T15713 16560-16561 -RRB- denotes )
T15714 16562-16570 NN denotes activity
T15715 16571-16572 -LRB- denotes (
T15716 16572-16573 NNP denotes D
T15717 16573-16574 -RRB- denotes )
T15718 16574-16575 . denotes .
T15719 16576-16583 NNP denotes Control
T15720 16584-16588 NNS denotes rats
T15721 16589-16590 -LRB- denotes (
T15722 16590-16595 JJ denotes white
T15723 16596-16600 NNS denotes bars
T15724 16600-16601 -RRB- denotes )
T15725 16601-16602 : denotes ;
T15726 16603-16605 CD denotes 10
T15727 16606-16609 NN denotes kDa
T15728 16610-16618 JJ denotes degraded
T15729 16619-16630 JJ denotes CGN-treated
T15730 16631-16635 NNS denotes rats
T15731 16636-16637 -LRB- denotes (
T15732 16637-16641 NN denotes grey
T15733 16642-16646 NNS denotes bars
T15734 16646-16647 -RRB- denotes )
T15735 16647-16648 : denotes ;
T15736 16649-16651 CD denotes 40
T15737 16652-16655 NN denotes kDa
T15738 16656-16664 JJ denotes degraded
T15739 16665-16676 JJ denotes CGN-treated
T15740 16677-16681 NNS denotes rats
T15741 16682-16683 -LRB- denotes (
T15742 16683-16688 JJ denotes black
T15743 16689-16693 NNS denotes bars
T15744 16693-16694 -RRB- denotes )
T15745 16694-16695 . denotes .
T15746 16696-16697 SYM denotes *
T15747 16698-16699 NN denotes p
T15748 16699-16700 NN denotes <
T15749 16700-16704 CD denotes 0.05
T15750 16705-16709 IN denotes from
T15751 16710-16717 NN denotes control
T15752 16717-16718 . denotes .
T15753 16719-16721 SYM denotes **
T15754 16722-16723 FW denotes p
T15755 16723-16724 FW denotes <
T15756 16724-16728 CD denotes 0.01
T15757 16729-16733 IN denotes from
T15758 16734-16741 NN denotes control
T15759 16741-16742 . denotes .
T15760 16743-16755 JJ denotes Histological
T15761 16756-16764 NN denotes analysis
T15762 16765-16767 IN denotes of
T15763 16768-16773 NN denotes colon
T15764 16774-16778 IN denotes from
T15765 16779-16786 NN denotes control
T15766 16787-16791 NNS denotes rats
T15767 16792-16793 -LRB- denotes (
T15768 16793-16794 NNP denotes E
T15769 16794-16795 -RRB- denotes )
T15770 16795-16796 , denotes ,
T15771 16797-16800 CC denotes and
T15772 16801-16805 IN denotes from
T15773 16806-16808 CD denotes 40
T15774 16809-16812 NNP denotes kDa
T15775 16813-16825 NNP denotes dCGN-treated
T15776 16826-16830 NNS denotes rats
T15777 16831-16832 -LRB- denotes (
T15778 16832-16833 NN denotes F
T15779 16833-16834 -RRB- denotes )
T15780 16834-16835 . denotes .
R397 T468 T469 nsubj CGN,inhibited
R398 T469 T469 ROOT inhibited,inhibited
R399 T470 T472 nummod THP-1,proliferation
R400 T471 T472 compound cell,proliferation
R401 T472 T469 dobj proliferation,inhibited
R402 T473 T469 prep in,inhibited
R403 T474 T473 pobj vitro,in
R404 T475 T469 punct ",",inhibited
R405 T476 T469 advcl arresting,inhibited
R406 T477 T478 det the,cells
R407 T478 T476 dobj cells,arresting
R408 T479 T476 prep in,arresting
R409 T480 T481 nummod G1,phase
R410 T481 T479 pobj phase,in
R411 T482 T469 punct .,inhibited
R412 T483 T487 prep In,increased
R413 T484 T483 pobj addition,In
R414 T485 T487 punct ",",increased
R415 T486 T487 nsubj dCGN,increased
R416 T487 T487 ROOT increased,increased
R417 T488 T489 compound ICAM-1,expression
R418 T489 T487 dobj expression,increased
R419 T490 T487 prep in,increased
R420 T491 T492 preconj both,PBM
R421 T492 T490 pobj PBM,in
R422 T493 T492 cc and,PBM
R423 T494 T492 conj THP-1,PBM
R424 T495 T487 conj cells,increased
R425 T496 T495 prep with,cells
R426 T497 T499 det a,effect
R427 T498 T499 amod major,effect
R428 T499 T496 pobj effect,with
R429 T500 T499 acl seen,effect
R430 T501 T500 prep after,seen
R431 T502 T505 nummod 40,exposure
R432 T503 T504 compound kDa,dCGN
R433 T504 T505 compound dCGN,exposure
R434 T505 T501 pobj exposure,after
R435 T506 T487 punct .,increased
R436 T507 T510 advmod Also,stimulated
R437 T508 T510 punct ",",stimulated
R438 T509 T510 nsubj dCGN,stimulated
R439 T510 T510 ROOT stimulated,stimulated
R440 T511 T512 compound monocyte,aggregation
R441 T512 T510 dobj aggregation,stimulated
R442 T513 T510 prep in,stimulated
R443 T514 T513 pobj vitro,in
R444 T515 T517 nsubjpass that,prevented
R445 T516 T517 auxpass was,prevented
R446 T517 T510 conj prevented,stimulated
R447 T518 T517 agent by,prevented
R448 T519 T518 pobj incubation,by
R449 T520 T517 prep with,prevented
R450 T521 T522 amod anti-ICAM-1,antibody
R451 T522 T520 pobj antibody,with
R452 T523 T510 punct .,stimulated
R453 T524 T527 advmod Finally,stimulated
R454 T525 T527 punct ",",stimulated
R455 T526 T527 nsubj dCGN,stimulated
R456 T527 T527 ROOT stimulated,stimulated
R457 T528 T529 amod TNF-α,expression
R458 T529 T527 dobj expression,stimulated
R459 T530 T529 cc and,expression
R460 T531 T529 conj secretion,expression
R461 T532 T527 prep by,stimulated
R462 T533 T534 preconj both,PBM
R463 T534 T537 nmod PBM,cells
R464 T535 T534 cc and,PBM
R465 T536 T534 conj THP-1,PBM
R466 T537 T532 pobj cells,by
R467 T538 T527 punct .,stimulated
R468 T539 T541 predet All,effects
R469 T540 T541 det these,effects
R470 T541 T543 nsubjpass effects,linked
R471 T542 T543 auxpass were,linked
R472 T543 T543 ROOT linked,linked
R473 T544 T543 prep to,linked
R474 T545 T546 compound NF-κB,activation
R475 T546 T544 pobj activation,to
R476 T547 T543 punct .,linked
R477 T548 T549 det These,data
R478 T549 T551 nsubj data,suggest
R479 T550 T551 advmod strongly,suggest
R480 T551 T551 ROOT suggest,suggest
R481 T552 T558 mark that,have
R482 T553 T555 det the,forms
R483 T554 T555 amod degraded,forms
R484 T555 T558 nsubj forms,have
R485 T556 T555 prep of,forms
R486 T557 T556 pobj CGN,of
R487 T558 T551 ccomp have,suggest
R488 T559 T561 det a,effect
R489 T560 T561 amod pronounced,effect
R490 T561 T558 dobj effect,have
R491 T562 T561 prep on,effect
R492 T563 T562 pobj monocytes,on
R493 T564 T563 punct ",",monocytes
R494 T565 T561 conj characteristic,effect
R495 T566 T565 prep of,characteristic
R496 T567 T569 det an,phenotype
R497 T568 T569 amod inflammatory,phenotype
R498 T569 T566 pobj phenotype,of
R499 T570 T551 punct .,suggest
R1155 T1344 T1348 nsubj Carrageenan,is
R1156 T1345 T1346 punct (,CGN
R1157 T1346 T1344 appos CGN,Carrageenan
R1158 T1347 T1344 punct ),Carrageenan
R1159 T1348 T1348 ROOT is,is
R1160 T1349 T1354 det a,polysaccharide
R1161 T1350 T1352 amod high,weight
R1162 T1351 T1352 amod molecular,weight
R1163 T1352 T1354 nmod weight,polysaccharide
R1164 T1353 T1354 amod sulphated,polysaccharide
R1165 T1354 T1348 attr polysaccharide,is
R1166 T1355 T1356 punct (,>
R1167 T1356 T1354 appos >,polysaccharide
R1168 T1357 T1358 nummod 200,kDa
R1169 T1358 T1354 appos kDa,polysaccharide
R1170 T1359 T1358 punct ),kDa
R1171 T1360 T1354 acl derived,polysaccharide
R1172 T1361 T1360 prep from,derived
R1173 T1362 T1363 amod red,algae
R1174 T1363 T1361 pobj algae,from
R1175 T1364 T1363 punct (,algae
R1176 T1365 T1363 appos Rhodophyceae,algae
R1177 T1366 T1363 punct ),algae
R1178 T1367 T1348 punct .,is
R1179 T1368 T1370 nummod Three,forms
R1180 T1369 T1370 amod main,forms
R1181 T1370 T1375 nsubjpass forms,identified
R1182 T1371 T1370 prep of,forms
R1183 T1372 T1371 pobj CGN,of
R1184 T1373 T1375 aux have,identified
R1185 T1374 T1375 auxpass been,identified
R1186 T1375 T1375 ROOT identified,identified
R1187 T1376 T1375 punct :,identified
R1188 T1377 T1375 npadvmod kappa,identified
R1189 T1378 T1377 punct ",",kappa
R1190 T1379 T1377 conj iota,kappa
R1191 T1380 T1379 punct ",",iota
R1192 T1381 T1379 cc and,iota
R1193 T1382 T1379 conj lambda,iota
R1194 T1383 T1375 punct .,identified
R1195 T1384 T1385 nsubj They,differ
R1196 T1385 T1385 ROOT differ,differ
R1197 T1386 T1385 prep from,differ
R1198 T1387 T1388 det each,other
R1199 T1388 T1386 pobj other,from
R1200 T1389 T1385 prep in,differ
R1201 T1390 T1391 compound sulphation,degree
R1202 T1391 T1389 pobj degree,in
R1203 T1392 T1391 cc and,degree
R1204 T1393 T1394 compound solubility,[
R1205 T1394 T1385 conj [,differ
R1206 T1395 T1396 nummod 1,]
R1207 T1396 T1394 conj ],[
R1208 T1397 T1396 punct ",",]
R1209 T1398 T1396 conj [,]
R1210 T1399 T1400 nummod 2,]
R1211 T1400 T1396 conj ],]
R1212 T1401 T1385 punct .,differ
R1213 T1402 T1403 amod Native,CGN
R1214 T1403 T1405 nsubjpass CGN,thought
R1215 T1404 T1405 auxpass is,thought
R1216 T1405 T1405 ROOT thought,thought
R1217 T1406 T1407 aux to,be
R1218 T1407 T1405 xcomp be,thought
R1219 T1408 T1407 acomp harmless,be
R1220 T1409 T1405 cc and,thought
R1221 T1410 T1412 auxpass is,used
R1222 T1411 T1412 advmod widely,used
R1223 T1412 T1405 conj used,thought
R1224 T1413 T1412 prep as,used
R1225 T1414 T1416 det a,additive
R1226 T1415 T1416 compound food,additive
R1227 T1416 T1413 pobj additive,as
R1228 T1417 T1418 aux to,improve
R1229 T1418 T1416 relcl improve,additive
R1230 T1419 T1418 dobj texture,improve
R1231 T1420 T1405 punct .,thought
R1232 T1421 T1424 nsubjpass It,used
R1233 T1422 T1424 auxpass is,used
R1234 T1423 T1424 advmod also,used
R1235 T1424 T1424 ROOT used,used
R1236 T1425 T1424 prep in,used
R1237 T1426 T1425 pobj cosmetics,in
R1238 T1427 T1426 cc and,cosmetics
R1239 T1428 T1426 conj pharmaceuticals,cosmetics
R1240 T1429 T1424 punct .,used
R1241 T1430 T1442 advmod However,triggers
R1242 T1431 T1442 punct ",",triggers
R1243 T1432 T1433 compound acid,treatment
R1244 T1433 T1442 nsubj treatment,triggers
R1245 T1434 T1433 prep at,treatment
R1246 T1435 T1436 amod high,temperature
R1247 T1436 T1434 pobj temperature,at
R1248 T1437 T1436 punct (,temperature
R1249 T1438 T1440 nummod 80,C
R1250 T1439 T1440 nummod °,C
R1251 T1440 T1433 npadvmod C,treatment
R1252 T1441 T1433 punct ),treatment
R1253 T1442 T1442 ROOT triggers,triggers
R1254 T1443 T1444 compound CGN,hydrolysis
R1255 T1444 T1442 dobj hydrolysis,triggers
R1256 T1445 T1446 aux to,lower
R1257 T1446 T1448 amod lower,weight
R1258 T1447 T1448 amod molecular,weight
R1259 T1448 T1444 appos weight,hydrolysis
R1260 T1449 T1450 punct (,<
R1261 T1450 T1452 nmod <,kDa
R1262 T1451 T1452 nummod 50,kDa
R1263 T1452 T1448 appos kDa,weight
R1264 T1453 T1448 punct ),weight
R1265 T1454 T1442 conj compounds,triggers
R1266 T1455 T1454 acl known,compounds
R1267 T1456 T1455 prep as,known
R1268 T1457 T1456 pobj poligeenan,as
R1269 T1458 T1457 cc or,poligeenan
R1270 T1459 T1460 amod degraded,CGN
R1271 T1460 T1457 conj CGN,poligeenan
R1272 T1461 T1460 punct (,CGN
R1273 T1462 T1460 appos dCGN,CGN
R1274 T1463 T1460 punct ),CGN
R1275 T1464 T1442 punct .,triggers
R1276 T1465 T1466 det These,dCGNs
R1277 T1466 T1467 nsubj dCGNs,induce
R1278 T1467 T1467 ROOT induce,induce
R1279 T1468 T1467 dobj inflammation,induce
R1280 T1469 T1467 cc and,induce
R1281 T1470 T1473 aux have,used
R1282 T1471 T1473 auxpass been,used
R1283 T1472 T1473 advmod widely,used
R1284 T1473 T1467 conj used,induce
R1285 T1474 T1473 prep as,used
R1286 T1475 T1474 pobj models,as
R1287 T1476 T1475 prep of,models
R1288 T1477 T1476 pobj colitis,of
R1289 T1478 T1475 prep in,models
R1290 T1479 T1480 amod several,species
R1291 T1480 T1478 pobj species,in
R1292 T1481 T1480 punct ",",species
R1293 T1482 T1480 prep including,species
R1294 T1483 T1484 nsubj rats,[
R1295 T1484 T1486 nmod [,]
R1296 T1485 T1486 nummod 3,]
R1297 T1486 T1482 pobj ],including
R1298 T1487 T1486 punct ",",]
R1299 T1488 T1486 conj rabbits,]
R1300 T1489 T1491 nmod [,]
R1301 T1490 T1491 nummod 4,]
R1302 T1491 T1494 nmod ],pigs
R1303 T1492 T1491 cc and,]
R1304 T1493 T1494 compound guinea,pigs
R1305 T1494 T1495 nsubj pigs,[
R1306 T1495 T1488 relcl [,rabbits
R1307 T1496 T1497 nummod 5,]
R1308 T1497 T1495 dobj ],[
R1309 T1498 T1467 punct .,induce
R1310 T1499 T1500 det The,role
R1311 T1500 T1507 nsubj role,remains
R1312 T1501 T1500 prep of,role
R1313 T1502 T1501 pobj dCGN,of
R1314 T1503 T1500 prep as,role
R1315 T1504 T1506 det a,factor
R1316 T1505 T1506 amod tumor-promoting,factor
R1317 T1506 T1503 pobj factor,as
R1318 T1507 T1507 ROOT remains,remains
R1319 T1508 T1507 acomp controversial,remains
R1320 T1509 T1518 nmod [,]
R1321 T1510 T1509 nummod 4,[
R1322 T1511 T1509 conj ],[
R1323 T1512 T1511 punct ",",]
R1324 T1513 T1515 quantmod [,]
R1325 T1514 T1515 nummod 6,]
R1326 T1515 T1509 appos ],[
R1327 T1516 T1518 nmod [,]
R1328 T1517 T1518 nummod 8,]
R1329 T1518 T1507 npadvmod ],remains
R1330 T1519 T1507 punct .,remains
R1331 T1520 T1525 mark Although,thought
R1332 T1521 T1523 det the,form
R1333 T1522 T1523 amod native,form
R1334 T1523 T1525 nsubjpass form,thought
R1335 T1524 T1525 auxpass is,thought
R1336 T1525 T1539 advcl thought,produced
R1337 T1526 T1527 aux to,be
R1338 T1527 T1525 xcomp be,thought
R1339 T1528 T1527 acomp harmless,be
R1340 T1529 T1527 prep for,be
R1341 T1530 T1531 amod human,consumption
R1342 T1531 T1529 pobj consumption,for
R1343 T1532 T1539 punct ",",produced
R1344 T1533 T1534 amod small,amounts
R1345 T1534 T1539 nsubjpass amounts,produced
R1346 T1535 T1534 prep of,amounts
R1347 T1536 T1535 pobj dCGN,of
R1348 T1537 T1539 auxpass are,produced
R1349 T1538 T1539 advmod probably,produced
R1350 T1539 T1539 ROOT produced,produced
R1351 T1540 T1539 agent by,produced
R1352 T1541 T1542 compound acid,hydrolysis
R1353 T1542 T1540 pobj hydrolysis,by
R1354 T1543 T1539 prep during,produced
R1355 T1544 T1545 amod gastric,digestion
R1356 T1545 T1543 pobj digestion,during
R1357 T1546 T1548 nmod [,]
R1358 T1547 T1548 nummod 9,]
R1359 T1548 T1545 appos ],digestion
R1360 T1549 T1548 punct ",",]
R1361 T1550 T1552 nmod [,]
R1362 T1551 T1552 nummod 10,]
R1363 T1552 T1548 conj ],]
R1364 T1553 T1552 cc or,]
R1365 T1554 T1552 conj interaction,]
R1366 T1555 T1554 prep with,interaction
R1367 T1556 T1557 amod intestinal,bacteria
R1368 T1557 T1555 pobj bacteria,with
R1369 T1558 T1560 compound [,]
R1370 T1559 T1560 compound 11,]
R1371 T1560 T1557 appos ],bacteria
R1372 T1561 T1560 punct ",",]
R1373 T1562 T1564 nmod [,]
R1374 T1563 T1564 nummod 12,]
R1375 T1564 T1560 appos ],]
R1376 T1565 T1539 punct .,produced
R1377 T1566 T1579 mark Whereas,analyzed
R1378 T1567 T1568 det the,effects
R1379 T1568 T1579 nsubjpass effects,analyzed
R1380 T1569 T1568 prep of,effects
R1381 T1570 T1573 quantmod native,on
R1382 T1571 T1570 cc and,native
R1383 T1572 T1570 conj dCGN,native
R1384 T1573 T1568 prep on,effects
R1385 T1574 T1575 amod intestinal,inflammation
R1386 T1575 T1573 pobj inflammation,on
R1387 T1576 T1579 aux have,analyzed
R1388 T1577 T1579 auxpass been,analyzed
R1389 T1578 T1579 advmod extensively,analyzed
R1390 T1579 T1589 advcl analyzed,conducted
R1391 T1580 T1579 prep in,analyzed
R1392 T1581 T1582 compound animal,models
R1393 T1582 T1580 pobj models,in
R1394 T1583 T1589 punct ",",conducted
R1395 T1584 T1586 advmod only,studies
R1396 T1585 T1586 amod few,studies
R1397 T1586 T1589 nsubjpass studies,conducted
R1398 T1587 T1589 aux have,conducted
R1399 T1588 T1589 auxpass been,conducted
R1400 T1589 T1589 ROOT conducted,conducted
R1401 T1590 T1589 advcl using,conducted
R1402 T1591 T1593 amod human,lines
R1403 T1592 T1593 compound cell,lines
R1404 T1593 T1590 dobj lines,using
R1405 T1594 T1589 punct .,conducted
R1406 T1595 T1596 amod Recent,studies
R1407 T1596 T1598 nsubj studies,shown
R1408 T1597 T1598 aux have,shown
R1409 T1598 T1598 ROOT shown,shown
R1410 T1599 T1600 det a,link
R1411 T1600 T1598 dobj link,shown
R1412 T1601 T1600 prep between,link
R1413 T1602 T1606 compound exposure,CGN
R1414 T1603 T1604 aux to,native
R1415 T1604 T1605 amod native,form
R1416 T1605 T1606 compound form,CGN
R1417 T1606 T1601 pobj CGN,between
R1418 T1607 T1606 cc and,CGN
R1419 T1608 T1609 amod IL-8,production
R1420 T1609 T1606 conj production,CGN
R1421 T1610 T1609 prep by,production
R1422 T1611 T1616 det the,line
R1423 T1612 T1615 amod human,cell
R1424 T1613 T1615 amod intestinal,cell
R1425 T1614 T1615 compound epithelial,cell
R1426 T1615 T1616 compound cell,line
R1427 T1616 T1610 pobj line,by
R1428 T1617 T1616 punct ",",line
R1429 T1618 T1616 appos NCM460,line
R1430 T1619 T1598 punct ",",shown
R1431 T1620 T1598 prep via,shown
R1432 T1621 T1622 compound Nuclear,Factor-κB
R1433 T1622 T1620 pobj Factor-κB,via
R1434 T1623 T1622 punct (,Factor-κB
R1435 T1624 T1622 appos NF-κB,Factor-κB
R1436 T1625 T1622 punct ),Factor-κB
R1437 T1626 T1627 compound activation,[
R1438 T1627 T1629 nmod [,]
R1439 T1628 T1629 nummod 13,]
R1440 T1629 T1598 dobj ],shown
R1441 T1630 T1629 punct ",",]
R1442 T1631 T1629 conj [,]
R1443 T1632 T1633 nummod 14,]
R1444 T1633 T1629 conj ],]
R1445 T1634 T1598 punct .,shown
R1446 T1635 T1636 nsubj NF-κB,is
R1447 T1636 T1636 ROOT is,is
R1448 T1637 T1639 det a,factor
R1449 T1638 T1639 compound transcription,factor
R1450 T1639 T1636 attr factor,is
R1451 T1640 T1641 nsubj that,regulates
R1452 T1641 T1639 relcl regulates,factor
R1453 T1642 T1643 det the,expression
R1454 T1643 T1641 dobj expression,regulates
R1455 T1644 T1643 prep of,expression
R1456 T1645 T1644 pobj genes,of
R1457 T1646 T1645 acl associated,genes
R1458 T1647 T1646 prep with,associated
R1459 T1648 T1647 pobj inflammation,with
R1460 T1649 T1651 nmod [,]
R1461 T1650 T1651 nummod 15,]
R1462 T1651 T1643 appos ],expression
R1463 T1652 T1651 punct ",",]
R1464 T1653 T1655 nmod [,]
R1465 T1654 T1655 nummod 16,]
R1466 T1655 T1651 appos ],]
R1467 T1656 T1636 punct .,is
R1468 T1657 T1658 compound Macrophage,infiltration
R1469 T1658 T1661 nsubj infiltration,is
R1470 T1659 T1658 cc and,infiltration
R1471 T1660 T1658 conj accumulation,infiltration
R1472 T1661 T1661 ROOT is,is
R1473 T1662 T1664 det a,characteristic
R1488 T1677 T1680 amod total,cells
R1489 T1678 T1680 compound lamina,cells
R1490 T1679 T1680 compound propria,cells
R1491 T1680 T1676 pobj cells,of
R1492 T1681 T1673 punct ",",represent
R1493 T1682 T1673 conj secrete,represent
R1494 T1683 T1685 det a,range
R1495 T1684 T1685 amod wide,range
R1496 T1685 T1682 dobj range,secrete
R1497 T1686 T1685 prep of,range
R1498 T1687 T1688 advmod biologically,active
R1499 T1688 T1689 amod active,compounds
R1500 T1689 T1686 pobj compounds,of
R1501 T1690 T1682 cc and,secrete
R1502 T1691 T1682 conj express,secrete
R1503 T1692 T1693 amod cell-adhesion,molecules
R1504 T1693 T1691 dobj molecules,express
R1505 T1694 T1673 punct .,represent
R1506 T1695 T1698 det The,response
R1507 T1696 T1698 amod immune,response
R1508 T1697 T1698 compound cell,response
R1509 T1698 T1703 nsubj response,seems
R1510 T1699 T1698 prep to,response
R1511 T1700 T1702 det an,stimulus
R1512 T1701 T1702 amod inflammatory,stimulus
R1513 T1702 T1699 pobj stimulus,to
R1514 T1703 T1703 ROOT seems,seems
R1515 T1704 T1706 aux to,amplified
R1516 T1705 T1706 auxpass be,amplified
R1517 T1706 T1703 xcomp amplified,seems
R1518 T1707 T1706 cc or,amplified
R1519 T1708 T1709 advmod directly,generated
R1520 T1709 T1706 conj generated,amplified
R1521 T1710 T1709 agent by,generated
R1522 T1711 T1710 pobj cells,by
R1523 T1712 T1711 acl exposed,cells
R1524 T1713 T1712 prep to,exposed
R1525 T1714 T1715 amod sulphated,polysaccharides
R1526 T1715 T1713 pobj polysaccharides,to
R1527 T1716 T1717 amod such,as
R1528 T1717 T1715 prep as,polysaccharides
R1529 T1718 T1717 pobj carrageenans,as
R1530 T1719 T1703 punct .,seems
R1531 T1720 T1727 advmod Indeed,associated
R1532 T1721 T1727 punct ",",associated
R1533 T1722 T1727 nsubjpass inflammation,associated
R1534 T1723 T1722 acl induced,inflammation
R1535 T1724 T1723 agent by,induced
R1536 T1725 T1724 pobj dCGN,by
R1537 T1726 T1727 auxpass was,associated
R1538 T1727 T1727 ROOT associated,associated
R1539 T1728 T1727 prep with,associated
R1540 T1729 T1728 pobj recruitment,with
R1541 T1730 T1729 prep of,recruitment
R1542 T1731 T1730 pobj macrophages,of
R1543 T1732 T1727 prep to,associated
R1544 T1733 T1734 compound inflammation,sites
R1545 T1734 T1732 pobj sites,to
R1546 T1735 T1737 nmod [,]
R1547 T1736 T1737 nummod 18,]
R1548 T1737 T1734 appos ],sites
R1549 T1738 T1737 punct ",",]
R1550 T1739 T1741 nmod [,]
R1551 T1740 T1741 nummod 19,]
R1552 T1741 T1737 conj ],]
R1553 T1742 T1727 punct .,associated
R1554 T1743 T1761 advmod Also,associated
R1555 T1744 T1761 punct ",",associated
R1556 T1745 T1761 nsubjpass inflammation,associated
R1557 T1746 T1745 acl induced,inflammation
R1558 T1747 T1746 agent by,induced
R1559 T1748 T1750 compound Dextran,Sodium
R1560 T1749 T1750 compound Sulphate,Sodium
R1561 T1750 T1761 nsubjpass Sodium,associated
R1562 T1751 T1752 punct (,DSS
R1563 T1752 T1750 appos DSS,Sodium
R1564 T1753 T1750 punct ),Sodium
R1565 T1754 T1750 punct ",",Sodium
R1566 T1755 T1757 det another,compound
R1567 T1756 T1757 amod sulphated,compound
R1568 T1757 T1750 appos compound,Sodium
R1569 T1758 T1761 punct ",",associated
R1570 T1759 T1761 auxpass was,associated
R1571 T1760 T1761 advmod directly,associated
R1572 T1761 T1761 ROOT associated,associated
R1573 T1762 T1761 prep with,associated
R1574 T1763 T1764 compound macrophages,recruitment
R1575 T1764 T1762 pobj recruitment,with
R1576 T1765 T1767 nmod [,]
R1577 T1766 T1767 nummod 20,]
R1578 T1767 T1764 appos ],recruitment
R1579 T1768 T1761 punct ",",associated
R1580 T1769 T1772 mark since,provoked
R1581 T1770 T1772 nsubj DSS,provoked
R1582 T1771 T1772 advmod still,provoked
R1583 T1772 T1761 advcl provoked,associated
R1584 T1773 T1772 dobj inflammation,provoked
R1585 T1774 T1772 prep after,provoked
R1586 T1775 T1774 pobj T-lymphocyte,after
R1587 T1776 T1775 cc and,T-lymphocyte
R1588 T1777 T1779 compound NK,depletion
R1589 T1778 T1779 compound cell,depletion
R1590 T1779 T1775 conj depletion,T-lymphocyte
R1591 T1780 T1782 nsubj [,]
R1592 T1781 T1782 nummod 20,]
R1593 T1782 T1782 ROOT ],]
R1594 T1783 T1761 punct .,associated
R1595 T1784 T1788 mark Although,induced
R1596 T1785 T1788 nsubjpass inflammation,induced
R1597 T1786 T1788 aux can,induced
R1598 T1787 T1788 auxpass be,induced
R1599 T1788 T1793 advcl induced,are
R1600 T1789 T1788 agent by,induced
R1601 T1790 T1789 pobj dCGN,by
R1602 T1791 T1793 punct ",",are
R1603 T1792 T1793 expl there,are
R1604 T1793 T1793 ROOT are,are
R1605 T1794 T1795 det no,data
R1606 T1795 T1793 attr data,are
R1607 T1796 T1795 prep on,data
R1608 T1797 T1799 amod human,responses
R1609 T1798 T1799 compound monocyte,responses
R1610 T1799 T1796 pobj responses,on
R1611 T1800 T1799 prep to,responses
R1612 T1801 T1802 compound dCGN,exposure
R1613 T1802 T1800 pobj exposure,to
R1614 T1803 T1793 punct .,are
R1615 T1804 T1829 advmod Therefore,exposed
R1616 T1805 T1829 punct ",",exposed
R1617 T1806 T1807 aux to,investigate
R1618 T1807 T1829 advcl investigate,exposed
R1619 T1808 T1809 det the,effects
R1620 T1809 T1807 dobj effects,investigate
R1621 T1810 T1809 prep of,effects
R1622 T1811 T1810 pobj dCGN,of
R1623 T1812 T1809 prep on,effects
R1624 T1813 T1814 amod human,monocytes
R1625 T1814 T1812 pobj monocytes,on
R1626 T1815 T1814 punct ",",monocytes
R1627 T1816 T1819 amod normal,Monocytes
R1628 T1817 T1818 compound Peripheral,Blood
R1629 T1818 T1819 compound Blood,Monocytes
R1630 T1819 T1827 nmod Monocytes,cells
R1631 T1820 T1821 punct (,PBM
R1632 T1821 T1819 appos PBM,Monocytes
R1633 T1822 T1821 punct ),PBM
R1634 T1823 T1819 cc and,Monocytes
R1635 T1824 T1825 amod tumoral,monocyte/macrophage
R1636 T1825 T1819 conj monocyte/macrophage,Monocytes
R1637 T1826 T1827 nummod THP-1,cells
R1638 T1827 T1829 nsubjpass cells,exposed
R1639 T1828 T1829 auxpass were,exposed
R1640 T1829 T1829 ROOT exposed,exposed
R1641 T1830 T1829 prep to,exposed
R1642 T1831 T1832 nummod 10,kDa
R1643 T1832 T1830 pobj kDa,to
R1644 T1833 T1832 cc and,kDa
R1645 T1834 T1836 nummod 40,dCGN
R1646 T1835 T1836 compound kDa,dCGN
R1647 T1836 T1832 conj dCGN,kDa
R1648 T1837 T1829 punct .,exposed
R1649 T1838 T1839 nsubj We,found
R1650 T1839 T1839 ROOT found,found
R1651 T1840 T1842 mark that,inhibited
R1652 T1841 T1842 nsubj dCGN,inhibited
R1653 T1842 T1839 ccomp inhibited,found
R1654 T1843 T1845 nummod THP-1,proliferation
R1655 T1844 T1845 compound cell,proliferation
R1656 T1845 T1842 dobj proliferation,inhibited
R1657 T1846 T1842 prep in,inhibited
R1658 T1847 T1846 pobj vitro,in
R1659 T1848 T1842 punct ",",inhibited
R1660 T1849 T1851 amod increased,expression
R1661 T1850 T1851 compound ICAM-1,expression
R1662 T1851 T1839 dobj expression,found
R1663 T1852 T1851 punct ",",expression
R1664 T1853 T1851 acl stimulated,expression
R1665 T1854 T1856 amod ICAM-1-dependent,aggregation
R1666 T1855 T1856 compound monocyte,aggregation
R1667 T1856 T1853 dobj aggregation,stimulated
R1668 T1857 T1839 punct ",",found
R1669 T1858 T1839 cc and,found
R1670 T1859 T1839 conj stimulated,found
R1671 T1860 T1861 compound TNF-α,expression
R1672 T1861 T1859 dobj expression,stimulated
R1673 T1862 T1861 cc and,expression
R1674 T1863 T1861 conj secretion,expression
R1675 T1864 T1839 punct .,found
R1676 T1865 T1866 det These,responses
R1677 T1866 T1867 nsubj responses,were
R1678 T1867 T1867 ROOT were,were
R1679 T1868 T1869 advmod more,pronounced
R1680 T1869 T1867 acomp pronounced,were
R1681 T1870 T1869 prep after,pronounced
R1682 T1871 T1874 nummod 40,exposure
R1683 T1872 T1874 compound kDa,exposure
R1684 T1873 T1874 compound dCGN,exposure
R1685 T1874 T1870 pobj exposure,after
R1686 T1875 T1867 cc and,were
R1687 T1876 T1877 auxpass were,linked
R1688 T1877 T1867 conj linked,were
R1689 T1878 T1877 prep to,linked
R1690 T1879 T1880 compound NF-κB,activation
R1691 T1880 T1878 pobj activation,to
R1692 T1881 T1867 punct .,were
R1693 T1882 T1896 prep In,induced
R1694 T1883 T1882 pobj addition,In
R1695 T1884 T1896 punct ",",induced
R1696 T1885 T1888 det the,dCGN
R1697 T1886 T1888 nummod 40,dCGN
R1698 T1887 T1888 compound kDa,dCGN
R1699 T1888 T1896 nsubj dCGN,induced
R1700 T1889 T1888 punct ",",dCGN
R1701 T1890 T1896 cc but,induced
R1702 T1891 T1896 neg not,induced
R1703 T1892 T1895 det the,dCGN
R1704 T1893 T1895 nummod 10,dCGN
R1705 T1894 T1895 compound kDa,dCGN
R1706 T1895 T1896 nsubj dCGN,induced
R1707 T1896 T1896 ROOT induced,induced
R1708 T1897 T1896 prep in,induced
R1709 T1898 T1899 compound vivo,colitis
R1710 T1899 T1897 pobj colitis,in
R1711 T1900 T1901 mark as,shown
R1712 T1901 T1896 advcl shown,induced
R1713 T1902 T1901 agent by,shown
R1714 T1903 T1905 det the,response
R1715 T1904 T1905 amod inflammatory,response
R1716 T1905 T1902 pobj response,by
R1717 T1906 T1905 prep in,response
R1718 T1907 T1909 det the,colon
R1719 T1908 T1909 compound rat,colon
R1720 T1909 T1906 pobj colon,in
R1721 T1910 T1896 punct .,induced
R1722 T1911 T1912 det These,results
R1723 T1912 T1913 nsubj results,suggest
R1724 T1913 T1913 ROOT suggest,suggest
R1725 T1914 T1920 mark that,have
R1726 T1915 T1917 det the,forms
R1727 T1916 T1917 amod degraded,forms
R1728 T1917 T1920 nsubj forms,have
R1729 T1918 T1917 prep of,forms
R1730 T1919 T1918 pobj CGN,of
R1731 T1920 T1913 ccomp have,suggest
R1732 T1921 T1923 det an,effect
R1733 T1922 T1923 amod important,effect
R1734 T1923 T1920 dobj effect,have
R1735 T1924 T1923 prep on,effect
R1736 T1925 T1924 pobj monocytes,on
R1737 T1926 T1923 acl resulting,effect
R1738 T1927 T1926 prep in,resulting
R1739 T1928 T1930 det an,phenotype
R1740 T1929 T1930 amod inflammatory,phenotype
R1741 T1930 T1927 pobj phenotype,in
R1742 T1931 T1913 punct .,suggest
R2132 T2416 T2449 nsubjpass Preparation,prepared
R2133 T2417 T2416 prep of,Preparation
R2134 T2418 T2419 compound Degraded,Carrageenan
R2135 T2419 T2417 pobj Carrageenan,of
R2136 T2420 T2421 nummod Two,preparations
R2137 T2421 T2447 nmod preparations,weight
R2138 T2422 T2421 prep of,preparations
R2139 T2423 T2424 amod degraded,carrageenan
R2140 T2424 T2422 pobj carrageenan,of
R2141 T2425 T2424 prep with,carrageenan
R2142 T2426 T2425 pobj low,with
R2143 T2427 T2421 punct ",",preparations
R2144 T2428 T2429 punct (,∼
R2145 T2429 T2421 appos ∼,preparations
R2146 T2430 T2431 nummod 10,kDa
R2147 T2431 T2429 npadvmod kDa,∼
R2148 T2432 T2421 punct ;,preparations
R2149 T2433 T2421 appos C10,preparations
R2150 T2434 T2421 punct ),preparations
R2151 T2435 T2421 punct ",",preparations
R2152 T2436 T2421 cc and,preparations
R2180 T2464 T2463 punct ",",Industry
R2181 T2465 T2463 conj Boulogne-Billancourt,Industry
R2182 T2466 T2465 punct ",",Boulogne-Billancourt
R2183 T2467 T2465 conj France,Boulogne-Billancourt
R2184 T2468 T2452 punct ),iota-carrageenan
R2185 T2469 T2449 punct .,prepared
R2186 T2470 T2471 amod Native,carrageenan
R2187 T2471 T2473 nsubjpass carrageenan,dissolved
R2188 T2472 T2473 auxpass was,dissolved
R2189 T2473 T2473 ROOT dissolved,dissolved
R2190 T2474 T2473 prep in,dissolved
R2191 T2475 T2476 amod distilled,water
R2192 T2476 T2474 pobj water,in
R2193 T2477 T2480 punct (,w/v
R2194 T2478 T2479 nummod 5,%
R2195 T2479 T2480 compound %,w/v
R2196 T2480 T2473 parataxis w/v,dissolved
R2197 T2481 T2480 punct ),w/v
R2198 T2482 T2473 prep under,dissolved
R2199 T2483 T2484 amod vigorous,stirring
R2200 T2484 T2482 pobj stirring,under
R2201 T2485 T2484 cc and,stirring
R2202 T2486 T2484 conj heated,stirring
R2203 T2487 T2486 prep to,heated
R2204 T2488 T2489 nummod 60,°
R2232 T2516 T2518 amod low,weight
R2233 T2517 T2518 amod molecular,weight
R2234 T2518 T2519 compound weight,fraction
R2235 T2519 T2514 pobj fraction,for
R2236 T2520 T2524 punct ",",hydrolyzed
R2237 T2521 T2522 compound carrageenan,solution
R2238 T2522 T2524 nsubjpass solution,hydrolyzed
R2239 T2523 T2524 auxpass was,hydrolyzed
R2240 T2524 T2524 ROOT hydrolyzed,hydrolyzed
R2241 T2525 T2524 prep with,hydrolyzed
R2242 T2526 T2527 nummod 0.3,%
R2243 T2527 T2525 pobj %,with
R2244 T2528 T2529 punct (,v/v
R2245 T2529 T2527 appos v/v,%
R2246 T2530 T2524 punct ),hydrolyzed
R2247 T2531 T2533 nmod concentrated,acid
R2248 T2532 T2533 compound sulphuric,acid
R2249 T2533 T2524 dobj acid,hydrolyzed
R2250 T2534 T2524 prep for,hydrolyzed
R2251 T2535 T2536 nummod 15,min
R2252 T2536 T2534 pobj min,for
R2253 T2537 T2524 prep at,hydrolyzed
R2254 T2538 T2540 nummod 80,C
R2255 T2539 T2540 nummod °,C
R2256 T2540 T2537 pobj C,at
R2257 T2541 T2524 punct .,hydrolyzed
R2258 T2542 T2552 prep After,filtered
R2259 T2543 T2542 pobj neutralization,After
R2260 T2544 T2543 prep with,neutralization
R2261 T2545 T2546 compound NaOH,4N
R2262 T2546 T2544 pobj 4N,with
R2263 T2547 T2552 punct ",",filtered
R2264 T2548 T2549 det the,solution
R2265 T2549 T2552 nsubjpass solution,filtered
R2266 T2550 T2552 auxpass was,filtered
R2267 T2551 T2552 advmod ultra,filtered
R2268 T2552 T2552 ROOT filtered,filtered
R2269 T2553 T2552 prep through,filtered
R2270 T2554 T2557 det a,cartridge
R2271 T2555 T2557 amod hollow,cartridge
R2272 T2556 T2557 compound fibre,cartridge
R2273 T2557 T2553 pobj cartridge,through
R2274 T2558 T2557 prep with,cartridge
R2275 T2559 T2562 nmod MW,kDa
R2276 T2560 T2562 nmod cut-off,kDa
R2277 T2561 T2562 nummod 5,kDa
R2278 T2562 T2558 pobj kDa,with
R2279 T2563 T2562 punct ",",kDa
R2280 T2564 T2566 punct (,Inc
R2281 T2565 T2566 compound Amicon,Inc
R2282 T2566 T2562 conj Inc,kDa
R2283 T2567 T2566 punct ",",Inc
R2284 T2568 T2566 conj Beverly,Inc
R2285 T2569 T2568 punct ",",Beverly
R2286 T2570 T2568 conj USA,Beverly
R2287 T2571 T2566 punct ),Inc
R2288 T2572 T2552 punct .,filtered
R2289 T2573 T2584 prep For,hydrolyzed
R2290 T2574 T2578 det the,fraction
R2291 T2575 T2578 compound medium,fraction
R2292 T2576 T2577 amod molecular,weight
R2293 T2577 T2578 compound weight,fraction
R2294 T2578 T2573 pobj fraction,For
R2295 T2579 T2584 punct ",",hydrolyzed
R2296 T2580 T2582 det the,solution
R2297 T2581 T2582 compound carrageenan,solution
R2298 T2582 T2584 nsubjpass solution,hydrolyzed
R2299 T2583 T2584 auxpass was,hydrolyzed
R2300 T2584 T2584 ROOT hydrolyzed,hydrolyzed
R2301 T2585 T2584 prep with,hydrolyzed
R2302 T2586 T2587 nummod 0.3,%
R2303 T2587 T2585 pobj %,with
R2304 T2588 T2589 punct (,v/v
R2305 T2589 T2587 appos v/v,%
R2306 T2590 T2584 punct ),hydrolyzed
R2307 T2591 T2591 ROOT concentrated,concentrated
R2308 T2592 T2593 amod sulphuric,acid
R2309 T2593 T2591 dobj acid,concentrated
R2310 T2594 T2591 prep for,concentrated
R2311 T2595 T2596 nummod 30,min
R2312 T2596 T2594 pobj min,for
R2313 T2597 T2591 prep at,concentrated
R2314 T2598 T2599 nummod 60,°
R2315 T2599 T2597 pobj °,at
R2316 T2600 T2591 npadvmod C,concentrated
R2317 T2601 T2584 punct .,hydrolyzed
R2318 T2602 T2609 prep After,filtered
R2319 T2603 T2602 pobj neutralization,After
R2320 T2604 T2609 punct ",",filtered
R2321 T2605 T2606 det the,supernatant
R2322 T2606 T2609 nsubjpass supernatant,filtered
R2323 T2607 T2609 auxpass was,filtered
R2324 T2608 T2609 advmod ultra,filtered
R2325 T2609 T2609 ROOT filtered,filtered
R2326 T2610 T2614 punct (,kDa
R2327 T2611 T2614 prt MW,kDa
R2328 T2612 T2614 nmod cut-off,kDa
R2329 T2613 T2614 nummod 100,kDa
R2330 T2614 T2609 dobj kDa,filtered
R2331 T2615 T2614 punct ),kDa
R2332 T2616 T2609 punct .,filtered
R2333 T2617 T2618 det The,filtrate
R2334 T2618 T2620 nsubjpass filtrate,submitted
R2335 T2619 T2620 auxpass was,submitted
R2336 T2620 T2620 ROOT submitted,submitted
R2337 T2621 T2620 prep to,submitted
R2338 T2622 T2625 det a,filtration
R2339 T2623 T2625 amod second,filtration
R2340 T2624 T2625 compound ultra,filtration
R2341 T2625 T2621 pobj filtration,to
R2342 T2626 T2630 punct (,kDa
R2343 T2627 T2630 nmod MW,kDa
R2344 T2628 T2630 punct cut-off,kDa
R2345 T2629 T2628 nummod 5,cut-off
R2346 T2630 T2625 appos kDa,filtration
R2347 T2631 T2630 punct ),kDa
R2348 T2632 T2620 punct .,submitted
R2349 T2633 T2634 det Both,preparations
R2350 T2634 T2638 nsubjpass preparations,precipitated
R2351 T2635 T2634 prep of,preparations
R2352 T2636 T2635 pobj dCGN,of
R2353 T2637 T2638 auxpass were,precipitated
R2354 T2638 T2638 ROOT precipitated,precipitated
R2355 T2639 T2638 prep with,precipitated
R2356 T2640 T2641 nummod 4,volumes
R2357 T2641 T2639 pobj volumes,with
R2358 T2642 T2641 prep of,volumes
R2359 T2643 T2644 nummod 95,%
R2360 T2644 T2645 compound %,ethanol
R2361 T2645 T2642 pobj ethanol,of
R2362 T2646 T2638 punct ",",precipitated
R2363 T2647 T2638 conj dried,precipitated
R2364 T2648 T2647 prep at,dried
R2365 T2649 T2650 compound room,temperature
R2366 T2650 T2648 pobj temperature,at
R2367 T2651 T2650 cc and,temperature
R2368 T2652 T2650 conj ground,temperature
R2369 T2653 T2647 prep to,dried
R2370 T2654 T2655 amod small,particles
R2371 T2655 T2653 pobj particles,to
R2372 T2656 T2658 punct (,mm
R2373 T2657 T2658 nummod 1,mm
R2374 T2658 T2655 parataxis mm,particles
R2375 T2659 T2658 prep in,mm
R2376 T2660 T2659 pobj diameter,in
R2377 T2661 T2658 punct ),mm
R2378 T2662 T2638 punct .,precipitated
R2379 T2663 T2663 ROOT Using,Using
R2380 T2664 T2665 compound gel-permeation,chromatography
R2381 T2665 T2663 dobj chromatography,Using
R2382 T2666 T2665 prep in,chromatography
R2383 T2667 T2666 pobj combination,in
R2384 T2668 T2663 prep with,Using
R2385 T2669 T2671 amod light,measurements
R2386 T2670 T2671 compound scattering,measurements
R2387 T2671 T2668 pobj measurements,with
R2388 T2672 T2673 punct (,see
R2389 T2673 T2663 parataxis see,Using
R2390 T2674 T2684 nsubj Viebke,confirmed
R2391 T2675 T2676 compound et,al.
R2392 T2676 T2684 prep al.,confirmed
R2393 T2677 T2676 appos [,al.
R2394 T2678 T2679 nummod 21,]
R2395 T2679 T2677 appos ],[
R2396 T2680 T2677 punct ),[
R2397 T2681 T2684 punct ",",confirmed
R2398 T2682 T2684 nsubjpass it,confirmed
R2399 T2683 T2684 auxpass was,confirmed
R2400 T2684 T2673 ccomp confirmed,see
R2401 T2685 T2689 mark that,had
R2402 T2686 T2688 det the,fraction
R2403 T2687 T2688 amod low,fraction
R2404 T2688 T2689 nsubj fraction,had
R2405 T2689 T2684 ccomp had,confirmed
R2406 T2690 T2693 det an,weight
R2407 T2691 T2693 amod average,weight
R2408 T2692 T2693 amod molecular,weight
R2409 T2693 T2689 dobj weight,had
R2410 T2694 T2693 prep of,weight
R2411 T2695 T2696 nummod 10,kDa
R2412 T2696 T2694 pobj kDa,of
R2413 T2697 T2689 punct ",",had
R2414 T2698 T2689 cc and,had
R2415 T2699 T2701 det the,fraction
R2416 T2700 T2701 compound medium,fraction
R2417 T2701 T2689 conj fraction,had
R2418 T2702 T2701 prep of,fraction
R2419 T2703 T2704 nummod 40,kDa
R2420 T2704 T2702 pobj kDa,of
R2421 T2705 T2673 punct .,see
R2422 T2706 T2708 det The,content
R2423 T2707 T2708 compound sulphate,content
R2424 T2708 T2715 nsubjpass content,measured
R2425 T2709 T2708 prep of,content
R2426 T2710 T2709 pobj polysaccharides,of
R2427 T2711 T2710 prep in,polysaccharides
R2428 T2712 T2713 det both,fractions
R2429 T2713 T2711 pobj fractions,in
R2430 T2714 T2715 auxpass was,measured
R2431 T2715 T2715 ROOT measured,measured
R2432 T2716 T2715 prep following,measured
R2433 T2717 T2718 det the,method
R2434 T2718 T2716 pobj method,following
R2435 T2719 T2718 prep of,method
R2436 T2720 T2723 compound Quemener,[
R2437 T2721 T2722 compound et,al.
R2438 T2722 T2723 compound al.,[
R2439 T2723 T2719 pobj [,of
R2440 T2724 T2725 nummod 22,]
R2441 T2725 T2718 npadvmod ],method
R2442 T2726 T2715 punct .,measured
R2443 T2727 T2740 advmod Finally,confirmed
R2444 T2728 T2740 punct ",",confirmed
R2445 T2729 T2730 det the,absence
R2446 T2730 T2740 nsubjpass absence,confirmed
R2447 T2731 T2730 prep of,absence
R2448 T2732 T2733 compound polysaccharide,structure
R2449 T2733 T2734 compound structure,modifications
R2450 T2734 T2731 pobj modifications,of
R2451 T2735 T2734 prep in,modifications
R2452 T2736 T2738 det the,fractions
R2453 T2737 T2738 nummod two,fractions
R2454 T2738 T2735 pobj fractions,in
R2455 T2739 T2740 auxpass was,confirmed
R2456 T2740 T2740 ROOT confirmed,confirmed
R2457 T2741 T2740 advcl using,confirmed
R2458 T2742 T2743 amod 2H-NMR,spectroscopy
R2459 T2743 T2741 dobj spectroscopy,using
R2460 T2744 T2740 punct .,confirmed
R2461 T2745 T2746 det The,absence
R2462 T2746 T2755 nsubjpass absence,confirmed
R2463 T2747 T2746 prep of,absence
R2464 T2748 T2749 compound LPS,contamination
R2465 T2749 T2747 pobj contamination,of
R2466 T2750 T2749 prep in,contamination
R2467 T2751 T2753 det the,fractions
R2468 T2752 T2753 nummod two,fractions
R2469 T2753 T2750 pobj fractions,in
R2470 T2754 T2755 auxpass was,confirmed
R2471 T2755 T2755 ROOT confirmed,confirmed
R2472 T2756 T2755 advcl using,confirmed
R2473 T2757 T2760 det the,kit
R2474 T2758 T2760 amod e-Toxate,kit
R2475 T2759 T2760 compound ®,kit
R2476 T2760 T2756 dobj kit,using
R2477 T2761 T2760 punct (,kit
R2478 T2762 T2760 appos Sigma,kit
R2479 T2763 T2762 punct ",",Sigma
R2480 T2764 T2766 compound St,Fallavier
R2481 T2765 T2766 compound Quentin,Fallavier
R2482 T2766 T2762 conj Fallavier,Sigma
R2483 T2767 T2766 punct ",",Fallavier
R2484 T2768 T2766 conj France,Fallavier
R2485 T2769 T2766 punct ),Fallavier
R2486 T2770 T2755 punct .,confirmed
R2487 T2771 T2781 prep Before,dissolved
R2488 T2772 T2771 pobj use,Before
R2489 T2773 T2772 prep in,use
R2490 T2774 T2775 compound cell,culture
R2491 T2775 T2773 pobj culture,in
R2492 T2776 T2781 punct ",",dissolved
R2647 T2959 T2960 auxpass were,housed
R2648 T2960 T2960 ROOT housed,housed
R2649 T2961 T2960 prep under,housed
R2650 T2962 T2963 amod standard,conditions
R2651 T2963 T2961 pobj conditions,under
R2652 T2964 T2960 cc and,housed
R2653 T2965 T2960 conj fed,housed
R2654 T2966 T2967 compound ad,libitum
R2655 T2967 T2965 dobj libitum,fed
R2656 T2968 T2967 prep with,libitum
R2657 T2969 T2972 amod standard,chow
R2658 T2970 T2972 compound rodent,chow
R2659 T2971 T2972 compound laboratory,chow
R2660 T2972 T2968 pobj chow,with
R2661 T2973 T2960 punct .,housed
R2662 T2974 T2975 amod Degraded,iota-carrageenans
R2663 T2975 T2977 nsubjpass iota-carrageenans,administered
R2664 T2976 T2977 auxpass were,administered
R2665 T2977 T2977 ROOT administered,administered
R2666 T2978 T2977 prep in,administered
R2667 T2979 T2981 det the,water
R2669 T2981 T2978 pobj water,in
R2730 T3042 T3038 punct ),water
R2731 T3043 T3034 punct .,maintained
R2732 T3044 T3045 aux To,increase
R2733 T3045 T3051 advcl increase,added
R2734 T3046 T3049 amod palatability,sucrose
R2735 T3047 T3048 nummod 0.2,%
R2736 T3048 T3049 compound %,sucrose
R2737 T3049 T3051 nsubjpass sucrose,added
R2738 T3050 T3051 auxpass was,added
R2739 T3051 T3051 ROOT added,added
R2740 T3052 T3051 prep to,added
R2741 T3053 T3055 det the,water
R2742 T3054 T3055 compound drinking,water
R2743 T3055 T3052 pobj water,to
R2744 T3056 T3055 prep of,water
R2745 T3057 T3058 det all,groups
R2746 T3058 T3056 pobj groups,of
R2747 T3059 T3062 punct (,Waaji
R2748 T3060 T3062 compound Van,Waaji
R2749 T3061 T3062 compound der,Waaji
R2750 T3062 T3064 compound Waaji,al.
R2751 T3063 T3064 compound et,al.
R2752 T3064 T3055 appos al.,water
R2753 T3065 T3055 punct ",",water
R2754 T3066 T3055 appos [,water
R2755 T3067 T3068 nummod 23,]
R2756 T3068 T3066 appos ],[
R2757 T3069 T3066 punct ),[
R2758 T3070 T3051 punct .,added
R2759 T3071 T3073 amod Fresh,solutions
R2760 T3072 T3073 compound carrageenan,solutions
R2761 T3073 T3075 nsubjpass solutions,prepared
R2762 T3074 T3075 auxpass were,prepared
R2763 T3075 T3075 ROOT prepared,prepared
R2764 T3076 T3075 advmod daily,prepared
R2765 T3077 T3075 punct .,prepared
R3012 T3368 T3390 nsubjpass Evaluation,recorded
R3013 T3369 T3368 prep of,Evaluation
R3014 T3370 T3371 compound Colitis,Body
R3015 T3371 T3372 compound Body,weight
R3016 T3372 T3369 pobj weight,of
R3017 T3373 T3372 punct ",",weight
R3018 T3374 T3372 conj liquid,weight
R3019 T3375 T3374 cc and,liquid
R3020 T3376 T3377 compound food,consumption
R3021 T3377 T3374 conj consumption,liquid
R3022 T3378 T3377 punct ",",consumption
R3023 T3379 T3377 conj diarrhea,consumption
R3024 T3380 T3379 cc and,diarrhea
R3025 T3381 T3382 amod rectal,bleeding
R3026 T3382 T3379 conj bleeding,diarrhea
R3027 T3383 T3382 punct (,bleeding
R3028 T3384 T3382 acl detected,bleeding
R3029 T3385 T3384 agent by,detected
R3030 T3386 T3387 compound eye,inspection
R3031 T3387 T3385 pobj inspection,by
R3032 T3388 T3368 punct ),Evaluation
R3033 T3389 T3390 auxpass were,recorded
R3034 T3390 T3390 ROOT recorded,recorded
R3035 T3391 T3390 prep throughout,recorded
R3036 T3392 T3394 det the,period
R3037 T3393 T3394 compound feeding,period
R3038 T3394 T3391 pobj period,throughout
R3039 T3395 T3390 punct .,recorded
R3040 T3396 T3402 prep After,sacrificed
R3041 T3397 T3398 nummod 55,days
R3042 T3398 T3396 pobj days,After
R3043 T3399 T3402 punct ",",sacrificed
R3044 T3400 T3402 nsubjpass animals,sacrificed
R3045 T3401 T3402 auxpass were,sacrificed
R3046 T3402 T3402 ROOT sacrificed,sacrificed
R3047 T3403 T3402 agent by,sacrificed
R3048 T3404 T3405 amod cervical,dislocation
R3049 T3405 T3403 pobj dislocation,by
R3050 T3406 T3402 punct .,sacrificed
R3051 T3407 T3408 det The,length
R3052 T3408 T3413 nsubjpass length,measured
R3053 T3409 T3408 prep of,length
R3054 T3410 T3411 det the,colon
R3055 T3411 T3409 pobj colon,of
R3056 T3412 T3413 auxpass was,measured
R3057 T3413 T3413 ROOT measured,measured
R3058 T3414 T3415 mark as,described
R3059 T3415 T3413 advcl described,measured
R3060 T3416 T3415 agent by,described
R3061 T3417 T3416 pobj Okayashu,by
R3062 T3418 T3422 nmod et,]
R3063 T3419 T3422 nmod al.,]
R3064 T3420 T3422 nmod [,]
R3065 T3421 T3422 nummod 24,]
R3066 T3422 T3415 npadvmod ],described
R3067 T3423 T3413 punct .,measured
R3068 T3424 T3429 advmod Then,ligated
R3069 T3425 T3429 punct ",",ligated
R3070 T3426 T3427 det each,colon
R3071 T3427 T3429 nsubjpass colon,ligated
R3072 T3428 T3429 auxpass was,ligated
R3073 T3429 T3429 ROOT ligated,ligated
R3074 T3430 T3429 prep in,ligated
R3075 T3431 T3430 pobj sections,in
R3076 T3432 T3431 prep of,sections
R3077 T3433 T3434 nummod 2,cm
R3078 T3434 T3432 pobj cm,of
R3079 T3435 T3429 cc and,ligated
R3080 T3436 T3438 quantmod 1,2
R3081 T3437 T3438 quantmod to,2
R3082 T3438 T3439 nummod 2,ml
R3083 T3439 T3445 nsubjpass ml,infused
R3084 T3440 T3439 prep of,ml
R3085 T3441 T3442 nummod 10,%
R3086 T3442 T3443 compound %,formalin
R3087 T3443 T3440 pobj formalin,of
R3088 T3444 T3445 auxpass was,infused
R3089 T3445 T3429 conj infused,ligated
R3090 T3446 T3445 prep into,infused
R3091 T3447 T3449 det the,lumen
R3092 T3448 T3449 amod intestinal,lumen
R3093 T3449 T3446 pobj lumen,into
R3094 T3450 T3445 punct .,infused
R3095 T3451 T3454 det The,segment
R3096 T3452 T3453 advmod moderately,distended
R3097 T3453 T3454 amod distended,segment
R3098 T3454 T3455 nsubj segment,was
R3099 T3455 T3455 ROOT was,was
R3100 T3456 T3455 attr sectioned,was
R3101 T3457 T3456 cc and,sectioned
R3102 T3458 T3456 conj fixed,sectioned
R3103 T3459 T3458 prep in,fixed
R3104 T3460 T3461 nummod 10,%
R3105 T3461 T3462 compound %,formalin
R3106 T3462 T3459 pobj formalin,in
R3107 T3463 T3455 punct .,was
R3108 T3464 T3466 det The,day
R3109 T3465 T3466 amod following,day
R3110 T3466 T3472 npadvmod day,removed
R3111 T3467 T3472 punct ",",removed
R3112 T3468 T3470 det the,content
R3113 T3469 T3470 amod intestinal,content
R3114 T3470 T3472 nsubjpass content,removed
R3115 T3471 T3472 auxpass was,removed
R3116 T3472 T3472 ROOT removed,removed
R3117 T3473 T3472 agent by,removed
R3118 T3474 T3473 pobj vortexing,by
R3119 T3475 T3472 punct .,removed
R3120 T3476 T3478 det The,segment
R3121 T3477 T3478 amod fixed,segment
R3122 T3478 T3480 nsubjpass segment,kept
R3123 T3479 T3480 auxpass was,kept
R3124 T3480 T3480 ROOT kept,kept
R3125 T3481 T3480 oprd in,kept
R3126 T3482 T3483 nummod 10,%
R3127 T3483 T3484 compound %,formalin
R3128 T3484 T3481 pobj formalin,in
R3129 T3485 T3480 prep at,kept
R3130 T3486 T3487 nummod 4,°
R3131 T3487 T3488 nummod °,C
R3132 T3488 T3485 pobj C,at
R3133 T3489 T3480 prep until,kept
R3134 T3490 T3493 det the,procedure
R3135 T3491 T3492 compound paraffin,embedding
R3136 T3492 T3493 compound embedding,procedure
R3137 T3493 T3489 pobj procedure,until
R3138 T3494 T3480 punct .,kept
R3139 T3495 T3496 aux To,evaluate
R3140 T3496 T3507 advcl evaluate,opened
R3141 T3497 T3498 det the,degree
R3142 T3498 T3496 dobj degree,evaluate
R3143 T3499 T3498 prep of,degree
R3144 T3500 T3499 pobj inflammation,of
R3145 T3501 T3507 punct ",",opened
R3146 T3502 T3503 det this,segment
R3161 T3517 T3507 conj recorded,opened
R3162 T3518 T3520 mark as,described
R3163 T3519 T3520 advmod previously,described
R3164 T3520 T3517 advcl described,recorded
R3165 T3521 T3523 nmod [,]
R3166 T3522 T3523 nummod 25,]
R3167 T3523 T3520 dobj ],described
R3168 T3524 T3523 punct ",",]
R3169 T3525 T3527 nmod [,]
R3170 T3526 T3527 nummod 26,]
R3171 T3527 T3523 appos ],]
R3172 T3528 T3517 punct .,recorded
R3173 T3529 T3532 det The,staining
R3174 T3530 T3532 nmod toluidine,staining
R3175 T3531 T3532 amod blue,staining
R3176 T3532 T3534 nsubjpass staining,used
R3177 T3533 T3534 auxpass was,used
R3178 T3534 T3534 ROOT used,used
R3179 T3535 T3534 prep for,used
R3180 T3536 T3535 pobj identification,for
R3181 T3537 T3536 prep of,identification
R3182 T3538 T3539 amod sulphated,polysaccharides
R3183 T3539 T3537 pobj polysaccharides,of
R3184 T3540 T3539 prep in,polysaccharides
R3185 T3541 T3543 det the,mucosa
R3186 T3542 T3543 amod intestinal,mucosa
R3187 T3543 T3540 pobj mucosa,in
R3188 T3544 T3534 punct .,used
R3189 T3545 T3562 prep On,collected
R3190 T3546 T3547 det the,day
R3191 T3547 T3545 pobj day,On
R3192 T3548 T3547 prep of,day
R3193 T3549 T3548 pobj sacrifice,of
R3194 T3550 T3562 punct ",",collected
R3195 T3551 T3553 det a,sample
R3196 T3552 T3553 amod fresh,sample
R3197 T3553 T3562 nsubjpass sample,collected
R3198 T3554 T3553 prep of,sample
R3199 T3555 T3556 det each,colon
R3200 T3556 T3554 pobj colon,of
R3201 T3557 T3556 punct (,colon
R3202 T3558 T3559 nummod 50,mg
R3203 T3559 T3556 appos mg,colon
R3204 T3560 T3556 punct ),colon
R3205 T3561 T3562 auxpass was,collected
R3206 T3562 T3562 ROOT collected,collected
R3207 T3563 T3562 prep for,collected
R3208 T3564 T3563 pobj myeloperoxidase,for
R3209 T3565 T3566 punct (,MPO
R3210 T3566 T3564 appos MPO,myeloperoxidase
R3211 T3567 T3566 punct ),MPO
R3212 T3568 T3569 amod assay,according
R3213 T3569 T3562 prep according,collected
R3214 T3570 T3569 prep to,according
R3215 T3571 T3573 compound Krawisz,al.
R3216 T3572 T3573 nmod et,al.
R3217 T3573 T3570 pobj al.,to
R3218 T3574 T3573 punct ",",al.
R3219 T3575 T3577 nmod [,]
R3220 T3576 T3577 nummod 27,]
R3221 T3577 T3573 appos ],al.
R3222 T3578 T3562 punct .,collected
R3223 T3579 T3580 det The,level
R3224 T3580 T3589 nsubj level,indicates
R3234 T3590 T3591 det the,rate
R3235 T3591 T3589 dobj rate,indicates
R3236 T3592 T3591 prep of,rate
R3237 T3593 T3592 pobj recruitment,of
R3238 T3594 T3593 prep of,recruitment
R3239 T3595 T3594 pobj neutrophils,of
R3240 T3596 T3589 prep to,indicates
R3241 T3597 T3599 det the,mucosa
R3242 T3598 T3599 amod intestinal,mucosa
R3243 T3599 T3596 pobj mucosa,to
R3244 T3600 T3589 punct .,indicates
R3245 T3601 T3602 nummod One,unit
R3246 T3602 T3606 nsubj unit,corresponds
R3247 T3603 T3602 prep of,unit
R3248 T3604 T3605 compound MPO,activity
R3249 T3605 T3603 pobj activity,of
R3250 T3606 T3606 ROOT corresponds,corresponds
R3251 T3607 T3606 prep to,corresponds
R3252 T3608 T3609 det the,degradation
R3253 T3609 T3607 pobj degradation,to
R3254 T3610 T3609 prep of,degradation
R3255 T3611 T3612 nummod 1,µmol
R3256 T3612 T3610 pobj µmol,of
R3257 T3613 T3612 prep of,µmol
R3258 T3614 T3613 pobj peroxide,of
R3259 T3615 T3614 prep per,peroxide
R3260 T3616 T3615 pobj minute,per
R3261 T3617 T3609 prep at,degradation
R3262 T3618 T3619 nummod 25,°
R3263 T3619 T3620 nummod °,C.
R3264 T3620 T3617 pobj C.,at
R3413 T3816 T3818 compound Cell,All
R3414 T3817 T3818 compound Culture,All
R3415 T3818 T3821 det All,reagents
R3416 T3819 T3820 compound tissue,culture
R3417 T3820 T3821 compound culture,reagents
R3418 T3821 T3822 nsubj reagents,were
R3419 T3822 T3822 ROOT were,were
R3420 T3823 T3822 prep from,were
R3421 T3824 T3823 pobj Invitrogen,from
R3422 T3825 T3824 punct (,Invitrogen
R3423 T3826 T3827 compound Cergy,Pontoise
R3424 T3827 T3824 appos Pontoise,Invitrogen
R3425 T3828 T3827 punct ",",Pontoise
R3426 T3829 T3827 npadvmod France,Pontoise
R3427 T3830 T3824 punct ),Invitrogen
R3428 T3831 T3822 punct .,were
R3429 T3832 T3835 nummod THP-1,cells
R3430 T3833 T3835 amod human,cells
R3431 T3834 T3835 amod monocytic,cells
R3432 T3835 T3837 nsubjpass cells,maintained
R3433 T3836 T3837 auxpass were,maintained
R3434 T3837 T3837 ROOT maintained,maintained
R3435 T3838 T3837 prep in,maintained
R3436 T3839 T3838 pobj RPMI-1640,in
R3437 T3840 T3837 conj supplemented,maintained
R3438 T3841 T3840 prep with,supplemented
R3439 T3842 T3843 nummod 10,%
R3440 T3843 T3844 compound %,FCS
R3441 T3844 T3841 pobj FCS,with
R3442 T3845 T3844 punct ",",FCS
R3443 T3846 T3850 nummod 2,glutamine
R3444 T3847 T3850 compound mM,glutamine
R3445 T3848 T3850 compound L,glutamine
R3446 T3849 T3850 punct -,glutamine
R3447 T3850 T3844 appos glutamine,FCS
R3448 T3851 T3850 punct ",",glutamine
R3449 T3852 T3854 nummod 50,penicillin
R3450 T3853 T3854 compound U/ml,penicillin
R3451 T3854 T3862 nsubj penicillin,C
R3452 T3855 T3854 cc and,penicillin
R3453 T3856 T3857 nummod 50,mg/ml
R3454 T3857 T3858 compound mg/ml,streptomycin
R3455 T3858 T3854 conj streptomycin,penicillin
R3456 T3859 T3854 prep at,penicillin
R3457 T3860 T3861 nummod 37,°
R3458 T3861 T3859 pobj °,at
R3459 T3862 T3844 conj C,FCS
R3460 T3863 T3837 prep in,maintained
R3461 T3864 T3868 det a,incubator
R3462 T3865 T3866 nummod 5,%
R3463 T3866 T3868 nmod %,incubator
R3464 T3867 T3868 nummod CO2,incubator
R3465 T3868 T3863 pobj incubator,in
R3466 T3869 T3837 punct .,maintained
R3467 T3870 T3874 amod Human,cells
R3468 T3871 T3874 amod peripheral,cells
R3469 T3872 T3874 compound blood,cells
R3470 T3873 T3874 compound mononuclear,cells
R3471 T3874 T3876 nsubjpass cells,obtained
R3472 T3875 T3876 auxpass were,obtained
R3473 T3876 T3876 ROOT obtained,obtained
R3474 T3877 T3876 prep from,obtained
R3475 T3878 T3879 amod heparinized,blood
R3476 T3879 T3877 pobj blood,from
R3477 T3880 T3876 agent by,obtained
R3478 T3881 T3883 amod Ficoll-Hypaque,gradient
R3479 T3882 T3883 compound density,gradient
R3480 T3883 T3880 pobj gradient,by
R3481 T3884 T3876 punct .,obtained
R3482 T3885 T3888 nsubjpass Monocytes,isolated
R3483 T3886 T3888 auxpass were,isolated
R3484 T3887 T3888 advmod then,isolated
R3485 T3888 T3888 ROOT isolated,isolated
R3486 T3889 T3888 agent by,isolated
R3487 T3890 T3889 pobj adherence,by
R3488 T3891 T3890 prep to,adherence
R3489 T3892 T3893 compound culture,flasks
R3490 T3893 T3891 pobj flasks,to
R3491 T3894 T3895 mark as,described
R3492 T3895 T3888 advcl described,isolated
R3493 T3896 T3898 compound [,]
R3494 T3897 T3898 compound 28,]
R3495 T3898 T3895 dobj ],described
R3496 T3899 T3888 punct .,isolated
R3497 T3900 T3906 prep For,cultured
R3498 T3901 T3902 compound cell,aggregation
R3499 T3902 T3900 pobj aggregation,For
R3500 T3903 T3906 punct ",",cultured
R3501 T3904 T3906 nsubjpass monocytes,cultured
R3502 T3905 T3906 auxpass were,cultured
R3503 T3906 T3906 ROOT cultured,cultured
R3504 T3907 T3906 prep in,cultured
R3505 T3908 T3909 det the,presence
R3506 T3909 T3907 pobj presence,in
R3507 T3910 T3909 cc or,presence
R3508 T3911 T3909 conj absence,presence
R3509 T3912 T3911 prep of,absence
R3510 T3913 T3912 pobj C10,of
R3511 T3914 T3913 cc or,C10
R3512 T3915 T3913 conj C40,C10
R3513 T3916 T3906 prep for,cultured
R3514 T3917 T3920 nummod 72,colonies
R3515 T3918 T3920 compound h.,colonies
R3516 T3919 T3920 compound Cell,colonies
R3517 T3920 T3916 pobj colonies,for
R3518 T3921 T3922 auxpass were,monitored
R3519 T3922 T3906 conj monitored,cultured
R3520 T3923 T3922 prep under,monitored
R3521 T3924 T3928 det an,microscope
R3522 T3925 T3926 amod inverted,phase
R3523 T3926 T3928 compound phase,microscope
R3524 T3927 T3928 compound contrast,microscope
R3525 T3928 T3923 pobj microscope,under
R3526 T3929 T3928 acl coupled,microscope
R3527 T3930 T3929 prep through,coupled
R3528 T3931 T3933 det a,camera
R3529 T3932 T3933 compound video,camera
R3530 T3933 T3930 pobj camera,through
R3531 T3934 T3929 prep to,coupled
R3532 T3935 T3936 det a,computer
R3533 T3936 T3934 pobj computer,to
R3534 T3937 T3906 punct .,cultured
R3535 T3938 T3964 prep In,added
R3536 T3939 T3940 det some,wells
R3537 T3940 T3938 pobj wells,In
R3538 T3941 T3964 punct ",",added
R3539 T3942 T3964 csubj neutralizing,added
R3540 T3943 T3944 amod monoclonal,antibody
R3541 T3944 T3942 dobj antibody,neutralizing
R3542 T3945 T3942 prep to,neutralizing
R3543 T3946 T3945 pobj ICAM-1,to
R3544 T3947 T3951 punct (,ml
R3545 T3948 T3949 nummod 2.5,µg
R3546 T3949 T3951 nmod µg,ml
R3547 T3950 T3951 compound /,ml
R3548 T3951 T3946 appos ml,ICAM-1
R3549 T3952 T3946 punct ),ICAM-1
R3550 T3953 T3964 punct (,added
R3551 T3954 T3964 dep Tebu,added
R3552 T3955 T3954 punct ",",Tebu
R3553 T3956 T3957 compound Le,Perray
R3554 T3957 T3964 nsubjpass Perray,added
R3555 T3958 T3957 prep en,Perray
R3556 T3959 T3958 pobj Yvelines,en
R3557 T3960 T3959 punct ",",Yvelines
R3558 T3961 T3959 appos France,Yvelines
R3559 T3962 T3959 punct ),Yvelines
R3560 T3963 T3964 auxpass was,added
R3561 T3964 T3964 ROOT added,added
R3562 T3965 T3964 punct .,added
R3680 T4100 T4102 compound Cell,Analysis
R3681 T4101 T4102 compound Cycle,Analysis
R3682 T4102 T4104 compound Analysis,cells
R3683 T4103 T4104 compound THP-1,cells
R3684 T4104 T4110 nsubjpass cells,exposed
R3685 T4105 T4104 prep in,cells
R3686 T4106 T4108 amod exponential,phase
R3687 T4107 T4108 compound growth,phase
R3688 T4108 T4105 pobj phase,in
R3689 T4109 T4110 auxpass were,exposed
R3690 T4110 T4110 ROOT exposed,exposed
R3691 T4111 T4112 aux to,complete
R3692 T4112 T4110 xcomp complete,exposed
R3693 T4113 T4112 dobj medium,complete
R3694 T4114 T4112 prep in,complete
R3695 T4115 T4116 det the,presence
R3696 T4116 T4114 pobj presence,in
R3697 T4117 T4116 cc or,presence
R3698 T4118 T4116 conj absence,presence
R3699 T4119 T4118 prep of,absence
R3700 T4120 T4119 pobj carrageenans,of
R3701 T4121 T4112 prep for,complete
R3702 T4122 T4123 nummod 24,h
R3703 T4123 T4121 pobj h,for
R3704 T4124 T4112 prep before,complete
R3705 T4125 T4126 auxpass being,stained
R3706 T4126 T4124 pcomp stained,before
R3707 T4127 T4126 prep with,stained
R3708 T4128 T4129 compound propidium,iodide
R3709 T4129 T4127 pobj iodide,with
R3710 T4130 T4124 pcomp using,before
R3711 T4131 T4134 det the,kit
R3712 T4132 T4133 compound DNA-Prep,Coulter
R3713 T4133 T4134 compound Coulter,kit
R3714 T4134 T4130 dobj kit,using
R3715 T4135 T4110 prep according,exposed
R3716 T4136 T4135 prep to,according
R3717 T4137 T4138 det the,manufacturer
R3718 T4138 T4140 poss manufacturer,instruction
R3719 T4139 T4138 case 's,manufacturer
R3720 T4140 T4136 pobj instruction,to
R3721 T4141 T4142 punct (,Beckman-Coulter
R3722 T4142 T4140 appos Beckman-Coulter,instruction
R3723 T4143 T4142 punct ",",Beckman-Coulter
R3724 T4144 T4142 conj Villepinte,Beckman-Coulter
R3725 T4145 T4144 punct ",",Villepinte
R3726 T4146 T4144 conj France,Villepinte
R3727 T4147 T4110 punct ),exposed
R3728 T4148 T4110 punct .,exposed
R3729 T4149 T4150 compound Cell,DNA
R3730 T4150 T4151 compound DNA,content
R3731 T4151 T4154 nsubjpass content,analyzed
R3732 T4152 T4154 auxpass was,analyzed
R3733 T4153 T4154 advmod then,analyzed
R3734 T4154 T4154 ROOT analyzed,analyzed
R3735 T4155 T4154 agent by,analyzed
R3736 T4156 T4157 compound flow,cytometry
R3737 T4157 T4155 pobj cytometry,by
R3738 T4158 T4154 advcl using,analyzed
R3739 T4159 T4161 det an,XL2
R3740 T4160 T4161 compound EPICS,XL2
R3741 T4161 T4158 dobj XL2,using
R3742 T4162 T4163 punct (,Beckman-Coulter
R3743 T4163 T4161 appos Beckman-Coulter,XL2
R3744 T4164 T4161 punct ),XL2
R3745 T4165 T4154 punct .,analyzed
R3746 T4166 T4167 amod Raw,data
R3747 T4167 T4182 nsubjpass data,expressed
R3748 T4168 T4167 prep for,data
R3749 T4169 T4170 det the,distribution
R3750 T4170 T4168 pobj distribution,for
R3751 T4171 T4170 prep of,distribution
R3752 T4172 T4173 compound DNA,content
R3753 T4173 T4171 pobj content,of
R3754 T4174 T4173 prep of,content
R3755 T4175 T4176 nummod "30,000",cells
R3756 T4176 T4174 pobj cells,of
R3757 T4177 T4170 acl retrieved,distribution
R3758 T4178 T4177 prep from,retrieved
R3759 T4179 T4180 det the,cytometer
R3760 T4180 T4178 pobj cytometer,from
R3761 T4181 T4182 auxpass were,expressed
R3762 T4182 T4182 ROOT expressed,expressed
R3763 T4183 T4182 prep as,expressed
R3764 T4184 T4185 det the,percentage
R3765 T4185 T4183 pobj percentage,as
R3766 T4186 T4185 prep of,percentage
R3767 T4187 T4186 pobj G0/G1,of
R3768 T4188 T4182 prep through,expressed
R3769 T4189 T4190 compound G2/M,populations
R3770 T4190 T4188 pobj populations,through
R3771 T4191 T4182 punct .,expressed
R3772 T4192 T4194 amod Multicycle,software
R3773 T4193 T4194 compound AV,software
R3774 T4194 T4206 nsubjpass software,used
R3775 T4195 T4198 punct (,Systems
R3776 T4196 T4198 compound Phoenix,Systems
R3777 T4197 T4198 compound Flow,Systems
R3778 T4198 T4194 appos Systems,software
R3779 T4199 T4198 punct ",",Systems
R3780 T4200 T4201 compound San,Diego
R3781 T4201 T4198 npadvmod Diego,Systems
R3782 T4202 T4201 punct ",",Diego
R3783 T4203 T4201 conj CA,Diego
R3784 T4204 T4198 punct ),Systems
R3785 T4205 T4206 auxpass was,used
R3786 T4206 T4206 ROOT used,used
R3787 T4207 T4208 aux to,generate
R3788 T4208 T4206 xcomp generate,used
R3789 T4209 T4212 compound DNA,histograms
R3790 T4210 T4211 amod content,frequency
R3791 T4211 T4212 compound frequency,histograms
R3792 T4212 T4208 dobj histograms,generate
R3793 T4213 T4208 cc and,generate
R3794 T4214 T4208 conj facilitate,generate
R3795 T4215 T4216 compound data,analysis
R3796 T4216 T4214 dobj analysis,facilitate
R3797 T4217 T4206 punct .,used
R3936 T4382 T4384 compound Cell,Antigen
R3937 T4383 T4384 compound Surface,Antigen
R3938 T4384 T4389 compound Antigen,Monocytes
R3939 T4385 T4386 compound Expression,Analysis
R3953 T4399 T4400 det the,presence
R3954 T4400 T4398 pobj presence,in
R3955 T4401 T4400 cc or,presence
R3956 T4402 T4400 conj absence,presence
R3957 T4403 T4402 prep of,absence
R3958 T4404 T4403 pobj carrageenan,of
R3959 T4405 T4396 prep for,complete
R3960 T4406 T4407 nummod 36,h
R3961 T4407 T4405 pobj h,for
R3962 T4408 T4394 punct .,exposed
R3963 T4409 T4423 prep After,incubated
R3964 T4410 T4411 nummod two,washes
R3965 T4411 T4409 pobj washes,After
R3966 T4412 T4411 prep in,washes
R3967 T4413 T4412 pobj PBS,in
R3968 T4414 T4411 prep without,washes
R3969 T4415 T4416 compound Ca2,+
R3970 T4416 T4414 pobj +,without
R3971 T4417 T4416 cc and,+
R3972 T4418 T4419 compound Mg2,+
R3973 T4419 T4416 conj +,+
R3974 T4420 T4423 punct ",",incubated
R3975 T4421 T4423 nsubjpass cells,incubated
R3976 T4422 T4423 auxpass were,incubated
R3977 T4423 T4423 ROOT incubated,incubated
R3978 T4424 T4423 prep in,incubated
R3979 T4425 T4424 pobj PBS,in
R3980 T4426 T4423 advcl containing,incubated
R3981 T4427 T4428 nummod 0.1,%
R3982 T4428 T4429 compound %,gelatin
R3983 T4429 T4426 dobj gelatin,containing
R3984 T4430 T4429 cc and,gelatin
R3985 T4431 T4432 nummod 8,%
R3986 T4432 T4429 conj %,gelatin
R3987 T4433 T4435 nmod AB,serum
R3988 T4434 T4435 amod human,serum
R3989 T4435 T4429 conj serum,gelatin
R3990 T4436 T4437 aux to,prevent
R3991 T4437 T4435 relcl prevent,serum
R3992 T4438 T4437 advmod binding,prevent
R3993 T4439 T4438 prep to,binding
R3994 T4440 T4441 compound Fc,receptors
R3995 T4441 T4439 pobj receptors,to
R3996 T4442 T4423 punct .,incubated
R3997 T4443 T4450 advmod Then,incubated
R3998 T4444 T4450 punct ",",incubated
R3999 T4445 T4448 nummod 5,cells
R4000 T4446 T4448 nmod ×,cells
R4001 T4447 T4448 nummod 105,cells
R4002 T4448 T4450 nsubjpass cells,incubated
R4003 T4449 T4450 auxpass were,incubated
R4004 T4450 T4450 ROOT incubated,incubated
R4005 T4451 T4450 prep with,incubated
R4006 T4452 T4453 amod primary,antibodies
R4007 T4453 T4451 pobj antibodies,with
R4008 T4454 T4450 prep at,incubated
R4009 T4455 T4456 nummod 4,°
R4010 T4456 T4454 pobj °,at
R4011 T4457 T4450 conj C,incubated
R4012 T4458 T4457 prep for,C
R4013 T4459 T4460 nummod 30,min
R4014 T4460 T4458 pobj min,for
R4015 T4461 T4450 punct .,incubated
R4016 T4462 T4464 nummod Two,washes
R4017 T4463 T4464 amod other,washes
R4018 T4464 T4467 nsubj washes,preceded
R4019 T4465 T4464 prep in,washes
R4020 T4466 T4465 pobj PBS,in
R4021 T4467 T4467 ROOT preceded,preceded
R4022 T4468 T4467 dobj incubation,preceded
R4023 T4469 T4467 prep with,preceded
R4024 T4470 T4475 advmod FITC-conjugated,diluted
R4025 T4471 T4472 compound goat,antibody
R4026 T4472 T4475 nsubj antibody,diluted
R4027 T4473 T4474 compound anti-mouse,IgG
R4028 T4474 T4472 appos IgG,antibody
R4029 T4475 T4469 pcomp diluted,with
R4030 T4476 T4475 dobj 1/1000,diluted
R4031 T4477 T4475 prep at,diluted
R4032 T4478 T4479 nummod 4,°
R4033 T4479 T4477 pobj °,at
R4034 T4480 T4475 npadvmod C,diluted
R4035 T4481 T4475 prep for,diluted
R4036 T4482 T4483 nummod 30,min
R4037 T4483 T4481 pobj min,for
R4038 T4484 T4483 punct (,min
R4039 T4485 T4483 appos Tebu,min
R4040 T4486 T4483 punct ),min
R4041 T4487 T4467 punct .,preceded
R4042 T4488 T4498 mark After,performed
R4043 T4489 T4491 nummod two,washes
R4044 T4490 T4491 amod additional,washes
R4045 T4491 T4488 pobj washes,After
R4046 T4492 T4491 punct ",",washes
R4047 T4493 T4491 conj analysis,washes
R4048 T4494 T4493 prep of,analysis
R4049 T4495 T4496 amod stained,cells
R4050 T4496 T4494 pobj cells,of
R4051 T4497 T4498 auxpass was,performed
R4052 T4498 T4498 ROOT performed,performed
R4053 T4499 T4498 prep on,performed
R4054 T4500 T4502 det an,XL2
R4055 T4501 T4502 compound EPICS,XL2
R4056 T4502 T4499 pobj XL2,on
R4057 T4503 T4502 punct (,XL2
R4058 T4504 T4502 appos Beckman-Coulter,XL2
R4059 T4505 T4502 punct ),XL2
R4060 T4506 T4498 punct .,performed
R4061 T4507 T4509 det The,population
R4062 T4508 T4509 compound cell,population
R4063 T4509 T4511 nsubjpass population,gated
R4064 T4510 T4511 auxpass was,gated
R4065 T4511 T4511 ROOT gated,gated
R4066 T4512 T4511 prep according,gated
R4067 T4513 T4512 prep to,according
R4068 T4514 T4519 poss its,scattering
R4069 T4515 T4519 amod forward,scattering
R4070 T4516 T4515 cc and,forward
R4071 T4517 T4515 conj wide-angle,forward
R4072 T4518 T4519 amod light,scattering
R4073 T4519 T4513 pobj scattering,to
R4074 T4520 T4511 punct .,gated
R4075 T4521 T4523 nsubjpass Data,expressed
R4076 T4522 T4523 auxpass were,expressed
R4077 T4523 T4523 ROOT expressed,expressed
R4078 T4524 T4523 prep as,expressed
R4079 T4525 T4528 amod mean,intensity
R4080 T4526 T4528 amod relative,intensity
R4081 T4527 T4528 compound fluorescence,intensity
R4082 T4528 T4524 pobj intensity,as
R4083 T4529 T4530 punct (,MFI
R4084 T4530 T4528 appos MFI,intensity
R4085 T4531 T4530 punct ),MFI
R4086 T4532 T4528 prep of,intensity
R4087 T4533 T4534 nummod 3000,cells
R4088 T4534 T4532 pobj cells,of
R4089 T4535 T4523 punct .,expressed
R4150 T4648 T4651 compound TNF,Monocytes
R4151 T4649 T4651 compound Activity,Monocytes
R4152 T4650 T4651 compound Bioassay,Monocytes
R4153 T4651 T4656 nsubjpass Monocytes,cultured
R4154 T4652 T4651 cc or,Monocytes
R4155 T4653 T4654 nummod THP-1,cells
R4156 T4654 T4651 conj cells,Monocytes
R4157 T4655 T4656 auxpass were,cultured
R4158 T4656 T4656 ROOT cultured,cultured
R4159 T4657 T4656 prep with,cultured
R4160 T4658 T4657 cc or,with
R4161 T4659 T4657 conj without,with
R4162 T4660 T4661 amod different,concentrations
R4163 T4661 T4659 pobj concentrations,without
R4164 T4662 T4661 prep of,concentrations
R4165 T4663 T4662 pobj CGNs,of
R4166 T4664 T4663 cc or,CGNs
R4167 T4665 T4663 conj LPS,CGNs
R4168 T4666 T4668 punct (,typhosa
R4169 T4667 T4668 compound Salmonella,typhosa
R4170 T4668 T4665 appos typhosa,LPS
R4171 T4669 T4668 punct ",",typhosa
R4172 T4670 T4668 npadvmod Sigma,typhosa
R4173 T4671 T4668 punct ),typhosa
R4174 T4672 T4665 prep for,LPS
R4175 T4673 T4674 nummod 24,h
R4176 T4674 T4672 pobj h,for
R4177 T4675 T4674 cc or,h
R4178 T4676 T4678 det the,time
R4179 T4677 T4678 amod indicated,time
R4180 T4678 T4674 conj time,h
R4181 T4679 T4656 punct .,cultured
R4182 T4680 T4684 advmod Biologically,β
R4183 T4681 T4684 amod active,β
R4184 T4682 T4684 compound TNF-α,β
R4185 T4683 T4684 compound /,β
R4186 T4684 T4690 nsubjpass β,measured
R4187 T4685 T4684 prep in,β
R4188 T4686 T4687 compound tissue,culture
R4189 T4687 T4685 pobj culture,in
R4190 T4688 T4690 nsubjpass supernatant,measured
R4191 T4689 T4690 auxpass was,measured
R4192 T4690 T4690 ROOT measured,measured
R4193 T4691 T4690 advcl using,measured
R4194 T4692 T4698 det the,assay
R4195 T4693 T4696 nmod WEHI,13-cell
R4196 T4694 T4693 nummod 164,WEHI
R4197 T4695 T4696 compound clone,13-cell
R4198 T4696 T4698 compound 13-cell,assay
R4199 T4697 T4698 compound killing,assay
R4200 T4698 T4691 dobj assay,using
R4201 T4699 T4701 nmod [,]
R4202 T4700 T4701 nummod 29,]
R4203 T4701 T4698 appos ],assay
R4204 T4702 T4690 punct .,measured
R4205 T4703 T4704 compound TNF,concentrations
R4206 T4704 T4706 nsubjpass concentrations,expressed
R4207 T4705 T4706 auxpass are,expressed
R4208 T4706 T4706 ROOT expressed,expressed
R4209 T4707 T4706 prep as,expressed
R4210 T4708 T4707 pobj pg/ml,as
R4211 T4709 T4706 punct .,expressed
R4414 T4951 T4952 compound RT-PCR,Analysis
R4415 T4952 T4958 nsubjpass Analysis,isolated
R4416 T4953 T4954 compound Total,RNA
R4417 T4954 T4952 appos RNA,Analysis
R4418 T4955 T4954 prep from,RNA
R4419 T4956 T4955 pobj monocytes,from
R4420 T4957 T4958 auxpass was,isolated
R4421 T4958 T4958 ROOT isolated,isolated
R4422 T4959 T4958 advcl using,isolated
R4423 T4960 T4962 compound TRIzol,™
R4424 T4961 T4962 compound Reagent,™
R4425 T4962 T4959 dobj ™,using
R4426 T4963 T4964 punct (,Invitrogen
R4427 T4964 T4962 appos Invitrogen,™
R4428 T4965 T4962 punct ),™
R4429 T4966 T4958 punct .,isolated
R4430 T4967 T4969 nsubjpass cDNA,generated
R4431 T4968 T4969 auxpass was,generated
R4432 T4969 T4969 ROOT generated,generated
R4433 T4970 T4969 prep on,generated
R4434 T4971 T4972 nummod 1,µg
R4435 T4972 T4970 pobj µg,on
R4436 T4973 T4972 prep of,µg
R4437 T4974 T4975 amod total,RNA
R4438 T4975 T4973 pobj RNA,of
R4439 T4976 T4969 prep in,generated
R4440 T4977 T4979 det a,volume
R4441 T4978 T4979 compound reaction,volume
R4442 T4979 T4976 pobj volume,in
R4443 T4980 T4979 prep of,volume
R4444 T4981 T4982 nummod 20,µl
R4445 T4982 T4980 pobj µl,of
R4446 T4983 T4969 punct ",",generated
R4447 T4984 T4969 advcl using,generated
R4448 T4985 T4986 compound M-MLV,reverse
R4449 T4986 T4984 dobj reverse,using
R4450 T4987 T4986 npadvmod transcriptase,reverse
R4451 T4988 T4987 punct (,transcriptase
R4452 T4989 T4987 appos Invitrogen,transcriptase
R4453 T4990 T4987 punct ),transcriptase
R4454 T4991 T4969 punct .,generated
R4455 T4992 T4994 nsubjpass PCR,done
R4456 T4993 T4994 auxpass was,done
R4457 T4994 T4994 ROOT done,done
R4458 T4995 T4994 prep in,done
R4459 T4996 T4998 det the,range
R4460 T4997 T4998 amod linear,range
R4461 T4998 T4995 pobj range,in
R4462 T4999 T4998 prep of,range
R4463 T5000 T4999 pobj amplification,of
R4464 T5001 T4998 punct (,range
R4465 T5002 T4994 conj determined,done
R4466 T5003 T5002 prep for,determined
R4467 T5004 T5007 det each,combination
R4468 T5005 T5006 compound primer,pair-cDNA
R4469 T5006 T5007 compound pair-cDNA,combination
R4470 T5007 T5003 pobj combination,for
R4471 T5008 T5002 punct ),determined
R4472 T5009 T4994 punct .,done
R4473 T5010 T5012 amod Standard,reactions
R4474 T5011 T5012 compound PCR,reactions
R4475 T5012 T5014 nsubjpass reactions,performed
R4476 T5013 T5014 auxpass were,performed
R4477 T5014 T5014 ROOT performed,performed
R4478 T5015 T5014 prep with,performed
R4479 T5016 T5017 nummod 1,µl
R4480 T5017 T5015 pobj µl,with
R4481 T5018 T5017 prep of,µl
R4482 T5019 T5021 det the,solution
R4483 T5020 T5021 compound cDNA,solution
R4484 T5021 T5018 pobj solution,of
R4485 T5022 T5021 punct ",",solution
R4486 T5023 T5024 nummod 50,µM
R4487 T5024 T5021 appos µM,solution
R4488 T5025 T5024 prep of,µM
R4489 T5026 T5028 det each,solution
R4490 T5027 T5028 compound primer,solution
R4491 T5028 T5025 pobj solution,of
R4492 T5029 T5024 punct ",",µM
R4493 T5030 T5031 nummod 10,mM
R4494 T5031 T5024 appos mM,µM
R4495 T5032 T5031 prep of,mM
R4496 T5033 T5034 det each,dNTP
R4497 T5034 T5032 pobj dNTP,of
R4498 T5035 T5024 punct ",",µM
R4499 T5036 T5038 nummod 25,MgCl2
R4500 T5037 T5038 compound mM,MgCl2
R4501 T5038 T5024 appos MgCl2,µM
R4502 T5039 T5038 punct ",",MgCl2
R4503 T5040 T5042 compound 10X,DNA
R4504 T5041 T5042 compound Goldstar,DNA
R4505 T5042 T5045 compound DNA,buffer
R4506 T5043 T5045 compound polymerase,buffer
R4507 T5044 T5045 compound reaction,buffer
R4508 T5045 T5038 appos buffer,MgCl2
R4509 T5046 T5038 punct ",",MgCl2
R4510 T5047 T5038 cc and,MgCl2
R4511 T5048 T5049 nummod 0.5,units
R4512 T5049 T5024 appos units,µM
R4513 T5050 T5049 prep of,units
R4514 T5051 T5053 compound Goldstar,polymerase
R4515 T5052 T5053 compound DNA,polymerase
R4516 T5053 T5050 pobj polymerase,of
R4517 T5054 T5055 punct (,Eurogentec
R4518 T5055 T5053 appos Eurogentec,polymerase
R4519 T5056 T5055 punct ",",Eurogentec
R4520 T5057 T5055 conj Seraing,Eurogentec
R4521 T5058 T5057 punct ",",Seraing
R4522 T5059 T5057 conj Belgium,Seraing
R4523 T5060 T5024 punct ),µM
R4524 T5061 T5014 punct .,performed
R4525 T5062 T5064 amod First,cycle
R4526 T5063 T5064 compound PCR,cycle
R4527 T5064 T5065 nsubj cycle,consisted
R4528 T5065 T5092 ccomp consisted,consisted
R4529 T5066 T5065 prep of,consisted
R4530 T5067 T5068 nummod 1,min
R4531 T5068 T5066 pobj min,of
R4532 T5069 T5065 prep at,consisted
R4533 T5070 T5071 nummod 92,°
R4534 T5071 T5069 pobj °,at
R4535 T5072 T5065 npadvmod C,consisted
R4536 T5073 T5072 punct ",",C
R4537 T5074 T5075 nummod 1,min
R4538 T5075 T5072 appos min,C
R4539 T5076 T5075 prep at,min
R4540 T5077 T5078 nummod 58,°
R4541 T5078 T5079 nummod °,C
R4542 T5079 T5076 pobj C,at
R4543 T5080 T5075 cc and,min
R4544 T5081 T5082 nummod 1,min
R4545 T5082 T5075 conj min,min
R4546 T5083 T5082 prep at,min
R4547 T5084 T5085 nummod 72,°
R4548 T5085 T5086 nummod °,C
R4549 T5086 T5083 pobj C,at
R4550 T5087 T5092 punct ;,consisted
R4551 T5088 T5092 advmod then,consisted
R4552 T5089 T5091 det each,cycle
R4553 T5090 T5091 compound PCR,cycle
R4554 T5091 T5092 nsubj cycle,consisted
R4555 T5092 T5092 ROOT consisted,consisted
R4556 T5093 T5092 prep of,consisted
R4557 T5094 T5095 nummod 40,sec
R4558 T5095 T5093 pobj sec,of
R4559 T5096 T5092 prep at,consisted
R4560 T5097 T5098 nummod 92,°
R4561 T5098 T5096 pobj °,at
R4562 T5099 T5092 dobj C,consisted
R4563 T5100 T5099 punct ",",C
R4564 T5101 T5102 nummod 40,sec
R4565 T5102 T5099 appos sec,C
R4566 T5103 T5102 prep at,sec
R4567 T5104 T5105 nummod 58,°
R4568 T5105 T5106 nummod °,C
R4569 T5106 T5103 pobj C,at
R4570 T5107 T5102 cc and,sec
R4571 T5108 T5109 nummod 50,sec
R4572 T5109 T5102 conj sec,sec
R4573 T5110 T5109 prep at,sec
R4574 T5111 T5112 nummod 72,°
R4575 T5112 T5114 nummod °,cDNA
R4576 T5113 T5114 compound C.,cDNA
R4577 T5114 T5110 pobj cDNA,at
R4578 T5115 T5109 prep for,sec
R4579 T5116 T5115 pobj β-actin,for
R4580 T5117 T5118 auxpass was,amplified
R4581 T5118 T5092 conj amplified,consisted
R4582 T5119 T5118 prep for,amplified
R4583 T5120 T5121 nummod 28,cycles
R4584 T5121 T5119 pobj cycles,for
R4585 T5122 T5118 advcl using,amplified
R4586 T5123 T5124 det the,oligos
R4587 T5124 T5122 dobj oligos,using
R4588 T5125 T5124 punct :,oligos
R4589 T5126 T5124 appos sense,oligos
R4590 T5127 T5131 nummod 5,′
R4591 T5128 T5130 punct ′,GGCATCGTGATGGACTCCG-3
R4592 T5129 T5130 punct -,GGCATCGTGATGGACTCCG-3
R4593 T5130 T5131 compound GGCATCGTGATGGACTCCG-3,′
R4594 T5131 T5124 appos ′,oligos
R4595 T5132 T5131 cc and,′
R4596 T5133 T5131 conj antisense,′
R4597 T5134 T5133 nummod 5,antisense
R4598 T5135 T5137 nummod ′,′
R4599 T5136 T5137 compound GCTGGAAGGTGGACAGCGA-3,′
R4600 T5137 T5133 appos ′,antisense
R4601 T5138 T5092 punct .,consisted
R4602 T5139 T5143 nsubjpass cDNA,amplified
R4603 T5140 T5139 prep for,cDNA
R4604 T5141 T5140 pobj TNF-α,for
R4605 T5142 T5143 auxpass was,amplified
R4606 T5143 T5143 ROOT amplified,amplified
R4607 T5144 T5143 prep for,amplified
R4608 T5145 T5146 nummod 35,cycles
R4609 T5146 T5144 pobj cycles,for
R4610 T5147 T5143 advcl using,amplified
R4611 T5148 T5149 det the,oligos
R4612 T5149 T5147 dobj oligos,using
R4613 T5150 T5149 punct :,oligos
R4614 T5151 T5149 appos sense,oligos
R4615 T5152 T5155 nummod 5,AAGCCTGTAGCCCATGTTGT-3
R4616 T5153 T5155 punct ′,AAGCCTGTAGCCCATGTTGT-3
R4617 T5154 T5155 punct -,AAGCCTGTAGCCCATGTTGT-3
R4618 T5155 T5156 compound AAGCCTGTAGCCCATGTTGT-3,′
R4619 T5156 T5149 appos ′,oligos
R4620 T5157 T5156 cc and,′
R4621 T5158 T5163 nmod antisense,′
R4622 T5159 T5158 nummod 5,antisense
R4623 T5160 T5162 punct ′,CAGATAGATGGGCTCATACC-3
R4624 T5161 T5162 punct -,CAGATAGATGGGCTCATACC-3
R4637 T5174 T5176 det the,sense
R4638 T5175 T5176 nsubj oligos,sense
R4639 T5176 T5173 dobj sense,using
R4640 T5177 T5181 nummod 5,′
R4641 T5178 T5180 punct ′,GTAGCAGCCGCAGTCATAATGG-3
R4642 T5179 T5180 punct -,GTAGCAGCCGCAGTCATAATGG-3
R4643 T5180 T5181 compound GTAGCAGCCGCAGTCATAATGG-3,′
R4644 T5181 T5176 appos ′,sense
R4645 T5182 T5181 cc and,′
R4646 T5183 T5181 conj antisense,′
R4647 T5184 T5183 nummod 5,antisense
R4648 T5185 T5189 punct ′,′
R4649 T5186 T5189 punct -,′
R4650 T5187 T5189 det A,′
R4651 T5188 T5189 amod TGCTGTTGTATCTGACTGAGG-3,′
R4652 T5189 T5181 appos ′,′
R4653 T5190 T5169 punct .,amplified
R4748 T5335 T5337 compound NF-kB,Reporter
R4749 T5336 T5337 compound Transcription,Reporter
R4750 T5337 T5363 nsubj Reporter,contains
R4751 T5338 T5339 compound Gene,Assay
R4752 T5339 T5340 compound Assay,The
R4753 T5340 T5342 det The,3XMHC-luc
R4754 T5341 T5342 compound plasmid,3XMHC-luc
R4755 T5342 T5363 nsubj 3XMHC-luc,contains
R4756 T5343 T5346 punct (,gift
R4757 T5344 T5346 det a,gift
R4758 T5345 T5346 amod generous,gift
R4759 T5346 T5342 appos gift,3XMHC-luc
R4760 T5347 T5346 prep from,gift
R4761 T5348 T5350 compound Drs.,Westwick
R4762 T5349 T5350 compound J.,Westwick
R4763 T5350 T5347 pobj Westwick,from
R4764 T5351 T5350 cc and,Westwick
R4765 T5352 T5353 compound D.A.,Brenner
R4766 T5353 T5350 conj Brenner,Westwick
R4767 T5354 T5353 punct ",",Brenner
R4768 T5355 T5353 appos University,Brenner
R4769 T5356 T5355 prep of,University
R4770 T5357 T5358 compound North,Carolina
R4771 T5358 T5356 pobj Carolina,of
R4772 T5359 T5353 punct ",",Brenner
R4773 T5360 T5361 compound Chapel,Hill
R4774 T5361 T5353 appos Hill,Brenner
R4775 T5362 T5353 punct ),Brenner
R4776 T5363 T5363 ROOT contains,contains
R4777 T5364 T5365 nummod three,copies
R4778 T5365 T5363 dobj copies,contains
R4779 T5366 T5365 prep of,copies
R4780 T5367 T5368 amod NF-κB-responsive,element
R4781 T5368 T5366 pobj element,of
R4782 T5369 T5368 prep from,element
R4783 T5370 T5372 det the,class
R4784 T5371 T5372 compound MHC,class
R4785 T5372 T5369 pobj class,from
R4786 T5373 T5374 compound I,locus
R4787 T5374 T5366 pobj locus,of
R4788 T5375 T5365 punct ",",copies
R4789 T5376 T5365 acl placed,copies
R4790 T5377 T5376 advmod upstream,placed
R4791 T5378 T5377 prep of,upstream
R4792 T5379 T5381 det the,gene
R4793 T5380 T5381 compound luciferase,gene
R4794 T5381 T5378 pobj gene,of
R4795 T5382 T5363 punct .,contains
R4796 T5383 T5386 amod Human,cells
R4797 T5384 T5386 amod monocytic,cells
R4798 T5385 T5386 compound THP-1,cells
R4799 T5386 T5389 nsubjpass cells,transfected
R4800 T5387 T5389 auxpass were,transfected
R4801 T5388 T5389 advmod transiently,transfected
R4802 T5389 T5389 ROOT transfected,transfected
R4803 T5390 T5392 mark as,described
R4804 T5391 T5392 advmod previously,described
R4805 T5392 T5389 advcl described,transfected
R4806 T5393 T5395 compound [,]
R4807 T5394 T5395 compound 30,]
R4808 T5395 T5392 dobj ],described
R4809 T5396 T5389 punct ",",transfected
R4810 T5397 T5389 cc and,transfected
R4811 T5398 T5399 advmod then,cultured
R4812 T5399 T5389 conj cultured,transfected
R4813 T5400 T5399 prep for,cultured
R4814 T5401 T5402 nummod 4,h
R4815 T5402 T5400 pobj h,for
R4816 T5403 T5402 advmod alone,h
R4817 T5404 T5400 cc or,for
R4818 T5405 T5400 conj with,for
R4819 T5406 T5407 amod increasing,concentration
R4820 T5407 T5405 pobj concentration,with
R4821 T5408 T5407 prep of,concentration
R4822 T5409 T5410 det either,C10
R4823 T5410 T5408 pobj C10,of
R4824 T5411 T5410 cc or,C10
R4825 T5412 T5410 conj C40,C10
R4826 T5413 T5389 punct .,transfected
R4827 T5414 T5415 amod Luciferase,activity
R4828 T5415 T5417 nsubjpass activity,determined
R4829 T5416 T5417 auxpass was,determined
R4830 T5417 T5417 ROOT determined,determined
R4831 T5418 T5417 advcl using,determined
R4832 T5419 T5420 det a,luminometer
R4833 T5420 T5418 dobj luminometer,using
R4834 T5421 T5420 punct (,luminometer
R4835 T5422 T5420 appos Monolight,luminometer
R4836 T5423 T5424 nummod 2010,Luminometer
R4837 T5424 T5424 ROOT Luminometer,Luminometer
R4838 T5425 T5424 punct ",",Luminometer
R4839 T5426 T5427 compound Ann,Arbor
R4840 T5427 T5424 conj Arbor,Luminometer
R4841 T5428 T5427 punct ",",Arbor
R4842 T5429 T5427 conj MI,Arbor
R4843 T5430 T5422 punct ),Monolight
R4844 T5431 T5417 punct .,determined
R5294 T5933 T5937 amod Western,cells
R5295 T5934 T5936 compound Blot,THP-1
R5296 T5935 T5936 compound Analysis,THP-1
R5297 T5936 T5937 compound THP-1,cells
R5298 T5937 T5939 nsubjpass cells,stimulated
R5299 T5938 T5939 auxpass were,stimulated
R5300 T5939 T5939 ROOT stimulated,stimulated
R5301 T5940 T5939 prep for,stimulated
R5302 T5941 T5942 amod various,lengths
R5303 T5942 T5940 pobj lengths,for
R5304 T5943 T5942 prep of,lengths
R5305 T5944 T5943 pobj time,of
R5306 T5945 T5942 prep with,lengths
R5307 T5946 T5947 nummod 0.1,mg/ml
R5308 T5947 T5945 pobj mg/ml,with
R5309 T5948 T5947 nummod C10,mg/ml
R5310 T5949 T5948 cc or,C10
R5311 T5950 T5948 conj C40,C10
R5312 T5951 T5950 punct ",",C40
R5313 T5952 T5950 cc or,C40
R5314 T5953 T5954 nummod 10,µg
R5315 T5954 T5957 compound µg,LPS
R5316 T5955 T5957 compound /,LPS
R5317 T5956 T5957 compound ml,LPS
R5318 T5957 T5950 conj LPS,C40
R5319 T5958 T5939 punct .,stimulated
R5320 T5959 T5962 nsubjpass Cells,pelleted
R5321 T5960 T5962 auxpass were,pelleted
R5322 T5961 T5962 advmod then,pelleted
R5323 T5962 T5962 ROOT pelleted,pelleted
R5324 T5963 T5962 punct ",",pelleted
R5325 T5964 T5962 conj washed,pelleted
R5326 T5965 T5964 cc and,washed
R5327 T5966 T5964 conj homogenised,washed
R5328 T5967 T5966 prep in,homogenised
R5329 T5968 T5969 compound lysis,buffer
R5330 T5969 T5967 pobj buffer,in
R5331 T5970 T5973 punct (,Hepes
R5332 T5971 T5973 nummod 10,Hepes
R5333 T5972 T5973 compound mM,Hepes
R5334 T5973 T5966 appos Hepes,homogenised
R5335 T5974 T5973 punct ",",Hepes
R5336 T5975 T5973 conj pH,Hepes
R5337 T5976 T5975 nummod 7.9,pH
R5338 T5977 T5975 punct ",",pH
R5339 T5978 T5980 nummod 150,NaCl
R5340 T5979 T5980 compound mM,NaCl
R5341 T5980 T5975 conj NaCl,pH
R5342 T5981 T5980 punct ",",NaCl
R5343 T5982 T5984 nummod 1,EDTA
R5344 T5983 T5984 compound mM,EDTA
R5345 T5984 T5980 conj EDTA,NaCl
R5346 T5985 T5984 punct ",",EDTA
R5347 T5986 T5987 nummod 0.6,%
R5348 T5987 T5988 compound %,NP-40
R5349 T5988 T5984 conj NP-40,EDTA
R5350 T5989 T5988 punct ",",NP-40
R5351 T5990 T5988 cc and,NP-40
R5352 T5991 T5993 nummod 0.5,PMSF
R5353 T5992 T5993 compound mM,PMSF
R5354 T5993 T5988 conj PMSF,NP-40
R5355 T5994 T5993 punct ),PMSF
R5356 T5995 T5988 prep on,NP-40
R5357 T5996 T5995 pobj ice,on
R5358 T5997 T5962 punct .,pelleted
R5359 T5998 T6000 nsubjpass Homogenates,sonicated
R5360 T5999 T6000 auxpass were,sonicated
R5361 T6000 T6000 ROOT sonicated,sonicated
R5362 T6001 T6000 punct ",",sonicated
R5363 T6002 T6000 conj centrifuged,sonicated
R5364 T6003 T6002 prep at,centrifuged
R5365 T6004 T6005 nummod "10,000",rpm
R5366 T6005 T6003 pobj rpm,at
R5367 T6006 T6007 aux to,remove
R5368 T6007 T6002 advcl remove,centrifuged
R5369 T6008 T6009 amod cellular,debris
R5370 T6009 T6007 dobj debris,remove
R5371 T6010 T6007 punct ",",remove
R5372 T6011 T6007 cc and,remove
R5373 T6012 T6013 amod supernatant,collected
R5374 T6013 T6002 conj collected,centrifuged
R5375 T6014 T6000 punct .,sonicated
R5376 T6015 T6016 compound Protein,concentration
R5377 T6016 T6018 nsubjpass concentration,determined
R5378 T6017 T6018 auxpass was,determined
R5379 T6018 T6018 ROOT determined,determined
R5380 T6019 T6018 advcl using,determined
R5381 T6020 T6023 det the,Assay
R5382 T6021 T6023 compound DC,Assay
R5383 T6022 T6023 compound Protein,Assay
R5384 T6023 T6019 dobj Assay,using
R5385 T6024 T6023 punct (,Assay
R5386 T6025 T6023 appos Bio-Rad,Assay
R5387 T6026 T6023 punct ),Assay
R5388 T6027 T6018 punct .,determined
R5389 T6028 T6038 nsubjpass Proteins,resolved
R5390 T6029 T6028 prep in,Proteins
R5391 T6030 T6029 pobj samples,in
R5392 T6031 T6030 punct (,samples
R5393 T6032 T6033 nummod 15,µg
R5394 T6033 T6038 nsubjpass µg,resolved
R5395 T6034 T6035 amod total,proteins
R5396 T6035 T6033 appos proteins,µg
R5397 T6036 T6033 punct ),µg
R5398 T6037 T6038 auxpass were,resolved
R5399 T6038 T6038 ROOT resolved,resolved
R5400 T6039 T6038 prep in,resolved
R5401 T6040 T6045 det a,gel
R5402 T6041 T6045 amod denaturing,gel
R5403 T6042 T6043 nummod 12,%
R5404 T6043 T6045 compound %,gel
R5405 T6044 T6045 compound polyacrylamide,gel
R5406 T6045 T6039 pobj gel,in
R5407 T6046 T6038 cc and,resolved
R5408 T6047 T6038 conj transferred,resolved
R5409 T6048 T6047 prep to,transferred
R5410 T6049 T6051 det a,membrane
R5411 T6050 T6051 amod nitrocellulose,membrane
R5412 T6051 T6048 pobj membrane,to
R5413 T6052 T6038 punct .,resolved
R5414 T6053 T6054 amod I-κBα,protein
R5415 T6054 T6056 nsubjpass protein,detected
R5416 T6055 T6056 auxpass was,detected
R5417 T6056 T6056 ROOT detected,detected
R5418 T6057 T6056 advcl using,detected
R5419 T6058 T6061 det a,antibody
R5420 T6059 T6061 nmod rabbit,antibody
R5421 T6060 T6061 compound polyclonal,antibody
R5422 T6061 T6057 dobj antibody,using
R5423 T6062 T6061 punct (,antibody
R5424 T6063 T6064 compound Santa,Cruz
R5425 T6064 T6065 compound Cruz,Biotechnology
R5426 T6065 T6061 appos Biotechnology,antibody
R5427 T6066 T6065 punct ",",Biotechnology
R5428 T6067 T6065 appos CA,Biotechnology
R5429 T6068 T6061 punct ),antibody
R5430 T6069 T6056 advcl followed,detected
R5431 T6070 T6069 agent by,followed
R5432 T6071 T6076 det a,antibody
R5433 T6072 T6076 amod horseradish,antibody
R5434 T6073 T6076 amod peroxidase-coupled,antibody
R5435 T6074 T6076 compound goat,antibody
R5436 T6075 T6076 compound polyclonal,antibody
R5437 T6076 T6070 pobj antibody,by
R5438 T6077 T6076 prep against,antibody
R5439 T6078 T6079 compound rabbit,Ig
R5440 T6079 T6077 pobj Ig,against
R5441 T6080 T6082 punct (,Laboratories
R5442 T6081 T6082 compound Caltag,Laboratories
R5443 T6082 T6079 appos Laboratories,Ig
R5444 T6083 T6082 punct ),Laboratories
R5445 T6084 T6056 punct .,detected
R5446 T6085 T6090 advmod Finally,revealed
R5447 T6086 T6090 punct ",",revealed
R5448 T6087 T6088 compound IκB,bands
R5449 T6088 T6090 nsubjpass bands,revealed
R5450 T6089 T6090 auxpass were,revealed
R5451 T6090 T6090 ROOT revealed,revealed
R5452 T6091 T6090 advcl using,revealed
R5453 T6092 T6096 det the,system
R5454 T6093 T6096 nmod ECL,system
R5455 T6094 T6096 compound ™,system
R5456 T6095 T6096 compound detection,system
R5457 T6096 T6091 dobj system,using
R5458 T6097 T6096 punct (,system
R5459 T6098 T6100 compound Amersham,Biotech
R5460 T6099 T6100 compound Pharmacia,Biotech
R5461 T6100 T6096 appos Biotech,system
R5462 T6101 T6096 punct ",",system
R5463 T6102 T6103 compound Les,Ullis
R5464 T6103 T6096 appos Ullis,system
R5465 T6104 T6103 punct ",",Ullis
R5466 T6105 T6103 appos France,Ullis
R5467 T6106 T6096 punct ),system
R5468 T6107 T6090 prep according,revealed
R5469 T6108 T6107 prep to,according
R5470 T6109 T6110 det the,manufacturers
R5471 T6110 T6112 poss manufacturers,instruction
R5472 T6111 T6110 case ',manufacturers
R5473 T6112 T6108 pobj instruction,to
R5474 T6113 T6090 punct .,revealed
R5475 T6114 T6121 nsubj Antibody,was
R5476 T6115 T6114 prep to,Antibody
R5477 T6116 T6115 pobj α-Tubulin,to
R5478 T6117 T6119 punct (,Cruz
R5479 T6118 T6119 compound Santa,Cruz
R5480 T6119 T6116 appos Cruz,α-Tubulin
R5481 T6120 T6116 punct ),α-Tubulin
R5482 T6121 T6121 ROOT was,was
R5483 T6122 T6121 attr use,was
R5484 T6123 T6122 prep as,use
R5485 T6124 T6123 pcomp loading,as
R5486 T6125 T6124 dobj control,loading
R5487 T6126 T6121 punct .,was
R5488 T6127 T6134 prep For,stimulated
R5489 T6128 T6129 amod nuclear,NF-κB
R5490 T6129 T6127 pobj NF-κB,For
R5491 T6130 T6134 punct ",",stimulated
R5492 T6131 T6132 compound THP-1,cells
R5493 T6132 T6134 nsubjpass cells,stimulated
R5494 T6133 T6134 auxpass were,stimulated
R5495 T6134 T6151 ccomp stimulated,pelleted
R5496 T6135 T6134 prep with,stimulated
R5497 T6136 T6137 nummod 1,mg/ml
R5498 T6137 T6135 pobj mg/ml,with
R5499 T6138 T6137 nummod C10,mg/ml
R5500 T6139 T6138 cc or,C10
R5501 T6140 T6138 conj C40,C10
R5502 T6141 T6140 prep for,C40
R5503 T6142 T6143 nummod 30,minutes
R5504 T6143 T6141 pobj minutes,for
R5505 T6144 T6143 prep at,minutes
R5506 T6145 T6146 nummod 37,°
R5507 T6146 T6148 compound °,Cells
R5508 T6147 T6148 compound C.,Cells
R5509 T6148 T6144 pobj Cells,at
R5510 T6149 T6151 auxpass were,pelleted
R5511 T6150 T6151 advmod then,pelleted
R5512 T6151 T6151 ROOT pelleted,pelleted
R5513 T6152 T6151 cc and,pelleted
R5514 T6153 T6151 conj nuclei,pelleted
R5515 T6154 T6151 conj separated,pelleted
R5516 T6155 T6156 mark as,described
R5517 T6156 T6151 advcl described,pelleted
R5518 T6157 T6159 nmod [,]
R5519 T6158 T6159 nummod 31,]
R5520 T6159 T6156 dobj ],described
R5521 T6160 T6151 punct .,pelleted
R5522 T6161 T6163 nsubjpass Nuclei,washed
R5523 T6162 T6163 auxpass were,washed
R5524 T6163 T6163 ROOT washed,washed
R5525 T6164 T6163 cc and,washed
R5526 T6165 T6163 conj homogenized,washed
R5527 T6166 T6165 advmod directly,homogenized
R5528 T6167 T6165 prep in,homogenized
R5529 T6168 T6167 pcomp loading,in
R5530 T6169 T6172 punct (,buffer
R5531 T6170 T6172 nmod Laemli,buffer
R5532 T6171 T6172 punct ),buffer
R5533 T6172 T6168 dobj buffer,loading
R5534 T6173 T6172 cc and,buffer
R5535 T6174 T6163 conj heated,washed
R5536 T6175 T6174 prep for,heated
R5537 T6176 T6177 nummod 5,minutes
R5538 T6177 T6175 pobj minutes,for
R5539 T6178 T6174 prep at,heated
R5540 T6179 T6182 nummod 100,Proteins
R5541 T6180 T6182 nummod °,Proteins
R5542 T6181 T6182 compound C.,Proteins
R5543 T6182 T6178 pobj Proteins,at
R5544 T6183 T6182 prep in,Proteins
R5545 T6184 T6183 pobj samples,in
R5546 T6185 T6186 auxpass were,resolved
R5547 T6186 T6163 conj resolved,washed
R5548 T6187 T6186 prep in,resolved
R5549 T6188 T6193 det a,gel
R5550 T6189 T6193 amod denaturing,gel
R5551 T6190 T6191 nummod 8,%
R5552 T6191 T6193 compound %,gel
R5553 T6192 T6193 compound polyacrylamide,gel
R5554 T6193 T6187 pobj gel,in
R5555 T6194 T6186 cc and,resolved
R5556 T6195 T6186 conj transferred,resolved
R5557 T6196 T6195 prep to,transferred
R5558 T6197 T6199 det a,fluoride
R5559 T6198 T6199 compound polyvinylidine,fluoride
R5560 T6199 T6196 pobj fluoride,to
R5561 T6200 T6199 punct (,fluoride
R5562 T6201 T6199 appos PVDF,fluoride
R5563 T6202 T6199 punct ),fluoride
R5564 T6203 T6195 dobj membrane,transferred
R5565 T6204 T6205 punct (,Immobilon-P
R5566 T6205 T6203 appos Immobilon-P,membrane
R5567 T6206 T6163 punct ;,washed
R5568 T6207 T6207 ROOT Millipore,Millipore
R5569 T6208 T6207 punct ",",Millipore
R5570 T6209 T6207 conj Bedford,Millipore
R5571 T6210 T6209 punct ",",Bedford
R5572 T6211 T6209 conj MA,Bedford
R5573 T6212 T6209 punct ),Bedford
R5574 T6213 T6207 punct .,Millipore
R5575 T6214 T6216 nsubjpass Membranes,incubated
R5576 T6215 T6216 auxpass were,incubated
R5577 T6216 T6216 ROOT incubated,incubated
R5578 T6217 T6216 prep in,incubated
R5579 T6218 T6217 pcomp blocking,in
R5580 T6219 T6218 dobj buffer,blocking
R5581 T6220 T6223 punct (,BSA
R5582 T6221 T6222 nummod 1,%
R5583 T6222 T6223 npadvmod %,BSA
R5584 T6223 T6218 dobj BSA,blocking
R5585 T6224 T6218 punct ",",blocking
R5586 T6225 T6224 oprd in,","
R5587 T6226 T6225 pobj PBS,in
R5588 T6227 T6225 punct ),in
R5589 T6228 T6216 prep for,incubated
R5590 T6229 T6230 nummod two,hours
R5591 T6230 T6228 pobj hours,for
R5592 T6231 T6230 prep at,hours
R5593 T6232 T6233 compound room,temperature
R5594 T6233 T6231 pobj temperature,at
R5595 T6234 T6216 punct .,incubated
R5596 T6235 T6238 nsubjpass Membranes,probed
R5597 T6236 T6238 auxpass were,probed
R5598 T6237 T6238 advmod subsequently,probed
R5599 T6238 T6238 ROOT probed,probed
R5600 T6239 T6238 prep with,probed
R5601 T6240 T6242 det the,antibody
R5602 T6241 T6242 amod corresponding,antibody
R5603 T6242 T6239 pobj antibody,with
R5604 T6243 T6238 prep in,probed
R5605 T6244 T6243 pcomp blocking,in
R5606 T6245 T6244 dobj buffer,blocking
R5607 T6246 T6247 punct ",",overnight
R5608 T6247 T6244 npadvmod overnight,blocking
R5609 T6248 T6238 punct .,probed
R5610 T6249 T6254 nmod Rabbit,subunit
R5611 T6250 T6254 amod polyclonal,subunit
R5612 T6251 T6254 nmod antibody,subunit
R5613 T6252 T6254 nmod anti-NF-κB,subunit
R5614 T6253 T6254 amod p50,subunit
R5615 T6254 T6272 nsubjpass subunit,used
R5616 T6255 T6254 punct (,subunit
R5617 T6256 T6257 nmod #,sc-114
R5618 T6257 T6272 nsubjpass sc-114,used
R5619 T6258 T6257 punct ),sc-114
R5620 T6259 T6257 cc or,sc-114
R5621 T6260 T6261 amod anti-NF-κB,p65
R5622 T6261 T6257 conj p65,sc-114
R5623 T6262 T6261 acl subunit,p65
R5624 T6263 T6261 punct (,p65
R5625 T6264 T6265 nmod #,sc-109
R5626 T6265 T6261 appos sc-109,p65
R5627 T6266 T6261 punct ),p65
R5628 T6267 T6261 prep from,p65
R5629 T6268 T6269 compound Santa,Cruz
R5630 T6269 T6270 compound Cruz,Biotechnology
R5631 T6270 T6267 pobj Biotechnology,from
R5632 T6271 T6272 auxpass were,used
R5633 T6272 T6272 ROOT used,used
R5634 T6273 T6272 punct .,used
R5635 T6274 T6276 nsubjpass Membranes,washed
R5636 T6275 T6276 auxpass were,washed
R5637 T6276 T6276 ROOT washed,washed
R5638 T6277 T6278 nummod six,times
R5639 T6278 T6276 npadvmod times,washed
R5640 T6279 T6276 prep in,washed
R5641 T6280 T6279 pobj PBS,in
R5642 T6281 T6276 prep with,washed
R5643 T6282 T6283 nummod 0.05,%
R5644 T6283 T6288 nummod %,minutes
R5645 T6284 T6288 nummod Tween,minutes
R5646 T6285 T6288 nummod 20,minutes
R5647 T6286 T6288 punct ",",minutes
R5648 T6287 T6288 nummod 5,minutes
R5649 T6288 T6281 pobj minutes,with
R5650 T6289 T6290 det each,time
R5651 T6290 T6276 npadvmod time,washed
R5652 T6291 T6276 punct ",",washed
R5653 T6292 T6276 cc and,washed
R5654 T6293 T6276 conj incubated,washed
R5655 T6294 T6293 prep with,incubated
R5656 T6295 T6297 det a,dilution
R5657 T6296 T6297 nummod 1/3000,dilution
R5658 T6297 T6294 pobj dilution,with
R5659 T6298 T6297 prep of,dilution
R5660 T6299 T6300 amod HRP-conjugated,F
R5661 T6300 T6298 pobj F,of
R5662 T6301 T6308 punct (,IgG
R5663 T6302 T6306 nmod ab,goat
R5664 T6303 T6302 case ',ab
R5665 T6304 T6302 punct ),ab
R5666 T6305 T6302 nummod 2,ab
R5667 T6306 T6308 nmod goat,IgG
R5668 T6307 T6308 compound anti-rabbit,IgG
R5669 T6308 T6314 nmod IgG,milk
R5670 T6309 T6308 prep in,IgG
R5671 T6310 T6311 nummod 5,%
R5672 T6311 T6314 nmod %,milk
R5673 T6312 T6314 amod nonfat,milk
R5674 T6313 T6314 amod dry,milk
R5675 T6314 T6297 appos milk,dilution
R5676 T6315 T6314 cc and,milk
R5677 T6316 T6317 nummod 0.05,%
R5678 T6317 T6314 conj %,milk
R5679 T6318 T6319 nummod Tween,20
R5680 T6319 T6322 dep 20,for
R5681 T6320 T6319 prep in,20
R5682 T6321 T6320 pobj PBS,in
R5683 T6322 T6293 conj for,incubated
R5684 T6323 T6324 nummod 1,hour
R5685 T6324 T6322 pobj hour,for
R5686 T6325 T6322 prep at,for
R5687 T6326 T6327 compound room,temperature
R5688 T6327 T6325 pobj temperature,at
R5689 T6328 T6276 punct .,washed
R5690 T6329 T6345 prep After,detected
R5691 T6330 T6329 pcomp washing,After
R5692 T6331 T6333 nummod six,times
R5693 T6332 T6331 amod more,six
R5694 T6333 T6330 dobj times,washing
R5695 T6334 T6330 prep in,washing
R5696 T6335 T6334 pobj PBS,in
R5697 T6336 T6330 prep with,washing
R5698 T6337 T6338 nummod 0.05,%
R5699 T6338 T6345 nsubj %,detected
R5700 T6339 T6340 nummod Tween,20
R5701 T6340 T6345 dep 20,detected
R5702 T6341 T6345 punct ",",detected
R5703 T6342 T6343 amod antibody-reactive,proteins
R5704 T6343 T6345 nsubjpass proteins,detected
R5705 T6344 T6345 auxpass were,detected
R5706 T6345 T6345 ROOT detected,detected
R5707 T6346 T6345 advcl using,detected
R5708 T6347 T6349 det a,substrate
R5709 T6348 T6349 compound chemiluminescence,substrate
R5710 T6349 T6346 dobj substrate,using
R5711 T6350 T6359 punct (,according
R5712 T6351 T6359 dep SuperSignal,according
R5713 T6352 T6351 punct ;,SuperSignal
R5714 T6353 T6351 conj Pierce,SuperSignal
R5715 T6354 T6353 punct ",",Pierce
R5716 T6355 T6353 conj Rockford,Pierce
R5717 T6356 T6355 punct ",",Rockford
R5718 T6357 T6355 conj IL,Rockford
R5719 T6358 T6359 punct ),according
R5720 T6359 T6345 prep according,detected
R5721 T6360 T6359 prep to,according
R5722 T6361 T6362 det the,manufacturer
R5723 T6362 T6364 poss manufacturer,instructions
R5724 T6363 T6362 case 's,manufacturer
R5725 T6364 T6360 pobj instructions,to
R5726 T6365 T6345 punct .,detected
R5727 T6366 T6367 aux To,confirm
R5728 T6367 T6380 advcl confirm,were
R5729 T6368 T6374 mark that,loaded
R5730 T6369 T6370 amod equivalent,amounts
R5731 T6370 T6374 nsubjpass amounts,loaded
R5732 T6371 T6370 prep of,amounts
R5733 T6372 T6371 pobj protein,of
R5734 T6373 T6374 auxpass were,loaded
R5735 T6374 T6367 ccomp loaded,confirm
R5736 T6375 T6374 prep in,loaded
R5737 T6376 T6377 det each,line
R5738 T6377 T6375 pobj line,in
R5739 T6378 T6380 punct ",",were
R5740 T6379 T6380 nsubj membranes,were
R5741 T6380 T6380 ROOT were,were
R5742 T6381 T6380 advmod also,were
R5743 T6382 T6383 amod Western,blotted
R5744 T6383 T6380 attr blotted,were
R5745 T6384 T6383 prep for,blotted
R5746 T6385 T6384 pobj ERK,for
R5747 T6386 T6387 mark as,described
R5748 T6387 T6380 advcl described,were
R5749 T6388 T6390 nmod [,]
R5750 T6389 T6390 nummod 32,]
R5751 T6390 T6387 dobj ],described
R5752 T6391 T6380 punct .,were
R5773 T6486 T6503 nsubjpass Analysis,performed
R5774 T6487 T6486 prep of,Analysis
R5775 T6488 T6489 compound NF-κB,Activation
R5776 T6489 T6487 pobj Activation,of
R5777 T6490 T6489 prep by,Activation
R5778 T6491 T6494 compound Flow,activation
R5779 T6492 T6494 compound Cytometry,activation
R5780 T6493 T6494 compound Nuclear,activation
R5781 T6494 T6490 pobj activation,by
R5782 T6495 T6494 prep of,activation
R5783 T6496 T6498 compound NF,κΒ
R5784 T6497 T6498 compound −,κΒ
R5785 T6498 T6495 pobj κΒ,of
R5786 T6499 T6486 prep by,Analysis
R5787 T6500 T6501 compound flow,cytometry
R5788 T6501 T6499 pobj cytometry,by
R5789 T6502 T6503 auxpass was,performed
R5790 T6503 T6503 ROOT performed,performed
R5791 T6504 T6505 mark as,described
R5792 T6505 T6503 advcl described,performed
R5793 T6506 T6508 nmod [,]
R5794 T6507 T6508 nummod 31,]
R5795 T6508 T6505 dobj ],described
R5796 T6509 T6503 punct .,performed
R5837 T6558 T6559 compound Statistical,Analysis
R5838 T6559 T6563 nsubjpass Analysis,expressed
R5839 T6560 T6561 det The,results
R5840 T6561 T6563 nsubjpass results,expressed
R5841 T6562 T6563 auxpass were,expressed
R5842 T6563 T6563 ROOT expressed,expressed
R5843 T6564 T6563 prep as,expressed
R5844 T6565 T6569 det the,S.E.M.
R5845 T6566 T6569 amod mean,S.E.M.
R5846 T6567 T6568 compound value,±
R5847 T6568 T6569 compound ±,S.E.M.
R5848 T6569 T6564 pobj S.E.M.,as
R5849 T6570 T6569 prep of,S.E.M.
R5850 T6571 T6572 amod individual,experiments
R5851 T6572 T6570 pobj experiments,of
R5852 T6573 T6563 punct .,expressed
R5853 T6574 T6576 det The,significance
R5854 T6575 T6576 amod statistical,significance
R5855 T6576 T6584 nsubjpass significance,assessed
R5856 T6577 T6576 prep of,significance
R5857 T6578 T6579 det the,differences
R5858 T6579 T6577 pobj differences,of
R5859 T6580 T6579 prep between,differences
R5860 T6581 T6582 amod mean,values
R5861 T6582 T6580 pobj values,between
R5862 T6583 T6584 auxpass was,assessed
R5863 T6584 T6584 ROOT assessed,assessed
R5864 T6585 T6584 agent by,assessed
R5865 T6586 T6587 det the,Student
R5866 T6587 T6591 poss Student,analysis
R5867 T6588 T6587 case 's,Student
R5868 T6589 T6591 amod t-test,analysis
R5869 T6590 T6589 cc and,t-test
R5870 T6591 T6585 pobj analysis,by
R5871 T6592 T6591 prep of,analysis
R5872 T6593 T6592 pobj variance,of
R5873 T6594 T6595 punct (,ANOVA
R5874 T6595 T6593 appos ANOVA,variance
R5875 T6596 T6584 punct ),assessed
R5876 T6597 T6584 punct .,assessed
R6115 T6857 T6859 amod Degraded,Induce
R6116 T6858 T6859 compound CGN,Induce
R6117 T6859 T6863 nmod Induce,rats
R6118 T6860 T6863 compound Colonic,rats
R6119 T6861 T6863 compound Inflammation,rats
R6120 T6862 T6863 compound All,rats
R6121 T6863 T6864 nsubj rats,developed
R6122 T6864 T6864 ROOT developed,developed
R6123 T6865 T6864 dobj diarrhea,developed
R6124 T6866 T6864 prep during,developed
R6125 T6867 T6869 amod degraded,administration
R6126 T6868 T6869 compound carrageenan,administration
R6127 T6869 T6866 pobj administration,during
R6128 T6870 T6869 cc and,administration
R6129 T6871 T6872 amod gross,evidence
R6130 T6872 T6869 conj evidence,administration
R6131 T6873 T6872 prep of,evidence
R6132 T6874 T6873 pobj blood,of
R6133 T6875 T6877 auxpass was,detected
R6134 T6876 T6877 advmod frequently,detected
R6135 T6877 T6864 conj detected,developed
R6136 T6878 T6877 prep in,detected
R6137 T6879 T6880 det the,stools
R6138 T6880 T6878 pobj stools,in
R6139 T6881 T6864 punct .,developed
R6140 T6882 T6883 compound Colon,length
R6141 T6883 T6885 nsubj length,decreased
R6142 T6884 T6885 advmod dramatically,decreased
R6143 T6885 T6885 ROOT decreased,decreased
R6144 T6886 T6885 prep in,decreased
R6145 T6887 T6889 det all,rats
R6146 T6888 T6889 amod treated,rats
R6147 T6889 T6886 pobj rats,in
R6148 T6890 T6889 prep with,rats
R6149 T6891 T6894 det a,effect
R6150 T6892 T6893 advmod more,pronounced
R6151 T6893 T6894 amod pronounced,effect
R6152 T6894 T6890 pobj effect,with
R6153 T6895 T6896 auxpass being,observed
R6154 T6896 T6894 acl observed,effect
R6155 T6897 T6896 prep in,observed
R6156 T6898 T6901 det the,dCGN
R6157 T6899 T6901 nummod 40,dCGN
R6158 T6900 T6901 compound kDa,dCGN
R6159 T6901 T6897 pobj dCGN,in
R6160 T6902 T6885 conj treated,decreased
R6161 T6903 T6902 dobj group,treated
R6162 T6904 T6906 punct (,1A
R6163 T6905 T6906 compound Fig.,1A
R6164 T6906 T6903 appos 1A,group
R6165 T6907 T6906 punct ),1A
R6166 T6908 T6885 punct .,decreased
R6167 T6909 T6917 advmod Furthermore,resulted
R6168 T6910 T6917 punct ",",resulted
R6169 T6911 T6912 amod prolonged,exposure
R6170 T6912 T6917 nsubj exposure,resulted
R6171 T6913 T6912 prep to,exposure
R6172 T6914 T6916 nummod 40,dCGN
R6173 T6915 T6916 compound kDa,dCGN
R6174 T6916 T6917 nsubj dCGN,resulted
R6175 T6917 T6917 ROOT resulted,resulted
R6176 T6918 T6917 prep in,resulted
R6177 T6919 T6923 amod high,scores
R6178 T6920 T6923 amod macroscopic,scores
R6179 T6921 T6920 cc and,macroscopic
R6180 T6922 T6920 conj histological,macroscopic
R6181 T6923 T6918 pobj scores,in
R6182 T6924 T6923 prep of,scores
R6183 T6925 T6924 pobj inflammation,of
R6184 T6926 T6930 punct (,C
R6185 T6927 T6928 compound Fig.,1B
R6186 T6928 T6930 dep 1B,C
R6187 T6929 T6930 punct ",",C
R6188 T6930 T6923 appos C,scores
R6189 T6931 T6923 punct ),scores
R6190 T6932 T6917 punct .,resulted
R6191 T6933 T6936 amod Only,activity
R6192 T6934 T6936 amod weak,activity
R6193 T6935 T6936 compound myeloperoxidase,activity
R6194 T6936 T6938 nsubjpass activity,detected
R6195 T6937 T6938 auxpass was,detected
R6196 T6938 T6938 ROOT detected,detected
R6197 T6939 T6938 prep in,detected
R6198 T6940 T6941 det both,control
R6199 T6941 T6939 pobj control,in
R6200 T6942 T6941 cc and,control
R6201 T6943 T6944 amod dCGN-treated,groups
R6202 T6944 T6941 conj groups,control
R6203 T6945 T6944 punct (,groups
R6204 T6946 T6947 compound Fig.,1D
R6205 T6947 T6944 appos 1D,groups
R6206 T6948 T6944 punct ),groups
R6207 T6949 T6938 punct ",",detected
R6208 T6950 T6938 advcl indicating,detected
R6209 T6951 T6955 mark that,play
R6210 T6952 T6955 nsubj granulocytes,play
R6211 T6953 T6955 aux did,play
R6212 T6954 T6955 neg not,play
R6213 T6955 T6950 ccomp play,indicating
R6214 T6956 T6958 det a,role
R6215 T6957 T6958 amod major,role
R6216 T6958 T6955 dobj role,play
R6217 T6959 T6955 prep in,play
R6218 T6960 T6961 det the,inflammation
R6219 T6961 T6959 pobj inflammation,in
R6220 T6962 T6955 prep at,play
R6221 T6963 T6964 det that,stage
R6222 T6964 T6962 pobj stage,at
R6223 T6965 T6938 punct .,detected
R6224 T6966 T6967 amod Histological,examination
R6225 T6967 T6968 nsubj examination,revealed
R6226 T6968 T6968 ROOT revealed,revealed
R6227 T6969 T6970 amod various,degrees
R6228 T6970 T6968 dobj degrees,revealed
R6229 T6971 T6970 prep of,degrees
R6230 T6972 T6973 amod mucosal,inflammation
R6231 T6973 T6971 pobj inflammation,of
R6232 T6974 T6968 punct .,revealed
R6233 T6975 T6976 nsubj Rats,treated
R6234 T6976 T6976 ROOT treated,treated
R6235 T6977 T6976 prep with,treated
R6236 T6978 T6980 nummod 10,dCGN
R6237 T6979 T6980 compound kDa,dCGN
R6238 T6980 T6977 pobj dCGN,with
R6239 T6981 T6976 conj showed,treated
R6240 T6982 T6981 dobj edema,showed
R6241 T6983 T6981 punct ",",showed
R6242 T6984 T6985 compound epithelium,atrophy
R6243 T6985 T6985 ROOT atrophy,atrophy
R6244 T6986 T6985 cc and,atrophy
R6245 T6987 T6989 amod slight,infiltration
R6246 T6988 T6989 compound lymphocyte,infiltration
R6247 T6989 T6985 conj infiltration,atrophy
R6248 T6990 T6993 punct (,shown
R6249 T6991 T6993 nsubjpass data,shown
R6250 T6992 T6993 neg not,shown
R6251 T6993 T6985 parataxis shown,atrophy
R6252 T6994 T6993 punct ),shown
R6253 T6995 T6976 punct .,treated
R6254 T6996 T6997 det These,symptoms
R6255 T6997 T6998 nsubj symptoms,were
R6256 T6998 T6998 ROOT were,were
R6257 T6999 T7000 advmod totally,absent
R6258 T7000 T6998 acomp absent,were
R6259 T7001 T7000 prep in,absent
R6260 T7002 T7003 det the,colon
R6261 T7003 T7001 pobj colon,in
R6262 T7004 T7003 prep of,colon
R6263 T7005 T7006 compound control,rats
R6264 T7006 T7004 pobj rats,of
R6265 T7007 T7003 punct (,colon
R6266 T7008 T7009 compound Fig.,1E
R6267 T7009 T7003 appos 1E,colon
R6268 T7010 T7003 punct ),colon
R6269 T7011 T6998 punct .,were
R6270 T7012 T7013 advmod More,severe
R6271 T7013 T7015 amod severe,injuries
R6272 T7014 T7015 amod mucosal,injuries
R6273 T7015 T7031 nsubjpass injuries,observed
R6274 T7016 T7015 prep including,injuries
R6275 T7017 T7016 pobj ulceration,including
R6276 T7018 T7017 punct ",",ulceration
R6277 T7019 T7020 amod hyperplastic,epithelium
R6278 T7020 T7017 conj epithelium,ulceration
R6279 T7021 T7020 punct ",",epithelium
R6280 T7022 T7023 compound crypt,distortion
R6281 T7023 T7020 conj distortion,epithelium
R6282 T7024 T7023 cc and,distortion
R6283 T7025 T7028 det a,infiltration
R6284 T7026 T7028 amod strong,infiltration
R6285 T7027 T7028 compound macrophage,infiltration
R6286 T7028 T7023 conj infiltration,distortion
R6287 T7029 T7031 punct ",",observed
R6288 T7030 T7031 auxpass were,observed
R6289 T7031 T7031 ROOT observed,observed
R6290 T7032 T7031 prep in,observed
R6291 T7033 T7037 det the,rats
R6292 T7034 T7037 nummod 40,rats
R6293 T7035 T7036 compound kDa,dCGN-treated
R6294 T7036 T7037 compound dCGN-treated,rats
R6295 T7037 T7032 pobj rats,in
R6296 T7038 T7031 punct (,observed
R6297 T7039 T7040 compound Fig.,1F
R6298 T7040 T7031 npadvmod 1F,observed
R6299 T7041 T7031 punct ),observed
R6300 T7042 T7031 punct .,observed
R6301 T7043 T7045 det No,polysaccharides
R6302 T7044 T7045 amod sulphated,polysaccharides
R6303 T7045 T7047 nsubjpass polysaccharides,detected
R6304 T7046 T7047 auxpass were,detected
R6305 T7047 T7047 ROOT detected,detected
R6306 T7048 T7047 agent by,detected
R6307 T7049 T7051 nmod toluidine,staining
R6308 T7050 T7051 amod blue,staining
R6309 T7051 T7048 pobj staining,by
R6310 T7052 T7051 prep of,staining
R6311 T7053 T7054 compound colon,mucosa
R6312 T7054 T7052 pobj mucosa,of
R6313 T7055 T7051 prep from,staining
R6314 T7056 T7055 pobj rats,from
R6315 T7057 T7056 acl treated,rats
R6316 T7058 T7057 prep with,treated
R6317 T7059 T7065 preconj either,dCGN
R6318 T7060 T7065 det the,dCGN
R6319 T7061 T7065 nummod 10,dCGN
R6320 T7062 T7061 cc or,10
R6321 T7063 T7061 conj 40,10
R6322 T7064 T7065 compound kDa,dCGN
R6323 T7065 T7058 pobj dCGN,with
R6324 T7066 T7068 punct (,shown
R6325 T7067 T7068 neg not,shown
R6326 T7068 T7065 parataxis shown,dCGN
R6327 T7069 T7068 punct ),shown
R6328 T7070 T7047 punct .,detected
R6329 T7071 T7075 mark Although,exclude
R6330 T7072 T7075 nsubj we,exclude
R6331 T7073 T7075 aux can,exclude
R6332 T7074 T7075 neg not,exclude
R6333 T7075 T7091 advcl exclude,indicates
R6334 T7076 T7078 det that,mat
R6335 T7077 T7078 compound dCGN,mat
R6336 T7078 T7075 dobj mat,exclude
R6337 T7079 T7081 neg not,retained
R6338 T7080 T7081 aux have,retained
R6339 T7081 T7078 acl retained,mat
R6340 T7082 T7081 prep in,retained
R6341 T7083 T7084 det the,section
R6342 T7084 T7082 pobj section,in
R6343 T7085 T7081 prep during,retained
R6344 T7086 T7088 det the,procedure
R6345 T7087 T7088 compound histology,procedure
R6346 T7088 T7085 pobj procedure,during
R6347 T7089 T7091 punct ",",indicates
R6348 T7090 T7091 nsubj this,indicates
R6349 T7091 T7091 ROOT indicates,indicates
R6350 T7092 T7099 mark that,phagocytosed
R6351 T7093 T7094 det these,polymers
R6352 T7094 T7099 nsubjpass polymers,phagocytosed
R6353 T7095 T7099 aux may,phagocytosed
R6354 T7096 T7099 neg not,phagocytosed
R6355 T7097 T7099 aux have,phagocytosed
R6356 T7098 T7099 auxpass been,phagocytosed
R6357 T7099 T7091 ccomp phagocytosed,indicates
R6358 T7100 T7091 punct .,indicates
R6776 T7658 T7659 compound CGN,"inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced-TNF-α"
R13449 T15673 T15674 amod Degraded,CGN
R13450 T15674 T15675 nsubj CGN,induced
R13451 T15675 T15675 ROOT induced,induced
R13452 T15676 T15677 compound colon,inflammation
R13453 T15677 T15675 dobj inflammation,induced
R13454 T15678 T15677 prep in,inflammation
R13455 T15679 T15678 pobj rats,in
R13456 T15680 T15675 punct .,induced
R13457 T15681 T15706 dep Histograms,score
R13458 T15682 T15681 acl showing,Histograms
R13459 T15683 T15684 det the,effect
R13460 T15684 T15682 dobj effect,showing
R13461 T15685 T15684 prep of,effect
R13462 T15686 T15687 amod degraded,CGN
R13463 T15687 T15685 pobj CGN,of
R13464 T15688 T15684 prep on,effect
R13465 T15689 T15688 punct :,on
R13466 T15690 T15691 compound colon,length
R13467 T15691 T15688 pobj length,on
R13468 T15692 T15693 punct (,A
R13469 T15693 T15691 appos A,length
R13470 T15694 T15693 punct ),A
R13471 T15695 T15706 punct ;,score
R13472 T15696 T15706 amod macroscopic,score
R13473 T15697 T15698 punct (,B
R13474 T15698 T15696 appos B,macroscopic
R13475 T15699 T15698 punct ),B
R13476 T15700 T15696 cc and,macroscopic
R13477 T15701 T15696 conj histological,macroscopic
R13478 T15702 T15701 punct (,histological
R13479 T15703 T15701 appos C,histological
R13480 T15704 T15701 punct ),histological
R13481 T15705 T15706 compound inflammation,score
R13482 T15706 T15706 ROOT score,score
R13483 T15707 T15706 prep of,score
R13484 T15708 T15707 pobj colon,of
R13485 T15709 T15706 punct ;,score
R13486 T15710 T15714 nmod Myeloperoxidase,activity
R13487 T15711 T15712 punct (,MPO
R13488 T15712 T15710 appos MPO,Myeloperoxidase
R13489 T15713 T15712 punct ),MPO
R13490 T15714 T15706 conj activity,score
R13491 T15715 T15716 punct (,D
R13492 T15716 T15714 appos D,activity
R13493 T15717 T15714 punct ),activity
R13494 T15718 T15706 punct .,score
R13495 T15719 T15720 compound Control,rats
R13496 T15720 T15720 ROOT rats,rats
R13497 T15721 T15723 punct (,bars
R13498 T15722 T15723 amod white,bars
R13499 T15723 T15720 appos bars,rats
R13500 T15724 T15723 punct ),bars
R13501 T15725 T15723 punct ;,bars
R13502 T15726 T15727 nummod 10,kDa
R13503 T15727 T15720 appos kDa,rats
R13504 T15728 T15730 amod degraded,rats
R13505 T15729 T15730 amod CGN-treated,rats
R13506 T15730 T15727 appos rats,kDa
R13507 T15731 T15733 punct (,bars
R13508 T15732 T15733 compound grey,bars
R13509 T15733 T15730 appos bars,rats
R13510 T15734 T15730 punct ),rats
R13511 T15735 T15727 punct ;,kDa
R13512 T15736 T15737 nummod 40,kDa
R13513 T15737 T15738 npadvmod kDa,degraded
R13514 T15738 T15740 amod degraded,rats
R13515 T15739 T15740 amod CGN-treated,rats
R13516 T15740 T15727 appos rats,kDa
R13517 T15741 T15743 punct (,bars
R13518 T15742 T15743 amod black,bars
R13519 T15743 T15740 appos bars,rats
R13520 T15744 T15740 punct ),rats
R13521 T15745 T15720 punct .,rats
R13522 T15746 T15748 punct *,<
R13523 T15747 T15748 compound p,<
R13524 T15748 T15748 ROOT <,<
R13525 T15749 T15748 nummod 0.05,<
R13526 T15750 T15748 prep from,<
R13527 T15751 T15750 pobj control,from
R13528 T15752 T15748 punct .,<
R13529 T15753 T15748 punct **,<
R13530 T15754 T15756 nmod p,0.01
R13531 T15755 T15756 nmod <,0.01
R13532 T15756 T15753 nummod 0.01,**
R13533 T15757 T15753 prep from,**
R13534 T15758 T15757 pobj control,from
R13535 T15759 T15753 punct .,**
R13536 T15760 T15761 amod Histological,analysis
R13537 T15761 T15761 ROOT analysis,analysis
R13538 T15762 T15761 prep of,analysis
R13539 T15763 T15762 pobj colon,of
R13540 T15764 T15761 prep from,analysis
R13541 T15765 T15766 compound control,rats
R13542 T15766 T15764 pobj rats,from
R13543 T15767 T15766 punct (,rats
R13544 T15768 T15766 appos E,rats
R13545 T15769 T15766 punct ),rats
R13546 T15770 T15761 punct ",",analysis
R13547 T15771 T15761 cc and,analysis
R13548 T15772 T15761 conj from,analysis
R13549 T15773 T15776 nummod 40,rats
R13550 T15774 T15775 compound kDa,dCGN-treated
R13551 T15775 T15776 compound dCGN-treated,rats
R13552 T15776 T15772 pobj rats,from
R13553 T15777 T15776 punct (,rats
R13554 T15778 T15776 appos F,rats
R13555 T15779 T15776 punct ),rats
R13556 T15780 T15761 punct .,analysis
R1474 T1663 T1664 amod common,characteristic
R1475 T1664 T1661 attr characteristic,is
R1476 T1665 T1664 prep of,characteristic
R1477 T1666 T1667 amod intestinal,diseases
R1478 T1667 T1665 pobj diseases,of
R1479 T1668 T1670 nmod [,]
R1480 T1669 T1670 nummod 17,]
R1481 T1670 T1664 appos ],characteristic
R1482 T1671 T1661 punct .,is
R1483 T1672 T1673 nsubj Macrophages,represent
R1484 T1673 T1673 ROOT represent,represent
R1485 T1674 T1675 nummod 10,%
R1486 T1675 T1673 dobj %,represent
R1487 T1676 T1675 prep of,%
R2153 T2437 T2421 conj medium,preparations
R2154 T2438 T2437 punct ",",medium
R2155 T2439 T2437 punct (,medium
R2156 T2440 T2437 appos ∼,medium
R2157 T2441 T2442 nummod 40,kDa
R2158 T2442 T2440 appos kDa,∼
R2159 T2443 T2437 punct ;,medium
R2160 T2444 T2421 appos C40,preparations
R2161 T2445 T2421 punct ),preparations
R2162 T2446 T2447 amod molecular,weight
R2163 T2447 T2416 conj weight,Preparation
R2164 T2448 T2449 auxpass were,prepared
R2165 T2449 T2449 ROOT prepared,prepared
R2166 T2450 T2449 prep from,prepared
R2167 T2451 T2452 amod native,iota-carrageenan
R2168 T2452 T2450 pobj iota-carrageenan,from
R2169 T2453 T2452 acl extracted,iota-carrageenan
R2170 T2454 T2453 prep from,extracted
R2171 T2455 T2456 compound Euchema,spinosum
R2172 T2456 T2454 pobj spinosum,from
R2173 T2457 T2459 punct (,provided
R2174 T2458 T2459 advmod generously,provided
R2175 T2459 T2452 acl provided,iota-carrageenan
R2176 T2460 T2459 agent by,provided
R2177 T2461 T2463 compound Sanofi,Industry
R2178 T2462 T2463 compound Biosystems,Industry
R2179 T2463 T2460 pobj Industry,by
R2205 T2489 T2487 pobj °,to
R2206 T2490 T2491 compound C.,Then
R2207 T2491 T2497 advmod Then,submitted
R2208 T2492 T2497 punct ",",submitted
R2209 T2493 T2495 det the,solution
R2210 T2494 T2495 compound carrageenan,solution
R2211 T2495 T2497 nsubjpass solution,submitted
R2212 T2496 T2497 auxpass was,submitted
R2213 T2497 T2497 ROOT submitted,submitted
R2214 T2498 T2497 prep to,submitted
R2215 T2499 T2501 nummod two,treatments
R2216 T2500 T2501 amod different,treatments
R2217 T2501 T2498 pobj treatments,to
R2218 T2502 T2503 aux to,obtain
R2219 T2503 T2497 xcomp obtain,submitted
R2220 T2504 T2505 advmod both,low
R2221 T2505 T2510 amod low,fractions
R2222 T2506 T2505 cc and,low
R2223 T2507 T2505 conj medium,low
R2224 T2508 T2509 amod molecular,weight
R2225 T2509 T2510 compound weight,fractions
R2226 T2510 T2503 dobj fractions,obtain
R2227 T2511 T2497 punct .,submitted
R2228 T2512 T2524 advmod Briefly,hydrolyzed
R2229 T2513 T2524 punct ",",hydrolyzed
R2230 T2514 T2524 prep for,hydrolyzed
R2231 T2515 T2519 det the,fraction
R2493 T2777 T2779 det the,fractions
R2494 T2778 T2779 nummod two,fractions
R2495 T2779 T2781 nsubjpass fractions,dissolved
R2496 T2780 T2781 auxpass were,dissolved
R2497 T2781 T2781 ROOT dissolved,dissolved
R2498 T2782 T2781 prep in,dissolved
R2499 T2783 T2784 amod complete,medium
R2500 T2784 T2782 pobj medium,in
R2501 T2785 T2781 prep during,dissolved
R2502 T2786 T2787 nummod 30,min
R2503 T2787 T2785 pobj min,during
R2504 T2788 T2787 prep at,min
R2505 T2789 T2790 nummod 56,°
R2506 T2790 T2791 nummod °,C.
R2507 T2791 T2788 pobj C.,at
R2633 T2945 T2960 nsubjpass Animals,housed
R2634 T2946 T2945 punct ",",Animals
R2635 T2947 T2952 nmod Chemicals,rats
R2636 T2948 T2947 cc and,Chemicals
R2637 T2949 T2947 conj Diet,Chemicals
R2638 T2950 T2951 compound Male,Wistar
R2639 T2951 T2947 conj Wistar,Chemicals
R2640 T2952 T2945 appos rats,Animals
R2641 T2953 T2952 punct (,rats
R2642 T2954 T2957 nummod 150,weight
R2643 T2955 T2957 amod g,weight
R2644 T2956 T2957 amod average,weight
R2645 T2957 T2952 appos weight,rats
R2646 T2958 T2952 punct ),rats
R2668 T2980 T2981 compound drinking,water
R2670 T2982 T2985 punct (,w/v
R2671 T2983 T2984 nummod 5,%
R2672 T2984 T2985 compound %,w/v
R2673 T2985 T2977 parataxis w/v,administered
R2674 T2986 T2985 punct ),w/v
R2675 T2987 T2977 prep for,administered
R2676 T2988 T2989 nummod 55,days
R2677 T2989 T2987 pobj days,for
R2678 T2990 T2989 prep to,days
R2679 T2991 T2992 nummod 2,groups
R2680 T2992 T2990 pobj groups,to
R2681 T2993 T2992 prep of,groups
R2682 T2994 T2995 nummod six,animals
R2683 T2995 T2993 pobj animals,of
R2684 T2996 T2992 npadvmod each,groups
R2685 T2997 T2977 punct .,administered
R2686 T2998 T3000 det The,group
R2687 T2999 T3000 amod first,group
R2688 T3000 T3001 nsubj group,received
R2689 T3001 T3001 ROOT received,received
R2690 T3002 T3006 det the,carrageenan
R2691 T3003 T3005 amod low,weight
R2692 T3004 T3005 amod molecular,weight
R2693 T3005 T3006 compound weight,carrageenan
R2694 T3006 T3001 dobj carrageenan,received
R2695 T3007 T3006 punct (,carrageenan
R2696 T3008 T3010 nummod 10,dCGN
R2697 T3009 T3010 compound kDa,dCGN
R2698 T3010 T3006 appos dCGN,carrageenan
R2699 T3011 T3010 punct ),dCGN
R2700 T3012 T3006 cc and,carrageenan
R2701 T3013 T3014 det the,second
R2702 T3014 T3015 nsubj second,received
R2703 T3015 T3001 conj received,received
R2704 T3016 T3020 det the,carrageenan
R2705 T3017 T3019 nmod medium,weight
R2706 T3018 T3019 amod molecular,weight
R2707 T3019 T3020 compound weight,carrageenan
R2708 T3020 T3015 dobj carrageenan,received
R2709 T3021 T3020 punct (,carrageenan
R2710 T3022 T3024 nummod 40,dCGN
R2711 T3023 T3024 compound kDa,dCGN
R2712 T3024 T3020 appos dCGN,carrageenan
R2713 T3025 T3024 punct ),dCGN
R2714 T3026 T3015 punct .,received
R2715 T3027 T3029 det An,group
R2716 T3028 T3029 amod additional,group
R2717 T3029 T3034 nsubjpass group,maintained
R2718 T3030 T3029 prep of,group
R2719 T3031 T3032 nummod four,rats
R2720 T3032 T3030 pobj rats,of
R2721 T3033 T3034 auxpass were,maintained
R2722 T3034 T3034 ROOT maintained,maintained
R2723 T3035 T3034 prep on,maintained
R2724 T3036 T3038 amod regular,water
R2725 T3037 T3038 compound tap,water
R2726 T3038 T3035 pobj water,on
R2727 T3039 T3041 punct (,group
R2728 T3040 T3041 compound control,group
R2729 T3041 T3038 appos group,water
R3147 T3503 T3507 nsubjpass segment,opened
R3148 T3504 T3503 prep of,segment
R3149 T3505 T3504 pobj colon,of
R3150 T3506 T3507 auxpass was,opened
R3151 T3507 T3507 ROOT opened,opened
R3152 T3508 T3507 advmod longitudinally,opened
R3153 T3509 T3508 cc and,longitudinally
R3154 T3510 T3508 conj macroscopic,longitudinally
R3155 T3511 T3510 cc and,macroscopic
R3156 T3512 T3510 conj histological,macroscopic
R3157 T3513 T3517 nsubjpass scores,recorded
R3158 T3514 T3513 prep of,scores
R3159 T3515 T3514 pobj inflammation,of
R3160 T3516 T3517 auxpass were,recorded
R3225 T3581 T3580 prep of,level
R3226 T3582 T3581 pobj MPO,of
R3227 T3583 T3580 punct ",",level
R3228 T3584 T3585 advmod mainly,expressed
R3229 T3585 T3580 acl expressed,level
R3230 T3586 T3585 agent by,expressed
R3231 T3587 T3586 pobj neutrophils,by
R3232 T3588 T3589 punct ",",indicates
R3233 T3589 T3589 ROOT indicates,indicates
R3940 T4386 T4389 compound Analysis,Monocytes
R3941 T4387 T4388 compound Peripheral,Blood
R3942 T4388 T4389 compound Blood,Monocytes
R3943 T4389 T4394 nsubjpass Monocytes,exposed
R3944 T4390 T4389 cc or,Monocytes
R3945 T4391 T4392 nummod THP-1,cells
R3946 T4392 T4389 conj cells,Monocytes
R3947 T4393 T4394 auxpass were,exposed
R3948 T4394 T4394 ROOT exposed,exposed
R3949 T4395 T4396 aux to,complete
R3950 T4396 T4394 xcomp complete,exposed
R3951 T4397 T4396 dobj medium,complete
R3952 T4398 T4396 prep in,complete
R4625 T5162 T5163 compound CAGATAGATGGGCTCATACC-3,′
R4626 T5163 T5156 conj ′,′
R4627 T5164 T5143 punct .,amplified
R4628 T5165 T5169 nsubjpass cDNA,amplified
R4629 T5166 T5169 prep for,amplified
R4630 T5167 T5166 pobj ICAM-1,for
R4631 T5168 T5169 auxpass was,amplified
R4632 T5169 T5169 ROOT amplified,amplified
R4633 T5170 T5169 prep for,amplified
R4634 T5171 T5172 nummod 35,cycles
R4635 T5172 T5170 pobj cycles,for
R4636 T5173 T5169 advcl using,amplified

UBERON-AE

Id Subject Object Predicate Lexical cue
T731 1614-1624 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T732 1688-1698 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T733 1924-1934 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T734 2201-2211 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T735 1722-1733 http://purl.obolibrary.org/obo/UBERON_2000106 denotes extensively
T736 1924-1945 http://purl.obolibrary.org/obo/UBERON_0001277 denotes intestinal epithelial
T737 2262-2268 http://purl.obolibrary.org/obo/UBERON_0000957 denotes lamina
T738 2262-2276 http://purl.obolibrary.org/obo/UBERON_0000030 denotes lamina propria
T739 3085-3090 http://purl.obolibrary.org/obo/UBERON_0000178 denotes Blood
T740 3609-3614 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3100 6565-6568 http://purl.obolibrary.org/obo/UBERON_0000970 denotes eye
T3101 6708-6713 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3102 6776-6781 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3103 7171-7176 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3104 7468-7473 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3105 6865-6875 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T3665 7805-7811 http://purl.obolibrary.org/obo/UBERON_0000479 denotes tissue
T3666 8072-8077 http://purl.obolibrary.org/obo/UBERON_0000178 denotes blood
T3667 8127-8132 http://purl.obolibrary.org/obo/UBERON_0000178 denotes blood
T4232 9332-9337 http://purl.obolibrary.org/obo/UBERON_0000178 denotes Blood
T4233 9562-9567 http://purl.obolibrary.org/obo/UBERON_0001977 denotes serum
T4578 10297-10303 http://purl.obolibrary.org/obo/UBERON_0000479 denotes tissue
T5473 14099-14103 http://purl.obolibrary.org/obo/UBERON_0001913 denotes milk
T6603 14912-14919 http://purl.obolibrary.org/obo/UBERON_0001155 denotes Colonic
T6604 15768-15773 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T6605 16062-16067 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T6606 15026-15031 http://purl.obolibrary.org/obo/UBERON_0000178 denotes blood
T6607 15654-15664 http://purl.obolibrary.org/obo/UBERON_0000483 denotes epithelium
T6608 15865-15875 http://purl.obolibrary.org/obo/UBERON_0000483 denotes epithelium
T6609 16062-16074 http://purl.obolibrary.org/obo/UBERON_0000317 denotes colon mucosa
T6610 16068-16074 http://purl.obolibrary.org/obo/UBERON_0000344 denotes mucosa
T15563 16378-16383 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T15564 16456-16461 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T15565 16533-16538 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T15566 16768-16773 http://purl.obolibrary.org/obo/UBERON_0001155 denotes colon
T3106 6980-6990 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T3107 7401-7411 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T3108 7670-7680 http://purl.obolibrary.org/obo/UBERON_0000160 denotes intestinal
T3109 7401-7418 http://purl.obolibrary.org/obo/UBERON_0001242 denotes intestinal mucosa
T3110 7670-7687 http://purl.obolibrary.org/obo/UBERON_0001242 denotes intestinal mucosa
T3111 7412-7418 http://purl.obolibrary.org/obo/UBERON_0000344 denotes mucosa
T3112 7681-7687 http://purl.obolibrary.org/obo/UBERON_0000344 denotes mucosa

sentences

Id Subject Object Predicate Lexical cue
T79 82-208 Sentence denotes In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure.
T80 209-320 Sentence denotes Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody.
T81 321-405 Sentence denotes Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells.
T82 406-456 Sentence denotes All these effects were linked to NF-κB activation.
T83 457-599 Sentence denotes These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype.
T753 614-733 Sentence denotes Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae).
T754 734-804 Sentence denotes Three main forms of CGN have been identified: kappa, iota, and lambda.
T755 805-878 Sentence denotes They differ from each other in sulphation degree and solubility [1], [2].
T756 879-973 Sentence denotes Native CGN is thought to be harmless and is widely used as a food additive to improve texture.
T757 974-1023 Sentence denotes It is also used in cosmetics and pharmaceuticals.
T758 1024-1188 Sentence denotes However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN).
T759 1189-1340 Sentence denotes These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5].
T760 1341-1421 Sentence denotes The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8].
T761 1422-1645 Sentence denotes Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12].
T762 1646-1821 Sentence denotes Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines.
T763 1822-2017 Sentence denotes Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14].
T764 2018-2129 Sentence denotes NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16].
T765 2130-2226 Sentence denotes Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17].
T766 2227-2374 Sentence denotes Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules.
T767 2375-2539 Sentence denotes The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans.
T768 2540-2657 Sentence denotes Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19].
T769 2658-2890 Sentence denotes Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20].
T770 2891-3000 Sentence denotes Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure.
T771 3001-3190 Sentence denotes Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN.
T772 3191-3380 Sentence denotes We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion.
T773 3381-3481 Sentence denotes These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation.
T774 3482-3615 Sentence denotes In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon.
T775 3616-3746 Sentence denotes These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype.
T2051 3771-3806 Sentence denotes Preparation of Degraded Carrageenan
T2052 3807-4075 Sentence denotes Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France).
T2053 4076-4311 Sentence denotes Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions.
T2054 4312-4610 Sentence denotes Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA).
T2055 4611-4835 Sentence denotes For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa).
T2056 4836-4911 Sentence denotes The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa).
T2057 4912-5062 Sentence denotes Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter).
T2058 5063-5291 Sentence denotes Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa.
T2059 5292-5408 Sentence denotes The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22].
T2060 5409-5533 Sentence denotes Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy.
T2061 5534-5664 Sentence denotes The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France).
T2062 5665-5767 Sentence denotes Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C.
T2810 5769-5796 Sentence denotes Animals, Chemicals and Diet
T2811 5797-5931 Sentence denotes Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow.
T2812 5932-6052 Sentence denotes Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each.
T2813 6053-6207 Sentence denotes The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN).
T2814 6208-6294 Sentence denotes An additional group of four rats were maintained on regular tap water (control group).
T2815 6295-6408 Sentence denotes To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]).
T2816 6409-6457 Sentence denotes Fresh carrageenan solutions were prepared daily.
T3117 6459-6480 Sentence denotes Evaluation of Colitis
T3118 6481-6625 Sentence denotes Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period.
T3119 6626-6689 Sentence denotes After 55 days, animals were sacrificed by cervical dislocation.
T3120 6690-6764 Sentence denotes The length of the colon was measured as described by Okayashu et al. [24].
T3121 6765-6882 Sentence denotes Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen.
T3122 6883-6956 Sentence denotes The moderately distended segment was sectioned and fixed in 10% formalin.
T3123 6957-7024 Sentence denotes The following day, the intestinal content was removed by vortexing.
T3124 7025-7114 Sentence denotes The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure.
T3125 7115-7308 Sentence denotes To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26].
T3126 7309-7419 Sentence denotes The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa.
T3127 7420-7562 Sentence denotes On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27].
T3128 7563-7688 Sentence denotes The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa.
T3129 7689-7786 Sentence denotes One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C.
T3668 7788-7800 Sentence denotes Cell Culture
T3669 7801-7875 Sentence denotes All tissue culture reagents were from Invitrogen (Cergy Pontoise, France).
T3670 7876-8054 Sentence denotes THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator.
T3671 8055-8168 Sentence denotes Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient.
T3672 8169-8247 Sentence denotes Monocytes were then isolated by adherence to culture flasks as described [28].
T3673 8248-8344 Sentence denotes For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h.
T3674 8345-8463 Sentence denotes Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer.
T3675 8464-8582 Sentence denotes In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added.
T3981 8584-8603 Sentence denotes Cell Cycle Analysis
T3982 8604-8884 Sentence denotes THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France).
T3983 8885-8975 Sentence denotes Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter).
T3984 8976-9133 Sentence denotes Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations.
T3985 9134-9278 Sentence denotes Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis.
T4234 9280-9320 Sentence denotes Cell Surface Antigen Expression Analysis
T4235 9321-9446 Sentence denotes Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h.
T4236 9447-9603 Sentence denotes After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors.
T4237 9604-9679 Sentence denotes Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min.
T4238 9680-9814 Sentence denotes Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu).
T4239 9815-9918 Sentence denotes After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter).
T4240 9919-10006 Sentence denotes The cell population was gated according to its forward and wide-angle light scattering.
T4241 10007-10087 Sentence denotes Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells.
T4583 10089-10110 Sentence denotes TNF Activity Bioassay
T4584 10111-10265 Sentence denotes Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time.
T4585 10266-10389 Sentence denotes Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29].
T4586 10390-10432 Sentence denotes TNF concentrations are expressed as pg/ml.
T4745 10434-10449 Sentence denotes RT-PCR Analysis
T4746 10450-10642 Sentence denotes Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen).
T4747 10643-10744 Sentence denotes PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination).
T4748 10745-10996 Sentence denotes Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium).
T4749 10997-11569 Sentence denotes First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′.
T5235 11571-11610 Sentence denotes NF-kB Transcription Reporter Gene Assay
T5236 11611-11850 Sentence denotes The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene.
T5237 11851-12024 Sentence denotes Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40.
T5238 12025-12124 Sentence denotes Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI).
T5475 12126-12147 Sentence denotes Western Blot Analysis
T5476 12148-12247 Sentence denotes THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS.
T5477 12248-12395 Sentence denotes Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice.
T5478 12396-12503 Sentence denotes Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected.
T5479 12504-12578 Sentence denotes Protein concentration was determined using the DC Protein Assay (Bio-Rad).
T5480 12579-12720 Sentence denotes Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane.
T5481 12721-12924 Sentence denotes I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories).
T5482 12925-13082 Sentence denotes Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction.
T5483 13083-13145 Sentence denotes Antibody to α-Tubulin (Santa Cruz) was use as loading control.
T5484 13146-13305 Sentence denotes For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31].
T5485 13306-13583 Sentence denotes Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA).
T5486 13584-13679 Sentence denotes Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature.
T5487 13680-13777 Sentence denotes Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight.
T5488 13778-13916 Sentence denotes Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used.
T5489 13917-14161 Sentence denotes Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature.
T5490 14162-14377 Sentence denotes After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions.
T5491 14378-14512 Sentence denotes To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32].
T6463 14514-14560 Sentence denotes Analysis of NF-κB Activation by Flow Cytometry
T6464 14561-14639 Sentence denotes Nuclear activation of NF−κΒ by flow cytometry was performed as described [31].
T6517 14641-14661 Sentence denotes Statistical Analysis
T6518 14662-14742 Sentence denotes The results were expressed as the mean value ± S.E.M. of individual experiments.
T6519 14743-14881 Sentence denotes The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA).
T6612 14892-14932 Sentence denotes Degraded CGN Induce Colonic Inflammation
T6613 14933-15070 Sentence denotes All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools.
T6614 15071-15215 Sentence denotes Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A).
T6615 15216-15345 Sentence denotes Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C).
T6616 15346-15534 Sentence denotes Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage.
T6617 15535-15609 Sentence denotes Histological examination revealed various degrees of mucosal inflammation.
T6618 15610-15725 Sentence denotes Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown).
T6619 15726-15800 Sentence denotes These symptoms were totally absent in the colon of control rats (Fig. 1E).
T6620 15801-15988 Sentence denotes More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F).
T6621 15989-16139 Sentence denotes No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown).
T6622 16140-16312 Sentence denotes Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed.
T15567 16357-16405 Sentence denotes Degraded CGN induced colon inflammation in rats.
T15568 16406-16575 Sentence denotes Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D).
T15569 16576-16742 Sentence denotes Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control.
T15570 16743-16835 Sentence denotes Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F).
T10 82-208 Sentence denotes In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure.
T11 209-320 Sentence denotes Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody.
T12 321-405 Sentence denotes Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells.
T13 406-456 Sentence denotes All these effects were linked to NF-κB activation.
T14 457-599 Sentence denotes These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype.
T15 601-613 Sentence denotes Introduction
T16 614-733 Sentence denotes Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae).
T17 734-804 Sentence denotes Three main forms of CGN have been identified: kappa, iota, and lambda.
T18 805-878 Sentence denotes They differ from each other in sulphation degree and solubility [1], [2].
T19 879-973 Sentence denotes Native CGN is thought to be harmless and is widely used as a food additive to improve texture.
T20 974-1023 Sentence denotes It is also used in cosmetics and pharmaceuticals.
T21 1024-1188 Sentence denotes However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN).
T22 1189-1340 Sentence denotes These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5].
T23 1341-1421 Sentence denotes The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8].
T24 1422-1645 Sentence denotes Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12].
T25 1646-1821 Sentence denotes Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines.
T26 1822-2017 Sentence denotes Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14].
T27 2018-2129 Sentence denotes NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16].
T28 2130-2226 Sentence denotes Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17].
T29 2227-2374 Sentence denotes Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules.
T30 2375-2539 Sentence denotes The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans.
T31 2540-2657 Sentence denotes Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19].
T32 2658-2890 Sentence denotes Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20].
T33 2891-3000 Sentence denotes Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure.
T34 3001-3190 Sentence denotes Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN.
T35 3191-3380 Sentence denotes We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion.
T36 3381-3481 Sentence denotes These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation.
T37 3482-3615 Sentence denotes In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon.
T38 3616-3746 Sentence denotes These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype.
T39 3748-3769 Sentence denotes Materials and Methods
T40 3771-3806 Sentence denotes Preparation of Degraded Carrageenan
T41 3807-4075 Sentence denotes Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France).
T42 4076-4180 Sentence denotes Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C.
T43 4181-4311 Sentence denotes Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions.
T44 4312-4459 Sentence denotes Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C.
T45 4460-4610 Sentence denotes After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA).
T46 4611-4756 Sentence denotes For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C.
T47 4757-4835 Sentence denotes After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa).
T48 4836-4911 Sentence denotes The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa).
T49 4912-5062 Sentence denotes Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter).
T50 5063-5291 Sentence denotes Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa.
T51 5292-5408 Sentence denotes The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22].
T52 5409-5533 Sentence denotes Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy.
T53 5534-5664 Sentence denotes The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France).
T54 5665-5767 Sentence denotes Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C.
T55 5769-5796 Sentence denotes Animals, Chemicals and Diet
T56 5797-5931 Sentence denotes Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow.
T57 5932-6052 Sentence denotes Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each.
T58 6053-6207 Sentence denotes The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN).
T59 6208-6294 Sentence denotes An additional group of four rats were maintained on regular tap water (control group).
T60 6295-6408 Sentence denotes To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]).
T61 6409-6457 Sentence denotes Fresh carrageenan solutions were prepared daily.
T62 6459-6480 Sentence denotes Evaluation of Colitis
T63 6481-6625 Sentence denotes Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period.
T64 6626-6689 Sentence denotes After 55 days, animals were sacrificed by cervical dislocation.
T65 6690-6764 Sentence denotes The length of the colon was measured as described by Okayashu et al. [24].
T66 6765-6882 Sentence denotes Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen.
T67 6883-6956 Sentence denotes The moderately distended segment was sectioned and fixed in 10% formalin.
T68 6957-7024 Sentence denotes The following day, the intestinal content was removed by vortexing.
T69 7025-7114 Sentence denotes The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure.
T70 7115-7308 Sentence denotes To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26].
T71 7309-7419 Sentence denotes The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa.
T72 7420-7562 Sentence denotes On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27].
T73 7563-7688 Sentence denotes The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa.
T74 7689-7786 Sentence denotes One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C.
T75 7788-7800 Sentence denotes Cell Culture
T76 7801-7875 Sentence denotes All tissue culture reagents were from Invitrogen (Cergy Pontoise, France).
T77 7876-8054 Sentence denotes THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator.
T78 8055-8168 Sentence denotes Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient.
T79 8169-8247 Sentence denotes Monocytes were then isolated by adherence to culture flasks as described [28].
T80 8248-8344 Sentence denotes For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h.
T81 8345-8463 Sentence denotes Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer.
T82 8464-8582 Sentence denotes In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added.
T83 8584-8603 Sentence denotes Cell Cycle Analysis
T84 8604-8884 Sentence denotes THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France).
T85 8885-8975 Sentence denotes Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter).
T86 8976-9133 Sentence denotes Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations.
T87 9134-9278 Sentence denotes Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis.
T88 9280-9320 Sentence denotes Cell Surface Antigen Expression Analysis
T89 9321-9446 Sentence denotes Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h.
T90 9447-9603 Sentence denotes After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors.
T91 9604-9679 Sentence denotes Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min.
T92 9680-9814 Sentence denotes Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu).
T93 9815-9918 Sentence denotes After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter).
T94 9919-10006 Sentence denotes The cell population was gated according to its forward and wide-angle light scattering.
T95 10007-10087 Sentence denotes Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells.
T96 10089-10110 Sentence denotes TNF Activity Bioassay
T97 10111-10265 Sentence denotes Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time.
T98 10266-10389 Sentence denotes Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29].
T99 10390-10432 Sentence denotes TNF concentrations are expressed as pg/ml.
T100 10434-10449 Sentence denotes RT-PCR Analysis
T101 10450-10642 Sentence denotes Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen).
T102 10643-10744 Sentence denotes PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination).
T103 10745-10996 Sentence denotes Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium).
T104 10997-11569 Sentence denotes First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′.
T105 11571-11610 Sentence denotes NF-kB Transcription Reporter Gene Assay
T106 11611-11659 Sentence denotes The plasmid 3XMHC-luc (a generous gift from Drs.
T107 11660-11662 Sentence denotes J.
T108 11663-11680 Sentence denotes Westwick and D.A.
T109 11681-11850 Sentence denotes Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene.
T110 11851-12024 Sentence denotes Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40.
T111 12025-12124 Sentence denotes Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI).
T112 12126-12147 Sentence denotes Western Blot Analysis
T113 12148-12247 Sentence denotes THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS.
T114 12248-12395 Sentence denotes Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice.
T115 12396-12503 Sentence denotes Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected.
T116 12504-12578 Sentence denotes Protein concentration was determined using the DC Protein Assay (Bio-Rad).
T117 12579-12720 Sentence denotes Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane.
T118 12721-12924 Sentence denotes I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories).
T119 12925-13082 Sentence denotes Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction.
T120 13083-13145 Sentence denotes Antibody to α-Tubulin (Santa Cruz) was use as loading control.
T121 13146-13240 Sentence denotes For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C.
T122 13241-13305 Sentence denotes Cells were then pelleted and nuclei separated as described [31].
T123 13306-13411 Sentence denotes Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C.
T124 13412-13583 Sentence denotes Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA).
T125 13584-13679 Sentence denotes Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature.
T126 13680-13777 Sentence denotes Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight.
T127 13778-13916 Sentence denotes Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used.
T128 13917-14161 Sentence denotes Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature.
T129 14162-14377 Sentence denotes After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions.
T130 14378-14512 Sentence denotes To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32].
T131 14514-14560 Sentence denotes Analysis of NF-κB Activation by Flow Cytometry
T132 14561-14639 Sentence denotes Nuclear activation of NF−κΒ by flow cytometry was performed as described [31].
T133 14641-14661 Sentence denotes Statistical Analysis
T134 14662-14742 Sentence denotes The results were expressed as the mean value ± S.E.M. of individual experiments.
T135 14743-14881 Sentence denotes The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA).
T136 14883-14890 Sentence denotes Results
T137 14892-14932 Sentence denotes Degraded CGN Induce Colonic Inflammation
T138 14933-15070 Sentence denotes All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools.
T139 15071-15215 Sentence denotes Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A).
T140 15216-15345 Sentence denotes Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C).
T141 15346-15534 Sentence denotes Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage.
T142 15535-15609 Sentence denotes Histological examination revealed various degrees of mucosal inflammation.
T143 15610-15725 Sentence denotes Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown).
T144 15726-15800 Sentence denotes These symptoms were totally absent in the colon of control rats (Fig. 1E).
T145 15801-15988 Sentence denotes More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F).
T146 15989-16139 Sentence denotes No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown).
T147 16140-16312 Sentence denotes Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed.
T148 16313-16405 Sentence denotes 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats.
T149 16406-16575 Sentence denotes Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D).
T150 16576-16742 Sentence denotes Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control.
T151 16743-16835 Sentence denotes Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F).

events-check-again

Id Subject Object Predicate Lexical cue
T654 100-109 Positive_regulation denotes increased
T655 110-116 Protein denotes ICAM-1
T656 117-127 Gene_expression denotes expression
T657 335-345 Positive_regulation denotes stimulated
T658 335-345 Positive_regulation denotes stimulated
T659 346-351 Protein denotes TNF-α
T660 352-362 Gene_expression denotes expression
T661 367-376 Localization denotes secretion
T2022 1895-1899 Protein denotes IL-8
T2023 1900-1910 Gene_expression denotes production
T2024 1965-1968 Positive_regulation denotes via
T2025 3255-3264 Positive_regulation denotes increased
T2026 3265-3271 Protein denotes ICAM-1
T2027 3272-3282 Gene_expression denotes expression
T2028 3295-3301 Protein denotes ICAM-1
T2029 3338-3348 Positive_regulation denotes stimulated
T2030 3338-3348 Positive_regulation denotes stimulated
T2031 3349-3354 Protein denotes TNF-α
T2032 3355-3365 Gene_expression denotes expression
T2033 3370-3379 Localization denotes secretion
T3652 7500-7515 Protein denotes myeloperoxidase
T3653 7517-7520 Protein denotes MPO
T3654 7576-7579 Protein denotes MPO
T3655 7588-7597 Gene_expression denotes expressed
T4725 10089-10092 Protein denotes TNF
T4726 10286-10291 Protein denotes TNF-α
T4727 10292-10293 Protein denotes β
T4728 10390-10393 Protein denotes TNF
T5210 11167-11174 Protein denotes β-actin
T5211 11302-11307 Protein denotes TNF-α
T5212 11438-11444 Protein denotes ICAM-1
T5452 11834-11844 Protein denotes luciferase
T5453 12025-12035 Protein denotes Luciferase
T6440 12721-12726 Protein denotes I-κBα
T7124 15356-15371 Protein denotes myeloperoxidase
T8141 4-16857 Positive_regulation denotes inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced
T8142 16858-16863 Protein denotes TNF-α
R530 T655 T656 themeOf ICAM-1,expression
R531 T656 T654 themeOf expression,increased
R532 T659 T660 themeOf TNF-α,expression
R533 T659 T661 themeOf TNF-α,secretion
R534 T660 T658 themeOf expression,stimulated
R535 T661 T657 themeOf secretion,stimulated
R1759 T2022 T2023 themeOf IL-8,production
R1760 T2023 T2024 themeOf production,via
R1761 T2026 T2027 themeOf ICAM-1,expression
R1762 T2027 T2025 themeOf expression,increased
R1763 T2031 T2032 themeOf TNF-α,expression
R1764 T2031 T2033 themeOf TNF-α,secretion
R1765 T2032 T2029 themeOf expression,stimulated
R1766 T2033 T2030 themeOf secretion,stimulated
R3269 T3653 T3652 equivalentTo MPO,myeloperoxidase
R3270 T3654 T3655 themeOf MPO,expressed

bionlp-st-ge-2016-reference-tees

Id Subject Object Predicate Lexical cue
T671 117-127 Gene_expression denotes expression
T672 100-109 Positive_regulation denotes increased
T673 215-219 Protein denotes dCGN
T674 299-319 Protein denotes anti-ICAM-1 antibody
T675 330-334 Protein denotes dCGN
T676 346-351 Protein denotes TNF-α
T677 352-362 Gene_expression denotes expression
T678 335-345 Positive_regulation denotes stimulated
T679 439-444 Protein denotes NF-κB
T680 445-455 Positive_regulation denotes activation
T681 512-515 Protein denotes CGN
T682 494-502 Protein_catabolism denotes degraded
T2034 1969-1986 Protein denotes Nuclear Factor-κB
T2035 1988-1993 Protein denotes NF-κB
T2036 1995-2005 Positive_regulation denotes activation
T2037 1995-2005 Positive_regulation denotes activation
T2038 2018-2023 Protein denotes NF-κB
T2039 2055-2064 Regulation denotes regulates
T2040 3265-3271 Protein denotes ICAM-1
T2041 3295-3301 Protein denotes ICAM-1
T2042 3349-3354 Protein denotes TNF-α
T2043 3272-3282 Gene_expression denotes expression
T2044 3355-3365 Gene_expression denotes expression
T2045 3255-3264 Positive_regulation denotes increased
T2046 3338-3348 Positive_regulation denotes stimulated
T2047 3464-3469 Protein denotes NF-κB
T2048 3470-3480 Positive_regulation denotes activation
T2049 3665-3668 Protein denotes CGN
T2050 3647-3655 Protein_catabolism denotes degraded
T2808 5627-5632 Protein denotes Sigma
T2809 5634-5644 Protein denotes St Quentin
T3656 7500-7515 Protein denotes myeloperoxidase
T3657 7517-7520 Protein denotes MPO
T3658 7486-7495 Positive_regulation denotes collected
T3659 7486-7495 Positive_regulation denotes collected
T3660 7576-7579 Protein denotes MPO
T3661 7588-7597 Gene_expression denotes expressed
T3662 7701-7704 Protein denotes MPO
T3663 7733-7744 Protein_catabolism denotes degradation
T4230 8947-8956 Protein denotes EPICS XL2
T4558 9285-9300 Protein denotes Surface Antigen
T4559 9301-9311 Gene_expression denotes Expression
T4560 9590-9602 Protein denotes Fc receptors
T4561 9579-9586 Binding denotes binding
T4562 9770-9773 Protein denotes IgG
T4563 9890-9899 Protein denotes EPICS XL2
T4729 10089-10101 Protein denotes TNF Activity
T4730 10286-10293 Protein denotes TNF-α/β
T4731 10390-10393 Protein denotes TNF
T4732 10413-10422 Gene_expression denotes expressed
T5213 10601-10628 Protein denotes M-MLV reverse transcriptase
T5214 10883-10906 Protein denotes Goldstar DNA polymerase
T5215 10941-10964 Protein denotes Goldstar DNA polymerase
T5216 11167-11174 Protein denotes β-actin
T5217 11302-11307 Protein denotes TNF-α
T5218 11438-11444 Protein denotes ICAM-1
T5454 11758-11763 Protein denotes NF-κB
T5455 11792-11809 Protein denotes MHC class I locus
T5456 11834-11849 Protein denotes luciferase gene
T5457 12025-12035 Protein denotes Luciferase
T6515 14561-14568 Entity denotes Nuclear
T6441 12573-12576 Protein denotes Rad
T6442 12721-12734 Protein denotes I-κBα protein
T6443 12828-12850 Protein denotes horseradish peroxidase
T6444 12934-12937 Protein denotes IκB
T6445 13095-13104 Protein denotes α-Tubulin
T6446 13150-13163 Protein denotes nuclear NF-κB
T6447 13778-13809 Protein denotes Rabbit polyclonal antibody anti
T6448 13810-13815 Protein denotes NF-κB
T6449 13816-13827 Protein denotes p50 subunit
T6450 13842-13864 Protein denotes anti-NF-κB p65 subunit
T6451 14038-14081 Protein denotes HRP-conjugated F(ab')2 goat anti-rabbit IgG
T6452 14490-14493 Protein denotes ERK
T6512 14526-14531 Protein denotes NF-κB
T6513 14532-14542 Positive_regulation denotes Activation
T6514 14583-14588 Protein denotes NF−κΒ
T6516 14569-14579 Positive_regulation denotes activation
T7125 15356-15371 Protein denotes myeloperoxidase
T8185 0-16863 Protein denotes CGN inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced-TNF-α
T15793 16540-16555 Protein denotes Myeloperoxidase
T15794 16557-16560 Protein denotes MPO
T670 110-116 Protein denotes ICAM-1
R543 T671 T672 themeOf expression,increased
R544 T675 T678 causeOf dCGN,stimulated
R545 T676 T677 themeOf TNF-α,expression
R546 T677 T678 themeOf expression,stimulated
R547 T679 T680 themeOf NF-κB,activation
R548 T681 T682 themeOf CGN,degraded
R555 T670 T671 themeOf ICAM-1,expression
R1767 T2034 T2036 themeOf Nuclear Factor-κB,activation
R1768 T2035 T2037 themeOf NF-κB,activation
R1769 T2038 T2039 themeOf NF-κB,regulates
R1770 T2040 T2043 themeOf ICAM-1,expression
R1771 T2042 T2044 themeOf TNF-α,expression
R1772 T2043 T2045 themeOf expression,increased
R1773 T2044 T2046 themeOf expression,stimulated
R1774 T2047 T2048 themeOf NF-κB,activation
R1775 T2049 T2050 themeOf CGN,degraded
R3271 T3656 T3658 themeOf myeloperoxidase,collected
R3272 T3657 T3659 themeOf MPO,collected
R3273 T3660 T3661 themeOf MPO,expressed
R3274 T3662 T3663 themeOf MPO,degradation
R4090 T4558 T4559 themeOf Surface Antigen,Expression
R4091 T4560 T4561 themeOf Fc receptors,binding
R4212 T4731 T4732 themeOf TNF,expressed
R5797 T6512 T6513 themeOf NF-κB,Activation
R5798 T6514 T6516 themeOf NF−κΒ,activation
R5799 T6515 T6516 Site Nuclear,activation

bionlp-st-ge-2016-reference

Id Subject Object Predicate Lexical cue
T63 100-109 Positive_regulation denotes increased
T64 110-116 Protein denotes ICAM-1
T65 117-127 Gene_expression denotes expression
T66 335-345 Positive_regulation denotes stimulated
T67 335-345 Positive_regulation denotes stimulated
T68 346-351 Protein denotes TNF-α
T69 352-362 Gene_expression denotes expression
T70 367-376 Localization denotes secretion
T741 1895-1899 Protein denotes IL-8
T742 1900-1910 Gene_expression denotes production
T743 1965-1968 Positive_regulation denotes via
T744 3255-3264 Positive_regulation denotes increased
T745 3265-3271 Protein denotes ICAM-1
T746 3272-3282 Gene_expression denotes expression
T747 3295-3301 Protein denotes ICAM-1
T748 3338-3348 Positive_regulation denotes stimulated
T749 3338-3348 Positive_regulation denotes stimulated
T750 3349-3354 Protein denotes TNF-α
T751 3355-3365 Gene_expression denotes expression
T752 3370-3379 Localization denotes secretion
T3114 7517-7520 Protein denotes MPO
T3115 7576-7579 Protein denotes MPO
T3116 7588-7597 Gene_expression denotes expressed
T4581 10292-10293 Protein denotes β
T4582 10390-10393 Protein denotes TNF
T5474 12721-12726 Protein denotes I-κBα
T7226 4-16857 Positive_regulation denotes inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced
T7227 16858-16863 Protein denotes TNF-α
T4579 10089-10092 Protein denotes TNF
T4580 10286-10291 Protein denotes TNF-α
T6611 15356-15371 Protein denotes myeloperoxidase
T5233 11834-11844 Protein denotes luciferase
T5234 12025-12035 Protein denotes Luciferase
T4742 11167-11174 Protein denotes β-actin
T4743 11302-11307 Protein denotes TNF-α
T4744 11438-11444 Protein denotes ICAM-1
T3113 7500-7515 Protein denotes myeloperoxidase
R571 T741 T742 themeOf IL-8,production
R572 T742 T743 themeOf production,via
R573 T745 T746 themeOf ICAM-1,expression
R574 T746 T744 themeOf expression,increased
R575 T750 T751 themeOf TNF-α,expression
R576 T750 T752 themeOf TNF-α,secretion
R577 T751 T748 themeOf expression,stimulated
R578 T752 T749 themeOf secretion,stimulated
R22 T64 T65 themeOf ICAM-1,expression
R23 T65 T63 themeOf expression,increased
R24 T68 T69 themeOf TNF-α,expression
R25 T68 T70 themeOf TNF-α,secretion
R26 T69 T67 themeOf expression,stimulated
R27 T70 T66 themeOf secretion,stimulated
R2770 T3114 T3113 equivalentTo MPO,myeloperoxidase
R2771 T3115 T3116 themeOf MPO,expressed

bionlp-st-ge-2016-uniprot

Id Subject Object Predicate Lexical cue
T322 110-116 P05362 denotes ICAM-1
T323 304-310 P05362 denotes ICAM-1
T324 346-349 P01375 denotes TNF
T325 346-351 P01375 denotes TNF-α
T1339 1895-1899 P10145 denotes IL-8
T1340 3265-3271 P05362 denotes ICAM-1
T1341 3295-3301 P05362 denotes ICAM-1
T1342 3349-3352 P01375 denotes TNF
T1343 3349-3354 P01375 denotes TNF-α
T2944 6380-6383 Q15025 denotes Van
T3815 8515-8521 P05362 denotes ICAM-1
T4381 10035-10043 Q04864 denotes relative
T4644 10089-10092 P01375 denotes TNF
T4645 10286-10289 P01375 denotes TNF
T4646 10286-10291 P01375 denotes TNF-α
T4647 10390-10393 P01375 denotes TNF
T4947 11167-11174 P60709 denotes β-actin
T4948 11302-11305 P01375 denotes TNF
T4949 11302-11307 P01375 denotes TNF-α
T4950 11438-11444 P05362 denotes ICAM-1
T5333 11834-11844 P08659 denotes luciferase
T5334 12025-12035 P08659 denotes Luciferase
T5928 12327-12329 P0A7Z4 denotes pH
T5929 13816-13819 P19838 denotes p50
T5930 13816-13827 P19838 denotes p50 subunit
T5931 13853-13856 Q04206 denotes p65
T5932 13853-13856 P21579 denotes p65
T7627 16858-16861 P01375 denotes TNF
T7628 16858-16863 P01375 denotes TNF-α

test2

Id Subject Object Predicate Lexical cue
T43 100-109 Positive_regulation denotes increased
T44 110-116 Protein denotes ICAM-1
T45 117-127 Gene_expression denotes expression
T46 335-345 Positive_regulation denotes stimulated
T47 346-351 Protein denotes TNF-α
T48 352-362 Gene_expression denotes expression
T49 367-376 Localization denotes secretion
T719 1895-1899 Protein denotes IL-8
T720 1900-1910 Gene_expression denotes production
T721 3255-3264 Positive_regulation denotes increased
T722 3265-3271 Protein denotes ICAM-1
T723 3272-3282 Gene_expression denotes expression
T724 3284-3294 Positive_regulation denotes stimulated
T725 3295-3301 Protein denotes ICAM-1
T726 3302-3311 Regulation denotes dependent
T727 3338-3348 Positive_regulation denotes stimulated
T728 3349-3354 Protein denotes TNF-α
T729 3355-3365 Gene_expression denotes expression
T730 3370-3379 Localization denotes secretion
T3096 7500-7515 Protein denotes myeloperoxidase
T3097 7517-7520 Protein denotes MPO
T3098 7576-7579 Protein denotes MPO
T3099 7588-7597 Gene_expression denotes expressed
T4231 9579-9586 Binding denotes binding
T4574 10089-10092 Protein denotes TNF
T4575 10286-10291 Protein denotes TNF-α
T4576 10292-10293 Protein denotes β
T4577 10390-10393 Protein denotes TNF
T4739 11167-11174 Protein denotes β-actin
T4740 11302-11307 Protein denotes TNF-α
T4741 11438-11444 Protein denotes ICAM-1
T5231 11834-11844 Protein denotes luciferase
T5232 12025-12035 Protein denotes Luciferase
T5472 12721-12726 Protein denotes I-κBα
T6602 15356-15371 Protein denotes myeloperoxidase
T7185 4-16857 Positive_regulation denotes inhibited THP-1 cell proliferation in vitro, arresting the cells in G1 phase. In addition, dCGN increased ICAM-1 expression in both PBM and THP-1 cells with a major effect seen after 40 kDa dCGN exposure. Also, dCGN stimulated monocyte aggregation in vitro that was prevented by incubation with anti-ICAM-1 antibody. Finally, dCGN stimulated TNF-α expression and secretion by both PBM and THP-1 cells. All these effects were linked to NF-κB activation. These data strongly suggest that the degraded forms of CGN have a pronounced effect on monocytes, characteristic of an inflammatory phenotype. Introduction Carrageenan (CGN) is a high molecular weight sulphated polysaccharide (>200 kDa) derived from red algae (Rhodophyceae). Three main forms of CGN have been identified: kappa, iota, and lambda. They differ from each other in sulphation degree and solubility [1], [2]. Native CGN is thought to be harmless and is widely used as a food additive to improve texture. It is also used in cosmetics and pharmaceuticals. However, acid treatment at high temperature (80°C) triggers CGN hydrolysis to lower molecular weight (<50 kDa) compounds known as poligeenan or degraded CGN (dCGN). These dCGNs induce inflammation and have been widely used as models of colitis in several species, including rats [3], rabbits [4] and guinea pigs [5]. The role of dCGN as a tumor-promoting factor remains controversial [4], [6]–[8]. Although the native form is thought to be harmless for human consumption, small amounts of dCGN are probably produced by acid hydrolysis during gastric digestion [9], [10] or interaction with intestinal bacteria [11], [12]. Whereas the effects of native and dCGN on intestinal inflammation have been extensively analyzed in animal models, only few studies have been conducted using human cell lines. Recent studies have shown a link between exposure to native form CGN and IL-8 production by the human intestinal epithelial cell line, NCM460, via Nuclear Factor-κB (NF-κB) activation [13], [14]. NF-κB is a transcription factor that regulates the expression of genes associated with inflammation [15], [16]. Macrophage infiltration and accumulation is a common characteristic of intestinal diseases [17]. Macrophages represent 10% of total lamina propria cells, secrete a wide range of biologically active compounds and express cell-adhesion molecules. The immune cell response to an inflammatory stimulus seems to be amplified or directly generated by cells exposed to sulphated polysaccharides such as carrageenans. Indeed, inflammation induced by dCGN was associated with recruitment of macrophages to inflammation sites [18], [19]. Also, inflammation induced by Dextran Sulphate Sodium (DSS), another sulphated compound, was directly associated with macrophages recruitment [20], since DSS still provoked inflammation after T-lymphocyte and NK cell depletion [20]. Although inflammation can be induced by dCGN, there are no data on human monocyte responses to dCGN exposure. Therefore, to investigate the effects of dCGN on human monocytes, normal Peripheral Blood Monocytes (PBM) and tumoral monocyte/macrophage THP-1 cells were exposed to 10 kDa and 40 kDa dCGN. We found that dCGN inhibited THP-1 cell proliferation in vitro, increased ICAM-1 expression, stimulated ICAM-1-dependent monocyte aggregation, and stimulated TNF-α expression and secretion. These responses were more pronounced after 40 kDa dCGN exposure and were linked to NF-κB activation. In addition, the 40 kDa dCGN, but not the 10 kDa dCGN induced in vivo colitis as shown by the inflammatory response in the rat colon. These results suggest that the degraded forms of CGN have an important effect on monocytes resulting in an inflammatory phenotype. Materials and Methods Preparation of Degraded Carrageenan Two preparations of degraded carrageenan with low, (∼10 kDa; C10), and medium, (∼40 kDa; C40) molecular weight were prepared from native iota-carrageenan extracted from Euchema spinosum (generously provided by Sanofi Biosystems Industry, Boulogne-Billancourt, France). Native carrageenan was dissolved in distilled water (5% w/v) under vigorous stirring and heated to 60°C. Then, the carrageenan solution was submitted to two different treatments to obtain both low and medium molecular weight fractions. Briefly, for the low molecular weight fraction, carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 15 min at 80°C. After neutralization with NaOH 4N, the solution was ultra filtered through a hollow fibre cartridge with MW cut-off 5 kDa, (Amicon Inc, Beverly, USA). For the medium molecular weight fraction, the carrageenan solution was hydrolyzed with 0.3% (v/v) concentrated sulphuric acid for 30 min at 60°C. After neutralization, the supernatant was ultra filtered (MW cut-off 100 kDa). The filtrate was submitted to a second ultra filtration (MW cut-off 5 kDa). Both preparations of dCGN were precipitated with 4 volumes of 95% ethanol, dried at room temperature and ground to small particles (1 mm in diameter). Using gel-permeation chromatography in combination with light scattering measurements (see Viebke et al. [21]), it was confirmed that the low fraction had an average molecular weight of 10 kDa, and the medium fraction of 40 kDa. The sulphate content of polysaccharides in both fractions was measured following the method of Quemener et al. [22]. Finally, the absence of polysaccharide structure modifications in the two fractions was confirmed using 2H-NMR spectroscopy. The absence of LPS contamination in the two fractions was confirmed using the e-Toxate® kit (Sigma, St Quentin Fallavier, France). Before use in cell culture, the two fractions were dissolved in complete medium during 30 min at 56°C. Animals, Chemicals and Diet Male Wistar rats (150 g average weight) were housed under standard conditions and fed ad libitum with standard rodent laboratory chow. Degraded iota-carrageenans were administered in the drinking water (5% w/v) for 55 days to 2 groups of six animals each. The first group received the low molecular weight carrageenan (10 kDa dCGN) and the second received the medium molecular weight carrageenan (40 kDa dCGN). An additional group of four rats were maintained on regular tap water (control group). To increase palatability 0.2% sucrose was added to the drinking water of all groups (Van der Waaji et al., [23]). Fresh carrageenan solutions were prepared daily. Evaluation of Colitis Body weight, liquid and food consumption, diarrhea and rectal bleeding (detected by eye inspection) were recorded throughout the feeding period. After 55 days, animals were sacrificed by cervical dislocation. The length of the colon was measured as described by Okayashu et al. [24]. Then, each colon was ligated in sections of 2 cm and 1 to 2 ml of 10% formalin was infused into the intestinal lumen. The moderately distended segment was sectioned and fixed in 10% formalin. The following day, the intestinal content was removed by vortexing. The fixed segment was kept in 10% formalin at 4°C until the paraffin embedding procedure. To evaluate the degree of inflammation, this segment of colon was opened longitudinally and macroscopic and histological scores of inflammation were recorded as previously described [25], [26]. The toluidine blue staining was used for identification of sulphated polysaccharides in the intestinal mucosa. On the day of sacrifice, a fresh sample of each colon (50 mg) was collected for myeloperoxidase (MPO) assay according to Krawisz et al., [27]. The level of MPO, mainly expressed by neutrophils, indicates the rate of recruitment of neutrophils to the intestinal mucosa. One unit of MPO activity corresponds to the degradation of 1 µmol of peroxide per minute at 25°C. Cell Culture All tissue culture reagents were from Invitrogen (Cergy Pontoise, France). THP-1 human monocytic cells were maintained in RPMI-1640 supplemented with 10% FCS, 2 mM L -glutamine, 50 U/ml penicillin and 50 mg/ml streptomycin at 37°C in a 5% CO2 incubator. Human peripheral blood mononuclear cells were obtained from heparinized blood by Ficoll-Hypaque density gradient. Monocytes were then isolated by adherence to culture flasks as described [28]. For cell aggregation, monocytes were cultured in the presence or absence of C10 or C40 for 72 h. Cell colonies were monitored under an inverted phase contrast microscope coupled through a video camera to a computer. In some wells, neutralizing monoclonal antibody to ICAM-1 (2.5 µg/ml) (Tebu, Le Perray en Yvelines, France) was added. Cell Cycle Analysis THP-1 cells in exponential growth phase were exposed to complete medium in the presence or absence of carrageenans for 24 h before being stained with propidium iodide using the DNA-Prep Coulter kit according to the manufacturer's instruction (Beckman-Coulter, Villepinte, France). Cell DNA content was then analyzed by flow cytometry using an EPICS XL2 (Beckman-Coulter). Raw data for the distribution of DNA content of 30,000 cells retrieved from the cytometer were expressed as the percentage of G0/G1 through G2/M populations. Multicycle AV software (Phoenix Flow Systems, San Diego, CA) was used to generate DNA content frequency histograms and facilitate data analysis. Cell Surface Antigen Expression Analysis Peripheral Blood Monocytes or THP-1 cells were exposed to complete medium in the presence or absence of carrageenan for 36 h. After two washes in PBS without Ca2+ and Mg2+, cells were incubated in PBS containing 0.1% gelatin and 8% AB human serum to prevent binding to Fc receptors. Then, 5×105 cells were incubated with primary antibodies at 4°C for 30 min. Two other washes in PBS preceded incubation with FITC-conjugated goat antibody anti-mouse IgG diluted 1/1000 at 4°C for 30 min (Tebu). After two additional washes, analysis of stained cells was performed on an EPICS XL2 (Beckman-Coulter). The cell population was gated according to its forward and wide-angle light scattering. Data were expressed as mean relative fluorescence intensity (MFI) of 3000 cells. TNF Activity Bioassay Monocytes or THP-1 cells were cultured with or without different concentrations of CGNs or LPS (Salmonella typhosa, Sigma) for 24 h or the indicated time. Biologically active TNF-α/β in tissue culture supernatant was measured using the WEHI 164 clone 13-cell killing assay [29]. TNF concentrations are expressed as pg/ml. RT-PCR Analysis Total RNA from monocytes was isolated using TRIzol Reagent™ (Invitrogen). cDNA was generated on 1 µg of total RNA in a reaction volume of 20 µl, using M-MLV reverse transcriptase (Invitrogen). PCR was done in the linear range of amplification (determined for each primer pair-cDNA combination). Standard PCR reactions were performed with 1 µl of the cDNA solution, 50 µM of each primer solution, 10 mM of each dNTP, 25 mM MgCl2, 10X Goldstar DNA polymerase reaction buffer, and 0.5 units of Goldstar DNA polymerase (Eurogentec, Seraing, Belgium). First PCR cycle consisted of 1 min at 92°C, 1 min at 58°C and 1 min at 72°C; then each PCR cycle consisted of 40 sec at 92°C, 40 sec at 58°C and 50 sec at 72°C. cDNA for β-actin was amplified for 28 cycles using the oligos: sense 5′-GGCATCGTGATGGACTCCG-3′ and antisense 5′GCTGGAAGGTGGACAGCGA-3′. cDNA for TNF-α was amplified for 35 cycles using the oligos: sense 5′-AAGCCTGTAGCCCATGTTGT-3′ and antisense 5′-CAGATAGATGGGCTCATACC-3′. cDNA for ICAM-1 was amplified for 35 cycles using the oligos sense 5′-GTAGCAGCCGCAGTCATAATGG-3′ and antisense 5′-A TGCTGTTGTATCTGACTGAGG-3′. NF-kB Transcription Reporter Gene Assay The plasmid 3XMHC-luc (a generous gift from Drs. J. Westwick and D.A. Brenner, University of North Carolina, Chapel Hill) contains three copies of NF-κB-responsive element from the MHC class I locus, placed upstream of the luciferase gene. Human monocytic THP-1 cells were transiently transfected as previously described [30], and then cultured for 4 h alone or with increasing concentration of either C10 or C40. Luciferase activity was determined using a luminometer (Monolight 2010 Luminometer, Ann Arbor, MI). Western Blot Analysis THP-1 cells were stimulated for various lengths of time with 0.1 mg/ml C10 or C40, or 10 µg/ml LPS. Cells were then pelleted, washed and homogenised in lysis buffer (10 mM Hepes, pH 7.9, 150 mM NaCl, 1 mM EDTA, 0.6% NP-40, and 0.5 mM PMSF) on ice. Homogenates were sonicated, centrifuged at 10,000 rpm to remove cellular debris, and supernatant collected. Protein concentration was determined using the DC Protein Assay (Bio-Rad). Proteins in samples (15 µg total proteins) were resolved in a denaturing 12% polyacrylamide gel and transferred to a nitrocellulose membrane. I-κBα protein was detected using a rabbit polyclonal antibody (Santa Cruz Biotechnology, CA) followed by a horseradish peroxidase-coupled goat polyclonal antibody against rabbit Ig (Caltag Laboratories). Finally, IκB bands were revealed using the ECL™ detection system (Amersham Pharmacia Biotech, Les Ullis, France) according to the manufacturers' instruction. Antibody to α-Tubulin (Santa Cruz) was use as loading control. For nuclear NF-κB, THP-1 cells were stimulated with 1 mg/ml C10 or C40 for 30 minutes at 37°C. Cells were then pelleted and nuclei separated as described [31]. Nuclei were washed and homogenized directly in loading (Laemli) buffer and heated for 5 minutes at 100°C. Proteins in samples were resolved in a denaturing 8% polyacrylamide gel and transferred to a polyvinylidine fluoride (PVDF) membrane (Immobilon-P; Millipore, Bedford, MA). Membranes were incubated in blocking buffer (1% BSA, in PBS) for two hours at room temperature. Membranes were subsequently probed with the corresponding antibody in blocking buffer, overnight. Rabbit polyclonal antibody anti-NF-κB p50 subunit (# sc-114) or anti-NF-κB p65 subunit (# sc-109) from Santa Cruz Biotechnology were used. Membranes were washed six times in PBS with 0.05% Tween 20, 5 minutes each time, and incubated with a 1/3000 dilution of HRP-conjugated F(ab')2 goat anti-rabbit IgG in 5% nonfat dry milk and 0.05% Tween 20 in PBS for 1 hour at room temperature. After washing six more times in PBS with 0.05% Tween 20, antibody-reactive proteins were detected using a chemiluminescence substrate (SuperSignal; Pierce, Rockford, IL) according to the manufacturer's instructions. To confirm that equivalent amounts of protein were loaded in each line, membranes were also Western blotted for ERK as described [32]. Analysis of NF-κB Activation by Flow Cytometry Nuclear activation of NF−κΒ by flow cytometry was performed as described [31]. Statistical Analysis The results were expressed as the mean value ± S.E.M. of individual experiments. The statistical significance of the differences between mean values was assessed by the Student's t-test and analysis of variance (ANOVA). Results Degraded CGN Induce Colonic Inflammation All rats developed diarrhea during degraded carrageenan administration and gross evidence of blood was frequently detected in the stools. Colon length dramatically decreased in all treated rats with a more pronounced effect being observed in the 40 kDa dCGN treated group (Fig. 1A). Furthermore, prolonged exposure to 40 kDa dCGN resulted in high macroscopic and histological scores of inflammation (Fig. 1B, C). Only weak myeloperoxidase activity was detected in both control and dCGN-treated groups (Fig. 1D), indicating that granulocytes did not play a major role in the inflammation at that stage. Histological examination revealed various degrees of mucosal inflammation. Rats treated with 10 kDa dCGN showed edema, epithelium atrophy and slight lymphocyte infiltration (data not shown). These symptoms were totally absent in the colon of control rats (Fig. 1E). More severe mucosal injuries including ulceration, hyperplastic epithelium, crypt distortion and a strong macrophage infiltration, were observed in the 40 kDa dCGN-treated rats (Fig. 1F). No sulphated polysaccharides were detected by toluidine blue staining of colon mucosa from rats treated with either the 10 or 40 kDa dCGN (not shown). Although we cannot exclude that dCGN mat not have retained in the section during the histology procedure, this indicates that these polymers may not have been phagocytosed. 10.1371/journal.pone.0008666.g001 Figure 1 Degraded CGN induced colon inflammation in rats. Histograms showing the effect of degraded CGN on: colon length (A); macroscopic (B) and histological (C) inflammation score of colon; Myeloperoxidase (MPO) activity (D). Control rats (white bars); 10 kDa degraded CGN-treated rats (grey bars); 40 kDa degraded CGN-treated rats (black bars). * p<0.05 from control. ** p<0.01 from control. Histological analysis of colon from control rats (E), and from 40 kDa dCGN-treated rats (F). Degraded CGN Induced
T7186 16858-16863 Protein denotes TNF-α
R17 T44 T45 themeOf ICAM-1,expression
R18 T45 T43 themeOf expression,increased
R19 T47 T48 themeOf TNF-α,expression
R20 T48 T46 themeOf expression,stimulated
R21 T49 T46 themeOf secretion,stimulated
R564 T719 T720 themeOf IL-8,production
R565 T722 T723 themeOf ICAM-1,expression
R566 T723 T721 themeOf expression,increased
R567 T726 T724 themeOf dependent,stimulated
R568 T728 T729 themeOf TNF-α,expression
R569 T729 T727 themeOf expression,stimulated
R570 T730 T727 themeOf secretion,stimulated
R2768 T3097 T3096 equivalentTo MPO,myeloperoxidase
R2769 T3098 T3099 themeOf MPO,expressed