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T*
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T*
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T*
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T*
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T*
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T*
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T*
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[(Each)-249.3(of)-248.2(the)-245.2(two)-246.1(experimental)-249.6(blocks)-245(was)-252(technically)-248(duplicated)]TJ
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T*
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T*
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T*
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(94)Tj
47.4914 0 TD
[(THELIN)-334.5(ET)-338.7(AL.)]TJ
ET
endstream
endobj
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T*
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T*
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T*
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T*
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T*
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[(Human)-462.1(Cytokine/Chemokine)-457.5(39)-460.6(\(Thermosher,)-461.8(Waltham,)-460.4(MA\))]TJ
T*
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(17)Tj
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T*
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T*
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T*
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[(parameter)-283.6(logistical)-286.8(standard)-280.6(curve)-283.2(for)-282(each)-277.2(cytokine.)]TJ
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[(For)-455(immunocytochemistry,)-461.3(astrocytes)-458(were)-464(plated)-452.6(onto)-457.3(clear)]TJ
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[(washed)-215.1(three)-217.7(times)-215.1(with)-211.7(phosphate-buffered)-212.5(saline)-213.5(\(PBS\).)-216.2(Samples)]TJ
T*
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T*
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[(primary)-308.9(antibody)-303.7(in)-300.2(0.1%)-300.7(Triton/PBS/5%)-307(goat)-301.7(serum)-304.5(at)-302.6(4)]TJ
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[(mary)-322.7(antibodies)-323(used)-317.8(were)-324.9(mouse)-314.5(monoclonal)-321.4(anti-GFAP)-322.3(\(Clone)]TJ
T*
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[(PBS,)-332.1(a)-330.7(secondary)-329.1(antibody)-329(\(goat)-335.4(anti-mouse,)-328.1(Alexa)-332.2(Fluor)-331.6(488)-327.3(or)]TJ
T*
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T*
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[(\(100)-171.4(ng/mL\).)]TJ
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[(Images)-268.5(were)-268(acquired)-262.9(using)-268.3(the)-264.2(automated)-267.3(Opera)-261.2(Phenix)-267.7(High-)]TJ
-1.1128 -1.1128 TD
[(Content)-365.8(Screening)-362.1(System)-360.9(\(Perkin)-354.6(Elmer\).)-362.7(For)-360.1(automated)-362.2(image)]TJ
T*
[(analysis,)-203.9(the)-201(Columbus)-199.2(Image)-204.5(Analysis)-201.9(System)-202.8(\(Perkin)-196.5(Elmer\))-208.1(was)]TJ
28.0042 57.8357 TD
[(used.)-263.8(Five)-268.6(non-overlapping)-270(elds)-269.3(per)-269.1(well)-264.7(were)-268(acquired,)-272.1(and)-266.5(two)]TJ
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[(wells)-278.1(per)-281.8(condition)-282(\(total)]TJ
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10.4265 0 TD
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[(5000)-276.3(cells\))-285.6(were)-280.7(analyzed.)]TJ
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[(RNA)-332.3(extraction,)-336.2(sequencing,)-334(and)-335.7(analysis)]TJ
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1.1128 -1.7514 TD
[(Single-stranded)-225.6(RNAseq)-227(data)-228.2(was)-226.7(generated)-226.9(from)-228.1(iPSC-derived)]TJ
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[(astrocytes)-685.6(and)-690.1(was)-688.3(analyzed)-692.7(alongside)-686.2(previously)-685.9(published)]TJ
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[(RNAseq)-543.1(data)-544.4(from)-537.9(iPSC-derived)-545.2(MN.)]TJ
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(19)Tj
8.9663 0 0 8.9663 470.0975 655.8235 Tm
[(Both)-544.2(MN)-540.5(and)-544.7(astro-)]TJ
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[(cyte)-266.1(cultures)-272(were)-268(derived)-267.3(from)-266(the)-264.2(same)-266.8(control)-268.9(iPSC)-269.5(lines)-268.4(using)]TJ
T*
[(two)-353.6(biological)-363.8(repeats)-359.4(in)-357.1(technical)-358.8(duplicate)-362.7(and)-355(were)-362.9(processed)]TJ
T*
[(using)-394.8(the)-397(same)-393.3(workow)-396.4(described)-402.7(subsequently.)-391.7(In)-400(addition)-396.3(to)]TJ
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[(using)-312.6(two)-309.4(lines)-312.7(\(from)-312.4(two)-315.7(different)-313.5(people\),)-314.5(we)-314.6(additionally)-309.9(used)]TJ
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[(two)-271.4(independent)-281.8(neural)-274.1(inductions)-276.3(from)-272.4(each)-277.2(of)-273.5(these)-279.5(lines,)-277.7(as)-274.5(we)]TJ
T*
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T*
[(intra-patient)-298.1(variability)-300.4(while)-295.9(increasing)-305.1(the)-295.8(power)-296.7(by)-296.2(conducting)]TJ
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[(perimental)-501.1(blocks)-497.9(was)-505(technically)-500.9(duplicated)-501.3(\(i.e.,)-503.2(two)-499(culture)]TJ
T*
(wells\).)Tj
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[(A)-192.5(Promega)-202.1(Maxwell)-203.2(RSC)-196.2(simplyRNA)-203.1(cells)-195(kit)-202.6(including)-193.5(DNase)]TJ
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T*
[(extractions.)-298.9(A)-306.3(NanoDrop)-298.6(\(Thermo)-304.7(Scientic,)-299.5(Waltham,)-302.4(MA\))-302.3(was)]TJ
T*
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0 -1.1065 TD
[(Agilent)-279.9(Bioanalyser)-277.7(\(Agilent)-282(Technologies,)-278.9(Santa)-283.2(Clara,)-275.9(CA\))-281.1(was)]TJ
0 -1.1128 TD
[(used)-273.5(to)-274.9(assess)-272.2(quality.)-273.2(RNA)-275.4(integrity)-276(\(RIN\))-279(scores)-272.4(were)]TJ
/F7 1 Tf
22.8889 0 TD
(>)Tj
/F4 1 Tf
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[(8)-271.4(for)-275.6(all)]TJ
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[(samples)-226.3(used)-229.3(in)-230.7(this)-228.6(work.)-228.2(RNAseq)-227(libraries)-233.9(were)-230.1(prepared)-229.9(using)-224.1(a)]TJ
T*
[(Truseq)-331.9(stranded)-337.5(mRNA)-335.4(kit)-329(\(Illumina\))-337.1(with)-331.9(1)]TJ
/F6 1 Tf
18.6399 0 TD
(l)Tj
/F4 1 Tf
.5501 0 TD
[(g)-334.6(input)-328.1(RNA.)-335.2(The)]TJ
-19.19 -1.1065 TD
[(products)-366.7(were)-369.2(then)-371.2(puried)-370.9(and)-367.7(enriched)-370.4(with)-369.8(polymerase)-367.8(chain)]TJ
0 -1.1128 TD
[(reaction)-406(\(PCR\))-402.8(amplication)-411.6(to)-401.4(create)-411.7(the)-403.3(nal)-407.7(cDNA)-402.4(libraries.)]TJ
T*
[(Libraries)-432(were)-432.4(sent)-435.6(for)-427.4(high)-432(throughput)-435.7(sequencing,)-427.8(run)-431.1(for)-433.7(75)]TJ
T*
[(cycles)-395.2(on)-397.4(a)-400.3(rapid)-392.3(ow)-398.8(cell,)-399.5(at)-397.5(the)-397(Institute)-397.3(of)-400(Neurologys)-396.5(next)]TJ
0 -1.1065 TD
[(generation)-386.1(sequencing)-387(\(NGS\))-393.7(core)-384.9(facility)-386.3(using)-388.5(the)-390.7(Hiseq2500.)]TJ
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[(RNA-seq)-292.3(data)-297.8(were)-293.3(generated)-296.4(from)-291.3(poly\(A\))]TJ
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17.8749 0 TD
(+)Tj
/F4 1 Tf
.8409 0 TD
[(RNA.)-297.3(After)-290.7(remov-)]TJ
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[(ing)-464.1(the)-466.5(adapter)-472(sequences)-469.2(and)-468.8(quality)-466.3(checking)-468.9(the)-472.9(data)-468.5(using)]TJ
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[(FastQC,)-437.9(the)-434.9(reads)-436.1(were)-438.8(initially)-439.5(aligned)-439.4(to)-433(ribosomal)-437(RNA)-439.8(se-)]TJ
0 -1.1128 TD
[(quences)-346.5(to)-350.8(lter)-350.6(out)-350.3(reads)-347.6(that)-349.9(might)-350.6(have)-350.6(come)-347.9(from)-348.2(ribosomal)]TJ
T*
[(RNA)-427.2(contamination)-431.5(using)-426.4(bowtie2)-427.7(\(-v)-433.7(0\).)-428.2(The)-424.4(remaining)-431.9(reads)]TJ
T*
[(were)-419.8(then)-421.8(aligned)-426.8(to)-420.4(the)-422.3(human)-420.8(genome)-422.8(\(h19\))-420.1(using)-426.4(the)-422.3(splice)]TJ
0 -1.1065 TD
[(aware)-541.8(aligner)-543.2(TopHat2)-543.6(with)-540.5(default)-543.2(parameters.)-546.5(The)-544.5(absolute)]TJ
0 -1.1128 TD
[(quantication)-255.8(of)-254.5(the)-257.9(genes)-256.5(was)-252(performed)-258.4(using)-255.7(HTSeqcount.)-255.2(The)]TJ
T*
[(raw)-221.8(read)-226.8(count)-225.3(les)-225.6(obtained)-223.9(were)-230.1(then)-225.8(processed)-222.8(for)-225.1(unsupervised)]TJ
T*
[(clustering)-262.4(analysis.)-267.2(The)-266.3(EdgeR)-263.7(package)-265.5(in)-262.3(R)-263.5(was)-264.7(used)-267.2(to)-262.3(generate)]TJ
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[(normalized)-389.6(reads)-391.9(per)-382.9(kilobase)-389.5(of)-387.3(transcript)-391.5(\(RPKM\))-389(values.)-389.4(For)]TJ
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[(corresponding)-249.2(cytokine)-249.2(receptors)-253.5(for)-250.3(the)-251.6(inducing)-246.8(cytokines,)-252.9(these)]TJ
T*
[(values)-392.8(were)-400.8(mean)-392.1(centralized)-400.8(and)-393(plotted)-396.8(using)-394.8(R)-396.3(version)-392.5(3.2.3)]TJ
0 -1.1065 TD
[(\(Table)-356.1(S1\).)-356.7(All)-357.6(libraries)-360.3(used)-355.7(had)]TJ
/F7 1 Tf
14.2455 0 TD
(<)Tj
/F4 1 Tf
.5501 0 TD
[(1%)-355.2(ribosomal)-361.2(RNA)-357.6(\(rRNA\),)]TJ
/F7 1 Tf
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(<)Tj
/F4 1 Tf
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[(1%)-184.5(mitochondrial)-185.1(DNA)-179.2(\(mtDNA\),)]TJ
/F7 1 Tf
13.9863 0 TD
(>)Tj
/F4 1 Tf
.5501 0 TD
[(90%)-184(strandedness,)-185.2(and)]TJ
/F7 1 Tf
9.1935 0 TD
(>)Tj
/F4 1 Tf
.5501 0 TD
(70%)Tj
-24.8301 -1.1128 TD
[(exonic)-277.8(reads)-284.4(\(data)-280.9(not)-280.8(shown\).)]TJ
/F10 1 Tf
0 -2.7188 TD
[(Clinical)-334.2(cytokine)-334.8(data)-332.9(from)-334.2(TBI)-335.9(patients)]TJ
/F4 1 Tf
1.1128 -1.7514 TD
[(We)-313.2(extracted)-313.1(data)-316.7(from)-310.3(two)-315.7(previously)-312.9(published)-310.8(studies)-312.9(from)]TJ
-1.1128 -1.1128 TD
[(our)-361.5(group)-360.6(describing)-359.5(the)-365.4(temporal)-362.9(proles)-365.8(and)-361.4(concentrations)-360.1(of)]TJ
T*
[(cytokines)-464.9(from)-468.4(severe)-463.4(TBI)-467.2(patients.)]TJ
6.7246 0 0 5.9768 447.1369 211.2377 Tm
(7,23)Tj
8.9663 0 0 8.9663 463.0676 207.2125 Tm
[(In)-463.2(a)-469.8(pilot)-464.9(trial)-463.6(of)-469.5(IL-1)]TJ
ET
59.754 177.165 490.167 .22676 re
f
59.754 106.923 490.167 .28348 re
f
59.754 155.395 490.167 .28348 re
f
BT
/F9 1 Tf
8.9663 0 0 8.9663 128.296 182.8346 Tm
(Table)Tj
/F4 1 Tf
3.0034 0 TD
(1)Tj
/F9 1 Tf
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[(.)-496.1(Cytokine)-335.5(Concentrations)-339.7(Used)-329.9(to)-332.7(Induce)-339.3(a)-333.2(Downstream)-333.6(Cytokine)-335.5(Response)]TJ
/F5 1 Tf
-11.1473 -2.3332 TD
[(Concentration)-9949.8(IL-1b)-7102(IL-6)-7292.2(TNF)-6672.3(IL-4)-6476.5(IL-10)]TJ
/F4 1 Tf
0 -2.0929 TD
[(Range)-335.9(seen)-339.2(physiologically)-2851.5(0.0221)-5197.4(0.154990)-5443(0.0523)-5191.1(0.137)-5697.4(0.8173)]TJ
0 -1.1128 TD
[(Low)-12217.5(1)-163.9(pg/mL)-5107.5(100)-162.9(pg/mL)-5354.1(1)-163.9(pg/mL)-5101.2(1)-170.2(pg/mL)-5101.2(1)-163.9(pg/mL)]TJ
0 -1.1065 TD
[(Medium)-10655.5(100)-169.3(pg/mL)-4102.2(10,000)-164.9(pg/mL)-4102.2(100)-162.9(pg/mL)-4102.2(100)-169.3(pg/mL)-4102.2(100)-162.9(pg/mL)]TJ
0 -1.1128 TD
[(High)-12050.5(10,000)-164.9(pg/mL)-2856.6(1,000,000)-160.5(pg/mL)-2856.6(10,000)-164.9(pg/mL)-2850.2(10,000)-164.9(pg/mL)-2856.6(10,000)-164.9(pg/mL)]TJ
7.9702 0 0 7.9702 67.748 95.1307 Tm
[(IL,)-332.3(interleukin)12(;)-330.7(TNF,)-334.3(tumor)-322.2(necrosis)-328.4(factor.)]TJ
0 -1.1239 TD
[(Cytokine)-237.2(concentrations)-239.9(used)-245(to)-243.3(stimulate)-242.5(the)-244.4(enriched)-236.7(astrocyte)-241.5(cultures.)-246.4(The)-238.6(concentrations)-239.9(are)-245.4(within)-241.6(range)-241.3(to)-250.4(what)-244.9(is)-244.5(seen)-241.1(in)-250.4(the)-244.4(extracellula)11.5(r)]TJ
58.2779 0 TD
(space)Tj
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[(as)-334.6(seen)-333.6(in)-328.7(the)-329.8(aftermath)-325.4(of)-333.6(TBI)-328.9(as)-334.6(measured)-321.2(by)-337.3(microdia)13.4(lysis.)]TJ
5.9774 0 0 5.3127 263.2251 80.7307 Tm
(1)Tj
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[(1.)-315(Helmy,)-315.5(A.,)-310.4(Guilfoyl)-12.3(e,)-311.1(M.R.,)-316.7(Carpenter,)-316.1(K.L.,)-316.2(Pickard)-13(,)-310.9(J.D.,)-309.5(Menon,)-316.2(D.K.)-315.9(and)-317.1(Hutchinson,)-316.6(P.J.)-319(\(2014\).)-311.5(Recom)-12.5(binant)-312(human)-320.6(interleukin-1)-319.8(receptor)-321.6(a)]TJ
61.4908 0 TD
[(ntagonist)-316.8(in)-316.5(severe)]TJ
-62.6371 -1.1463 TD
[(traumat)-11.1(ic)-337(brain)-337.2(injury:)-340.1(a)-333.4(phase)-338.7(II)-333.9(randomized)-345(control)-336.7(trial)-11.1(.)-327.2(J)-338.5(Cereb)-338.5(Blood)-341.5(Flow)-331(Metab)-342.3(34,)-335.3(845-851.)]TJ
/F3 1 Tf
8.9663 0 0 8.9663 59.7543 749.7069 Tm
[(ASTROCYTIC)-341.1(CYTOKINE)-339(RESPONSE)]TJ
/F11 1 Tf
18.2163 0 TD
[(IN)-337.4(VITRO)]TJ
/F3 1 Tf
4.6157 0 TD
[(TO)-331.2(SIMULATED)-336.9(TBI)-21231.7(95)]TJ
ET
endstream
endobj
62 0 obj
<>stream
/GS1 gs
BT
/F4 1 Tf
8.9663 0 0 8.9663 62.022 725.7825 Tm
0 0 0 rg
0 Tc
0 Tw
[(receptor)-493.1(antagonist)-487.4(treatment)-489(for)-490.6(TBI,)-489.1(we)-485.3(used)-488.5(brain)-493.5(cytokine)]TJ
0 -1.1128 TD
[(concentrations)-240(from)-234.4(the)-232.6(placebo)-235.5(group.)]TJ
6.7246 0 0 5.9768 202.5637 719.7731 Tm
(23)Tj
8.9663 0 0 8.9663 211.3511 715.8046 Tm
[(Demographic)-236.6(data)-234.5(for)-237.7(all)]TJ
-16.6545 -1.1128 TD
[(patients)-203.9(are)-208.3(available)-207(in)-199.1(Table)-208.6(S2,)-202.9(including)-206.1(age,)-202.3(gender,)-203.4(admission)]TJ
0 -1.1065 TD
[(Glasgow)-295.3(Coma)-293.2(Scale)-291.9(\(GCS\),)-290.5(and)-291.8(head)-293.7(computerized)-296.2(tomography)]TJ
0 -1.1128 TD
[(\(CT\))-273.4(veriable)-277.6(injury)-267(as)-274.5(dened)-272.4(by)-270.9(the)-270.5(Marshall)-279(CT)-269.1(classication)]TJ
T*
[(\(Grades)-390.5(IIIV)]TJ
/F7 1 Tf
5.9372 0 TD
(=)Tj
/F4 1 Tf
.7208 0 TD
[(diffuse)-388.8(and)-386.7(Grade)-394(VI)]TJ
/F7 1 Tf
9.0544 0 TD
(=)Tj
/F4 1 Tf
.7208 0 TD
(focal\).)Tj
6.7246 0 0 5.9768 232.611 679.9746 Tm
(24)Tj
8.9663 0 0 8.9663 242.8157 675.9495 Tm
[(As)-389.2(described)-390(in)]TJ
-20.1638 -1.1065 TD
[(detail)-425(in)-426.7(the)-422.3(previous)-423.6(studies,)]TJ
6.7246 0 0 5.9768 173.4236 669.9967 Tm
(7,23)Tj
8.9663 0 0 8.9663 189.0141 666.0282 Tm
[(patients)-425.2(were)-426.1(monitored)-423.2(for)-427.4(a)]TJ
-14.1633 -1.1128 TD
[(maximum)-266.5(of)-267.2(5)-271.4(consecutive)-266.4(days)-267.2(and)-272.8(cytokine/chemokine)-269.5(samples)]TJ
T*
[(were)-198.5(pooled)-193.2(from)-190.1(6)-170.2(h)-189.2(periods.)-199.4(Although)-190.8(there)-198.7(was)-195.1(some)-188.5(disparity)-197.2(in)]TJ
T*
[(commencement)-318(of)-311.4(monitoring)-310.5(following)-317.1(ictus,)-309.3(all)-312.5(data)-316.7(have)-312.7(been)]TJ
0 -1.1065 TD
[(adjusted)-269.4(to)-274.9(the)-270.5(trauma)-271.3(time)-265(to)-274.9(enable)-267.6(comparison)-270.5(with)-275(the)]TJ
/F5 1 Tf
23.7236 0 TD
[(in)-268.6(vitro)]TJ
/F4 1 Tf
-23.7236 -1.1128 TD
(experiments.)Tj
/F10 1 Tf
0 -2.605 TD
[(Statistical)-336.1(analysis)]TJ
/F4 1 Tf
1.1128 -1.7514 TD
[(The)-253.7(statistical)-256(program)-254.3(R,)-247.4(with)-256(the)-251.6(interface)-258.8(R-studio,)-249.5(was)-258.4(used)]TJ
-1.1128 -1.1065 TD
[(for)-193.4(the)-194.7(analyses.)]TJ
6.7246 0 0 5.9768 119.6787 571.181 Tm
(25)Tj
8.9663 0 0 8.9663 128.1259 567.2125 Tm
[(The)-196.8(effect)-196.9(of)-191.3(each)-195(of)-197.6(the)-194.7(added)-192.1(cytokines)-193(\(IL-1)]TJ
/F6 1 Tf
18.4945 0 TD
(b)Tj
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.5501 0 TD
(,)Tj
-26.4171 -1.1128 TD
[(IL-4,)-309.5(IL-6,)-309.5(IL-10,)-315.3(and)-310.8(TNF\))-312.2(on)-308.8(the)-308.5(measured)-314.2(cytokines)-313.2(\(39)-304.6(cyto-)]TJ
T*
[(kine)-308(panel\))-299.4(was)-308.9(analyzed)-307(using)-299.9(a)-305.4(two)-303(way)-307.8(mixed)-304.7(analysis)-308.5(of)-298.8(var-)]TJ
T*
[(iance)-533.6(\(ANOVA\))-536.8(\(to)-530(compare)-534.8(concentrations)-537.2(of)-532.8(each)-536.4(cytokine)]TJ
0 -1.1065 TD
[(analyzed\).)-267.7(The)-266.3(time)-265(at)-264.7(which)-266.5(the)-264.2(sample)-263.6(was)-264.7(taken)-265.7(\()52.7(time)53.3(;)-264.9(24)-163.4(h,)]TJ
0 -1.1128 TD
[(48)-169.7(h,)-280.6(72)-163.4(h\))-286.2(was)-283.6(used)-279.9(as)-280.8(the)-283.2(repeated)-284.3(measure)-289.4(\(within-subject\))-282.4(var-)]TJ
T*
[(iable.)-304.2(The)-297.9(concentration)-308.8(of)-298.8(added)-305.9(cytokine)-299.8(\()52.7(concentration)53.6(;)-302.8(un-)]TJ
0 -1.1065 TD
[(treated,)-542.2(and)-538.4(three)-533.8(concentrations)-543.5(of)-539.1(added)-539.8(cytokine)-533.8(shown)-539.5(in)]TJ
0 -1.1128 TD
[(Table)-316(1\))-317.8(was)-315.3(used)-311.5(as)-318.8(the)-314.8(independent)-319.8(\(between-subject\))-319(variable.)]TJ
T*
[(A)-432.8()61.1(time)-431.5(and)-430.9(concentration)49.4()-425.7(relationship)-429.8(was)-435.4(also)-429.3(generated)-429.2(to)]TJ
T*
[(ascertain)-333.3(whether)-327.5(time)-328.2(and)-329.7(concentration)-334.1(as)-325.1(an)-330.2(interactive)-332.8(model)]TJ
0 -1.1065 TD
[(affected)-286.9(downstream)-280.2(cytokine)-280.9(generation.)]TJ
1.1128 -1.1128 TD
[(For)-385.4(the)-384.3(immunocytochemistry)-382.5(analyses,)-385.7(data)-386.3(are)-385.3(presented)-390(as)]TJ
-1.1128 -1.1128 TD
(mean)Tj
/F7 1 Tf
2.3458 0 TD
()Tj
/F4 1 Tf
.7145 0 TD
[(standard)-565.2(deviation)-556.3(\(SD\))-561.6(from)-556.9(two)-562.3(technical)-561.1(replicates.)]TJ
-3.0603 -1.1128 TD
[(Two)-311.5(way)-314.1(ANOVA)-311.3(with)-312.9(Dunnetts)-310.3(multiple)-315.3(comparison)-308.5(tests)-313.9(was)]TJ
0 -1.1065 TD
(performed.)Tj
1.1128 -1.1128 TD
[(Descriptively,)-387.4(we)-384.1(compared)-388.9(the)-384.3(temporal)-381.8(proles)-384.8(of)-387.3(cytokine)]TJ
-1.1128 -1.1128 TD
[(release)-304.9(as)-306.1(well)-302.7(as)-306.1(concentrations)]TJ
/F5 1 Tf
13.55 0 TD
[(in)-306.5(vivo)]TJ
/F4 1 Tf
2.7631 0 TD
[(,)-300.1(including)-307.3(from)-304(enriched)]TJ
-16.3131 -1.1065 TD
[(neuronal)-241.5(cultures)]TJ
/F5 1 Tf
7.1639 0 TD
[(in)-243.3(vitro)]TJ
/F4 1 Tf
2.9212 0 TD
(,)Tj
6.7246 0 0 5.9768 154.6582 391.8613 Tm
(17)Tj
8.9663 0 0 8.9663 163.559 387.8928 Tm
[(from)-240.7(previous)-240.3(studies)-243.4(and)-241.2(compared)-243.4(it)]TJ
-11.3243 -1.1128 TD
[(with)-281.3(our)-279.3(results.)]TJ
6.7246 0 0 5.9768 120.6425 381.8834 Tm
(7,23)Tj
/F3 1 Tf
8.9663 0 0 8.9663 62.022 354.5574 Tm
(Results)Tj
/F10 1 Tf
0 -1.8336 TD
[(RNAseq)-334.4(data)-332.9(of)-336.7(iPSC-derived)-336.5(astrocyte)]TJ
0 -1.2203 TD
[(and)-335.7(MN)-333.2(cultures)]TJ
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[(,)-432.9(IL-6,)-435.9(and)-437.2(TNF,)-433.1(whereas)-434.6(the)-434.9(MN)-433(counter-)]TJ
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T*
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T*
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[(\(27)-374.2(cytokines,)]TJ
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(p)Tj
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[(majority)-313.9(of)-305.1(cases)-310.7(\(Table)-305.5(2\).)-308(IL-6)-306.6(\(13)-304.6(cytokines\))-309(and)-310.8(TNF)-303.7(\(11)-304.6(cy-)]TJ
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T*
[(duction,)-281.8(although)-278.4(fewer)-282.8(were)-280.7(detected)-279.4(if)-283.9(stimulated)-280(with)-274.9(IL-4)-281.3(and)]TJ
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[(ANOVA)-311.3(output)-308.6(for)-307.2(the)-314.8(experimental)-306.5(conditions)-314.3(showing)-308.6(a)-305.4(statis-)]TJ
T*
[(tically)-346.3(signicant)-340.6(response)-341(with)-338.2(increasing)-343(levels)-339.9(of)-343.1(cytokines)-338.5(as)]TJ
T*
[(compared)-287.7(with)-281.3(unstimulated)-279(cultures.)]TJ
1.1128 -1.2266 TD
[(We)-300.5(next)-301.7(considered)-301(the)-295.8(underlying)-299(principles)-300.3(of)-298.8(secondary)-297.5(cy-)]TJ
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[(tokine)-260.4(release.)-257.2(These)-256.3(are)-252.6(exemplied)-259.1(by)-251.9(Figure)-257.6(3)-252.4(and)-253.9(summarized)]TJ
T*
[(in)-388.7(Tables)-386.3(2)-385.2(and)-386.6(3.)-381.8(Three)-390.5(patterns)-385.8(were)-388.2(evident.)-385.4(\(1\))-383.1(A)-388.5(subset)-385.3(of)]TJ
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[(cytokines)-521.8(was)-523.9(generated)-517.7(in)-521.5(a)-520.4(time-dependent)-520.5(fashion,)-521.9(without)]TJ
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[(signicance)-385.2(to)-382.4(escalating)-384.8(concentrations)-385.4(\(29)-380.5(cytokines,)-379.3(Fig.)-383.4(3A\))]TJ
T*
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[(macrophage)-289.1(colony-stimulating)-280.4(factor)-283(\()-84.3(M-CSF\).)-279.7(\(2\))-282(Another)-280.8(sub-)]TJ
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T*
[(variation)-356.5(over)-350.8(time)-353.5(\(21)-348.9(cytokines,)-354(Fig.)-351.8(3B\))-347.3(\(Tables)-356.8(2)-353.6(and)-348.7(3\),)-352.3(in-)]TJ
T*
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[(secreted)-215.9(\(RANTES\))-213.2(and)-209.6(serum)-209.6(soluble)-210.6(CD40)-208.4(ligand)-209.9(\(sCD40L\).)-215.6(\(3\))]TJ
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T*
[(concentration)-214(dependency)-207.1(\(17)-209.8(cytokines,)-208.6(Fig.)-206.4(3C\))-208.2(\(Tables)-205.1(2)-208.2(and)-209.6(3\),)]TJ
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[(such)-406.3(as)-401(IL-8,)-404.3(vascular)-403.2(endothelial)-403.7(growth)-405.3(factor)-403.2(\(VEGF\)-A)-404.2(and)]TJ
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(IL-1)Tj
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[(The)-443.4(levels)-447.4(of)-444.2(exogenously)-441(added)-445(cytokines)-445.9(were)-445.1(maintained)]TJ
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[(throughout)-284(all)-280.9(time)-283.9(points)-278.2(during)-281.9(the)-276.9(experiment)-282.1(\(Fig.)-284.4(4\).)]TJ
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[(FIG.)-322.1(1.)]TJ
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[(Immunocytochemistry)-324.2(illustrating)-321.6(the)-321.1(enriched)-319.8(astrocytic)-321.7(cell)-320.7(cultures.)]TJ
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29.1486 0 TD
(\(A\))Tj
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1.7009 0 TD
[(A)-319(co-staining)-325.1(of)-317.8(DAPI)-322(and)-317.1(glial)-321.9(brillary)-323.7(acidic)]TJ
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[(protein)-249.9(\(GFAP\))-252.6(showing)-251.7(a)-254.8(representative)-257.7(image)-250.2(of)-254.5(the)-251.6(cultures.)]TJ
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[(\(B,)-255.9(C\))]TJ
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[(Highlights)-249.6(the)-251.6(presence)-256.1(of)-248.2(cleaved)-256.9(Caspase)-257.5(3)-246.1(a)-254.8(sign)-249.8(of)-254.5(apoptosis.)]TJ
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[(Scale)-336.2(bar)-332.3(50)]TJ
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5.2923 0 TD
(l)Tj
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[(m.)-334.8(Color)-331.4(image)-338.7(is)-333(available)-333.5(online.)]TJ
/F3 1 Tf
-5.8424 77.2217 TD
(96)Tj
47.4914 0 TD
[(THELIN)-334.5(ET)-338.7(AL.)]TJ
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