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doi:10.1371/journal.ppat.1009871
Gila Lustig, Sandile Cele, Farina Karim, Anne Derache, Abigail Ngoepe, Khadija Khan, Bernadett I. Gosnell, Mahomed-Yunus S. Moosa, Ntombi Ntshuba, Suzaan Marais, Prakash M. Jeena, Katya Govender, John Adamson, Henrik Kløverpris, Ravindra K. Gupta, Rohen Harrichandparsad, Vinod B. Patel, Alex Sigal
T cell derived HIV-1 is present in the CSF in the face of suppressive antiretroviral therapy
10.1371/journal.ppat.1009871
http://dx.doi.org/10.1371/journal.ppat.1009871
2021-09-23
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11.9999 0 0 11.9999 200.012 706.053 cm
BT
/F12 1 Tf
-0.0096 Tc
1 TL
0 0 Td
(Detection)Tj
4.252 0 Td
(of)Tj
1.0582 0 Td
(ART)Tj
2.1969 0 Td
(concentrations)Tj
6.3402 0 Td
(in)Tj
1.0536 0 Td
(CSF)Tj
1.9228 0 Td
(and)Tj
1.7717 0 Td
(matched)Tj
3.7748 0 Td
(plasma)Tj
3.1607 0 Td
(by)Tj
1.2 0 Td
(LC-MS/MS)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 689.102 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
0 0 Td
(Sample)Tj
3.135 0 Td
(analysis)Tj
3.3279 0 Td
(was)Tj
1.6951 0 Td
(performed)Tj
4.4731 0 Td
(using)Tj
2.3867 0 Td
(an)Tj
1.2019 0 Td
(Agilent)Tj
3.1578 0 Td
(High)Tj
2.245 0 Td
(Pressure)Tj
3.617 0 Td
(Liquid)Tj
2.8573 0 Td
(Chromatography)Tj
-28.0968 -1.3039 Td
(\(HPLC\))Tj
3.4242 0 Td
(system)Tj
2.9423 0 Td
(coupled)Tj
3.3902 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.4683 0 Td
(AB)Tj
1.4854 0 Td
(Sciex)Tj
2.2733 0 Td
(5500,)Tj
2.3471 0 Td
(triple)Tj
2.3471 0 Td
(quadrupole)Tj
4.8075 0 Td
(mass)Tj
2.194 0 Td
(spectrometer)Tj
5.4595 0 Td
(equipped)Tj
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(with)Tj
1.9955 0 Td
(an)Tj
1.2019 0 Td
(electrospray)Tj
5.0513 0 Td
(ionization)Tj
4.2803 0 Td
(\(ESI\))Tj
2.2847 0 Td
(TurboIonSpray)Tj
6.3893 0 Td
(source.)Tj
3.0614 0 Td
(The)Tj
1.7858 0 Td
(LC-MS/MS)Tj
4.8359 0 Td
(method)Tj
3.3221 0 Td
(was)Tj
-34.2082 -1.2983 Td
(developed)Tj
4.2689 0 Td
(and)Tj
1.7291 0 Td
(optimised)Tj
4.2123 0 Td
(for)Tj
1.3833 0 Td
(the)Tj
1.4683 0 Td
(quantitation)Tj
5.159 0 Td
(of)Tj
1.0148 0 Td
(tenofovir,)Tj
4.1102 0 Td
(emtricitabine,)Tj
5.794 0 Td
(efavirenz,)Tj
4.0705 0 Td
(lopinavir,)Tj
-33.2104 -1.3039 Td
(ritonavir,)Tj
3.9628 0 Td
(nevirapine,)Tj
4.6998 0 Td
(zidovudine,)Tj
4.9095 0 Td
(lamivudine,)Tj
4.9663 0 Td
(abacavir,)Tj
3.7701 0 Td
(atazanvir)Tj
3.8721 0 Td
(and)Tj
1.7291 0 Td
(dolutegravir)Tj
5.1193 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.4684 0 Td
(same)Tj
-35.5292 -1.2983 Td
(sample.)Tj
3.2484 0 Td
(A)Tj
0.9014 0 Td
(protein)Tj
3.1522 0 Td
(precipitation)Tj
5.3631 0 Td
(extraction)Tj
4.2576 0 Td
(method)Tj
3.3222 0 Td
(using)Tj
2.3867 0 Td
(acetonitrile)Tj
4.7282 0 Td
(was)Tj
1.7008 0 Td
(used)Tj
2.0579 0 Td
(to)Tj
1.0261 0 Td
(process)Tj
3.1862 0 Td
(clini-)Tj
-35.3308 -1.2982 Td
(cal)Tj
1.3209 0 Td
(plasma)Tj
3.033 0 Td
(and)Tj
1.7292 0 Td
(CSF)Tj
1.8878 0 Td
(samples.)Tj
3.6057 0 Td
(The)Tj
1.7858 0 Td
(procedure)Tj
4.3143 0 Td
(was)Tj
1.6951 0 Td
(performed)Tj
4.473 0 Td
(using)Tj
2.3868 0 Td
(50)Tj
/F13 1 Tf
1.1735 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2643 0 Td
(of)Tj
1.0148 0 Td
(plasma)Tj
3.033 0 Td
(or)Tj
1.0942 0 Td
(CSF.)Tj
2.1146 0 Td
(50)Tj
/F13 1 Tf
-35.926 -1.304 Td
()Tj
/F7 1 Tf
(L)Tj
1.2642 0 Td
(of)Tj
1.0148 0 Td
(water)Tj
2.4321 0 Td
(and)Tj
1.7291 0 Td
(50)Tj
/F13 1 Tf
1.1736 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2642 0 Td
(of)Tj
1.0148 0 Td
(ISTD)Tj
2.3811 0 Td
(solution)Tj
3.4809 0 Td
(was)Tj
1.7008 0 Td
(added)Tj
2.6589 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
1.4683 0 Td
(sample)Tj
3.0217 0 Td
(was)Tj
1.7008 0 Td
(briefly)Tj
2.7779 0 Td
(mixed.)Tj
2.948 0 Td
(150)Tj
/F13 1 Tf
1.6441 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2643 0 Td
(of)Tj
-36.6744 -1.2982 Td
(acetonitrile)Tj
4.7281 0 Td
(was)Tj
1.7008 0 Td
(subsequently)Tj
5.4028 0 Td
(added)Tj
2.6589 0 Td
(to)Tj
1.0261 0 Td
(facilitate)Tj
3.6 0 Td
(protein)Tj
3.1521 0 Td
(precipitation,)Tj
5.5899 0 Td
(vortex)Tj
2.7382 0 Td
(mixed)Tj
2.7213 0 Td
(and)Tj
1.7348 0 Td
(cen-)Tj
-35.053 -1.2983 Td
(trifuged)Tj
3.3902 0 Td
(at)Tj
0.9524 0 Td
(16000)Tj
2.5965 0 Td
(g)Tj
0.686 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1735 0 Td
(min)Tj
1.8539 0 Td
(at)Tj
0.9524 0 Td
(4C.)Tj
1.9786 0 Td
(170)Tj
/F13 1 Tf
1.6441 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2642 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(clear)Tj
2.1146 0 Td
(supernatant)Tj
4.9833 0 Td
(was)Tj
1.7008 0 Td
(then)Tj
2.0183 0 Td
(transferred)Tj
4.6374 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
-36.8444 -1.2983 Td
(clean)Tj
2.296 0 Td
(micro-centrifuge)Tj
6.9959 0 Td
(tube)Tj
1.9729 0 Td
(and)Tj
1.7348 0 Td
(dried)Tj
2.3357 0 Td
(down)Tj
2.4888 0 Td
(using)Tj
2.3867 0 Td
(a)Tj
0.652 0 Td
(SpeedVac)Tj
4.1442 0 Td
(dryer)Tj
2.3641 0 Td
(set)Tj
1.3039 0 Td
(at)Tj
0.9525 0 Td
(40C.)Tj
2.4548 0 Td
(The)Tj
1.7801 0 Td
(dried)Tj
-33.8624 -1.3039 Td
(samples)Tj
3.3788 0 Td
(were)Tj
2.126 0 Td
(then)Tj
2.0183 0 Td
(reconstituted)Tj
5.4935 0 Td
(in)Tj
1.0374 0 Td
(100)Tj
/F13 1 Tf
1.6441 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2699 0 Td
(of)Tj
1.0148 0 Td
(0.02%)Tj
2.6249 0 Td
(sodium)Tj
3.2315 0 Td
(deoxycholate)Tj
5.4595 0 Td
(\(Sigma\))Tj
3.3618 0 Td
(in)Tj
1.0375 0 Td
(Millipore)Tj
-33.698 -1.2982 Td
(filtered)Tj
3.067 0 Td
(water,)Tj
2.6532 0 Td
(vortex)Tj
2.7383 0 Td
(mixed,)Tj
2.9537 0 Td
(briefly)Tj
2.7779 0 Td
(centrifuged,)Tj
5.0059 0 Td
(placed)Tj
2.795 0 Td
(in)Tj
1.0318 0 Td
(a)Tj
0.6576 0 Td
(small)Tj
2.3244 0 Td
(insert)Tj
2.4888 0 Td
(vial,)Tj
1.8482 0 Td
(capped,)Tj
3.2882 0 Td
(placed)Tj
2.7949 0 Td
(in)Tj
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(the)Tj
1.4683 0 Td
(auto)Tj
1.9842 0 Td
(sampler)Tj
3.3959 0 Td
(compartment)Tj
5.6806 0 Td
(\(maintained)Tj
5.1307 0 Td
(at)Tj
0.9524 0 Td
(4C\))Tj
2.092 0 Td
(and)Tj
1.7291 0 Td
(analyzed)Tj
3.702 0 Td
(using)Tj
2.3868 0 Td
(LC-MS/MS.)Tj
5.0626 0 Td
(The)Tj
1.7858 0 Td
(ana-)Tj
-35.3704 -1.3039 Td
(lytes)Tj
2.0012 0 Td
(were)Tj
2.126 0 Td
(separated)Tj
4.0081 0 Td
(on)Tj
1.2756 0 Td
(an)Tj
1.2019 0 Td
(Agilent)Tj
3.1521 0 Td
(Zorbax)Tj
3.1068 0 Td
(Eclipse)Tj
3.0273 0 Td
(Plus)Tj
1.9162 0 Td
(C18)Tj
1.8312 0 Td
(HPLC)Tj
2.7496 0 Td
(column)Tj
3.2938 0 Td
(using)Tj
2.3868 0 Td
(gradient)Tj
3.5603 0 Td
(elu-)Tj
-35.6369 -1.2983 Td
(tion.)Tj
2.0636 0 Td
(The)Tj
1.7858 0 Td
(column)Tj
3.2881 0 Td
(oven)Tj
2.1544 0 Td
(was)Tj
1.7007 0 Td
(set)Tj
1.304 0 Td
(at)Tj
0.9524 0 Td
(40C,)Tj
2.4491 0 Td
(a)Tj
0.6577 0 Td
(sample)Tj
3.0217 0 Td
(volume)Tj
3.1974 0 Td
(of)Tj
1.0148 0 Td
(2)Tj
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0.6973 0 Td
()Tj
/F7 1 Tf
(L)Tj
1.2643 0 Td
(was)Tj
1.6951 0 Td
(injected)Tj
3.3902 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.474 0 Td
(flow)Tj
1.9446 0 Td
(rate)Tj
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(was)Tj
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(set)Tj
1.3039 0 Td
(to)Tj
1.0261 0 Td
(0.2)Tj
1.3947 0 Td
(mL/min.)Tj
3.77 0 Td
(Mobile)Tj
3.0614 0 Td
(phase)Tj
2.4888 0 Td
(A)Tj
0.9014 0 Td
(consisted)Tj
3.9402 0 Td
(of)Tj
1.0148 0 Td
(water)Tj
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(with)Tj
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(0.1%)Tj
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(formic)Tj
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(acid)Tj
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(and)Tj
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(B)Tj
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(con-)Tj
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(sisted)Tj
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(of)Tj
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(acetonitrile)Tj
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(with)Tj
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(0.1%)Tj
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(formic)Tj
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(acid.)Tj
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(The)Tj
1.7801 0 Td
(drug)Tj
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(analytes)Tj
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(were)Tj
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(monitored)Tj
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(using)Tj
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(multiple-)Tj
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(reaction)Tj
3.4922 0 Td
(monitoring)Tj
4.8189 0 Td
(mode)Tj
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(for)Tj
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(positive)Tj
3.3165 0 Td
(ions)Tj
1.9049 0 Td
(except)Tj
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(for)Tj
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(efavirenz)Tj
3.8437 0 Td
(which)Tj
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(was)Tj
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(monitored)Tj
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(in)Tj
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(the)Tj
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(negative)Tj
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(ion)Tj
1.542 0 Td
(scan)Tj
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(mode.)Tj
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(Analyst)Tj
3.2485 0 Td
(software,)Tj
3.8097 0 Td
(version)Tj
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(1.6.2)Tj
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(was)Tj
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(used)Tj
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(for)Tj
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(quantitative)Tj
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(data)Tj
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(analysis.)Tj
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3.4752 0 Td
(values)Tj
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(for)Tj
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(EFV,)Tj
2.2394 0 Td
(FTC)Tj
2.0182 0 Td
(and)Tj
1.7292 0 Td
(TFV)Tj
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(were)Tj
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(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(range)Tj
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(of)Tj
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(3)Tj
0.6973 0 Td
(ng/mL)Tj
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(and)Tj
1.7348 0 Td
(this)Tj
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(was)Tj
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(set)Tj
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(as)Tj
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(the)Tj
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(detection)Tj
3.9344 0 Td
(limit.)Tj
ET
Q
q
1 j
1 J
0.0001 w
11.9999 0 0 11.9999 200.012 383.698 cm
BT
/F2 1 Tf
-0.0033 Tc
1 TL
0 0 Td
(Results)Tj
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0 -1.4976 Td
(Prevalence)Tj
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(of)Tj
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(CSF)Tj
1.9418 0 Td
(escape)Tj
ET
Q
q
1 j
1 J
0.0001 w
10 0 0 10 200.012 348.718 cm
BT
/F7 1 Tf
-0.004 Tc
1 TL
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(We)Tj
1.61 0 Td
(sampled)Tj
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(CSF)Tj
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(and)Tj
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(matched)Tj
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(blood)Tj
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(from)Tj
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(participants)Tj
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(living)Tj
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(with)Tj
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(HIV)Tj
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()Tj
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(their)Tj
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(diagnostic)Tj
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(workup)Tj
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(Durban,)Tj
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(South)Tj
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(Africa)Tj
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(Table)Tj
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(1)Tj
0 g
(\).)Tj
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(majority)Tj
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(\()Tj
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()Tj
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(80)Tj
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(or)Tj
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(51%\))Tj
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(of)Tj
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(participants)Tj
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(showed)Tj
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(an)Tj
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(undetectable)Tj
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(HIV)Tj
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(viral)Tj
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(load)Tj
ET
Q
q
0.45 0.36 0.35 0.01 k
1 j
1 J
0.0001 w
36 294.614 m
576 294.614 l
576 294.114 l
36 294.114 l
f*
99.868 294.114 m
99.868 272.114 l
99.3679 272.114 l
99.3679 294.114 l
f*
99.3679 272.114 m
36 272.114 l
36 272.614 l
99.3679 272.614 l
f*
139.493 294.114 m
139.493 272.114 l
138.993 272.114 l
138.993 294.114 l
f*
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99.868 272.114 l
99.868 272.614 l
138.993 272.614 l
f*
180.613 294.114 m
180.613 272.114 l
180.113 272.114 l
180.113 294.114 l
f*
180.113 272.114 m
139.493 272.114 l
139.493 272.614 l
180.113 272.614 l
f*
240.838 294.114 m
240.838 272.114 l
240.338 272.114 l
240.338 294.114 l
f*
240.338 272.114 m
180.613 272.114 l
180.613 272.614 l
240.338 272.614 l
f*
308.706 294.114 m
308.706 272.114 l
308.206 272.114 l
308.206 294.114 l
f*
308.206 272.114 m
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308.206 272.614 l
f*
361.385 294.114 m
361.385 272.114 l
360.885 272.114 l
360.885 294.114 l
f*
360.885 272.114 m
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360.885 272.614 l
f*
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413.73 294.114 l
f*
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f*
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489.669 294.114 l
f*
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489.669 272.614 l
f*
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490.169 272.614 l
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f*
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99.868 247.114 l
99.3679 247.114 l
99.3679 272.114 l
f*
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36 247.114 l
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99.3679 247.614 l
f*
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139.493 259.614 l
138.993 259.614 l
138.993 272.114 l
f*
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99.868 259.614 l
99.868 260.114 l
138.993 260.114 l
f*
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180.613 259.614 l
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f*
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139.493 259.614 l
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180.113 260.114 l
f*
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240.838 259.614 l
240.338 259.614 l
240.338 272.114 l
f*
240.338 259.614 m
180.613 259.614 l
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f*
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308.706 259.614 l
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f*
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f*
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361.385 259.614 l
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f*
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f*
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414.23 259.614 l
413.73 259.614 l
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f*
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f*
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490.169 259.614 l
489.669 259.614 l
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f*
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489.669 260.114 l
f*
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f*
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