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Emanuel Ott, Yuko Kawaguchi, Denise Kölbl, Palak Chaturvedi, Kazumichi Nakagawa, Akihiko Yamagishi, Wolfram Weckwerth, Tetyana Milojevic
Proteometabolomic response of Deinococcus radiodurans exposed to UVC and vacuum conditions: Initial studies prior to the Tanpopo space mission
www.plosone.org
www.plosone.org
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1.1735 0 Td
(mM)Tj
1.9332 0 Td
(phosphate)Tj
4.3313 0 Td
(buffer)Tj
2.6249 0 Td
(three)Tj
2.2677 0 Td
(times)Tj
2.3867 0 Td
(and)Tj
1.7292 0 Td
(finally)Tj
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(resuspended)Tj
5.2384 0 Td
(in)Tj
1.0318 0 Td
(ice-)Tj
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(cold)Tj
1.9218 0 Td
(1)Tj
0.6973 0 Td
(mL)Tj
1.5761 0 Td
(MCW)Tj
2.7439 0 Td
(\(methanol:chloroform:wa)Tj
10.284 0 Td
(ter)Tj
1.3096 0 Td
(2.5:1:0.5\).)Tj
4.0649 0 Td
(0.5)Tj
1.4003 0 Td
(g)Tj
0.6803 0 Td
(of)Tj
1.0148 0 Td
(FastPrepTM)Tj
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(lysing)Tj
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(matrix)Tj
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(B)Tj
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(\(MP)Tj
2.0125 0 Td
(Biomedicals,)Tj
5.3008 0 Td
(Santa)Tj
2.4094 0 Td
(Ana,)Tj
2.1147 0 Td
(USA\))Tj
2.4547 0 Td
(was)Tj
1.6952 0 Td
(added)Tj
2.6645 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(mixture)Tj
3.3902 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
1.4683 0 Td
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(homogenized)Tj
-31.7818 -1.2982 Td
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(a)Tj
0.652 0 Td
(FastPrepTM-24)Tj
6.5196 0 Td
(Instrument)Tj
4.7565 0 Td
(\(MP)Tj
2.0126 0 Td
(Biomedicals,)Tj
5.3008 0 Td
(Santa)Tj
2.4094 0 Td
(Ana,)Tj
2.1146 0 Td
(USA\))Tj
2.4548 0 Td
(at)Tj
0.9525 0 Td
(3x4.5)Tj
2.3414 0 Td
(m/s)Tj
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(30)Tj
1.1735 0 Td
(s)Tj
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1.9955 0 Td
(a)Tj
0.652 0 Td
(5)Tj
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(min)Tj
1.8539 0 Td
(cooldown)Tj
4.1725 0 Td
(on)Tj
1.2756 0 Td
(ice)Tj
1.3323 0 Td
(between)Tj
3.5319 0 Td
(the)Tj
1.4684 0 Td
(cycles.)Tj
2.7779 0 Td
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2.2847 0 Td
(centrifugation)Tj
5.8904 0 Td
(\(21000)Tj
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0.9128 0 Td
(15)Tj
1.1679 0 Td
(min,)Tj
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1.8425 0 Td
(a)Tj
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(new)Tj
1.8765 0 Td
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4.989 0 Td
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1.7008 0 Td
(stored)Tj
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0.9525 0 Td
(4C)Tj
1.7518 0 Td
(for)Tj
1.3889 0 Td
(the)Tj
1.4684 0 Td
(subsequent)Tj
4.7055 0 Td
(extraction.)Tj
-31.4927 -1.3039 Td
(Phase)Tj
2.5228 0 Td
(separation)Tj
4.3823 0 Td
(was)Tj
1.7008 0 Td
(induced)Tj
3.4639 0 Td
(by)Tj
1.1849 0 Td
(adding)Tj
2.9877 0 Td
(200)Tj
/F14 1 Tf
1.6497 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2643 0 Td
(of)Tj
1.0148 0 Td
(water.)Tj
2.6532 0 Td
(The)Tj
1.7858 0 Td
(phases)Tj
2.8403 0 Td
(were)Tj
2.126 0 Td
(separated)Tj
4.0081 0 Td
(in)Tj
1.0375 0 Td
(differ-)Tj
-34.6221 -1.2983 Td
(ent)Tj
1.491 0 Td
(tubes)Tj
2.3357 0 Td
(and)Tj
1.7291 0 Td
(dried)Tj
2.3358 0 Td
(in)Tj
1.0374 0 Td
(a)Tj
0.6577 0 Td
(vacuum)Tj
3.4072 0 Td
(concentrator.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 419.0173 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Derivatisation)Tj
6.1984 0 Td
(and)Tj
1.8662 0 Td
(analysis)Tj
3.7843 0 Td
(of)Tj
1.0441 0 Td
(the)Tj
1.5921 0 Td
(metabolites)Tj
5.2725 0 Td
(with)Tj
1.9795 0 Td
(GC-TOF-MS)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 402.0094 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Polar)Tj
2.33 0 Td
(metabolites)Tj
4.7962 0 Td
(were)Tj
2.1146 0 Td
(dissolved)Tj
3.8891 0 Td
(in)Tj
1.0262 0 Td
(10)Tj
/F14 1 Tf
1.1622 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2586 0 Td
(of)Tj
1.0034 0 Td
(40)Tj
1.1679 0 Td
(mg)Tj
1.4967 0 Td
(mL-1)Tj
2.3924 0 Td
(methoxyamine-hydroch)Tj
9.6831 0 Td
(loride)Tj
2.5511 0 Td
(in)Tj
1.0318 0 Td
(pyr-)Tj
-35.9033 -1.2982 Td
(idine)Tj
2.2506 0 Td
(through)Tj
3.447 0 Td
(shaking)Tj
3.3221 0 Td
(at)Tj
0.9525 0 Td
(650)Tj
1.6441 0 Td
(rpm)Tj
1.9275 0 Td
(at)Tj
0.9524 0 Td
(30C)Tj
2.2281 0 Td
(for)Tj
1.3889 0 Td
(90)Tj
1.1679 0 Td
(min.)Tj
2.0806 0 Td
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5.7486 0 Td
(40)Tj
/F14 1 Tf
1.1679 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2642 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.6577 0 Td
(silylation)Tj
3.8494 0 Td
(mix)Tj
-35.0643 -1.2983 Td
(\(1)Tj
1.0318 0 Td
(mL)Tj
1.5817 0 Td
(N-methyl-N-trimethylsilyltr)Tj
11.2478 0 Td
(ifluoroacetamid)Tj
6.5593 0 Td
(spiked)Tj
2.812 0 Td
(with)Tj
2.0012 0 Td
(30)Tj
/F14 1 Tf
1.1679 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2642 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.6576 0 Td
(mix)Tj
1.7689 0 Td
(of)Tj
1.0148 0 Td
(even-number)Tj
-32.122 -1.3039 Td
(alkanes)Tj
3.1634 0 Td
(\(C10-C40\)\))Tj
4.8189 0 Td
(was)Tj
1.7007 0 Td
(added)Tj
2.6589 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(mixture)Tj
3.3959 0 Td
(was)Tj
1.7007 0 Td
(incubated)Tj
4.1726 0 Td
(for)Tj
1.389 0 Td
(30)Tj
1.1735 0 Td
(min)Tj
1.8539 0 Td
(at)Tj
0.9524 0 Td
(an)Tj
1.2019 0 Td
(agitation)Tj
3.736 0 Td
(rate)Tj
-35.1153 -1.2983 Td
(of)Tj
1.0147 0 Td
(650)Tj
1.6441 0 Td
(rpm)Tj
1.9276 0 Td
(at)Tj
0.9524 0 Td
(37C.)Tj
2.4548 0 Td
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2.2847 0 Td
(centrifugation)Tj
5.8904 0 Td
(\(14000)Tj
2.9366 0 Td
(g,)Tj
0.9128 0 Td
(2)Tj
0.6916 0 Td
(min\),)Tj
2.4208 0 Td
(the)Tj
1.4684 0 Td
(supernatant)Tj
4.9832 0 Td
(was)Tj
1.7008 0 Td
(transferred)Tj
4.6375 0 Td
(into)Tj
-35.9204 -1.2982 Td
(a)Tj
0.6519 0 Td
(glass)Tj
2.0863 0 Td
(vial)Tj
1.6214 0 Td
(and)Tj
1.7348 0 Td
(1)Tj
/F14 1 Tf
0.6916 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2643 0 Td
(of)Tj
1.0148 0 Td
(it)Tj
0.788 0 Td
(was)Tj
1.7008 0 Td
(injected)Tj
3.3845 0 Td
(into)Tj
1.8425 0 Td
(the)Tj
1.4684 0 Td
(GC)Tj
1.5987 0 Td
(\(Agilent)Tj
/F16 1 Tf
(1)Tj
/F7 1 Tf
4.4901 0 Td
(6890)Tj
2.1259 0 Td
(gas)Tj
1.4797 0 Td
(chromatograph\))Tj
6.7634 0 Td
(in)Tj
-34.7071 -1.3039 Td
(splitless)Tj
3.2995 0 Td
(injection)Tj
3.7587 0 Td
(mode.)Tj
-5.862 -1.2983 Td
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1.627 0 Td
(separation)Tj
4.3824 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(metabolites,)Tj
5.0343 0 Td
(an)Tj
1.2019 0 Td
(Agilent)Tj
3.1521 0 Td
(HP-5MS)Tj
3.7417 0 Td
(column)Tj
3.2938 0 Td
(\(30)Tj
1.5081 0 Td
(m)Tj
1.0431 0 Td
(length,)Tj
2.9537 0 Td
(0.25)Tj
1.8708 0 Td
(mm)Tj
-33.4882 -1.2983 Td
(diameter)Tj
3.7813 0 Td
(and)Tj
1.7348 0 Td
(0.25)Tj
/F14 1 Tf
1.8709 0 Td
[()]TJ
/F7 1 Tf
(m)Tj
1.5477 0 Td
(film\))Tj
2.177 0 Td
(was)Tj
1.7008 0 Td
(used.)Tj
2.2847 0 Td
(Further)Tj
3.2711 0 Td
(parameters)Tj
4.6715 0 Td
(were)Tj
2.1203 0 Td
(set)Tj
1.3039 0 Td
(as)Tj
1.0148 0 Td
(following:)Tj
4.2066 0 Td
(flow)Tj
1.9446 0 Td
(rate)Tj
1.7461 0 Td
(1)Tj
0.6973 0 Td
(mL)Tj
-36.0734 -1.2982 Td
(min)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 216.3401 289.474 cm
BT
/F7 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 222.5196 285.0519 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(;)Tj
0.4478 0 Td
(injection)Tj
3.7587 0 Td
(temperature)Tj
5.1307 0 Td
(230C;)Tj
2.9253 0 Td
(column)Tj
3.2939 0 Td
(temperature)Tj
5.1307 0 Td
(started)Tj
2.931 0 Td
(at)Tj
0.9524 0 Td
(70C)Tj
2.228 0 Td
(for)Tj
1.3833 0 Td
(one)Tj
1.7008 0 Td
(minute,)Tj
3.3278 0 Td
(then)Tj
-35.461 -1.3039 Td
(heated)Tj
2.8515 0 Td
(up)Tj
1.2642 0 Td
(to)Tj
1.0262 0 Td
(330C)Tj
2.7042 0 Td
(in)Tj
1.0375 0 Td
(9)Tj
0.6916 0 Td
(min,)Tj
2.0806 0 Td
(where)Tj
2.6533 0 Td
(it)Tj
0.7823 0 Td
(was)Tj
1.7008 0 Td
(hold)Tj
2.0296 0 Td
(for)Tj
1.3833 0 Td
(8)Tj
0.6973 0 Td
(min;)Tj
2.0806 0 Td
(recorded)Tj
3.7984 0 Td
(masses)Tj
2.9764 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(LECO)Tj
2.7382 0 Td
(Pega-)Tj
-35.0018 -1.2982 Td
(sus)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 212.485 263.4519 cm
BT
/F16 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 222.1795 259.0299 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(4D)Tj
1.4286 0 Td
(GC)Tj
/F14 1 Tf
[()]TJ
/F7 1 Tf
(GC-TOF)Tj
5.7826 0 Td
(spectrometer)Tj
5.4595 0 Td
(were)Tj
2.126 0 Td
(set)Tj
1.2983 0 Td
(between)Tj
3.5319 0 Td
(40700)Tj
3.1124 0 Td
(m/z.)Tj
2.007 0 Td
(Apart)Tj
2.5284 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(samples,)Tj
3.6113 0 Td
(a)Tj
-36.7763 -1.2982 Td
(house)Tj
2.568 0 Td
(intern)Tj
2.6759 0 Td
(standard)Tj
3.7191 0 Td
(mix)Tj
1.7744 0 Td
(of)Tj
1.0148 0 Td
(certain)Tj
2.9877 0 Td
(metabolites)Tj
4.8019 0 Td
(was)Tj
1.7008 0 Td
(measured)Tj
4.1102 0 Td
(to)Tj
1.0261 0 Td
(get)Tj
1.406 0 Td
(level)Tj
2.0126 0 Td
(1)Tj
0.6973 0 Td
(identifications)Tj
5.947 0 Td
(of)Tj
-36.4418 -1.3039 Td
(common)Tj
3.8436 0 Td
(primary)Tj
3.4583 0 Td
(metabolites.)Tj
-6.1057 -1.2983 Td
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6.0206 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(metabolites)Tj
4.8075 0 Td
(were)Tj
2.1203 0 Td
(based)Tj
2.4775 0 Td
(on)Tj
1.2755 0 Td
(matching)Tj
4.0025 0 Td
(the)Tj
1.4684 0 Td
(obtained)Tj
3.736 0 Td
(MS-spectra)Tj
4.7679 0 Td
(and)Tj
-34.3556 -1.2983 Td
(retention)Tj
3.906 0 Td
(times)Tj
2.3867 0 Td
(with)Tj
2.0013 0 Td
(an)Tj
1.2019 0 Td
(in-house)Tj
3.7303 0 Td
(library)Tj
2.8744 0 Td
(\(extended)Tj
4.1952 0 Td
(gmd)Tj
2.0353 0 Td
(database\).)Tj
4.2009 0 Td
(Peak)Tj
2.1373 0 Td
(integration)Tj
4.6488 0 Td
(was)Tj
1.6951 0 Td
(per-)Tj
-35.0132 -1.3039 Td
(formed)Tj
3.1576 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.4684 0 Td
(LECO)Tj
2.7382 0 Td
(ChromaTOF)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 345.7133 198.4818 cm
BT
/F16 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 355.3511 194.0031 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(software.)Tj
3.8154 0 Td
(Metabolites)Tj
4.8755 0 Td
(which)Tj
2.6532 0 Td
(were)Tj
2.1203 0 Td
(also)Tj
1.7688 0 Td
(identified)Tj
4.0592 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
-35.8636 -1.2982 Td
(standard)Tj
3.719 0 Td
(mix)Tj
1.7745 0 Td
(were)Tj
2.1203 0 Td
(considered)Tj
4.6034 0 Td
(a)Tj
0.652 0 Td
(level)Tj
2.0183 0 Td
(1)Tj
0.6915 0 Td
(identification,)Tj
5.811 0 Td
(the)Tj
1.4683 0 Td
(ones)Tj
2.0636 0 Td
(which)Tj
2.6476 0 Td
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2.126 0 Td
(not)Tj
1.5703 0 Td
(present)Tj
3.1635 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
-35.4668 -1.2983 Td
(mix,)Tj
1.9956 0 Td
(but)Tj
1.5533 0 Td
(the)Tj
1.4684 0 Td
(retention)Tj
3.9061 0 Td
(index)Tj
2.4548 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(mass)Tj
2.1939 0 Td
(spectrum)Tj
3.9629 0 Td
(was)Tj
1.7007 0 Td
(similar)Tj
2.9877 0 Td
(to)Tj
1.0262 0 Td
(one)Tj
1.6951 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(database)Tj
3.6396 0 Td
(were)Tj
-34.2705 -1.2982 Td
(considered)Tj
4.5978 0 Td
(a)Tj
0.6576 0 Td
(level)Tj
2.0126 0 Td
(2)Tj
0.6973 0 Td
(identification.)Tj
5.811 0 Td
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1.78 0 Td
(measured)Tj
4.1102 0 Td
(areas)Tj
2.2451 0 Td
(were)Tj
2.1203 0 Td
(normalized)Tj
4.7791 0 Td
(against)Tj
3.0274 0 Td
(the)Tj
1.4684 0 Td
(number)Tj
3.4185 0 Td
(of)Tj
-36.7253 -1.304 Td
(cells,)Tj
2.143 0 Td
(used)Tj
2.0636 0 Td
(for)Tj
1.3833 0 Td
(the)Tj
1.4683 0 Td
(extraction.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 112.9889 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Protein)Tj
3.3496 0 Td
(extraction)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 95.9811 cm
BT
/F7 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(pellets)Tj
2.7382 0 Td
(were)Tj
2.1203 0 Td
(suspended)Tj
4.4391 0 Td
(in)Tj
1.0374 0 Td
(400)Tj
/F14 1 Tf
1.6441 0 Td
[()]TJ
/F7 1 Tf
(L)Tj
1.2643 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.6576 0 Td
(protein)Tj
3.1521 0 Td
(extraction)Tj
4.2576 0 Td
(buffer)Tj
2.6249 0 Td
(\(100)Tj
1.9842 0 Td
(mM)Tj
1.9332 0 Td
(NaCl,)Tj
2.5285 0 Td
(100)Tj
1.6441 0 Td
(mM)Tj
-34.8205 -1.2982 Td
(Tris-HCl)Tj
3.855 0 Td
(pH)Tj
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1 J
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11.9999 0 0 11.9999 200.0125 587.622 cm
BT
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1 TL
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(free)Tj
1.9228 0 Td
(quantification\))Tj
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q
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BT
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1 TL
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1.627 0 Td
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1 J
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1 TL
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4.4504 0 Td
(evaluation)Tj
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Q
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0 w
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BT
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1 TL
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(metabolite)Tj
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1.9956 0 Td
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1.4683 0 Td
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0.8447 0 Td
(packages)Tj
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1 TL
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ET
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1 TL
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(10)Tj
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[()]TJ
/F7 1 Tf
(1)Tj
ET
Q
q
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1 J
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BT
/F7 1 Tf
1 TL
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(s.d.\))Tj
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[( )20(\n)6()12()4()4()12()12( )20(\n)16()]TJ
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(scanning)Tj
-33.5959 -1.2983 Td
(electron)Tj
3.4582 0 Td
(microscopy)Tj
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0.83 0.64 0.02 0 k
(Fig)Tj
1.8198 0 Td
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[()]TJ
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(.)Tj
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[( )20(\n)6()12()4()4()12()12( )20(\n)16()]TJ
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5.0286 0 Td
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(shown)Tj
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(.)Tj
0.9241 0 Td
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