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Jo E. Lewis, John M. Brameld, Phil Hill, Cristina Cocco, Barbara Noli, Gian-Luca Ferri, Perry Barrett, Francis J. P. Ebling, Preeti H. Jethwa
Hypothalamic over-expression of VGF in the Siberian hamster increases energy expenditure and reduces body weight gain
www.plosone.org
www.plosone.org
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(cell)Tj
1.5591 0 Td
(line,)Tj
1.9275 0 Td
(the)Tj
-35.6652 -1.304 Td
(pAAV-VGF-GFP)Tj
7.2396 0 Td
(plasmid)Tj
3.3959 0 Td
(was)Tj
1.6951 0 Td
(packaged)Tj
3.9401 0 Td
(into)Tj
1.8425 0 Td
(AAV-2)Tj
3.1124 0 Td
(by)Tj
1.1792 0 Td
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(BioLabs)Tj
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(\(PA,)Tj
2.0296 0 Td
(USA\))Tj
2.4548 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
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(packaged)Tj
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(AAV-GFP)Tj
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(control)Tj
3.1238 0 Td
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1.7008 0 Td
(purchased)Tj
4.3313 0 Td
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2.2053 0 Td
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2.9424 0 Td
(Biolabs)Tj
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(\(PA,)Tj
2.0296 0 Td
(USA\).)Tj
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Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 555.137 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Infusion)Tj
3.6803 0 Td
(of)Tj
1.0441 0 Td
(viral)Tj
2.0268 0 Td
(constructs)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 538.1291 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Animal)Tj
3.2201 0 Td
(surgical)Tj
3.3165 0 Td
(procedures)Tj
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(described)Tj
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(16)Tj
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(,)Tj
0.83 0.64 0.02 0 k
1.7405 0 Td
(17)Tj
0 g
(].)Tj
1.7348 0 Td
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(a)Tj
0.6576 0 Td
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2.2054 0 Td
(stereotaxic)Tj
4.49 0 Td
(frame)Tj
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(\(David)Tj
2.982 0 Td
(Kopf)Tj
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5.3405 0 Td
(NY,)Tj
1.8141 0 Td
(USA\))Tj
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1.9956 0 Td
(the)Tj
1.4683 0 Td
(incisor)Tj
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(bar)Tj
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(positioned)Tj
4.4447 0 Td
(level)Tj
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(with)Tj
2.0013 0 Td
(the)Tj
1.4683 0 Td
(interaural)Tj
4.1386 0 Td
(line)Tj
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(under)Tj
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(general)Tj
3.1351 0 Td
(anaesthesia)Tj
4.7282 0 Td
(\(0.52.5%)Tj
4.1896 0 Td
(isoflurane\).)Tj
-31.9746 -1.3039 Td
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1.7008 0 Td
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1.3777 0 Td
(subcutaneous)Tj
5.6692 0 Td
(injection)Tj
3.7587 0 Td
(of)Tj
1.0148 0 Td
(carprofen)Tj
4.1046 0 Td
(\(50)Tj
1.508 0 Td
(mg/kg)Tj
2.8006 0 Td
(Rimadyl,)Tj
3.8154 0 Td
(Pfizer,)Tj
-34.6164 -1.2983 Td
(Kent,)Tj
2.3867 0 Td
(UK\).)Tj
2.1997 0 Td
(Using)Tj
2.5965 0 Td
(the)Tj
1.4683 0 Td
(sutures)Tj
3.0898 0 Td
(confluence)Tj
4.5807 0 Td
(bregma)Tj
3.2429 0 Td
(as)Tj
1.0147 0 Td
(a)Tj
0.652 0 Td
(landmark,)Tj
4.3257 0 Td
(a)Tj
0.6519 0 Td
(small)Tj
2.3244 0 Td
(hole)Tj
1.9276 0 Td
(was)Tj
1.7007 0 Td
(drilled)Tj
2.829 0 Td
(on)Tj
-34.9906 -1.2982 Td
(midline)Tj
3.3165 0 Td
(and)Tj
1.7348 0 Td
(the)Tj
1.4683 0 Td
(dura)Tj
2.0806 0 Td
(mater)Tj
2.5682 0 Td
(was)Tj
1.6951 0 Td
(pierced)Tj
3.1748 0 Td
(just)Tj
1.6667 0 Td
(lateral)Tj
2.6703 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(mid-sagittal)Tj
4.989 0 Td
(sinus.)Tj
2.5115 0 Td
(A)Tj
0.907 0 Td
(drawn)Tj
2.7836 0 Td
(glass)Tj
-34.0608 -1.3039 Td
(capillary)Tj
3.6113 0 Td
(microinjector)Tj
5.7089 0 Td
(\(30-micron)Tj
4.7962 0 Td
(tip)Tj
1.3039 0 Td
(diameter\))Tj
4.1216 0 Td
(was)Tj
1.7008 0 Td
(lowered)Tj
3.4015 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.474 0 Td
(correct)Tj
3.0331 0 Td
(location.)Tj
3.6793 0 Td
(Using)Tj
2.5966 0 Td
(a)Tj
-36.4533 -1.2983 Td
(Nanolitre)Tj
4.0535 0 Td
(Injection)Tj
3.8324 0 Td
(system)Tj
2.9367 0 Td
(\(WPI,)Tj
2.6475 0 Td
(Stevenage,)Tj
4.3937 0 Td
(UK\))Tj
1.9729 0 Td
(200nl)Tj
2.4378 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(viral)Tj
1.9899 0 Td
(vector)Tj
2.6985 0 Td
(\(AAV-GFP,)Tj
5.0003 0 Td
(n)Tj
0.771 0 Td
(=)Tj
0.7994 0 Td
(3)Tj
-36.0224 -1.2983 Td
(or)Tj
1.0941 0 Td
(AAV-VGF-GFP,)Tj
6.9505 0 Td
(n)Tj
0.771 0 Td
(=)Tj
0.7994 0 Td
(4\))Tj
1.0375 0 Td
(was)Tj
1.6951 0 Td
(directed)Tj
3.4866 0 Td
(towards)Tj
3.4015 0 Td
(the)Tj
1.4684 0 Td
(PVN)Tj
2.211 0 Td
(\(anteroposterior)Tj
6.7634 0 Td
(+0.03,)Tj
2.6816 0 Td
(mediolat-)Tj
-32.3601 -1.3039 Td
(eral)Tj
/F11 1 Tf
1.6951 0 Td
[()]TJ
/F5 1 Tf
0.788 0 Td
(0.03,)Tj
2.0976 0 Td
(dorsoventral)Tj
5.2724 0 Td
(-0.58)Tj
2.2224 0 Td
(\(co-ordinates)Tj
5.5785 0 Td
(from)Tj
2.2054 0 Td
([)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(]\).)Tj
2.4151 0 Td
(Infusions)Tj
3.9684 0 Td
(were)Tj
2.126 0 Td
(over)Tj
1.9729 0 Td
(two)Tj
1.7121 0 Td
(minutes,)Tj
-32.0539 -1.2983 Td
(however)Tj
3.6113 0 Td
(the)Tj
1.4683 0 Td
(glass)Tj
2.0863 0 Td
(microinjector)Tj
5.7033 0 Td
(was)Tj
1.7007 0 Td
(kept)Tj
1.9559 0 Td
(in)Tj
1.0375 0 Td
(place)Tj
2.262 0 Td
(for)Tj
1.389 0 Td
(an)Tj
1.2019 0 Td
(additional)Tj
4.2746 0 Td
(5)Tj
0.6973 0 Td
(minutes)Tj
3.4639 0 Td
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1.0262 0 Td
(allow)Tj
2.3414 0 Td
(for)Tj
1.3833 0 Td
(dif-)Tj
-35.6029 -1.2982 Td
(fusion)Tj
2.7155 0 Td
(and)Tj
1.7348 0 Td
(prevention)Tj
4.5751 0 Td
(of)Tj
1.0148 0 Td
(backflow)Tj
3.8097 0 Td
(through)Tj
3.4469 0 Td
(the)Tj
1.4684 0 Td
(cannula)Tj
3.3789 0 Td
(track,)Tj
2.4718 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(incision)Tj
3.4129 0 Td
(was)Tj
1.6951 0 Td
(closed)Tj
-32.9213 -1.3039 Td
(using)Tj
2.3867 0 Td
(Michel)Tj
3.0047 0 Td
(clips.)Tj
2.2564 0 Td
(The)Tj
1.7858 0 Td
(surgically-prepared)Tj
7.954 0 Td
(Siberian)Tj
3.5092 0 Td
(hamsters)Tj
3.8211 0 Td
(were)Tj
2.1203 0 Td
(allowed)Tj
3.2882 0 Td
(a)Tj
0.6519 0 Td
(seven)Tj
2.4321 0 Td
(day)Tj
1.6385 0 Td
(recov-)Tj
-34.8489 -1.2983 Td
(ery)Tj
1.4683 0 Td
(period,)Tj
3.0614 0 Td
(during)Tj
2.9253 0 Td
(which)Tj
2.6532 0 Td
(they)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(handled)Tj
3.4583 0 Td
(on)Tj
1.2755 0 Td
(a)Tj
0.652 0 Td
(daily)Tj
2.1487 0 Td
(basis,)Tj
2.3754 0 Td
(received)Tj
3.5376 0 Td
(analgesia)Tj
3.8324 0 Td
(and)Tj
1.7291 0 Td
(had)Tj
1.7121 0 Td
(access)Tj
-34.8715 -1.2983 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.6519 0 Td
(palatable)Tj
3.7588 0 Td
(diet)Tj
1.7404 0 Td
(consisting)Tj
4.2633 0 Td
(of)Tj
1.0148 0 Td
(soaked)Tj
2.9707 0 Td
(Teklab)Tj
2.9367 0 Td
(diet.)Tj
1.9615 0 Td
(Over-expression)Tj
6.8145 0 Td
(of)Tj
1.0148 0 Td
(VGF)Tj
2.16 0 Td
(mRNA)Tj
3.0784 0 Td
(and)Tj
1.7291 0 Td
(eGFP)Tj
-35.121 -1.2982 Td
(was)Tj
1.6951 0 Td
(previously)Tj
4.3539 0 Td
(described)Tj
4.0536 0 Td
(\(16\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 313.1716 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
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4.4457 0 Td
(gases)Tj
2.8582 0 Td
(and)Tj
1.8615 0 Td
(feeding)Tj
3.4583 0 Td
(behaviour)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 296.2204 cm
BT
/F5 1 Tf
1 TL
0.01 Tc
0 0 Td
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3.7473 0 Td
(respiratory)Tj
4.7509 0 Td
(and)Tj
1.7744 0 Td
(feeding)Tj
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(behaviour)Tj
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2.1827 0 Td
(measured)Tj
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(using)Tj
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(a)Tj
0.669 0 Td
(Comprehen-)Tj
-32.241 -1.3039 Td
(sive)Tj
1.7801 0 Td
(Lab)Tj
1.7405 0 Td
(Animal)Tj
3.3051 0 Td
(Monitoring)Tj
5.0343 0 Td
(System)Tj
3.1351 0 Td
(\(CLAMS;)Tj
4.1386 0 Td
(Linton)Tj
3.0104 0 Td
(Instrumentation,)Tj
7.2623 0 Td
(Linton,)Tj
3.2485 0 Td
(UK,)Tj
1.8992 0 Td
(and)Tj
-34.5541 -1.2982 Td
(Columbus)Tj
4.49 0 Td
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5.5105 0 Td
(Columbus,)Tj
4.7338 0 Td
(OH,)Tj
2.0013 0 Td
(USA\))Tj
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1.0431 0 Td
(described)Tj
4.1783 0 Td
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([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7915 0 Td
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.44 0 Td
(19)Tj
0 g
(].)Tj
1.7915 0 Td
(This)Tj
2.0409 0 Td
(is)Tj
-36.0224 -1.2983 Td
(an)Tj
1.2302 0 Td
(open-circuit)Tj
5.3121 0 Td
(calorimeter)Tj
4.9606 0 Td
(configured)Tj
4.7055 0 Td
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1.4286 0 Td
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2.3981 0 Td
(rodents,)Tj
3.5943 0 Td
(where)Tj
2.7213 0 Td
(the)Tj
1.5136 0 Td
(rodents)Tj
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(were)Tj
2.1826 0 Td
(individu-)Tj
-33.3975 -1.3039 Td
(ally)Tj
1.6554 0 Td
(housed)Tj
3.1804 0 Td
(with)Tj
2.0523 0 Td
(food)Tj
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(hoppers)Tj
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(in)Tj
1.0658 0 Td
(the)Tj
1.508 0 Td
(centre)Tj
2.795 0 Td
(of)Tj
1.0431 0 Td
(each)Tj
2.0863 0 Td
(cage)Tj
2.0182 0 Td
(containing)Tj
4.6715 0 Td
(chow)Tj
2.4151 0 Td
(ground)Tj
3.2542 0 Td
(into)Tj
1.8935 0 Td
(a)Tj
-35.2854 -1.2983 Td
(rough)Tj
2.6872 0 Td
(powder,)Tj
3.5773 0 Td
(and)Tj
1.7688 0 Td
(dropper-style)Tj
5.7713 0 Td
(water)Tj
2.5001 0 Td
(bottles.)Tj
3.1975 0 Td
(Metabolic)Tj
4.3483 0 Td
(parameters)Tj
4.8132 0 Td
(measured)Tj
4.2179 0 Td
(included)Tj
-32.8816 -1.2982 Td
(oxygen)Tj
3.1691 0 Td
(consumption)Tj
5.6976 0 Td
(\(VO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 306.8787 216.5102 cm
BT
/F5 1 Tf
1 TL
0.0133 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 310.6204 218.211 cm
BT
/F5 1 Tf
1 TL
0.01 Tc
0 0 Td
(\))Tj
0.5725 0 Td
(and)Tj
1.7745 0 Td
(carbon)Tj
3.0898 0 Td
(dioxide)Tj
3.3051 0 Td
(production)Tj
4.8586 0 Td
(\(VCO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 471.6283 216.5102 cm
BT
/F5 1 Tf
1 TL
0.0133 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 475.3133 218.211 cm
BT
/F5 1 Tf
1 TL
0.01 Tc
0 0 Td
(\),)Tj
0.8107 0 Td
(normalised)Tj
4.8698 0 Td
(over)Tj
2.0296 0 Td
(the)Tj
-35.24 -1.3039 Td
(estimated)Tj
4.1953 0 Td
(lean)Tj
1.9332 0 Td
(mass)Tj
2.2507 0 Td
(\(BW)Tj
2.143 0 Td
(0.75\))Tj
2.2847 0 Td
(due)Tj
1.7405 0 Td
(to)Tj
1.0544 0 Td
(the)Tj
1.5081 0 Td
(change)Tj
3.1294 0 Td
(in)Tj
1.0658 0 Td
(body)Tj
2.2677 0 Td
(composition,)Tj
5.6408 0 Td
(such)Tj
2.1147 0 Td
(that)Tj
1.8311 0 Td
(the)Tj
1.5137 0 Td
(energy)Tj
-34.6731 -1.2982 Td
(expenditure)Tj
5.1647 0 Td
(\(EE\))Tj
2.0977 0 Td
(and)Tj
1.7688 0 Td
(respiratory)Tj
4.7508 0 Td
(exchange)Tj
4.0479 0 Td
(ratio)Tj
2.1656 0 Td
(\(RER\))Tj
2.7836 0 Td
(could)Tj
2.5228 0 Td
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1.1736 0 Td
(calculated)Tj
4.3482 0 Td
(as)Tj
1.0431 0 Td
(previously)Tj
-31.8668 -1.2983 Td
(described)Tj
4.1783 0 Td
([)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(].)Tj
2.143 0 Td
(Feeding)Tj
3.5036 0 Td
(behaviour)Tj
4.3653 0 Td
(parameters)Tj
4.8132 0 Td
(measured)Tj
4.2179 0 Td
(included)Tj
3.8268 0 Td
(the)Tj
1.5136 0 Td
(timing)Tj
2.965 0 Td
(and)Tj
1.7745 0 Td
(duration)Tj
-33.3012 -1.2983 Td
(of)Tj
1.0432 0 Td
(feeding,)Tj
3.4979 0 Td
(individual)Tj
4.4277 0 Td
(meal)Tj
2.1997 0 Td
(size,)Tj
1.9785 0 Td
(and)Tj
1.7745 0 Td
(total)Tj
2.075 0 Td
(food)Tj
2.1033 0 Td
(intake)Tj
2.7722 0 Td
(per)Tj
1.5761 0 Td
(unit)Tj
1.9162 0 Td
(time.)Tj
2.3243 0 Td
(A)Tj
0.9184 0 Td
(meal)Tj
2.1997 0 Td
(equal)Tj
2.4264 0 Td
(to)Tj
1.0545 0 Td
(or)Tj
-34.2876 -1.3039 Td
(greater)Tj
3.1011 0 Td
(than)Tj
2.0863 0 Td
(0.02g)Tj
2.4094 0 Td
(is)Tj
0.8731 0 Td
(considered)Tj
4.7395 0 Td
(to)Tj
1.0545 0 Td
(be)Tj
1.1792 0 Td
(a)Tj
0.669 0 Td
(feeding)Tj
3.2598 0 Td
(bout.)Tj
2.3584 0 Td
(Ambulatory)Tj
5.21 0 Td
(\(locomotor\))Tj
5.2383 0 Td
(activity)Tj
3.2315 0 Td
(was)Tj
-35.4101 -1.2982 Td
(also)Tj
1.8199 0 Td
(measured)Tj
4.2236 0 Td
(continuously)Tj
5.6069 0 Td
(using)Tj
2.4548 0 Td
(two)Tj
1.7518 0 Td
(sets)Tj
1.7234 0 Td
(of)Tj
1.0431 0 Td
(infrared)Tj
3.5603 0 Td
(beams)Tj
2.8347 0 Td
(traversing)Tj
4.3539 0 Td
(each)Tj
2.0862 0 Td
(cage)Tj
2.024 0 Td
(that)Tj
1.8311 0 Td
(mea-)Tj
-35.3137 -1.2983 Td
(sure)Tj
1.9559 0 Td
(linear)Tj
2.5966 0 Td
(and)Tj
1.7744 0 Td
(vertical)Tj
3.2485 0 Td
(movement.)Tj
4.8642 0 Td
(The)Tj
1.8255 0 Td
(system)Tj
3.0218 0 Td
(was)Tj
1.7404 0 Td
(operated)Tj
3.8381 0 Td
(with)Tj
2.0523 0 Td
(an)Tj
1.2301 0 Td
(air)Tj
1.3323 0 Td
(intake)Tj
2.7722 0 Td
(of)Tj
1.0432 0 Td
(0.6)Tj
1.44 0 Td
(Lmin)Tj
-34.7355 -1.3039 Td
(for)Tj
1.4287 0 Td
(each)Tj
2.0806 0 Td
(chamber,)Tj
4.0649 0 Td
(and)Tj
1.7744 0 Td
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