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T*
[(ciency.)-201.9(First,)-198.7(the)-202.7(application)-197.4(of)-202.1(PEF)]TJ
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[(in)-202(vivo)]TJ
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(induces)Tj
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[(immune)-493.1(system)-490.8(responses,)]TJ
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(48)Tj
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T*
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[(in)-375.4(vitro)]TJ
/F6 1 Tf
3.8588 0 TD
[(experiments)-377.6(allowed)-373.1(us)-380.5(to)-377.8(test)-380.2(a)]TJ
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T*
[(ues.)-358.2(Using)-357.6(our)]TJ
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[(setup,)-355(we)-361.1(followed)-352.2(the)-359.9(3R)]TJ
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(principle)Tj
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(49)Tj
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[(of)-381(reduction)-383.6(and)-381.1(signicantly)-378.7(reduced)]TJ
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[(the)-365.3(number)-366(of)-364.7(animals)-363.4(that)-361.9(would)-369.8(be)-362.5(required)-364.6(to)]TJ
T*
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[(parameters)-511.8(on)-511.9(inactivation)-516.8(levels)-509.5(of)-511.1(bacteria)-512.8(in)]TJ
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[(future)-461.7(studies.)-463(The)-463.7(demonstration)-462.3(of)-462.3(the)-462.9(hurdle)]TJ
T*
[(approach)-805.9(for)-812(inactivation)-804.1(of)-809.1(potential)-805.2(wound)]TJ
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T*
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[(cal)-375.2(a).3(nd)-375.4(prophy)-15.2(l)14(a).3(ctic)-375.7(an)-2.8(tibiotics,)]TJ
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[(an)-376.3(enzymatic)-375.3(d)4.4(e-)]TJ
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[(tachment)-703.2(of)-701(biolms,)]TJ
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[(immu)-3.9(no)-4.1(pro)-4.1(p)-2.5(hylaxis)-701.8(a)-.8(n)-3.9(d)]TJ
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[(im)4(mu)3.9(no)3.8(the)3.8(r)-.5(apy,)]TJ
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[(the)-512.1(e)-2.7(mergence)-512.1(of)-511.1(multidrug-resis)4.1(t)-3.9(ant)-513.3(s)-1.3(trains,)]TJ
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T*
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T*
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[(wounds)-277.5(and)-282.7(s)-1.1(urgical)-277.2(m)-2.3(esh)-278.7(d)-.9(is)4.3(infection.)]TJ
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[(25)-7.2(2)-7.2(7)]TJ
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[(MATERIALS)-333.6(AND)-328.2(METHODS)]TJ
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[(Bact)-8.2(erial)-333.3(cu)-7(lture)]TJ
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/F7 1 Tf
5.127 0 TD
[(S)10.2(.)-363.9(ep)21.8(id)14.8(e)13.4(r)0(m)15(i)13.2(di)14.8(s)]TJ
/F6 1 Tf
7.0401 0 TD
[(RP)18.3(62)20.1(A)]TJ
-19.9767 -1.1869 TD
[(\(R)21.3(P6)14.9(2A)-393.8(ki)16.2(n)12.4(d)0(l)16.9(y)-407.4(p)13.8(r)0(o)14.8(v)0(i)18.4(d)13.8(ed)-396.7(by)-397.4(P)10.2(r)0(o)14.8(f)13.2(.)-407.3(M)0(i)17.9(c)0(h)20.4(a)-407.3(F)10.2(r)0(i)16.5(d)13.8(ma)17.1(n)12.4(,)]TJ
0 -1.1923 TD
[(Sc)20.6(ho)19.2(ol)-255.7(of)-256.3(Ch)15(e)12.2(m)12.4(is)16.7(t)11(r)0(y)17.9(,)-266.4(F)10.2(a)0(c)18.1(u)0(l)20.9(t)0(y)-260.9(o)12.2(f)-268.6(E)13.5(xa)19.9(ct)-257.3(Sc)15.2(i)13.9(e)0(n)19.2(c)0(e)14.8(s)13.6(,)-266.4(T)10.2(el)]TJ
T*
[(Av)17.9(iv)-165.9(Un)22.3(iv)18.4(e)12.2(r)0(s)16.2(i)0(t)19.6(y)0(\))-166.6(w)11.4(e)12.2(r)0(e)-164(g)0(r)17.9(o)12.2(wn)-165.8()12.4(r)0(s)16.2(t)-173.2(o)12.2(n)-171.8(el)15.3(e)12.2(c)0(t)19.1(r)0(o)20.3(p)0(o)15.2(r)0(a)18.1(t)11(io)20.7(n)]TJ
0 -1.1869 TD
[(lo)20.7(w)-481.8(s)0(a)18.2(l)0(t)-468.2(\()0(E)15.9(L)10.2(S)12.6(\))-479.9(me)19.3(di)16.9(a-)17.9(ba)20.1(s)13.6(e)0(d)-472.6(s)0(o)20.4(l)0(i)17(d)-479.4(ag)19.9(ar)18.1(.)-483.2(T)10.2(h)12.4(e)-481(EL)18.3(S)]TJ
0 -1.1923 TD
[(me)19.3(di)16.9(a)-277.2(c)0(o)20.3(m)0(p)15.4(o)12.2(si)16.7(t)11(i)0(o)20.7(n)-280.2(wa)16(s)-279.1(a)0(s)-269(f)0(o)14.6(l)13.9(lo)15.3(w)11.4(s)0(:)-269(0)0(.)20.1(1)-158(mg)16.9(/m)22.5(L)-282.4(N)0(a)19.9(C)0(l)]TJ
T*
[(\()-73.5(M)0(e)21.7(r)0(c)16(k)0(,)-540.5(D)0(a)16(r)13.4(ms)15.2(t)11(a)0(d)18.4(t)11(,)-553.6(G)11.4(e)12.2(r)0(m)15(a)0(n)22.5(y)0(\))17.7(,)-553.6(0.)20.1(01)-142.5(g/)14.5(m)12.4(L)-553.4(B)13.5(a)0(c)18.1(t)11(o-)]TJ
0 -1.1869 TD
[(t)11(r)0(y)17.9(p)0(t)19.4(o)0(n)19.2(e)-459.3(\()13.2(A)0(c)16.1(a)0(d)18.4(e)12.2(m)12.4(i)0(a)18.5(,)-456.1(Is)18.5(ra)18.1(e)12.2(l)0(\))16.3(,)-456.1(0.)20.1(00)20.1(5)-152.5(g)0(/)14.5(m)12.4(L)-455.9(y)0(e)16.7(a)0(s)23.6(t)-455(ex)16.7(-)]TJ
0 -1.1923 TD
[(t)11(r)0(a)18.1(c)0(t)-197.7(\()0(B)21.3(D)-210.8(e)12.2(x)0(t)20.9(r)0(a)18.1(c)0(t)-197.7(o)12.2(f)-209(a)0(u)22.5(t)0(o)17.9(l)0(y)18.4(z)0(e)19.7(d)-208.4(ye)22.1(as)18.2(t)11(,)-206.8(Is)18.5(ra)18.1(e)12.2(l)0(\))16.3(,)-206.8(0.)20.1(01)20.1(5)-152.5(g)0(/)]TJ
T*
[(mL)-492.2(ag)19.9(ar)-491.4(\(B)21.3(ac)18.1(te)17.9(ri)16.5(o)12.2(l)0(o)20.7(g)0(i)18.4(c)0(a)18.1(l)-500.9(Ag)17.9(ar)18.1(-)13.2(A)0(c)16.1(a)0(d)18.4(e)12.2(mi)15.5(a,)-489.4(Is)18.5(ra)18.1(e)12.2(l)0(\))21.7(,)]TJ
0 -1.1869 TD
[(0.)20.1(5)-158(m)12.4(g/)19.9(mL)-551.8(gl)18.4(uc)15(o)12.2(s)0(e)20.4(-)0(D)]TJ
/F9 1 Tf
9.7011 0 TD
(+)Tj
/F6 1 Tf
1.1164 0 TD
[(\()13.2(S)0(i)15.7(g)0(m)22.3(a)0(-)17.9(A)13.5(ld)16.9(ri)16.5(ch)20.4(,)-564.5(S)12.6(t)11(.)-564.5(L)10.2(o)12.2(ui)15.5(s)13.6(,)]TJ
-10.8175 -1.1923 TD
[(MO)20.8(\),)-697.5(an)22.5(d)-707(0)0(.)20.1(0)0(2)20.1(3)0(9)-147.9(g)0(/)19.9(m)12.4(L)-705.2(H)0(E)17.3(P)10.2(ES)-694.8(bu)17(f)13.2(f)0(e)20.1(r)-707.4(\(H)17(E)13.4(P)10.2(ES)]TJ
T*
[(10)20.1(0G)21.5(-H)-302.7(bu)22.5(ff)15.6(e)12.2(r)0(;)-301.7(S)12.6(ig)18.4(ma)17.1(-)13.2(A)0(l)16.5(d)0(r)16.4(i)13.9(ch)15(\).)-301.9(T)10.2(h)12.4(e)-312.9(r)13.4(ea)16.9(ge)22.1(nt)18.1(s)-311.6(w)11.4(e)12.2(r)0(e)]TJ
0 -1.1869 TD
[(di)16.9(s)13.6(s)0(o)15(l)13.9(ve)16.7(d)-490.2(i)0(n)-477.7(t)11(he)-479.4(do)15.2(u)12.4(b)0(l)18.5(e)12.2(-d)16.2(is)22.1(ti)14.1(l)13.9(l)0(e)15.3(d)-490.2(w)11.4(a)0(t)21.1(e)12.2(r)-490.6(a)0(n)17(d)-490.2(au)22.5(t)11(o)0(-)]TJ
0 -1.1923 TD
[(cl)16.5(av)19.9(e)12.2(d)-240.9(\(i)16.3(n)12.4(s)0(t)19.2(r)0(u)20.4(m)0(e)19.3(n)0(t)18.1(\))-241.5(f)13.2(o)0(r)-229.1(3)0(0)-147.9(m)0(i)15.5(n)-236.9(at)-228.2(12)20.1(1)]TJ
/F9 1 Tf
16.7032 0 TD
()Tj
/F6 1 Tf
.3848 0 TD
[(C.)-231.2(Ea)18.1(ch)-228.9(pl)16.9(at)21.1(e)]TJ
-17.088 -1.1869 TD
[(w)11.4(a)0(s)-344.9()12.4(ll)17(e)12.2(d)-354.8(w)11.4(i)0(t)19.6(h)-350.7(10)-147.9(mL)-340.4(EL)18.3(S)-350.5(m)0(e)19.3(d)0(i)16.9(a)0(.)-343.1(F)10.2(o)12.2(r)-355.1(s)13.6(t)0(a)15.7(r)0(t)19.1(e)12.2(r)-355.1(c)13.4(ul)15.5(-)]TJ
0 -1.1923 TD
[(t)11(u)0(r)15(e)-421.3(pr)16.4(e)12.2(p)0(a)18.4(r)0(a)18.1(t)11(io)15.3(n)12.4(,)-423.5(a)-428.9(s)13.6(in)15.5(gl)18.4(e)-421.3(c)0(o)14.8(l)13.9(on)19.2(y)-429.1(w)11.4(as)-415.3(cu)20.4(lt)19.6(ur)15(e)12.2(d)-425.2(in)]TJ
T*
[(2)-158(m)12.4(L)-645.5(o)0(f)-635.7(l)0(i)17(q)0(u)17(i)0(d)-633.5(E)0(L)18.3(S)-648.6(at)-634.7(co)20.2(nd)15.4(i)13.9(t)0(i)19.6(o)0(n)19.2(s)-647.6(of)-635.7(32)]TJ
/F9 1 Tf
19.6732 0 TD
()Tj
/F6 1 Tf
.3848 0 TD
.6477 Tc
[(Ca)647.7(n)664.7(d)]TJ
-20.058 -1.1869 TD
0 Tc
[(15)20.1(0)-158(r)0(p)21.8(m)-361.5(f)13.2(o)0(r)-348.3(8)-158(h)12.4(.)-358.5(O)11.4(n)0(e)-349.3(h)12.4(un)19.4(dr)16.4(ed)-347.9(m)12.4(i)0(c)16.5(r)0(o)20.3(l)0(i)17(t)11(er)14.8(s)-354.9(o)12.2(f)-360.7(l)13.9(iq)18.5(ui)15.5(d)]TJ
0 -1.1923 TD
[(st)19.2(ar)18.1(t)11(e)0(r)-407.9(w)11.4(it)14.1(h)-415.7(o)12.2(pt)14(i)13.9(c)0(a)18.1(l)-419.6(de)20.6(ns)20.6(it)14.1(y)-418.3(\()13.2(O)11.4(D)0(\))-408.9(0)0(.)20.1(2)0(2)20.1()0(0)20.1(.)0(2)20.1(6)-418.1(\(m)20.3(ea)16.9(-)]TJ
T*
[(su)20.6(re)14.8(d)-284.3(u)12.4(si)16.7(n)12.4(g)-288.2(T)10.2(e)12.2(ca)18.1(n)-285.7(i)0(n)20.9()0(n)19.4(i)0(t)19.6(e)-285.8(M2)19.5(00)-272.6(P)10.2(R)0(O)-278.6(w)11.4(it)19.6(h)-285.7(6)0(0)20.1(0)-158(n)12.4(m)]TJ
0 -1.1869 TD
[(w)11.4(a)0(v)19.9(e)0(\))-213(a)0(n)17(d)-219.3(pH)-215.5(7)-223(w)11.4(e)12.2(r)0(e)-212.8(s)0(p)16.5(r)0(e)20.3(a)0(d)-214.6(o)12.2(n)-220.6(s)0(o)15(l)13.9(id)-216.2(EL)18.3(S)-220.5(a)0(g)19.9(a)0(r)-215(w)11.4(it)19.6(h)]TJ
0 -1.1923 TD
[(D)11.4(r)0(e)14.8(g)0(a)19.9(l)13.9(sk)15.9(i)-230(s)13.6(ti)14.1(c)13.4(k)-230.8(an)17(d)-224.7(c)0(u)15(l)0(t)19.6(i)0(v)18.4(a)0(t)21.1(e)0(d)-217.9(a)0(t)-222.8(3)0(2)]TJ
/F9 1 Tf
15.9879 0 TD
()Tj
/F6 1 Tf
.3848 0 TD
[(C)-230.4(f)13.2(or)-223.6(8)-152.5(h)-231.5(be)22.3(fo)20.1(re)]TJ
-16.3727 -1.1923 TD
[(e)12.2(l)0(e)15.3(c)0(t)19.1(r)0(o)20.3(p)0(o)20.6(r)0(a)18.1(t)11(io)15.3(n)-269.4(e)12.2(xp)18.2(er)20.3(im)15.5(e)12.2(n)0(t)18.1(s)0(.)]TJ
/F1 1 Tf
9.9626 0 0 9.9626 41.8394 315.5527 Tm
[(PEF)-302.7(experim)-9.3(ental)-296.2(s)-6.5(etup)-299(for)-295.5(th)-8.2(e)-295.9(determin)-11.8(ation)]TJ
0 -1.2519 TD
[(of)-339.1(irreversi)-7.9(ble)-334.1(electr)-8.1(opora)-9.3(tion)-336.7(electr)-8.1(ic)-334.3(eld)]TJ
T*
[(strength)-344.1(thresho)-12.1(ld)-334.2(with)-338.8(a)-335.7(single)-339(step)]TJ
/F6 1 Tf
10.4607 0 0 10.4607 53.8015 278.1921 Tm
[(Concentric)-332.6(ring)-335.1(electroporation)-332.6(as)-334.1(described)-333.3(by)]TJ
-1.1435 -1.1923 TD
(Fernand)Tj
/F7 1 Tf
4.1785 0 TD
[(et)-212.5(al.)]TJ
/F6 1 Tf
2.4117 0 TD
[(was)-214.2(used.)]TJ
7.8454 0 0 6.9729 157.096 270.3684 Tm
(46)Tj
10.4607 0 0 10.4607 167.981 265.7196 Tm
[(The)-214.4(concentric)-213.4(electrode)]TJ
-12.0586 -1.1923 TD
[(design)-363.1(creates)-358.4(a)-363.9(gradient)-358.5(of)-359.3(disinfection)-364.1(from)-360.5(the)]TJ
0 -1.1869 TD
[(center)-217.8(outwards)-216.3(to)-215.2(the)-219(periphery.)-216.2(The)-214.4(local)-218.3(electric)]TJ
0 -1.1923 TD
[(eld)-587.1(strength)-586.8(at)-580.5(each)-585.9(point)-586.9(is)-584.9(described)-588.1(using)]TJ
T*
[(equation)-279.6(\(1\))-277.8(as)-279.9(follows:)]TJ
/F7 1 Tf
7.7609 -2.9212 TD
.3822 Tc
(Er)Tj
/F6 1 Tf
1.55 0 TD
0 Tc
(,)Tj
/F7 1 Tf
.4444 0 TD
(V)Tj
/F12 1 Tf
-1.279 0 TD
1.6745 Tc
[()1506.7()]TJ
ET
169.398 187.767 33.392 .39685 re
f
BT
/F15 1 Tf
10.4607 0 0 10.4607 178.5259 192.4724 Tm
0 Tc
(D)Tj
/F7 1 Tf
.6666 0 TD
(V)Tj
-1.5392 -1.4362 TD
(rln)Tj
ET
188.674 179.83 9.411 .39685 re
f
BT
7.3225 0 0 7.3225 188.674 181.2472 Tm
(R)Tj
/F6 1 Tf
.7355 0 TD
(2)Tj
/F7 1 Tf
-.7355 -1.0065 TD
(R)Tj
/F6 1 Tf
.7355 0 TD
(1)Tj
/F16 1 Tf
10.4607 0 0 10.4607 183.9684 185.896 Tm
.9025 Tc
()Tj
/F6 1 Tf
8.3408 -.0542 TD
0 Tc
(\(1\))Tj
-21.9278 -3.1759 TD
-.0151 Tc
(where)Tj
/F7 1 Tf
3.0512 0 TD
0 Tc
(E)Tj
/F6 1 Tf
.9864 0 TD
-.0152 Tc
[(\(V/mm\))-283.8(is)-283.4(eld)-283.2(s)3.8(t)-4.2(rength,)]TJ
/F7 1 Tf
11.2457 0 TD
0 Tc
(r)Tj
/F6 1 Tf
.7154 0 TD
-.0146 Tc
[(is)-282.8(d)4.6(i)-.7(stance)-278.8(from)]TJ
-15.9987 -1.1923 TD
-.0144 Tc
[(the)-191.9(c)-1(enter)-190.7(o)-2.2(f)-190.8(t)2.1(he)-191.9(central)-195.6(e)3.3(lectr)4.5(o)-2.2(de,)]TJ
/F17 1 Tf
15.1261 0 TD
0 Tc
(D)Tj
/F7 1 Tf
.6504 0 TD
(V)Tj
/F6 1 Tf
.8888 0 TD
-.0164 Tc
[(\(V\))-198.3(i)-2.5(s)-192.5(p)-2.6(ot)-5.4(en)-4(tial)]TJ
-16.6653 -1.1869 TD
-.0161 Tc
[(differe)-3.9(n)1.7(ce)-334.5(bet)-5.1(w).7(ee)-3.9(n)-334.3(t).4(h)-3.7(e)-334.5(ce)-3.9(ntral)-338.2(a)-.6(nd)-338.3(pe)-3.9(riph)-3.7(eral)-332.8(e)-3.9(l)-2.2(ec-)]TJ
0 -1.1923 TD
-.0131 Tc
[(trod)6.1(es)5.9(,)]TJ
/F7 1 Tf
3.2843 0 TD
0 Tc
(R)Tj
/F6 1 Tf
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-.0133 Tc
[(1)-241.7(\()-.1(m)4.6(m)-.9(\))-238.5(i).6(s)-243.6(t)3.2(he)-239.5(radiu)4.5(s)-243.6(o)4.4(f)-243.9(in)4.5(ne)4.4(r)-243.8(e)4.4(lec)5.6(t)-2.3(r)5.6(o)-1.1(de)4.4(,)-241.7(a)2.2(nd)]TJ
/F7 1 Tf
-4.0051 -1.1869 TD
0 Tc
(R)Tj
/F6 1 Tf
.7262 0 TD
-.0131 Tc
[(2)-469.1(\().1(m)4.8(m)-.7(\))-471.4(i).8(s)-471(t)3.4(he)-472.4(r)5.8(a)2.4(dius)-471(o)4.6(f)-471.4(ou)4.7(te)4.6(r)-471.2(e)-.9(le)4.6(ctro)4.6(de)4.6(.)-474.6(I)2.7(n)-472.2(t)3.4(his)]TJ
-.7262 -1.1923 TD
-.0137 Tc
[(stud)5.5(y)]TJ
/F7 1 Tf
2.7748 0 TD
0 Tc
(R)Tj
/F6 1 Tf
.7262 0 TD
-.0157 Tc
[(1)-276.6(w)-4.3(as)-278.5(0.75)-168.3(mm)-279.7(and)]TJ
/F7 1 Tf
8.8719 0 TD
0 Tc
(R)Tj
/F6 1 Tf
.7262 0 TD
-.0154 Tc
[(2)-276.3(w)-4(as)-278.2(11.95)-167.9(mm.)]TJ
-11.9557 -1.1923 TD
-.0172 Tc
[(Pu)-4.8(ls)-3.6(e)-5(s)-534.7(we)-5(re)-536.1(d)-3.4(e)-5(li)-3.3(ve)-5(r)-3.8(e).5(d)-534.5(u)-4.8(s)-3.6(i)2.1(n)-4.8(g)-538.4(a)-532.9(B)-3.7(TX)-539.5(830)-532.9(p)-3.4(u)-4.8(ls)-3.6(e)]TJ
-1.1435 -1.1869 TD
-.0149 Tc
[(generator)-218.3(\()-1.7(H)5.2(a).6(rvard)-217.9(A)-1.4(pparatus,)-216.2(Inc.,)-216.2(H)-.3(olliston,)-216.2(MA\).)]TJ
0 -1.1923 TD
-.0147 Tc
[(Currents)-331.7(w)2.1(ere)-327.6(m)-2.3(easured)]TJ
/F7 1 Tf
11.6359 0 TD
-.0174 Tc
[(in)-335.6(v)-6.8(i)1.3(vo)]TJ
/F6 1 Tf
3.3927 0 TD
-.0135 Tc
[(us)5.5(ing)-328.8(a)-334.1(Pi)5.8(coS)4.5(c)-.1(o)4.2(p).3(e)]TJ
-15.0286 -1.1923 TD
-.0153 Tc
[(4224)-287.1(Oscilloscope)-284.9(w)-3.9(ith)-284.7(a)-292.5(Pico)-290.3(Current)-286.1(Clamp)-288.8(\()-2.1(60)-167.9(A,)]TJ
25.4288 64.2712 TD
-.0148 Tc
[(AC)4.1(/)-14.8(D)12.1(C\))-446(and)-450.9(a).7(nal)4.5(y)-14.8(z)13.4(e)2.9(d)-450.9(w)2(ith)-452.2(P).8(ic)4(o)-452.4(S)3.2(cope)-447(6)-449.2(s)-1.2(oftware)]TJ
0 -1.1923 TD
-.0152 Tc
[(\(Pico)-664.2(t)-4.2(echnologies)3.8(,)-666.4(Inc.,)-666.4(C)-1.8(ambrid)4(g)-15.2(e)12.3(shire,)-666.4(United)]TJ
0 -1.1869 TD
(Kingdom\).)Tj
/F1 1 Tf
9.9626 0 0 9.9626 307.8424 668.7494 Tm
0 Tc
[(Taguc)-8.1(hi)-334.2(rob)-8.8(ust)-333.3(exper)-8.2(iment)-9.3(al)-333.6(design)]TJ
0 -1.2519 TD
[(to)-338.3(determin)-11.8(e)-330(t)0(h)-8.2(e)-330(indi)-8.2(vidual)-338.2(impac)-8.1(t)]TJ
0 -1.2462 TD
[(of)-339.1(a)-330(num)-10.2(ber)-332.6(of)-339.1(pulses,)-339.7(pulse)-341.2(length,)]TJ
0 -1.2519 TD
[(and)-336.9(freque)-11.5(ncy)-337.1(of)-333.4(deli)-7.6(very)-337.6(on)-336.9(minimum)]TJ
T*
[(electr)-8.1(ic)-334.3(eld)-340.4(strength)-344.1(require)-8.5(d)-330.6(t)0(o)-338.3(inactiv)-10.3(ate)]TJ
/F2 1 Tf
0 -1.2462 TD
[(P.)-334.2(aer)-7.6(uginosa)]TJ
/F1 1 Tf
6.8571 0 TD
(PAO1)Tj
/F6 1 Tf
10.4607 0 0 10.4607 319.8046 594.0282 Tm
-.0158 Tc
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[(the)-374.7(e)-.7(quivalent)]TJ
/F7 1 Tf
6.8233 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 140.6551 394.7527 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 147.8551 396.5101 Tm
-.0248 Tc
[(was)-379.7(c)-.5(alcula)-3.9(ted)-379.6(w)-2.5(ith)-380.9(e)-1.7(quation)-375.5(\()-.7(2\).)]TJ
-8.2053 -1.1923 TD
-.0239 Tc
[(Th)4.8(e)-607.8(t)3.4(otal)-606.1(n)4.8(umber)-606.6(o)4.6(f)-606.8(replicates)-606.5(pe)4.6(r)-606.6(e)-.8(xperimental)]TJ
T*
-.0244 Tc
[(condition)-277.5(w)-2.1(as)-281.8(69)-274.5(f)-5.7(or)-282(each)-283(dilution.)]TJ
/F1 1 Tf
9.9626 0 0 9.9626 62.022 346.9038 Tm
0 Tc
[(Sta)-7.4(tistical)-340.3(analysi)-8.3(s)]TJ
/F6 1 Tf
10.4607 0 0 10.4607 73.9842 334.4881 Tm
[(Statistical)-434.4(analysis)-439.1(was)-436.4(done)-437.1(using)-436.3(Minitab18)]TJ
-1.1435 -1.1923 TD
[(\()-84.3(Minitab,)-651.6(Inc.\),)-651.3(Matlab)-650.9(\(ver.)-651.1(2013;)-648.9(The)-653.4(Math-)]TJ
T*
[(Works,)-476.1(Inc.,)-480.3(Natick,)-476.7(MA\),)-479.6(and)-478.6(Excel)-479(\(ver.)-477.7(2013;)]TJ
0 -1.1869 TD
[(Microsoft)-597.4(Corporation\).)-591.8(For)-598.2(randomization,)-593.2(103)]TJ
0 -1.1923 TD
[(permutation)-298.5(simulations)-294(were)-297.5(done.)-297(Results)-294.5(show)]TJ
T*
[(mean)-771.2(and)-765.8(standard)-774.4(deviation.)-769.3(The)-767.2(minimum)]TJ
0 -1.1869 TD
[(number)-268.4(of)-267.2(repeats)-265.9(per)-264(experimental)-269.7(condition)-263.9(was)]TJ
0 -1.1923 TD
[(15.)-408.9(For)-413.9(the)-408.7(linear)-406.3(regression)-409.5(model)-412.3(developed)-406.2(in)]TJ
T*
[(this)-431.3(study,)-433.3(we)-431.6(calculated)-433.9(the)-430.4(total)-434.9(relative)-429(error)]TJ
0 -1.1869 TD
[(\(TRE\))-277.8(using)-284.6(equation)-274.1(\(4\):)]TJ
4.8993 -3.4523 TD
(TRE)Tj
/F12 1 Tf
2.2708 0 TD
()Tj
ET
148.535 188.674 17.348 .39685 re
f
BT
/F6 1 Tf
10.4607 0 0 10.4607 148.5354 193.3795 Tm
(100)Tj
/F7 1 Tf
.3848 -1.3657 TD
(m)Tj
/F16 1 Tf
2.4388 2.531 TD
[()129()]TJ
/F18 1 Tf
.2818 -1.8427 TD
(+)Tj
/F7 1 Tf
7.3225 0 0 7.3225 182.0976 196.7244 Tm
(m)Tj
-.5497 -2.8027 TD
(i)Tj
/F12 1 Tf
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()Tj
/F6 1 Tf
.9368 0 TD
(1)Tj
ET
201.94 188.674 38.778 .39685 re
f
BT
/F7 1 Tf
10.4607 0 0 10.4607 201.9401 193.3795 Tm
(S)Tj
7.3225 0 0 7.3225 208.9133 191.7921 Tm
(i)Tj
/F12 1 Tf
10.4607 0 0 10.4607 213.6188 193.3795 Tm
()Tj
/F7 1 Tf
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(PV)Tj
7.3225 0 0 7.3225 237.77 191.7921 Tm
(i)Tj
10.4607 0 0 10.4607 212.7117 179.0929 Tm
(PV)Tj
7.3225 0 0 7.3225 227.0551 177.5054 Tm
(i)Tj
/F16 1 Tf
10.4607 0 0 10.4607 194.3432 201.033 Tm
3.7042 Tc
()Tj
/F6 1 Tf
7.3225 0 0 7.3225 248.3149 198.5385 Tm
0 Tc
(2)Tj
/F16 1 Tf
10.4607 0 0 10.4607 167.5842 205.5685 Tm
(s)Tj
/F6 1 Tf
11.8364 -1.8427 TD
(\(4\))Tj
-21.9278 -3.1813 TD
(where)Tj
/F7 1 Tf
3.2355 0 TD
(m)Tj
/F6 1 Tf
1.2844 0 TD
[(is)-395.2(number)-398.5(of)-391.8(measurements,)]TJ
/F7 1 Tf
13.8742 0 TD
(S)Tj
7.8454 0 0 6.9729 261.4109 151.2567 Tm
(i)Tj
/F6 1 Tf
10.4607 0 0 10.4607 268.1574 153.0141 Tm
[(is)-400.6(mea-)]TJ
-19.7057 -1.1923 TD
[(sured)-281.4(value,)-278.3(and)]TJ
/F7 1 Tf
7.9668 0 TD
(PV)Tj
7.8454 0 0 6.9729 159.7606 138.7842 Tm
(i)Tj
/F6 1 Tf
10.4607 0 0 10.4607 165.2598 140.5417 Tm
[(is)-281.4(predicted)-283.4(value.)]TJ
/F1 1 Tf
10.959 0 0 10.959 62.022 103.6913 Tm
[(RESULTS)-329.7(AND)-333.4(DISCUSSION)]TJ
/F6 1 Tf
10.4607 0 0 10.4607 73.9842 89.7448 Tm
[(The)-534.2(experimental)-535.2(setup)-533.1(with)-536.5(concentric)-533.1(elec-)]TJ
-1.1435 -1.1923 TD
[(trodes)-286.8(for)-291.7(one-step)-284(determination)-288.6(of)-283.4(the)-289.5(minimum)]TJ
T*
[(electric)-351.6(eld)-348.7(required)-348.3(for)-351.3(irreversible)-351.7(electropora-)]TJ
0 -1.1869 TD
[(tion)-297.2(of)-299.7(bacteria)-296(with)-298(PEF)-302.1(is)-297.7(shown)-295.6(in)-298.8(Fig.)-297.4(1a.)-300.5(The)]TJ
0 -1.1923 TD
[(dynamic)-450.6(current)-453.5(of)-446(the)-452(individual)-451.8(pulse)-448.9(of)-451.4(60)]TJ
/F8 1 Tf
22.1337 0 TD
(l)Tj
/F6 1 Tf
.5528 0 TD
(s)Tj
2.7423 64.282 TD
[(duration)-323.4(is)-324.8(shown)-322.7(in)-325.9(Fig.)-319(1b.)-327.6(Figure)-323.5(1c)-323.4(shows)-321.5(the)]TJ
0 -1.1923 TD
[(PEF)-264.2(effect)-260.6(on)-268(the)-262.4(culture)-263.3(of)]TJ
/F7 1 Tf
12.8933 0 TD
[(P.)-261.6(aeruginosa)]TJ
/F6 1 Tf
6.2976 0 TD
[(.)-266.4(A)-263(clean)]TJ
-19.1909 -1.1869 TD
[(area)-339(with)-336(dead)-334.9(cells)-339.4(closer)-338.4(to)-334.4(the)-338.2(center)-337.1(is)-341(appar-)]TJ
0 -1.1923 TD
[(ent,)-566.7(and)-570.7(unaffected)-564.1(cells)-567(remain)-570.4(on)-566.1(the)-565.9(edges,)]TJ
T*
[(where)-339.3(the)-343.7(strength)-337.5(of)-343(the)-338.2(electric)-340.7(eld)-343.3(was)-338.9(insuf-)]TJ
0 -1.1869 TD
[(cient)-282.2(to)-280.2(kill)-275.7(the)-284(bacteria.)]TJ
1.1435 -1.1923 TD
-.0245 Tc
[(In)-201.8(the)-202(t)2.8(ested)-195(r)-5.7(anges,)-198.8(increasing)-204.3(p).1(ulse)-196.5(duration)-201.8(\()]TJ
/F7 1 Tf
20.6758 0 TD
0 Tc
(t)Tj
7.8454 0 0 6.9729 559.729 635.8676 Tm
(p)Tj
/F6 1 Tf
10.4607 0 0 10.4607 563.9243 637.625 Tm
-.0241 Tc
(\),)Tj
-22.551 -1.1923 TD
-.0239 Tc
[(increasing)-290.4(number)-292.3(of)-292.5(pulses,)-290.3(a)-3(n)4.8(d)-297.4(e)4.6(n)-.6(ergy)-295.8(invested)-291.9(in)]TJ
0 -1.1869 TD
[(the)-212.2(w)-1.6(h)4.8(o)-.8(le)-217.6(tre)4.6(a)-3(tment)-213.4(d).7(ecreased)]TJ
/F7 1 Tf
13.6412 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 477.9778 610.9794 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 483.477 612.7368 Tm
-.023 Tc
[(\(Fig)3.1(s.)-213.5(2a)3.3(,)-213.5(c)1.3(,)-218.9(d)-209.7(and)-215.2(3)]TJ
-14.8606 -1.1923 TD
-.0225 Tc
[(a)3.8(n).8(d)-404.4(T)4(ab)3.8(le)-400.5(1\).)-397.3(I)-1.3(ncrea)3.8(s)1.9(ing)-402.8(t)4.8(he)-400.5(pu)6.2(lse)-400.5(i)2.3(nt)4.8(erva)3.8(l)-404.2(h)6.2(ad)-398.9(al-)]TJ
T*
-.0229 Tc
[(most)-244.9(no)-243.7(effe)5.6(ct)-244.9(o)5.6(n)]TJ
/F7 1 Tf
7.6037 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 414.8219 586.0912 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 420.6613 587.792 Tm
-.0246 Tc
[(\(Fi)-5.3(g)1.5(s.)-247.6(2b)-247.6(and)-249.3(3)1.7(\))-5.9(.)-242.2(In)-245.2(studies)-249.5(u)4.1(si)-5.3(ng)]TJ
-8.8556 -1.1869 TD
-.0238 Tc
[(Taguchi)-503.1(o)-.7(rthogonal)-503.1(a)-2.9(rrays)-498(and)-503.2(t)-1.9(he)-499.3(individual)-497.6(p)-4.6(a)2.5(-)]TJ
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-.0228 Tc
[(ram)5.9(e).3(te)5.7(rs)-236.8(of)-237.2(a)-240.4(p)1.8(ulse)5.7(,)-240.4(p)1.8(ulse)-243.6(duration)-238(had)-242.1(t)-.9(h)5.9(e)-243.6(stro)5.7(nge)5.7(s)1.6(t)]TJ
T*
-.0237 Tc
[(effect)-348.7(on)]TJ
/F7 1 Tf
4.1189 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 378.4251 548.7306 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 381.6566 550.4313 Tm
-.0239 Tc
[(,)-349.9(f).2(ollowe)4.6(d)-351.6(b)-3(y)-350(p).7(ulse)-353.1(n)4.8(umber)-351.9(\(Fig.)-349.9(3)-3(\).)-349.9(The)]TJ
-5.127 -1.1869 TD
-.0247 Tc
[(inter)-5.9(v)1.4(al)-287.2(between)-283.3(p)-.1(ulses)-282.1(h)-1.4(ad)-281.9(the)-283.4(l).1(owest)-284.6(i).1(mpact)-284.6(o)3.8(n)]TJ
/F7 1 Tf
22.112 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 566.5888 536.2581 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 328.0251 525.5432 Tm
-.0239 Tc
[(\(Fig.)-241.5(3)2.4(\).)-236.1(The)-239.3(l).9(owest)]TJ
/F7 1 Tf
8.563 0 TD
0 Tc
(E)Tj
7.8454 0 0 6.9729 424.8566 523.7857 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 430.5826 525.5432 Tm
-.0244 Tc
[(was)-238.4(o)-1.3(bserv)-3.7(e)4.1(d)-243.7(w)3.3(hen)-239.6(200)-242(pulses)]TJ
-9.8041 -1.1923 TD
-.0247 Tc
[(of)-201.1(60)]TJ
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2.2058 0 TD
0 Tc
(l)Tj
/F6 1 Tf
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[(s)-202.5(d)-1.8(u)-3.1(r)-2.1(at)-4.5(io)-3.3(n)-203.7(w)-4.1(ere)-203.9(d)-7.2(eli)-7.1(v)-.3(e)-3.3(r)-2.1(e)-3.3(d)-202.3(w)-4.1(i)-1.6(th)-203.7(3)-5.5(5)-.1(0)-168.1(m)-3.1(s)-202.5(i)-1.6(n)-3.1(t)-4.5(er)-7.6(val.)]TJ
-1.5934 -1.1869 TD
0 Tc
[(Using)-368.4(a)-369.3(multivariable)-368.4(regression)-366.2(approach,)-367.7(we)]TJ
-1.1435 -1.1923 TD
[(constructed)-381.2(a)-374.8(linear)-379.3(regression)-377(model)-374.3(to)-377.8(describe)]TJ
T*
[(the)-370.8(dependence)-364.6(of)]TJ
/F7 1 Tf
8.8286 0 TD
(E)Tj
7.8454 0 0 6.9729 427.918 474.0093 Tm
(c)Tj
/F6 1 Tf
10.4607 0 0 10.4607 435.2881 475.7101 Tm
[(\(V/mm\))-364.1(of)]TJ
/F7 1 Tf
5.0023 0 TD
[(P.)-370(aeruginosa)]TJ
/F6 1 Tf
6.7745 0 TD
(on)Tj
-22.0307 -1.1869 TD
[(the)-229.8(tested)-224.1(parameters)-224.5(of)-229.2(the)-224.4(electric)-232.3(pulse)-226.7(\()]TJ
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19.6569 0 TD
(n)Tj
/F9 1 Tf
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(=)Tj
/F6 1 Tf
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(150\))Tj
-21.1582 -1.1923 TD
[(as)-279.9(in)-282.6(equation)-274.1(\(5\),)]TJ
/F7 1 Tf
5.1541 -2.2762 TD
(E)Tj
7.3225 0 0 7.3225 389.4235 425.4802 Tm
(c)Tj
/F12 1 Tf
10.4607 0 0 10.4607 394.9794 427.0109 Tm
()Tj
/F8 1 Tf
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(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 410.003 425.4802 Tm
(0)Tj
/F12 1 Tf
10.4607 0 0 10.4607 416.2393 427.0109 Tm
()Tj
/F8 1 Tf
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(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 431.2629 425.4802 Tm
(1)Tj
/F7 1 Tf
10.4607 0 0 10.4607 435.7983 427.0109 Tm
(t)Tj
7.3225 0 0 7.3225 439.54 425.4802 Tm
(p)Tj
/F12 1 Tf
10.4607 0 0 10.4607 445.9464 427.0109 Tm
()Tj
/F8 1 Tf
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(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 460.97 425.4802 Tm
(2)Tj
/F7 1 Tf
10.4607 0 0 10.4607 465.5054 427.0109 Tm
(N)Tj
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1.0406 0 TD
()Tj
/F8 1 Tf
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(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 491.4141 425.4802 Tm
(3)Tj
/F7 1 Tf
10.4607 0 0 10.4607 495.9495 427.0109 Tm
(I)Tj
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()Tj
/F19 1 Tf
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()Tj
/F6 1 Tf
4.3357 0 TD
(\(5\))Tj
-21.9278 -2.4551 TD
(where)Tj
/F8 1 Tf
3.1976 0 TD
(a)Tj
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7.3225 0 0 7.3225 366.6896 399.7983 Tm
(0)Tj
10.4607 0 0 10.4607 374.9101 401.329 Tm
[(is)-357.3(intercept,)-353.3(and)]TJ
/F8 1 Tf
8.0156 0 TD
(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 463.9747 399.7983 Tm
(1)Tj
10.4607 0 0 10.4607 468.5101 401.329 Tm
(,)Tj
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/F6 1 Tf
7.3225 0 0 7.3225 480.3589 399.7983 Tm
(2)Tj
10.4607 0 0 10.4607 484.8943 401.329 Tm
(,)Tj
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(a)Tj
/F6 1 Tf
7.3225 0 0 7.3225 496.7999 399.7983 Tm
(3)Tj
10.4607 0 0 10.4607 505.0204 401.329 Tm
[(are)-354.5(linear)-352.1(co-)]TJ
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[(efcients)-291.9(of)-294.3(pulse)-297.1(duration)-296.3(\()]TJ
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12.9583 0 TD
(t)Tj
7.8454 0 0 6.9729 467.2629 387.1558 Tm
(p)Tj
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(,)Tj
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(s\),)Tj
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[(is)-292.2(number)-295.5(of)]TJ
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(pulses,)Tj
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(I)Tj
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[(is)-351.9(interval)-352.9(between)-351.9(pulses)-354(\(ms\),)-354.8(and)]TJ
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[(model)-276.8(error.)]TJ
1.1435 -1.1869 TD
[(The)-458.3(determined)-457.7(coefcients)-455.9(were:)-460.9(962.15)]TJ
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(0)Tj
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10.4607 0 0 10.4607 549.3542 351.5527 Tm
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()Tj
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1.7018 0 TD
0 Tc
(,)Tj
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(-)Tj
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()Tj
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0 Tc
(,)Tj
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(2)Tj
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()Tj
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0 Tc
[(,)-239.3(0.77)]TJ
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3.0783 0 TD
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7.3225 0 0 7.3225 463.4644 337.4928 Tm
(3)Tj
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10.4607 0 0 10.4607 454.2235 339.0802 Tm
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()Tj
/F6 1 Tf
1.7018 0 TD
0 Tc
[(,)-233.9(indicating)-238.3(that)]TJ
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(t)Tj
7.8454 0 0 6.9729 555.0802 337.3228 Tm
(p)Tj
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10.4607 0 0 10.4607 562.0534 339.0802 Tm
(is)Tj
-22.3722 -1.1869 TD
[(the)-343.7(strongest)-340.3(predictor)-348.8(among)-338.6(measured)-344.2(parame-)]TJ
0 -1.1923 TD
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7.8454 0 0 6.9729 470.2676 312.4346 Tm
(c)Tj
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10.4607 0 0 10.4607 473.7825 314.192 Tm
[(.)-315.2(The)-322.8(model)]TJ
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[(2.2)-268(10)]TJ
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7.8454 0 0 6.9729 384.1511 306.3684 Tm
(-)Tj
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(16)Tj
10.4607 0 0 10.4607 397.1337 301.7196 Tm
[(,)-266.4(the)-262.4(adjusted)]TJ
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7.8454 0 0 6.9729 473.1022 306.3684 Tm
(2)Tj
10.4607 0 0 10.4607 480.1888 301.7196 Tm
[(was)-268.4(0.868,)-265(and)-267.2(the)]TJ
-14.5462 -1.1869 TD
[(corresponding)-283(TRE)-277.2(was)-279.3(0.78%.)]TJ
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[(Next,)-326.3(using)-322.5(the)-327.4(PEF)-323.8(protocol)-325.8(with)-325.1(the)-327.4(smallest)]TJ
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(E)Tj
7.8454 0 0 6.9729 335.5653 262.6015 Tm
(c)Tj
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10.4607 0 0 10.4607 342.992 264.3589 Tm
(\()Tj
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(t)Tj
7.8454 0 0 6.9729 350.192 262.6015 Tm
(p)Tj
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10.4607 0 0 10.4607 356.3716 264.3589 Tm
(=)Tj
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