L- ~~ ~ W A HUNT UNCLASSIFIED 12-79

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1 / AD AO76 Th8 ARMED FORCES RADIOBIOLOGY RESEARCH INST BETHESDA MD FIG 6/1 EFFECTS OF ACUTE AND CHRONIC ADMINISTRAT ION OF ETHANOL ON CYCLI ETC(U ) 1919 W A HUNT UNCLASSIFIED NL END 1279 DX L.

2 V V Fwm : lt!ii,l1 L..: ;. (:: ll! \ OL, 2 V I U ( UI ffects of Acute and Chronic 6 _. 8 1 Administration of Ethanol on Cyclic Nuc leotides and Related System I Wal ter A.\Hunt i!.. I. D c 4A 1. A. INTRODUCTION Over the past years the possible role of cyclic nucleotides in biological funct ion has received considerable attention. It is now believed that cyclic nucieotides act as a second messenger in the actions of a variety of hormones. Since a physiological response could result from the disruption of any one of a chain of events. the possible role of cyclic nucleotides in the actions of ethanol both after acute and chronic administration have been explored. It will be the purpose of this review to collate and analyze the existing data on ethanol cyc licnucleotide interactions and to determine if these effects could be responsible for some of the actions of ethanol. In order to gain perspective it will be necessary to summarize the present theorie s on the role of cyclic nucleotides in biological function and how they accomplish their actions. Many excellent reviews 24 have been written exhaustively detailing what has been reported on cyclic nucleotides. Hence, our purpose here will be only to present basic concepts in order to help understand the potential relevance of the actions of ethanol on this system. B. ETHANOL AND CYCLIC NUCLEOT IDES IN THE NERVOUS SYSTEM 1. General Aspects of Cyclic Nucleot ide Funct ion Evidence to date supports a role of cyclic nucleotides in synaptic transmission. Cyclic nucleotides have been implicated in the actions of a number of _j neurotransmitters. including norepinephrine, dopamine. serotonin. acetylcho Walte r A. Hunt Behavioral Sciences Department. Armed Forces Radiobiology Researc h Institute. Bethesda. Maryland ;9 IV.., V V L..

3 122 % iller IlunI line. (i \ U,ind glnt iinatc Ade nosinc3. e clue nkinopl pliate Ic \ Yst P1 appears to he ic Ltted to the et ec t s of note pinephrine. dopaititne. and set otonin, hue eti.tnosine cite inonophosp hate ie. \ die ( im Pt is I iclated to the ctti. cts of acei Icholine. (,\ HA. and glutau ate. l he etirien t belief concerning the possible role of e Iic uuc leotidcs in s naptie transmission rii I es th e init ial stunlu latit)n ola receptol b the neiui ot i ansnnt tel substance In the ease of hioge nic am ines. inter a ct ion it h the ii i eceptor in oi es ;iden late cy clas e hich convert s A I P to e clic A MP. hue.icet Icholine..\ BA. and glutamate act thi ough guan% late elase. L\ hich converts ( I I to c c Iic ( I\1 P: When these en. fli es are st inuulated. an aceti mul,ution of the coi ies nding c. c Ite nucleotide occurs. I hus ekvated c Iic iii uc leoti de Ic e i ni.. ie.uscs t he ac ti it of c c ic ntic leot idedc pe ude n I protein k i n.ises. hich a ppear to he separate entities, one specific lot c\ cite. MP and one c clu e I.,M P. these protein kinases eatalvie the phosphoivlation of IM o teins located in s miptosomal metnhi anes, and thi s process is believed to alter the permeahul ut ot t he membrane to ions. h Ic ii in t urn changes the lest ing me mhra ne te itt i.il the most detailed,icctirint of the lrlter ;ucf ion of neutoti ansmit tel s and e e tic uric k ot ides and the et Iki ml it tnt c h. i ulge s in the me ni bi a ne potential has been ith the sti pei iol cci v i al ganglit m it ide nec ace timtu lated he ic has been e; is ici to interpret because of it i c lat i el simp le st i uct uial oi gani /at ion when LOll1 pai ed to t he brain It no appeal s t hat dopaun inc is i eleased front intei neurons in the ganghon and stinuulates the loi mat ion of c elk A Ys t P posts\ napticall Fhese e ents coincide it h the de% elopnient of the slou iuhubitor posts vn uptic potentia l. :I ( )n t he other hand,.icet lc holine i ek ased front preganglionic tibeis.1 Imulates the format ton of e e lie (; NI P pt si s napt Id.11 I and iiia\ tie i elated to the sio esc itator posts\ naptic potential v I hus et fect scents to be mediated huough In use all hid receptors since it can he blocked b muscar inic antagonists but not b nicotinic antagonists. Most oft he i eseaich on the elkct of ethanol oil c die nucleot ides has been on c% die, \Ysf I and i elatcd syste ms in brain tissue. Measurements of brain c Liid A Ysil are ei Ii icks cspeciall in t he cei ehellum because of a rapid postmoitenl accumulation of this nucleotide. ( onseqtient l. rapid inactivation of brain en/s mes is ei\ inlpoi lant. l o accomp lish this. highintensit niicrowa e Iii a diation has been used, Initiafly. a conventional microwav e oven as tr ied, bitt it required 20 sec to inacti ate adenv iate c c lase and phosptiodiesieia e More recenf I a wier a e o efl m as developed which locuses the beam on the head of an iiiuii ) il. : Wit h this modification the en/v mes are inactivated in 2 sec ii. teiu te l rc*aliitci if 2. IlTecis of I thanol on ( elk A NI I In an earls studs Volicer and (ioid reporte d. using conventional mic ro a e irradiation fo r 46 si c. that a single I to get haiiol k g dose, given orall.

4 _ 7 on ( cik \urkoitdt s 123 depresse d cyclic AMP levels up to 6(Y. in whole brain of rats in a dosedependent manner This response was predominantly in the cere bellum. Orenb erg et at examined cyclic AMP levels in several areas of the mouse brain after 9 sec of conventional microwave irradiation. In the cere bra l cortex, cyclic AMP levels were depressed up to h5 1 hr after trea tment by 0.4 to 3.2gethano l/kg doses. given intraperitoneally. However, under similar conditions, subeort ical and cerebellar levels were significantly elevated by 60 and l 7(Y h respectively. 10 mm after treatment, When mice were killed by immersion in liquid nitroge n, ethanol had no effect on cyclic AMP levels in the cere b ra l hemispheres. tm Recently. using a 9sec exposure of focused microwave irradiati on. Volicer and Hurter 2 found a dosedep endent reduction in cyclic AMP levels after 1 6 g ethanol/kg, given orally, in the cerebral cortex, cerebellum, and brain stem of the rat I hr after trea tment. On the other hand. Redos et at. were unable to demonstrate any alteration in cyclic AMP levels in any are a of the brain 2 hr after treatment with a 6gethan ol/kg oral dose and using 3.5 sec of focused microwave i rradiation. The preceding discussion illustrates the uncertainty of the effect of sing le doses of ethanol on cyclic AMP levels in the brain. The sourc e of these ambiguities is not clear but may be related to methodological considerations. A summary of some of the data can be found in Tabl e 1. As pointed out earlier. rap id inactivation of the synthetic and degradative enzymes for cyclic AMP is erv import ant to minimize its postmortem accumulation in the brain and to reflect more closely the actual levels in vita In th e studies to date, exposure durations var fro m 3.5 to 9.0 sec for focused microwave irradiation and 9 to 45 5CC using conventional microwav e ovens. Although the enzymes can be inactivated fairl y qu uckls. it is unclear whether the existing technology does it fast enough. especially in the cerebellum, where postmortem accumulation is most pronounced. Also, ethanol inhibits this accumulation in ra ts 2 and could induce art i tlicts h making it appear that ethanoldepleted cyclic AMP, when in F Table I. ElTeet of a Single Dose of Ethanol on Cyclic AIP Levels in Brain Time Mode Cyclic AMP after of IAm c ircar men i kittin g Controt Treated Reference ( erebeftum 4 k g. pa 3 hr CMI gig. pa I hr FMI gig. pa 2 h r FMI gig. p 10 mm CMI Cerebral cortex 4 g k g pa 3 hr CMI gkg. po I hr FMI g g, pa 2 hr FMI g/kg. ip 10mm CMI t w kg. ip 1.5 hr ILN CMI, conventional microwave irradiation: FM1.focused microw ave irradiation: tln. immers,on in liquid niirogen. Values expressed as pmotelmg tissue. va lues expressed as pmoie mg protein.. _j i, H:.

5 I 21 % alit, S lio u rt I.uc t it might o nix has e blocked the pmms i nmoi te in ace um ii l.a ion t Ii,, I could take pl,ice xx ithun the fist tess seconds.ittei death. lit t uuih eu siip xti I of thus psissibulits is the u epoi t ot Jones.und St ax iimoha th at,u si gnif icant el xat uo ii of cx cli ANt I can oct ui in t he brain undeu these cure umstances l he compai ed t he cxc lie A NI I lex els in,mrlous pails of the brain of mice exposed eit her to 4 sec m it I ot 0., see of (s I. W mit focuse d iilictossa xe radiati on. In all east s t he tx clue A MI lesels xse ie higher xx it h the loxser intensitx. longer duration irradi ati on. In t he cci ebr lluin, for e s,unple, the x,t l it s inc teased bx lt P Ysto st of the studies using inucross as t ii i adiat ion expose t he brain longer than is necessai x to una ctu x ate en/ mt s. l he pote ntial problems of oxeu expmisu iie has t not been e xplored lot esa niple. does puo longed high heat decompose cxc lit N NI I lxmes the puogi essus e cellulai damage induced bx continued exp s msui u e.i l (ci the d iso ihut ion of S S l ie NlI in a inaum ncr i hat unig ht nit i ma icr afkc t its qtu.mt It,ttiofl ( )r does inte rae t ion of ci haumol and other di rigs xx jib the hi ai n.ul Ic m t he dx nanhics of rnactis ation I )ifl ei ent i espsmnse. in Ct lie A NI I lex els h.ix c been obtained iii dif lei cu lt,ri eas of t he brain attm i et hanol ti eatnmenf and,ii t a funct ion mi t the dose and tuutt e af tei cx msrii e, Most st tidies do not it ps m it blood ethanol coneenti at ions, making meaningful eompai uson s bet xx cc it studies ml to Ic tilt ti m suuunmai, in order to detei muui e xx hat single doses,it ethanol do to cx e lie ANt I It s els in th e brain, ii is necess,ui x to emp km methods of killing animals that.ii e xx eli rindei stmxmtl and stand,ii dii1 d I \ lx rintents need to be earned out eaiefu llx piox iding complete dose uespsmnse and timecourse relationsh ips accompanied hx blood e t hanoi conceit ii at Ions, Iii ial lx. the po ss ibi lit x of s pee ics dutfei ences should not he ignot ed b. ( hrunic I rt atu uenf Chronic treat ment xx it h ethanol has produced results on cx el he N NI P Ic els and i e lated s s te ills that,u e less e out io ci s ial. It an inmal s are I i eated fo r at least $ mt.ix s, cei ehral eselie AMP is elev ated 0,. However, aoer onlx 4 via s s of trea(ntent, no increases iii : t he N NI I art mm b serx emh, I hese increases in cx th e NM P.tl f t.ti to be related to an elevation ill adenvlate cx clase actix it V. K lit is :inma and Is, :im l found that at a time when cx tile N NI P lex els xx crc dcx ated. there xx as,il s mt an increase in adenx late cxc lase activit. N either i esponse xx as obsersed a fler a single dosc of ethanol. Also, phosphodiesterase act ixits xx,is unaffected,itlei eit her treatment, More intercst ung anti possibly nmoi e t unct ionail important findings ate the studies determining the sensiti x its of the cx t lit. NNI sx stein to neurt mti ans init tel s Present evidence is quite compelling that chronic administi ation of et hanol,iltei s the scil sit i si t x of cx eli A NI Pgenerating sx stt nis to noi cpinephrine. tn ear ls strudy demimonsti ated that treatment with ethanol for 14 dax s dt t irased the se it sitix it, of t his s stem to norepitmephrine. Ftenc h and co xx oi ket s have stubsequenti, reported some x crx interesting i esults on change s of sensitix its of... m.

6 I ITm mis tin 1 mm hm Sum Im m I iii, u 14 v tnt I,ie,, ii i iti ul.u t i,iit it, ilv li mmilm.iits itt i t t m l s iii i.iis.i Itt t I xx m ks m it t th,iitm il it a tine itt l hc i e suits ol se i md mit l limit,t,mn t he Ii tnt at It i xx ml ht ml i ass a I th e inc a stli t itlv flts 5% t t m lit,i m 1 d lit. i tuc,ui slice s,ilsi,u i it& mf Ii out,uitinl,ls xx ith du.ixm it foi hi. the scimsit is it s of the s itt ANtI geutci,iting x stein to itoucpiiicphi inc xx.us mtt l it ss m d, xx uth htt t m iose icspoits c iii xc sht i It i ui 4 1 told I. the tight I bis mle elopitmt i mt at itoh,idtt ttt i 5k s ii bst its it ix u t s em i ru lmt bc esph unem i f x f ix pm uat t u x it S at I1S I am li eneigit itci xc tei itunak I Ioxx cxci, it xx.ts lim it est.ii listted xx hm tht i.i single mim ist of t t hanoi in igli t pu mm mli let t he s,uit im it sti lt m ii x cite \ NI P.i ii i i latim imi I hrt e d,ix s it let xx ithdi axx,il,.i titti c eou it spm mnm luug to ti lt mlv s m Imipntciii at mldhii hum trettleits. t he op pos ite i espouse was obsei x m ml the st ut s it ix it x of the v S eli ANIl g em te u at u m tg sx stem 1mm nolepint Iihi inc ineix ast ml 1 4 Mml lit.iddith tt, th e sensitix it s I. tnst.umnne.tttd III xs,is also m nltanem mi the I es rx i utse to II I itt ugh I it s u I t ti m itt a cit i mit t it tic pa ss imi it mit se I m itoite i iv functi on l ilt sti, it i ttv,iitv m m it,u lie i :itim iit s in tilt se nsitix it S mit t ilt cx lic tnt l gcitc i it ing s x ster n to spe c rite.tspm ets mit ak oholisrtm has lim it been mteitmoust i atemi. At t Ime least t hex i ctk\ t e tm,in gcs induced t i s long term eonsuiti1ml ion of ethanol It is not L le,tm xx hethei i hex.u v i elated 1m m the tics el m iptuc itt at pbs sie,ri mtc pm h mv te rm et, xx hi5 ix xx as uniutimal xxi th Ott ntet homls tist m I. oi xx bet hei titt x ii c expie ssim mns m it s milt it I,. nm its pm c uf ie ite tit al tosicitx l it m xx hole vtues tton at.mvt.m ptmx e nxe c ts,sr x rsnxs t,ms Cs ls I um it i,ii lt 5 i5it it It 1 itt e I at t S plai it i itg I mml cia i tt c.u iid ph x s u,i I mit pm itmic nec It et hanol trt.u tineiit inmfue cs,iilm t.lttm iits in nctiix it iansn t it tt r fun ction tscc t haptci I I 1 cit ionic t ha uigc s ciii lead to cm i ittpc it satm ii x e ha utgm s in mit itt i paia mc tel s i it.iit a? t cittpl 1m m iii., hilt :1 ill fit hi lt m is i.is is I hc i ol c m t e t lie ituie k ot kit s in.imlapt u e Imi m i ct sses has mcccix Cml em mns imi eia hlc,ii tt itt umi pt,iit m f tt.is Iim t n conei.c lx m m s Ulxx t d urce it t lx lix I )i sun t u km s,inm l I Nih 13cc,l list mit I itt pot cit Ii.11 lil t P iu t,iltt t mit I best It itm ting s, it i. wipe i a lix e I )t.t t the st rid ut s mi t 1 u t itt It,iitmI xx mi rkeis be i m pm,itt mt ii sing,i it,ut m itt,i I lit m imic I I ha I I,ik m s minis.i Ic xx t,ix s tot i itd lie ing ph x sie,il mit pm it,lt ic C In t It is xx.i., Itm ilts pm e itlt utc rita I to x it ll x cail he mti %t m ruit tem i,,iltml tilt re Li I uoiiship t Ix x ecit neuixi tran snmtt tt i sensitix its,mnmt mis m t t, sl ii tt. t htt Otis xx t Ilt mii.i x x.ul s igns xx i l l he a unoic eoi tx lilting Possit i litx \ml.ip t u x m t h,inges c,iii takm p lace ito t onlx i tite seitsitu x its at i m ee itou s 1m m u tm Iiti Ii.i m tsi it i tt, is, hut po ssib lx.iism m xx lilt,il tc m tt im it s itt 11mm.ieti x its 01 tilt i m ecp tm i i tat x 5 Itc N NiP. tt t m cx lie N Nil ml epm m tmlt mtt Iul\ileiit kiutast \s i t tc ntm m ned e. ut I it i, I his Cii, x til t.it.i lx, t s I itt IS itm is pitoi x Ia t ion m if 1uix m tt iii hit sx ita it m sm iln.i I ntt itt l u;i itt. Rc cntlx ki u p i x,iiti,i. rt xiu lemi,i stfljx in xx hich th ex itte,i suit t,l cx lie N NI P mit pm itmi e mit pixitcin I i ll,ism at tix flx ut ittm iti5c em iv i i al m miite s i,ic ill m,i itm i viii au ic,iditt m ist ia I ioit m it m t tt,i itol, I itt x t.ii i mt m l t h.ut.rflei lix.u meitt tm ii 1 xx t m ks, sx itap lm ism int.ul cit ix inatie itt ix tl x xx,us t It x.ltt mi t i itmilm ild bit t it t uuit ett to control It xels mli x s at\t i xx ithdi,ixx,it Na,itiel,ltim mit in 1m ix m t t ih t kttt. is v,uettx its xx,is, bsm i x ed,itiei.i stngtm ml ast of m th,inol, itt pi oblent xx ilit thi s xx oi k is t hat t i e miit,1 i t im m i t s ii nmiei xx hi ie h I ftc, ti i ti i,i Is xx m iv km I led afl er e lt rv i itit m t h.i ut mil ti v.rt nlv nt xx et v not mtt ttncd.,tnd the,ippm.ii,uxec at possibl e xx tttmmti as.mi signs 55,15 not ix psii It d I It ups, it is ii ill Ic iult Im i miete mit t mm xx he t Itci t hi s,ittt t.ii tm iit is ir latm mi to the mtt m.clm iptttm nt ot ; lmx siv.il miepm ttmletict It,ip v.ti s, ho sxexer, t hat the dcx atloil I,, g.. 1L4 4

7 I lit 5% aiim, S linu S iii cx tilt A Nt I depcnmlent piotcilt kiit.ism.ic t ix its nmig hl he., emiitst qtuemt ec m it the iit v i cased,ti. Iix ax mit amlcnx late cx ci.,sm.umtd cx th e tnt P Icsels ant i itot lv i ait.idap tixe haitgc in t he s em tsi ti x mix mit Pi ot eiit kmnase to exel ie tnt I,un,ilmigous I, nm iepinephu inc iit mttic ctt ex 5 It5 N N I I aecumit tulatim mn. ;. I ff t r FIh iimm I arm ( miii ( Nif I itt possible ho le mit x die ( NI I in t he.iv t ioits m it m th.i nmil tt,is l ceii iccei it ix ie l s mm t em i Retim is 1 a/ It itui t mi that t t ft ai tm m l miep lete. t eic i cll,ui x d ie t,n t I u t ti lt t,i t u t a mimi s,, tic jie itmim itt mtt, m it net I lie mtt a xiii tn itt t tlt e I xx.i. mi hst i x cmi I hi.i t Im i mi r.i I,u mfi t u inst i;ut mon mi t ti g et ita itol kg, xx i ic ii iv suited itt.1 kiss mit Q, mit I ftc cx cliv intl In.i mid it im m it. t ft c degree of deple tion xt.ts tiiux et lx Pi mm Imorlioi ta l Imi tilt hloo t m tha,t, ii em iitet nli,l i im n xi lift eoi iri mi l x alii es mmiit :i i hteml xx iten eth anmil hami hecit chin inatcd, Si itt ui,u it stilts xx cmx iou utm l b \ mil ice r anti Iii u i ter, St tid it s m it I ftc d Ice Is of eth.uto l on cx cli ( i NI I has,, bt m it p u mt m Ieml 1m m mmim, Iude measulieni eitts in ot ltcr at t.is of the fmi a iit aitd,it tei Iit it it acute and m him it ic.uditt u nis t tat ion Sautt plc iv sum its c,u ii ix f oni nmi u mt l able II Si itg le ml mises of t I it., tim I xx etc touund to deplete cx cliv S. Nil il tii,, ccl t i i.ui cortex, e uiml ite it mie leus.mnd I italint u s,is xx eli.us in I he cci e bell uitt lit e hi on ic a il s Imc,i tem i t.i is it mt m tcu cl m I h.iital mlepm nd e itt, si ut t ii. i it sti lts xx m s c,i l st i x cii ii iht.inij ti, u s xx e i still,,fl,ix, c.itcmi II mm xx e xcr. in ti lt cc mm fit II tint,und titm hi a in sit itt, to lt i,uit e m vie x m lv i li. to I itt ef fect of cth,uim il m lt c x cliv ; NI I lex t is cm iru c spondi ng 1mm I itt mit S el m i pmite iit mif he ha x tmii,i I I olci a nec tu nmiei Si it il, uh cx iie ml mitt itt a I em m it mi it ioit i NV he it.ie tilt lx and e him mliv a lix t reatm d.mu t i ntal s.u c co in p.ii cmi.ui s iitt ml,ur filmi mid e t lt.u mt ol c m nee lit ma ti m ilis, v S c liv ( N1 P it x m is 5% ci& sigi ti fim,.iittl x highm i lit ti lt t h mo nie.illx tic.i tcmi :iniitta ls If5 x v lie t, NI P lexe ls,ui C mic prm ss eml tm i.i lm mit g I,, r im i ml, one utti gl t l m xpm m t to set a i mix ers hmimit aflei I hc c lint mn. m I ion m it ethanol lit is xx as not oim ser x cmi aft ci 4 tt,ix s mif Irm.utmtt eitt, S. x c Ii (INI P lexe ls in all,um v.is of the hi am st t um iied xx etc at v oltt rol lest Is dii i iitg titm e t itanol xx it lid r.uxx al s x nmlm m ittc NV heit it t.i Ii itt itt xx as cx teitmi em i 1m m S mi.i x s,, sigit lilt.ini ek x at ion at cci t hit I Lii v S eii (I N il mm.ms f oil tim t I im ixx CS cr. s ii tt t xx m thdi axx il sight s dm m im i p xx ith li mit it Ii cat it lt u it :x gm it meit.. m lc x a lion ofeei cbella r cx t he i NI P h ul s m li do lim it,ippm ai tm h t,i ; em equ iis itv toi lilt C\pi t ssi mii t 01.tn ethan mil xx ith d raxx al sx nmli mini m i iitil m II, I ffm t I i I lit iuutmil l rv iilnmm nt on ( m Ik I NIP I m x m Is lit Iti uitn It it. m him ist mm uiii,ii x,,,t,,,,t, x,,,ii,i mm ili,,i,,,mm ii K i,, ii, m m icl v Iliii ti I,i (t, I I 4 It I, 51 I i I I, ii m l i.it,,, tm s ii it I it, It 4 3o 4 Ii i it it ii i i, ii I m s, K t xii I,, I. sm, u,k lm i,ti,n, si I hi,itl, i i,ulii im C,. i tu,,, I tt,i, sm. is* ui,i dr x,,t,,.,,. i S,iI,. ;,ni, k iii i. tm i i,. piii. k i t t,ss, m,.

8 I IT m is nil ( mmii, Sum im nsi mit s u:. l he ntev itaitis utt lix xx hi h et h,un m i l dt plm l es 5 x v Itv (itt I is ui it knmi xx it Nit atte mpt to.hmmxx,t ti irt ct cocc i mit ethanol in cmmnet nti,ilitiits up to IINI itim on ui.ii ix l.itc v v i,msc mit v S lk (,ti I phm is phom iuc stei asc,set mx it\ at un, xx as 1101 su uee esst t ij li mixx cx er, h i m,issax s xx civ lierf orined uinmici miptinm.il m itl\ ittalie eminmlitmouts xx huv hi mimi limit nm v m ss ui ilx ieiicm, t xx ittt t mm Cv tii s ttt itt, 1 oi exam ple salt iii itt itia x regulate I he,sv lix it x mit gu i,u it x l.a c x v I ase, anm l v haitges iii t itc.idlimiits mit c.ileiuiitt ittighl mlcpiess the sx iitht si s of 5 x c lue ( Nil I tf ta nm i ht, m s been sht xx it to inmf uce sm xera l,ul teia iim iut s ut ealv i ui i it tftat ntigltt hes hi itt lit m mill macel I ui.ur conce lit rat im ih l of e a Iv in itt.mnt i stipp le ss gui.u n x Iatm cx c l.isv,ic lix its I th.imt mi l can bloc k the inxx,irmi e:uieimi nt v uu ureitts in i; /s vt: nculim ns, enlt,utv e calcium hiutmiung to ei th im t t c ghost nlent hlancs, autm i iemi u uv m hiaiit c,ilettitit Nl I hmi uig h all these.1 tid ies rit a s not lit id,, x ant Iv,i ii efl ec I mm gi ia it x late v x chase in biain. t itex tim mnducalc.i niecmt t mir iitm ic m imi ccl t x lx riil tm i tts mimi the pmissi hk rm lm of.,.ileiiiiit mit the.iv I im i tis mit e th,u tmil \ttei.ttimifls mix ne ti%tmlm,tnsnxtttel,iciix its might be tn m oh m mt tti the ethanol indumee mi mieple timin at cx cite ( Nt I N s umtdie.uted cam h er, v S die ( I Nil aplx ai s to lit ic iatem t tm the.iet toils mit,iv t t% lcht Iii tm. (it lit,,litm i glu taitt ate I Iit.until it.is ix t n tit ii)m its tr,ul cmi I mi in hi bit eom t iv.t l.u Utf i C tic tt l.ii.iee ix Ic ho ii tic iv lease in itt,.umd mtcp rcsses eei e belh ar glr itat iiat c hex ci., Witet bet I ftes t eitangm s v mitil d u. x p lau it,,ii Ic,ust tit p,mu t, t he tie plc t mm n mit cx cliv (; Nil has x e I I mi he tic I et iit inied Al tci.itit ils in s st enis iiix mil x mm ig v x cliv (,Nt I eomilti he iutxmilsed in t h e cx pie 551 mm m it a ii e I haitol xx it hd i ;ixx.ml sx liti immune, ( mi I lic i c i a! has m st ilt1 iem l I ftc m fiem, t mit,igc ht is xx hiefi base,iv t lm tm ils t e latt d It cxc ltv nuelcotim ics mitt etit.uutmil xx ithmli,,xx.ii heami lxx ut ef t e s it ttt tee Fiie t mi ii nvi thai dibut ix I v Sv I iv ( Ntl ailti Cd P. em irn poumnd s xx inch dcx.mlc bi.ui tt c die ( tnt P. iitv it asc t ite iiicmdenec mit heaml lxx utv h,, s mix er txx mit m i im l, xx bile dibtu t i I x c liv tnt P. NIP, and F tasi.ugl.u11 dmi is, v m itt pm mun ds xx hueh dcx ale hi aiit e die N NI F, aittagtini/c head lxx ite ltes I hcse i es ults supp mim t the x tess th at ti n alteration in tue Iialaticc of cxc liv S. N1 P.utd 5 x chi c N Nil in t.u x m it of t he t mirn icr might pi:u x a role u t tue ex pi essimii t mif Iii,, xx it htl i :ixx.uh.x nd tonic, I till hci. I hci e is a pai al Id bet xx ecu itartn:mi itte anti xx it i mit,ixx.mi intl uccml Irentimirs, Ilarniahine afyects the cei ehehluim lix stimulating li,, exeit.,im l x v Iimimimig fibers that s s ita i si, on the Purkinje cells 5 in a inaititer rese nif ihing.me li x,ii imilt mif di ulith lng ti hers, I h c ti cni mir induc ed h harnialiute is,it i. mint paitu eml b.itt v Im x.ii imin mu cc i ebeh lar v x v liv (1Nt I.mitt l c,uit l e hlmiv k cd f m Pu kit :udmi nm.t, tu tu o n mif heiii mimhi:u icpi nes, l Ilest m hli t 5s itax e hem it 115dm1 Im i Ireal th e m th.in m l xx ith mira xx.ii. nim li onte in mail, ( miti%m t t uelttl, tiitmiiv i t se.ii vii us xx ari a ntcmi 1m m I m x Ia tj nmi.1 possible l mi ld m it v S t h y Nt I mit i elalct i sx st t ni. in et hano l dependence. ptm..ihtx Ihu ough ai lei ti t ut ins i est ilting f i omt t ehl mimt ie e x v lt I Nil dephetimin r H (, I I ll t OI t\l) (. ( I,I( L ( I.Ffl l IIWS IN I HE S l ONI,t( II is kn mixx ii to sti mm i ltm te g.ist i ic,iv iti set i Ct tm t i in a numiibc r mit slx eiv s, inc luding man. l he response oiit.iincml is similar Im i t hat elieilcmi lix histaliiiuie, 1 but mubsequetit xx mir k rtilemi out a im ilc mit itist,uuittitc in lii,, effect mit, =

9 N w I Z$ 5% a lim i S I tiiiti elh.iltoi ( xc lue \Ntl ft,ms iit m tt uutlp lieatcti.us.t rllctiiatm i il l av iv1 sm s Iv I mmmii,,unt y f iii av Iv1 it im itt the g,i sti lt,il li d m s,t dm hit.0115,u C 5 C liv N NI I tt pm it dent pimiletni kmnt,isc xx hiefi iit,,x lx un ixm ixevi in I;tin sh,id:itimiit mit im lts : I tt,, i m tm im. it is J mmiss ifilt that c h.m ttge in s s Itt N Nil inc t,ulitm Ii silt ii 1.1 x fit it 1.u ted lvi mitt.ibil it s of eth att t l It.ilt ci.uc lvi sec t m t im ifl I he lit tm. si tilt uc l.ut imimi s lii p he Ii ecu lilt C fleet at ci h.u ilmil mill d.t Silt s iv tmf sct idt ioul.mnd 5 slid NNII niet,mtimmlis itt Il,us bm, eri e xplmii ed itt st s m r,ml species. univ hr mtt imt g ii i,tti lit Ibm I it, loyal.mp lilic,ulim irl at elftamtt l m int lilt d.isil lv iiltiv m s.i Ii) dtiii v em it m at mm ut s mit I I t I n emhmiv cmf at iii st s t v I im iit ill.u v m iitv m ul tr.ut ion vicps itdc lit tilti flhim t m m I0t,,,iliti vtc v ie.iscm f cx s it s t Nil Im x el, up to, lit the dime, dl hanmil has.u hip liasm e il ly e I mill,uc itt see let halt lit d0htv d lilt a iit miis lx lmixx tl, ct ii.urimih s(miiluil.ste s scdlelim n, xx hilt.mt emmil eenti.ut loui s :ili mix c, it dept esses sm cre tim m ti: I thanmil.mhso y Sm i Is the s,iiitm imip itasie m licv t mil l ttitiv mis,tl v s v lid N Nil Ic x els lit iit,i It, e th.tiim il v mi lid chit h.11 imiti s t tt tt.t liii I Life at id ses i m t imilt.11 C lest i panned fi x Uty tt.i st ni muv uis,ul cx Cliv N Ni P it, cis It xx mitml y i,mppm.um, ti teni, ttt,il altct :rtimiiis Ill,usti lv.me itt s,, v im, Iimiit.is.tfteelctl l s etitailmil ii v ducv tls corux laiett xx ith eii.ung m s in cx et ue \Ml dmilll etlt I nt dcl ei iiti mit ng m ftc milec h.utt isni b s xx lt ic It c t h.m iim il alters e x el mc N NIh hex c Is, Iii st ste 1i is tmi sltitlx the m i k v t at clit.tnol miii ttiuv misal.udeuix l,mte cxc l.tsm.tiltl phospitmiti i es tc rasc,mc mix it ic s I It time t,ut. t l ha rial iiihi iii Is limit i t it / 5 lilt s iii.1 em ittc cilli.ulim it l depcndcnt i.mshimin lint xx uth diticu emit sensitix itics, i at exam pit,.tt.u litmu it 0 ethanol, phm isplt mitl mm st e i :m.e tie mix its is rethiced t x co,, but.tyi ct lx late CSv l.ism us micarls em nt pk te ls bhmiekeml lit Ott dog and milan, m tii.t tal im u it us mnct Tcetise simni mii.iics.uy lemt x l.stc s s v I,tsm av t ix its at hm i sx dtihlv t ii t t,tti m it.ut itight emiilet nln xx lieu C, thud N NI I Ics ci. anti.iv it i sddi dt 1mm time r etir tecv h, \s xx mlii the i.iu, p hmssp fl mmmimesler:ise iii I ftc dm ig is iuihihitctt, I he se Iiultlings suig,gm si I hat e t hanoi iii., x dc x tile,ic Id scm m c I mmii f s st i rti ti hal m i title us Lit e v S d lasc, but he, do nim it m x pl: uui t iimixx cthtuitil e in dciii m s s Sm s lt t it tml t,miit i cxc liv N Nil lex m ls, Ns Pm iiti td ti mio t v ai I,ci,,i s.ix 5 in till,,iu e limit.ulxx.u ss i clmablc mm i reflecting th e rue st.iltis in I li e lix itig animal, ( it item I.ic l ii s mill m n y miflt ruh ut e t mi I ftc rat,, ot ss itl hcsts anti degm ad.utltin m m 1 a em mmit pv itind, I. x c ite N Nt I falls m u tt liii. datcgvirx It is xxelh kitx xx i tli. ut v St hiv N Nil lcxelx iii,u x,uicm s m f Itssiit s.it C :uti ev tt ml his a m umbem mit hm mi uiim itids, new m itr art sfl tut let s,,ufltl ions I I hamitil tti.ux nitl nec alicia I imilis in one of these tither t.i e Im it s One l.it lm ii th at :i tl cet s th e emmn tdult mit cx Cut N Nil ts the ax :m iiabih rt ol its piev ul u s m ii A Il if.umleutx h,ilm v S v l.i sm is u t mil sattiiatetl, Piimii uneui if,:/ tx rt uscmi t he stt imadli xx itl m to cthanm if,unmi mttm,i.umied iv id sm ei m ti m il aitm l s s die N I) anti N IF k s m Is in Iii,, xx timilm.isii he nt uet is.m,un mi in the su pt u Iiv mal iil uet s :m. xx 11cr,, ittos l oi t ilt.iciv t scv l ci iite p, it l l1 em II s Itt locateml, \ftv i 40 mutt, all I hm cc paranie te ls xx m Id sig ml ilic tint l x v m I iy m m I in lii,,.11 ix lily i.m l ;lltid m 55,i litti limit ill (lit xx it l,, niuv mis,*, \\ hen pci t usim ml xx as discontunied.,iv tvl sdv i dtltiit.iilml x elk N Nil.mm i m i N Il im tuil flt ml Iv v millii til les els i about (iii unit, Sine,, 110.mm iy f mt im iit,ih excite N NI I xx.is detected in him g.isi u tc lm v h t its,ilm, the rcvltudctl cxc lie N NI I uui (lie smipm u tie i.i h iittiv m daunmil lx explaiiicmt his leakage m i t cx mi t t N NI,i F h t ml the mit ii em us :mh v eils N u ctli im tim ii mit uiiuiemis:sl N II lexel s h.is.ih m hieeui m,li.m i x i d in lilt mlog,ii It i ethanol x rtu.im ii,,, ;,,,,. it,_>,,.

10 I (frvts mmmi ( S, liv \uv Ii imitmt,,, I 2 Ik ost agla nvf unx mnt t cra dt xx itft cs site N\ll itt eta bvih is ni t anti inhibit gastri c acid sec te non, In an attempt to detem n tune ml pi mmstagla nvtins might plas a role in elhafltil s depressant ct t eet mmli acid sccu elimmn i irt t he rat. Ktu rpp tii tcn and Puunittett pi etn catem i the ir tunlintals xx ith inmio nteth tmein i. till iuthihitor mit prosta glandmn s% nt hcsis, I ites t ounml t hat the tuhilit of et hanoi to dcpu css acid secretion xx as untagmmnu. cvi, suggesting that au increase in prosttuglanditt ss ntltesis nt,is ntt ediate, in part, this effect of ethan ol, Howexer, Ihes dud not measure cyc lic N NIP hcxchs, so coitc itisions tilt an interaction of pi mistaglandins and es cite NNIF mi t his ss steni cannttt he matte, althmmugh esehic. NMP lcxei v hax e been re pmmrteml to he dcx ated in rat h undic muiscle tiller intraperitoiteal injection of prmistagltundmn I, A lso, it is t uti t ortunti te that these experiments were limit done in dmigs, xx here the mechani sm of Ihe inhihitimint of acid secretim mnt at ttigh ethanol cot ice ntr tu tt mm ns is less ele,ur, in sunimars. t he eficcts tifcthtuntol miii gastric acid secretio n turd aceonipanied hs c hanges in supertiemal niucyisal cyclic,\tll levels. these changes reflect, in part. alterations in the actix miles of adens late es eltmsc and phosphodiestei ase. It is not clear boss cthtuiiol exerts its effect, hut con sidering t he high ethanol coneent ratimins used, pail al dentuhuirzmtion or structural altcratitiuis of the enzymes are possihle.. Nismi, part of the i eduet ion in acmd seeretioti that can he induced by et hanol in some spe c es might he mediated through a reduction in A UP Leve ls vi m an increase d synthesis anti release mif prmistag ha nditis. It xx tiulmi be interesting to determine xx hether ethanol aclutillv stimulates pi ostag landtn i c leasc and xx hether il ls more t I ect ixe in species s. here ethanol has a pronounced in hibitory effect for example. in the rat. Nt t his stage. no causeandeffect relationship has been estahlishcmh betxxec ii the ethanvmlinduced reduction of ac id secretion and prostaglandins, Further rcse:ii c h xx iii be required to charif s this point. Under normal circumstances where systentie effects of ethanol are conisidcred. t he high cmmncentratim,ins of i O used in the experiments with the stomach are far in excess of xx hat would he compatihlc sx ith life. However, we are dealing xx ith localized actions of ethanol under conmlitions similar Itt normal drinking of alcohmilic hexertiges, Ftmking into account the damage to the stomach known to result from high concentrations of ethanoi. t he pi eccding discussion further emphasize. the need for caution in drinking beverages with a high alcoholic content, I). ETHANOl. t Nt) ( Cl.I( Nt (, I.EOTII)ES IN OTHER TISSUES I. I.iser Compared to the central nervous s s tem tu nmi the stomach. x ery little research has been done on the possible involvement of cyclic nucleotides in the actions of ethanol on other mirgans.afe xx reports ex ist hut generally are not enlightening as to their possible significance, Ethanol has been reported to induce tmliertmtions in cyclic AM P metabolism in the liver, Shortehaimi aliphatic alctihmils stimulate glucagont esponsis e a,,ielv i i s. 1.

11 ethanol 1,10 S% Mli m F S iluuii.ti,, v S yi,t m the hxi tc rtv x iutc ie.is t xx itft t,iv it mtimlitimin.ml cai fit ut iii the v It.uiit I ivi5 d er. itmi d it tv i m t l ccs in polettes.tm e mihscu xcvi amttm itg p1 i i i i.ii, st v m llt h.ui,uitd t m u s t ti t,mitol s I he cith.mn iccm i lynx mt x at adens hate cx c i.msv hs el hanmil is limit.1 pp.m mm ut t ul t ftc s miitv cult n.11 1mm s.11 t.il least 2, xx it It the tit a xi mn ii mit e they I n e.mched.mi I ftcse alcohols,.iisvi. sti mitu l,utc amieiis hat,, c s v l.ism Jut tat, i tither tmss ud s, mnvi ud im i g kudmi m s, im itc sti it,,. t.a, utlv i hittiimi. lii Iftese st iimh im, s min is uutp hs simm logicalls high domtv cntu amiv mis coum lml c liv it the ucsportsc. Because at th,, peripheral.idt ciiei gte its per,mv tmx it s mm hserx cmi at tet y lti m ill d ci hanvil ad till mu st i ti t i v ilt I i d lid Ii it m 1 cx plmii cd ti le po s sibi lit s t hat s liii i I to trc.mtntenut might,mhi m i the sv ils i li s lix of the v s hic N NI I ss stcmt i to utvime pitit phruutm itt the lix cr I tic s t m tir td that at Ici IS xx ecks of ct itanmil mngcsliort, the I I ),, for utm mi eputtcp hm mttc stintu latitmit hati iti c i m..is emt smxf olm i itt Ii s er hontoge miate p,ut tm e tilate itiris, imimhic,utmtig the tics chmipitteitl of adm eutet gue subscitsitt x ml s I fi ms altcmm mi si Ii sit ix (1% i c III nicd Iv cm 1111m m I x aloes via s after xx i ltd na xx til mirth mkc t hi,ut obsei xemi lit ht ;mi ml, I Ioxx cx er. if the titeasu cute ills xv vu itiamie itt lix er it litm i chontiria. an tneuc,ms m d sensitix its xx as seen l:unalls, the etl cels of v hioutme m lh. In tmi h Irealltim nt appe.u lv i fit llic mli.mlt d I hi mmugh / i eem plmmu s. sih)t t /.lvf t t lt i, l t Iy ant.igm iitt s ts hlmick the ahi l it s m i t itmit epinepiti iitc lvi stmittu ilatc v s v Iiv, N%I l.iv y ullitul h,mtim mit. xx ithi no d ied bs iat ln enet gic tml tt.mg mitt ists 2. Skin (inc st iv l s has appeared studs mug the effect vit shvirt chamn alcohols on ese lm c tntp itt the skin N mi.hikaxx a it al i epmmi ted that v S chic NM) i sc hs ate ltidit.isy yt tip I mi 2,, t mil m I i.i evitic em utrt mlim muim icpemtticmt l mitatuner iii epithernual sltecs indmihal,, ml in I ethatitil, Sintilar i esiilts xx die f otmitd xx it it I pt opattmml atuvi acetone. but not xx ith nuethtttiol anti I htittun,,ii. I Iils effect appears Imt hu e related 1m m,uii.uv t I\.u tlan of epmderuiial, mter t s l.ul m v x chase, Ns x e, lit phs siologmcal smguiiticartce can he attributed tm i these linmi mngs. I. SL N1NtAR AN)) ( ON( I.L SIONS It us lt m mxx y h m,li t hat.mctit m iltyi y hi m tt id tm catntc nt xx itit ethanol m xv i is,u nuuii hm r mu cttccts mii t he Cs v hiv itucleotities mit sex eral v m u galt.5 stcnt s. li tue brain. t mm r ilisltihlv c, dlhtiiimil imllo\idtitititi i csuilts in Ioxx er lex els mit csc lic uiticlem i limit s, espt cialls cs cliv ( itt I. Iim m sx,, x ci,,it uer chu tmi umc amimiiiitisti atimimt ot et hanol, ln mhiv tmt m iu s mit v x v hv rtti cl em mt it le t urtct im n ate dcx atemf. l iim sm v hattgcs appear tm retlt dl 5 hanges iui the,iv Il x his mif the tierx ous s s sl m it i, I)ui ing i tito\ietmtiohi. the hramui is mlepre.scmi. xx hereas du rin g ethtinol xx mthdi a xx ttl ss utm hta ntc, fus pei ttct ixits can hit oh mscrxcd, It is et to be mletcn ntunt mi xx fuetfier chttmntges in t s c hiv numc leotidcii uedu.m teml 55 st m itu s arc u espmiuusil mle for intmixictmt imili. dependence. mmr other biological.ml tm tat im ins t mir xx hich th es has c been imp licatcmi after etltanol cmmnsumptimmn. Nt the prcsm nl si.i tm of i cs,, arc h. change s in cyclic micleotidcs ctm t i be sfioxx n to 1

12 ,u i I M..W. ar. d rc p I him s Is, ii i mm Ii Sum, h,ii,im i i,,ilts alil i t,uil t s at v m i taln s itv sis i i? s lh.miimil Iioxx cxci. ti m m u te h,ts bcv ii,i hlm Iv vlctii muiisth. vtc tii.mi i itcsm v ll.i tt L lii ti tm ui tst i scs s ill s \H,lltl v t h.ut im i l tt ittiis v,,l t mm. tu,ix im mi.v l.ubnm miiiu.i httim, s lit t,is I i t s.uiist u t tit,, et is,, t y th.mttm l has miii tit tu l t i il. illstllltl cis.mtt d their ielatl m ults tiip tti,,\v Its ilttv iv v ttt lv s, II 5 F mmss it lm that.m ltv t.t ti m uls iii,, s sits il(is iemiliyht s.mtm c,,, iivt, ui x it smu tt y u t it m t im s[im u hl s m at m Ifi,utivul I he es.is t im uis, I cx sit,, u uuls ic m u iitt es iii the tmv ttm tts at chh.u itm l.ult mi its pm ui e t t ti.i i sui, ili tlc.it ts m St i l i I m mjli tt t Intl hci v sl,ut I. KF, F FRFM Fs 1,., I.,.1 /,, I I,. t uu.,,,i I, S it. I i,,n, h. \.., i I r i I xi,,,,i i; I i.,,, I, s,... tj 1,. k I.. i i, it,t i ss I i,is I,, it. 5, t,,im f,,, I I S \. I,,.,i R. i s u \% utlt, u,,,! R R.,i a /,, S,,j s :si,i i. 5, I.i,aa It I I tt,. mud I I Ill,s,,i S u t,, lt litis i s, U..i, l I xx lm,i, / \,,,,. I,, up l s I I I, m,u,s.tiimt xt Si si. / i..... F, / r I. I i S i xx t.,,,t.,i t,,,l, I i R, P,Z, d, t, :,, 104 JU., t 1 I 0 u xx kai,t,, t I fti:, I,i,mn i P m,ie r u,ut, i /, S I a/ I xt 1,0,t0 iii I x \,ut h,,fls,,,mn,,t I (,,m v tiv,,t d I, Sal t a: S I t t a ii I x I, ut, p.1, h i, s I Il it SI, It,uui,,I I) Si K 1 ta ft, 5i if, t ( a. 41 ItIt,t to i I i I, I 1 I,, I I Is, ;, It I, xx,,ip, n I I i, t a,, iit ii, ill,, t I i tt,, tlm:,jt.i I i is,. I It Il I I t,,, I I K,,ut, t I i r, i i,,i I,, 5,1 t,,., S. I xx s,,,a, I ) S l.iu L i,.ini I h hii,, m a, i5 Si t I S I (mu: to : I m I, SI, 5 (,w,i, t(,,t umi 1, st,i. i f,! / a, j,;, I ii ) t. I Il I II, i, I, SI,, S t utimk tli,.uitd I i,, st,, i,, R, I, 1 I xx s :h, huh1. t ss i,ii m ud I SI, a, I /1 I /t, tu, it.: IS I t Ii,,,,i, u,,,u mud xx xx 5,1, i 1,1 1 it i a, a 44 u. 1* ) I I K is.mnmt I t i,, ip,i ml, / //, 1.1 Io ttl ii I SI i, hnsm n, II SI, i,in,$ I i.p,,, ptmi,,i, I / it. J,, 4(u t i I tq t I I Si l, Ittls, ih I t.,/ II SI,.,.uitd I t,i,, ium i.,i,i / / I.4 u i I 10, I I,.,l,s tl S SI,, xa SI a, a s,, i. i,/,i. p intl I & i,, ipmi,,:,i.1 i,.,,,,, i i, Ii,,, uss u i i 4, i xx K, P.?. ii i I n. S I s, ii i,uuu,,l I i u,,,t,ui d tj,,,, i ):i tn :i ii x SI. xi,,,mi l m t i t,,,, n,.,/ u,, i% t 1 l.1, I,,, tum,ui,i,it.i u S K,t,ii i,,i /,,j I, \ ij 11:111 ) i0 ii h 55 K, i,,t i.,,p I im i,um t t uml I i,li,, tum,,?,t.1 f,,,,, 1 Ii l/i ho _i 4,i t Si I iiit,, II Ii I 5, hu t, int l t, s R, la,ps, ii 5,,, I,i I 4, i 0 t,. s &,,i,, t I m SI t u.,t iiv,, ii, SI I\ t,, iii,, Ii xx,,iu.i R 5 II,,uu k is It \,,,, i,t.,,,d,, It. ti i I 4I t x li,, ui,mnml 14 I & 1.1 fr, 5, I.t i, u lit Si i4i fs, ( i,flns l,, I R, s, i,ifl I I II li.uts h, is, 5,,,,,, ui f,, I I I It Kt,pi,,irt,,,,it,i St S Is t,um I, ft Ii, p f aa,,,, I I i t0 I S, Iam, mud 14 P IIuiut,, i,.1 Ia,.,, I ti; I, 2(Nl:, if, t i s it I im Iim tkus xx li, i iii,itid 4.m,,Il,,S,,s it, x,u IS S, t i ti 14 Ii I Ji t uim,,,,l xx ii t.,,r, h,i F 5,,., 5 l pi 1 xl x l,, i Ii i.iiuii,,a intl Iv Ixtuuim,itui.i. F ui, t, i,,, S it :

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