Anti-radiation vaccine and specific substances for diagnosis and therapy of radiation injury. Slava Maliev, Dmitri Popov.

Abstract Multifactor fundamental research under experimental and industrial conditions on farm and laboratory animals (parabionts, gnotobionts, mice, rats, rabbits, dogs, sheep, pigs, cattle, and horses), and also on humans (blood serum of Chernobyl NPP clean-up workers) conducted between !"# and #$$# ha%e enabled us to establish the existence of two pre%iously unknown phenomena& the phenomenon of re%ersible redistribution of cytobiochemical parameters in the blood-interstices-lymph-blood system of irradiated animals, which supports compensatory maintenance of homeostasis'( the phenomenon of specific immunochemical reactions to the radiobiological effect, in%ol%ing the formation of high molecular mass glycoprotein (molecular mass-#$$#)$ k*a) + radiation antigens in the lymphoid system with epitopes specific to each form of radiation sickness , #, ,, -, after animals ha%e been irradiated in doses inducing the de%elopment of the cerebral ( ), toxic (#), gastrointestinal (,) and typical (-) forms of acute radiation sickness, ./, , , , 'a0'

1hese two phenomena allowed us to de%elop a methodology for producing specific means to pre%ent, diagnose, and treat radiation sickness (know-how) + an antiradiation %accine, a serum and a set of diagnostics for conducting 23456 analyses'

L S! "# A$$%&' A! "(S MM " $ % ()* ,- ,2 56 56 1 56 7 56 8 56 9:1 56 9:7 56 9:8 56 9:9 5 ;* >6? A, 5B , ; Chernobyl clean-up workers Molecular mass Deoxyribonucleic acid "-lymphocy#e $- lymphocy#e &n#ermedia#e 'orms o' lymphoid cells Cyclic adenosine monophospha#e Circula#in. lymphocy#e /olume !cu#e radia#ion sickness 4peci'ic radia#ion de#erminan# 43D 'or cerebral !34 syndrome 43D 'or #oxic !34 syndrome 43D 'or .as#roin#es#inal !34 syndrome 43D 'or mild #ypical !34 43D 'or modera#e #ypical !34 43D 'or se/ere #ypical !34 43D 'or ex#remely se/ere #ypical !34 coe''icien# o' speci'ici#y <n=yme linked immunosorben# assay @uinoid radio#oxins <x#racorporeal di/ersion o' lymph 4peci'ic an#iradia#ion /accine 3adia#ion dose &n'rared

D !

c!M+ C01 !34 43D 43D 1 43D 7 43D 8 43D 9:1 43D 9:7 43D 9:8 43D 9:9 C4 <0&4! @3" <D0 4!1 3D &3

!'#er analy=in. recen# a##ainmen#s in radiobiolo.y in #he 'ollowin. 'i/e areasC +a#ho.enesis o' acu#e and chronic radiobiolo.ical e''ec#sE #heir e''ec#i/e pre/en#ionE dia.nosis and #rea#men#F 7D 3adia#ion mu#a.enesis and carcino.enesisF 8D 3adia#ion a.in.E decrease in li'e spanF 9D &mmunode'iciencyE which is closely associa#ed wi#h decreased physiolo.ical resis#ance #o ad/erse en/ironmen#al 'ac#orsF GD 4#imula#ion o' physiolo.ical 'unc#ions a'#er exposure #o low radia#ion dosesF we came #o #he conclusion #ha# unders#andin. o' #he na#ure and or.anic in#errela#ionship o' #hese problems and #heir solu#ion are de'ined by #he me#hodolo.ical approaches and a##ainmen#s in #he s#udy o' #he 'irs# problemE iDeDE #he pa#ho.enic mechanisms underlyin. de/elopmen# o' acu#e and chronic radiobiolo.ical e''ec#sD ! li#era#ure and pa#en# search a##es#s #o #he 'ac# #ha# when any li/in. sys#em is irradia#edE #he induc#ion mechanism o' #he radiobiolo.ical e''ec#s is a 'unc#ion o' #he syner.ism o' #he direc# e''ec# o' radia#ion and #he biolo.ically ac#i/e radio#oxic subs#ances #ha# 'ormE #he e''icacy o' which are a 'unc#ion o' #he dura#ion and dose o' irradia#ion H88E 8IE 91E JKLD "hese s#udies 'ormed #he basis 'or #he me#hods o' dia.nosisE 1D

pre/en#ion and #rea#men# o' acu#e and chronic radia#ion inMuries in use #oday and were nonspeci'ic wi#h respec# #o #he pa#ho.en-'ormin. a.en#D Nowe/erE polyclonal experimen#al s#udies o' 'arm and labora#ory animals ha/e unambi.uously shown #ha# #he immunochemical processes #ha# occur in #he lymphoid sys#em are de'ini#i/e wi#h respec# #o pa#ho.enesis o' #he radiobiolo.ical e''ec# and are speci'ic #o le/el o' radia#ion o/er a wide spec#rum o' /alues HJE 81E 81DaLD Compara#i/e lo.ical analysis o' #he ex#ensi/e base o' da#a #ha# had been ob#ained by #hen by well-known radiobiolo.ical schoolsE .roups o' scien#is#s in rela#ed scien#i'ic areas in #he Oni#ed 4#a#esE P3QE Prance and Rapan ser/ed as #he #heore#ically-based prelude #o #hese s#udiesD Se will consider here #he mos# si.ni'ican# 'undamen#al radiobiolo.ical research s#udiesD &n par#icularE !DMD Tu=in e# al H7UL isola#ed re.ak#enon 'rom plan# #issueD "his subs#ance has a radioimuni=in. e''ec# on cell .enomesE accompanied by depression o' D ! syn#hesisE decreased cell di/ision and #issue .row#hE 'orma#ion o' pykno#ic nucleiE chromosome 'ra.men#a#ionE and 'orma#ion o' mu#an#sD !# #he same #imeE a'#er adminis#ra#ion o' re.ak#enon and o#her radio#oxins isola#ed 'rom irradia#ed #issuesE animals 'ailed #o display #he en#ire VbouWue#X o' radia#ion reac#ions Ysymp#oms o' acu#e radia#ion sickness o' /arious 'orms and se/eri#y could no# be modeled in experimen#al animalsZD "he broad spec#rum o' ordinary chemical compounds #ha# can ha/e analo.ous radiomime#ic e''ec#s #o #hose o' re.ak#enon demons#ra#es #he lack o' speci'ici#y in induc#ion o' #hese e''ec#s by #he main classes o' radio#oxinsC hydroperoxidesE peroxidesE polyphenolsE semiWuinines and WuininesE ke#oaldehydesE bio.enic aminesE pro#eins and polypep#idesD "hese compounds are always presen# in an un#rea#ed animal and play a role in #he biochemical processes o' ca#abolism and anabolism #ha# main#ain homeos#asisD 3adia#ion exposure induces a process in which #he le/els o' one or ano#her class o' #hese compounds increasesD "he s#udies o' 1D!D Topylo/a e# al H78L demons#ra#e #he cri#ical impor#ance o' elimina#in. Wuinoid radio#oxins 'rom #he body in #rea#men# by hemosorp#ion o' animals a'#er le#hal doses o' .amma irradia#ionD 1DQD 1ladimiro/ e# alD H[L de#ec#ed a 'rac#ion o' low molecular wei.h# D ! in #he blood plasma o' an animal exposed #o ioni=in. radia#ionD "here are #wo poin#s o' /iew on #he ori.in and role o' #his D !D %ne o' #hem links i#s ori.in wi#h #he pos#radia#ion 'ra.men#a#ion o' chroma#in 'rom lymphocy#e nucleiD "he o#her poin# o' /iew pos#ula#es an ac#i/e process ser/in. #o #ransmi# a Vhori=on#alX 'low o' .ene#ic in'orma#ion and #o correc# cell 'unc#ionin. by means o' in#ra.enome res#ruc#urin.sD </olu#ionary phylo.enesis o' #he lymphoid sys#em in animals Y'rom #he simples# or.anisms #o mammalsZE which e/ol/ed in #he presence o' a cons#an# radia#ion 'ac#orE made i# possible #o in#e.ra#e #he 'ollowin. ei.h# /i#al 'unc#ionsC immunolo.icalE me#abolicE #ranspor#E s#ora.eE hema#opoiesisE resorp#ionE excre#ion and barrier-'il#ra#ion H7E JE IE 77E 7GE 87E 9ILD "his in#e.ra#ion makes i# #he mos# radiosensi#i/e o' all #he o#her physiolo.ical sys#emsE as was no#ed as early as 1IU9 by ND NeinekeD H99LD "here are /ery many s#udies o' #he radia#ion pa#holo.y o' lymphoid or.an s#ruc#ure and 'unc#ion in mammals H1E 8E 9E K 11E 17E 19E 1GE 71E 79 7[E 7KE 81E 89E 8GE 91E

97E 98LD "hey ha/e helped #o disclose cer#ain mechanisms underlyin. #he essen#ial immunodepressi/e e''ec# o' ioni=in. radia#ionE emphasi=in. #he dea#h o' in#erphase lymphocy#es H17E 89E 97E 99LD "he reproduc#i/e dea#h o' lymphocy#es also makes some con#ribu#ion #o #his immunodepressi/e mechanism H1JE 97LD "hese au#hors s#ar#ed 'rom #he 'ac# #ha# an impor#an# proo' o' #he decisi/e role o' lymphocy#e dea#hs in radia#ion lymphopa#holo.y is #he correspondence o' #he 3DE ob#ained when #he whole body is immuni=edE 3D\UDK Qy and #ha# in #he sys#em o' adap#i/e #rans'er o' lymphoid cellsE 3D\UD[ QyD "his a priori circums#ance con/inced radiobiolo.is#s #ha# #he in#erphase and reproduc#i/e dea#h o' lymphocy#es is decisi/e in de/elopmen# o' radia#ion immunopa#holo.y Ywhich we do no# 'ind incon#ro/er#ibleE since #his occurred be'ore our disco/ery o' #he phenomenon o' re/ersible redis#ribu#ion o' biochemical parame#ers in #he blood-in#ers#ices-lymph-blood sys#em o' irradia#ed animalsZD Pur#hermoreE $DDD ]hi/o#o/skiy H1UL demons#ra#ed a Wuan#i#a#i/e associa#ion be#ween #he pykno#ic chan.es in #he cell nuclei o' #hymocy#es and produc#ion o' produc#s o' pos#radia#ion chroma#in decayD "he en=yme responsible 'or #he decomposi#ion o' chroma#in in irradia#ed cells is Ca:M.-dependen# endonucleaseD !reas o' endonuclease a##ack are dis#ribu#ed randomly and are no# associa#ed wi#h #he le/el o' repe#i#ion o' nucleo#ide seWuences o' D ! or #he #ranscrip#ional ac#i/i#y o' chroma#inD "his suppor#s #he conclusion #ha# #he radia#ion dea#h o' lymphoid cells and #he .eneral biolo.ical phenomenon o' pro.rammed cell dea#h are iden#ical in na#ureD &oni=in. radia#ionE #husE like o#her lympholy#ic 'ac#orsE can e/oke induc#ion o' .roups o' speci'ic .enes #ha# swi#ch on #he cell dea#h pro.ramD Nowe/er some researchers H1UE 9ULE whoE includin. ioni=in. radia#ion in #he .roup o' lympholy#ic a.en#sE wi#hou# su''icien# Mus#i'ica#ionE ar.ue #ha# .lucocor#icoidsE c!M+ and alkyla#in. compoundsE which are so di''eren# in chemical na#ureE mus# en#er in#o speci'ic chemical in#erac#ions o' #he epi#ope-idio#ope #ype wi#h #he speci'ic .ene loci responsible 'or cell dea#hE while ioni=in. radia#ion by na#ure is incapable o' direc#ly en#erin. in#o such in#erac#ion wi#hou# adduc#or-media#ors speci'ic #o radia#ionD "husE we ha/e pos#ula#ed #ha# hi.h-ener.y irradia#ionE like o#her .eno#oxinsE can e/oke #he induc#ion o' speci'ic .enes #o swi#ch on #he cell dea#h pro.ram only i' media#ed by a sys#em o' speci'ic adduc#orsD "hese hypo#heses ha/e been con'irmed in our researchE as well as elsewhereD 4#udyin. #he #ri..er mechanism 'or radia#ion-induced lymphocy#e dea#hE D&D 4orokina H8[L used #he resul#s o' numerous experimen#s #o show #ha# ioni=in. radia#ion induces chan.es in #he an#i.en pheno#ype o' imma#ure #hymocy#es in miceD "his has #he same #ype o' e''ec# as chemical di''eren#ia#ion induc#ors and #hymo#ropinE which indirec#ly a##es#s #o #he speci'ic modi'yin. e''ec# o' ioni=in. radia#ionD Pundamen#al a##ainmen#s in molecular radiobiolo.y ha/e 'os#ered si.ni'ican# pro.ress in explainin. #he mechanisms o' #his process and #his has permi##ed TD+D Thanson #o approach unders#andin. o' many aspec#s o' #he occurrenceE de/elopmen# and reali=a#ion o' radia#ion e''ec#s in lymphocy#es H9ULD Si#hin #he con#ex# o' #he proposed hypo#hesisE in#erphase dea#h o' irradia#ed lymphocy#es is considered one o' #he examples o' #he .eneral biolo.ical phenomenon o' pro.rammed cell dea#hD &n#erphase cell dea#h occurs as #he resul# o' swi#chin. on a speci'ic .ene#ic pro.ramE #he #ri..erin. o' whichE

#he au#hor asser#sE may occur as #he resul# o' many si.nalsE amon. which are /arious #ypes o' lympholy#ic a.en#sC .lucocor#icoidsE alkyla#in. compoundsE c!M+E di''eren#ia#ion induc#orsE and ioni=in. radia#ionD "husE in addi#ion #o con'irmin. #he radiobiolo.ical idea #ha# #he main reason 'or #he de/elopmen# o' lymphopenia is in#erphase and reproduc#i/e lymphocy#e dea#hE #he resul#s we ha/e ob#ained a##es# #ha# #he 'ac#or o' slowed lymph #ranspor# also makes an impor#an# con#ribu#ion #o de/elopin. lymphopenia YPi.ure 1ZE resul#in. in de/elopmen# o' a de'ici# in circula#in. lymphocy#es in #he lymph 'lowD "he ma.ni#ude o' #his con#ribu#ion can reach 8U-9G^ dependin. on 3D and elapsed #ime a'#er irradia#ion YPi.ure 7ZD
ml)min.

L*MP+ #L", - ( !+& ("%M

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#igure 0. 1hanges over time in lymph flo2 .A/ and in number of lymphocytes .$/ of rabbits 2ith severe A%S.

cells:hour

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#igure 3. 1hanges over time in circulation of lymphocytes and subpopulations in the lymph-blood systems of rabbits 2ith severe A%S. &'E alon. wi#h #he de/elopmen# o' #his processE we consider #he mi.ra#ion o' lymphoid cells #o #he wall o' lymphan.ions and an.ions Y#he 'i.ure shows #his #o in/ol/e 1U-1G^ o' #he lymphoid 'orms o' cellsZE iDeDE impairmen# o' #he speci'ici#y o' lymphocy#e Vhomin.XE #hen #he in#erphase and reproduc#i/e dea#h o' lymphocy#esE and #hus #his processE make a 9U-9G^ con#ribu#ion #o #he de/elopmen# o' lymphopeniaD "he increase in #he #i#er o' au#o-an#ibodiesE #he morpholo.ical subs#ra#e 'or #he syn#hesis o' which is lymphocy#e cellsE #he absence o' se/ere hyperpro#einemia in #he #ranspor# media in #he irradia#ed animal_s body Ya'#er all #he dea#h o' any cell 'orms should be accompanied by .enera#ion o' pro#einE pep#idesE e#cDZ and #he analysis o' #he chan.es o/er #ime in absolu#e numbers o' lymphocy#esE #heir subpopula#ions Yand #heir ac#i/e capaci#y #o 'orm rose##es wi#h sheep ery#hrocy#esE see Pi.ure 8Z and o' ery#hrocy#es and pla#ele#s all suppor# #his hypo#hesisD

#igure 4. !-theophylline tolerant lymphocytes of central lymph in cattle irradiated at a dose of 3 5y, Scanning image. &nlargement 6 7888. 3esearch per'ormed by 1D!D TirshinE winner o' #he O443 4#a#e +ri=eE and his associa#es be#ween 1IKJ and 1IIUE which unambi.uously demons#ra#ed #ha# black mo#ley ca##le exposed #o radia#ion 'rom Chernobyl 'allou# exhibi#ed enhanced radia#ion resis#anceE is par#icularly no#ewor#hyD "hese s#udies con'irm #he principle underlyin. radia#ion immuni=a#ion o' animals H18LD "he resul#s o' #his research ha/e si.ni'ican#ly con'irmed our hypo#hesis concernin. #he speci'ici#y o' induc#ion o' #he radiobiolo.ical e''ec#D %ur research be#ween 1IK7 and 7UU7 was de/o#ed #o s#udy o' #his areaD %esearch materials and procedures "hese s#udies were conduc#ed in #he $elarussian 4cien#i'ic and 3esearch &ns#i#u#e 'or 3adiobiolo.y in QomelE #he 4#D +e#ersbur. 1e#erinary &ns#i#u#eE #he Ta=an !ll-Onion 4cien#i'ic 3esearch 1e#erinary &ns#i#u#eE #he 0i#huanian 1e#erinary !cademy YTaunasZE and &4< Y$onnE P3QE "oron#oE CanadaZ on black mo#ley ca##leE 7DG-8DU years o' a.eE li/e wei.h# o' 8UU-8GU k. Yn\189ZF Okrainian pi.sE UDG-1DU years o' a.eE li/e wei.h# 8G-IU k. Yn\197ZF Vprekos X sheepE 8-17 mon#hs oldE li/e wei.h# 1K-78 k. Yn\1GJZF mixed breed do.sE 7-9 years o' a.eE li/e wei.h# JDU-JEG k. Yn\1J7ZE Chinchilla rabbi#sE 11-17 mon#hs oldE li/e wei.h# 8DG-8D[ Yn\1KUZE 0a#/ian dra'# horsesE 8-K years o' a.eE li/e wei.h# 8GUGGU k. Yn\87ZE $alp miceE 7-8 mon#hs oldE li/e wei.h# 7U-77 . Yn\7J8JZE Sis#ar ra#sE 8-9 mon#hs oldE li/e wei.h# 1KU-77U . Yn\9UU7ZD "o exclude #he microbial 'ac#or in #he pa#ho.enesis o' !34E #he experimen#s were per'ormed in .no#obio#ic animal labora#oriesD "o s#udy #he mi.ra#ional and #oxemic 'ac#or we used parabion#s wi#h crossed lymph and blood circula#ionD "he experimen#al .roup consis#ed o' animals wi#h normal blood pro'iles and body #empera#ureE in which #he homeos#a#ic cons#an# o' cen#ral lymph did no# exceed #he limi#s o' normal /ariabili#yE which we had pre/iously de#ermined 'or 'i/e species o' mammals Y"able 1Z since #his was o' 'undamen#al impor#ance #o our s#udyD

"able 1 +omeostatic constants of central lymph in animals

1onstant C01 Yl:k.Z 0ymph 'low Yml:min:k.Z 0eukocy#es Y#:`lZ 0ymphocy#es Y#:`lZ -::Y^Z ?a -lymphocy#es Y^Z ?y -lymphocy#es Y^Z ? clymphocy#es Y^Z B -lymphocy#es Y^Z Monocy#es Y^Z eu#rophils Y^Z +la#ele#s Y#:`lZ <ry#hrocy#esYkl:`lZ "o#al pro#ein Y.^Z +ro#ein 'rac#ionsC ^ 1D !lbumins 7D +os#albumins dD"rans'errins 9DNap#o.lobulins GD Qamma.lobulins JD Macro.lobulins !c#i/i#yC ly=osomal Y^Z bac#ericidal Y^Z be#a-ly=ines Y^Z pro#eoly#ic Ym.:lZ "riiodo#hyronineE nM:l ?hyroxinE nM:l Cor#isolE n):l &nsulin MCE uni#s :ml 1attle Pigs Dogs Sheep %abbits

UDU7[-UDUK7 UDU9U-UDUIU UDU81-UDU8J UDU99-UD 1JJ 9DI aUD7 9DK b UD7 IKDU b UD8 GJDU b 7DJ 7GDU b 7DI 1IDUa b 7D1 89DU b 8DU 1D8 b UD8 UD8bUD8 I[DU b 1DG 8UDU b1GDU 9D8 b UDUJ 87D9 b 1D7 [D7 b UD9 1[DI b UD[ 71DJ b UDJ 1JDI b UD8 9D 1 b UD 1 JDK b UDI G8DU b1JDU 9DK b UD7 81GDUb1KDU 1D1K b UDUG 8JD9 b 7D9J JD8 b UD9 JD1 b UD1 IKD9 b UDG J8DU b 9D8 8UDU b 8D8 JD 1 b 1D7 89DU b 9DK UDI b UD9 UD9 b UD7 K8DU b 1D7 1[UDU b 9GDU GD9 bUDU8 88D9 b 1DJ [DU b UD9 1[D 1 b UD[ 7UD 1 b 1D 1 1JD8 b UDK 9DI b UD9 KDI b UD9 JIDU b1UDU ID8 b UDI 8KUDU b1UDU 1DK8 b UD 1U 79D9 b 8D71

UDU8K-UDU91 UDU7U-UDU[U UDU9K-UDU98 UDU[1-UDU[[ UDU19-UDUJG UDU[J-UDUKU 7D[ b UD7 7DG b UD7 IJDU b 1DU G8DU b 7D9 1KDU b 7DU 17D 1 b 1D8 8UDU b 8D9 8DI b 1DU UD9 b UD7 JKDU b 7D7 17DU b17DU 8DUJbUDUJ 87D[ b 1D[ JDU b UD8 1KD 1 b 1D1 77DU b1D9 1GDI b UD7 GDU b UD7 18D[ b UD[ J7DU b IDU GDG b UD 1 7[7D Ub88D U 7DUU b UDUK [KD[ b [D81 8JD[ b 1DI1 79D8 b 1D77 GD8 b UD8 GD7 b UD7 IKDJ b UDI J1DU b 9DU 1[DU b 9D1 GD9 b 1D9 8KDU b GD1 UDJ b UD8 UD9 b UD8 1UUDU b 1DJ 1GDU b [DU 9D 1 b UDU7 81D8 b 1D9 JD9 b UD8 1KDI b UDK 7UD8 b 1D8 1JDJ b UD9 9DG b UD8 [DU b 1D 1 J8DU b ID9 GDI b UD9 7I1DUb17DU 1DU1 b UDU8 8DK b UD9 8D[ b UD8 IKDU b UD9 JJDU b 7D1 8KDU b 1DI 7JD 7 b 7D7 7KDU b 1DK UDI b UD7 UDJ b UD7 88DU b 1D[ 8KDU b17DU 8D81 bUDU8 87D7 b 1DK JD 1 b UDK 1KDG b UDG 71D 1 b 1DJ 19D8 b UDI 8D[ b UD 1 [DJ b UDG GJDU b 9DU 9D7 b UD1 1[IDUb7GDU 1DUU b UDU9

117DGb JDUJ KUDU b 8D77 7KD9 b 1D 1G 8KD9 b 1DUI

118D9b 9D 17 171D Ub [D97 91D9 b 7D 1[ 88D[ b 1DI8 7JDI e 1D11 7KD8 b 1D88

"hrou.hou# #he en#ire period o' each experimen#E ran.in. 'rom 1 day #o 7 yearsE #he con#rol and experimen#al animals were main#ained under iden#ical condi#ions o' 'eedin.E housin.E and careE correspondin. #o li/es#ock main#enance reWuiremen#s and s#andardsD MildE modera#eE se/ereE and ex#remely se/ere acu#e radia#ion sicknessE o' #he #ypical 'ormE as well as #he .as#roin#es#inalE #oxic and cerebral syndromes were induced

in #he experimen#al .roups o' animalsD "he animals were irradia#ed in 3OM-1[E +umaE and +anorama de/icesD "he exposure dose ra#e ran.ed 'rom 8 !:k. #o 7I !:k.D %n #he day precedin. radia#ion exposureE and also 1GE 8UE and 9G days pos#exposureE a lympho/enous anas#omosis was crea#ed in #he animals usin. #he me#hod described in H8ULD ! schema#ic dia.ram o' #he experimen#al procedure is pro/ided in Pi.ure 9D &6P&% M&(!AL M&!+"DS Morphological Surgical $iochemical mmunologic

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#igure 9. A schematic diagram of the e:perimental procedure 0 !horacic duct; 3. <ugular vein; 4. 1ephalic vein .humerus/; 9. 1istern; =. Lymph nodes. "hrou.hou# #he en#ire pos#radia#ion periodE samples o' cen#ral lymph and peripheral blood were #aken 'rom #he animals and used 'or measuremen# o' cy#obiochemical parame#ers usin. #he me#hod described in H8ULD "he me#hods o' immune deple#ionE a''ine immuno-lympho-plasmasorp#ion YPi.ure GZ as well as direc# ex#rac#ion YPi.ure JZ were used #o ex#rac# #he speci'ic immunochemical de#erminan# Y43DZ 'rom #he cen#ral lymph o' animals wi#h cerebral Y43D 1ZE #oxic Y43D 7ZE .as#roin#es#inal syndromes Y43D 8Z and #ypical Ymild-43D 9:1E modera#e 43D 9:7F se/ere 43D 9:8F and ex#remely se/ere 43D 9:9Z 'orms o' !34D

#igure = . Diagram of specific .affine/ immuno-lympho-plasmasorption> 0-animal lymphoplasma; 3, 9, = - membranes 2ith immobili?ed ligands; 4 - activated charcoal; @ - eluent.

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#igure @. Direct e:tract of S%D 4 from central lymph .(o. 9049. #irst hours after irradiation in dose of =8 5y/. "he speci'ici#y o' #he 43D was de#ermined #hrou.h #he reac#ions o' en=yme-linked immonosorben# assay Y<0&4!Z YPi.ure [Z in research subMec#sE as shown in Pi.ure KD

11

#igure A. &L SA method .A - diagnostic substance containing Bno2n positive and negative antigens; $ - antibodies marBed 2ith en?yme marBers; 1 - substrate; D product of color reaction; & - product of the color reaction; #- remaining liCuid; 5recording of result using automatic photometer. &' analysis resul#s were posi#i/eE blood was #akenE serum separa#ed ou# and &.Q isola#edD

17

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$L""D

L*MP+

#igure F. %esearch subjects Ya##en#ion was 'ocused on cen#ral lymph and peripheral bloodZ1D "his allowed us #o de#ec# #he presence and se/eri#y o' radia#ion inMury in #he animals by iden#i'yin. 43D-1F 7F 8 and 9 speci'ic #o Ydi''eren# le/els o'Z radia#ion ener.y and also #he radia#ion an#ibodies speci'ic #o each 'orm o' radia#ion sickness Y<0&4!-en=yme-linked immunosorben# assay me#hodZE H7ILD 3eac#ions were /isually assessedE based on #he di''erence be#ween #he color in#ensi#y o' #he reac#ion produc#s 'or #he experimen#al and con#rol .roupsE as well as bein. measured wi#h a scannin. spec#ropho#ome#er wi#h wa/e len.#h o' 9I7 nmD 1isual assessmen# u#ili=ed a 9 poin# scale YeeeeZD "he #es# was considered posi#i/e i' #he assessmen# was ee or hi.herD Shen #he reac#ion resul# was recorded spec#ropho#ome#rically #he coe''icien# o' speci'ici#y YC4Z was compu#edD ! reac#ion was considered posi#i/e i' #he C4 was 7DU or more and ne.a#i/e when i# was less #han 7DUD &mmunochemical reac#ions were induced be#ween #he 43D prepara#ions and an#i43D in order #o ob#ain a soluble immune complexD "he 'ollowin. procedure was usedC 9-G `l 4D3 an#ibody Yprecipi#a#ed 'rac#ion a# 88^ sa#ura#ion o' an ammonia sul'a#e solu#ion o' an#i-43D wi#h a #i#er o' 1C1UUUZ was added #o a UDG ml solu#ion o' 43D o' #he appropria#e epi#ope Y1 `MZ in UDU1 M #ris-NC0E pN KDU and incuba#ed 'or 7DG hours a# a #empera#ure o' 7UfCD "o de#ermine #he speci'ici#y o' #he bindin. o' 43D and an#i-43DE we incuba#ed #his mix#ure a# 7UfC 'or 9G minu#es and #hen #he reac#ion produc#s and #he ini#ial componen#s o' #he .el-'il#ra#ion sys#em were separa#ed on a "4T-#ele JU NSD "he <0&4! me#hod pro#o#ype and #he compara#i/e descrip#ion o' #he 43D prepara#ions were based on !DMD Tu=in_s me#hod H7ULE which was desi.ned 'or iden#i'ica#ion o' Wuinoid radio#oxins H9GF 9JLD "he compara#i/e analysis o' 43D samples
1

2ditor8s note& #he 'irs# par# o' #he sen#ence appears #o be missin.
18

o' /arious epi#opes used a 43-7U YP3QZ &3 spec#rome#er usin. #he me#hod described in H7J and 9KLD 3adiomime#ic proper#ies o' 43D prepara#ions were e/alua#ed on #he basis o' capaci#y #o induce radiobiolo.ical e''ec#s in #he animals a'#er #hey were adminis#ered paren#erallyD 0yophili=a#es o' 43D 1 and 43D 9:9 Yisola#ed 'rom #he lymph o' animals irradia#ed a# doses inducin. cerebral and ex#remely se/ere !34Z were dissol/ed in an iso#onic solu#ion o' aClD "he dose o' prepara#ion adminis#ered was based on compu#a#ion o' #he amoun# o' 43D per uni# /olume o' cen#ral lymph and absorbed dose o' radia#ionD 4ymp#oms o' !34 induced wi#h 43D 1 and 43D 9D9 were compared wi#h similar symp#oms induced by irradia#ion YPi.ure IZD

#igure 7. +orse (o. 09. &:tremely severe A%S .@.= 5y/ "he resul#s o' #he s#udies were analy=ed usin. ! %1!D %esearch %esults 4ubs#ances isola#ed 'rom #he cen#ral lymph o' animals su''erin. 'rom #ypicalE .as#roin#es#inalE #oxic and cerebral 'orms o' !34 ha/e #he appearance o' li.h# bei.e #o li.h# yellow powderE which adheres somewha# #o .lassE is soluble in wa#erE aWueous solu#ions o' aClE and alcoholE and reac#s sli.h#ly #o Wuinoid radio#oxinsD eu#ral aWueous solu#ions were usedD

19

"able 7 presen#s #he componen#s o' 43D 8 and 43D 9D9 isola#ed 'rom #he cen#ral lymph in #he .as#roin#es#inal and ex#remely se/ere 'orms o' !34D "he increase in le/el o' absorbed dose 'rom #he #ypical 'orm o' !34 #o #he .as#roin#es#inal 'orm is accompanied by a J-K^ .row#h in #he pro#ein and lipid componen#s and a s#able le/el o' carbohydra#esD "he mineral residue decreases by 7^D "he resul#s o' #he chroma#o.raphic analysis re/ealed #he rela#i/e homo.enei#y o' #he 43D prepara#ionsE and accordin. #o compu#a#ionsE #he molecular mass o' #he isola#ed de#erminan#s were on #he order o' 7UU-7GU kDaD "his circums#ance is responsible 'or 43D bein. #ranspor#ed #hrou.h #he lympho.enic rou#eD "able 7 1omponents of S%D 4 and S%D 9)9 Componen# Y^Z 1D +ro#eins 7D 0ipids 8D Carbohydra#es 9D Mineral residue &n#es#inal 43D 8 GUD 1 e UDUI 8KD7 e UDU9 1UD7 e UDU8 1D8 e UDU9 "ypical 43D 9:9 GJD7 e UD 17 8UD 1 e UDUI 1UD 1 e UDU[ 8D9 e UD 1[

"he &3-spec#ra o' #he prepara#ions showed bo#h #he .eneral and #he indi/idual 'ea#ures o' 43DD &n par#icularE #he &3-spec#ra o' 43D 8 and 43D 9:9 su..es# #ha# bo#h #hese de#erminan#s are compounds o' iden#ical #ypeD "his was con'irmed by <0&4! reac#ionsE where we obser/ed cross reac#ions be#ween 43D 9D9 and an#i.ens #o 43D 8 and /ice /ersaD "he resul#s o' immunochemical analysis o' 43D 8 and 43D 9:9 su..es# #ha# #heir molecules consis# o' #he iden#ical s#able hea/y chain macromoleculesE and rela#i/ely /ariable li.h# chain moleculesE which de'ine #he 43D 8 and 43D 9:9 epi#opesD "he hi.h immuno.enici#y and speci'ici#y o' #he isola#ed 43D 1-9 prepara#ions allowed us #o use #hem as an#i.ens 'or conduc#in. <0&4! analyses #o dia.nose radia#ion inMuries o' animalsD "his allowed us #o de#ec# Vradia#ionX an#i.ens Y43D 1-9Z in #he blood serumE urineE and or.ans o' animals YPi.ure KZ durin. #he 'irs# 7 mon#hs a'#er radia#ion exposure Y"able 8Z and #he Vradia#ionX an#ibodies #o #hem o/er #he subseWuen# 7 years Yobser/a#ion periodZD Si#h re.ard #o #he chemical na#ure o' isola#ed de#erminan#s o' 43D 8 and 43D 9:9E i# should be no#ed #ha# #hey are conMu.a#ed pro#eins and #heir pros#he#ic .roup con#ains lipid and carbohydra#e componen#sD "his #ype o' conMu.a#ed pro#ein is a componen# o' #he cell membranes and in#racellular biomembranes o' #he nucleusE mi#ochondriaE microsomesE and also is presen# in a 'ree s#a#e Yin physiolo.ical #ranspor# mediaZD &# has been es#ablished #ha# an analo.ous #ype o' conMu.a#ed pro#ein par#icipa#es in #he s#ruc#uralE complex or.ani=a#ion o' myelin shea#hsE ner/esE chloroplas#sE pho#orecep#or and elec#ron #ranspor# sys#emsE and #he rods and cones o' #he re#ina HGLD

1G

!s 'or #he mechanisms #hrou.h which #he conMu.a#ed pro#eins we ha/e isola#ed 'orm a'#er radia#ion exposureE we ha/e #wo workin. hypo#hesesC c 'reein. o' s#ored conMu.a#ed pro#eins o' #his class #ha# were presen# earlierE 'ollowed by #heir modi'ica#ionF c radia#ion-chemical and en=yma#ic processes responsible 'or new 'orma#ion o' lipo-.lyco-pro#einsD <xis#in. da#a a##es# #o #he 'ac# #ha# /arious nonco/alen# 'orcesE de#ermined by #he presence or absence o' lipid componen#s o' #he ioni=ed .roup o' a#omsE par#icipa#e in #he 'orma#ion o' conMu.a#ed pro#eins o' #his #ypeD !# #he same #imeE #he lack o' cross reac#ions in #he <0&4! #es# be#ween normal conMu.a#ed pro#eins isola#ed 'rom myelin shea#hs and ner/es and #he an#ibodies #o 43D 8 and 43D 9:9 and /ice /ersaE ar.ues in 'a/or o' radia#ion chemical and en=yma#ic processes bein. responsible 'or 'orma#ion o' Vradia#ionX .lycopro#eins #ha# may be classi'ied as lympholy#ic a.en#sE since #hey are adduc#s #ri..erin. #he induc#ion o' pro.rammed dea#hD &n #he nex# series o' experimen#s we s#udied #he 'orma#ion o' 4D3 in cen#ral lymph and peripheral blood in animals su''erin. 'rom di''eren# #ypes o' !34D "he resul#s o' #hese s#udies are presen#ed in "able 8D "able 8 Specific immunochemical reactions in different types of A%S

!ype of A%S

'isual and spectrophotometric assessment of &L SA .G and 1S/

SD%-0

SD%-3

SD%H-4

SD% 9)9

SD% 9)4

SD% 9)3

SD% 9)0

eeee Cerebral 7DK-8D8 ee "oxic 1DK-7D 1

eeee 7D[-8DU eeee 7DG-8D 1

eeee 7DJ-7DI eee 7D7-7D9

eee #'#-#'! eee 7D7-7D9

eee 7D1-7D7 eee 7D1-7D8

ee 7DU-7D1 ee 1DK-7DU

ee 1DK-7DU ee 1DK-7DU

Qas#roin#es#inal

c g7DU

e99 7D7-7DG

eeee 7DJ-8D 1

eee #'#-#')

eee #'#-#')

ee 1DK-7DU

ee 1DK-7DU

1J

"ypicalC c g7DU c g7DU eee 7D 1-7D7 eeee 7DG-8DU eee 7D 1-7D7 ee 1DK-7DU ee 1DK-7DU

hZ ex#remely se/ere

bZ se/ere

c g7DU

c g7DU

c g7DU

eee 7D7-7DG

eeee 7DJ-8DU

eee #'#-#')

ee 1DK-7DU

cZ modera#e

c g7DU

c g7DU &

c g7DU

c g7DU

eee 17DU-7D1

eeee 7DG-7DJ

e99 7DU-7D1

dZ mild

g7DU

g7DU

g7DU

g7DU

c g7DU

ee 1DK-7DU

eee9 7DG-8DU

"hree #o 'i/e hours a'#er irradia#ion Y"able 9Z #ha# induced a mild 'orm o' !34E #here was an increase in le/el o' 43D 9D1 in #he lymph and blood Ya'#er J-7U hoursZ o' irradia#ed animals #o #he maximum le/elD 43D 9D1 remained a# #ha# pla#eau 'or G-7U hoursD "he an#ibody #i#er #o 43D 9:1 in lymphoplasma and blood serum pre-irradia#ion was 1CKU-1C1UUE a##es#in. #o #he lack o' 43D 9:1 in #he animals_ bodiesD !'#er GU-JU hoursE #he an#ibody #i#er #o 43D 9:1 increased and reached a maximum on day 7G-9GD "he coe''icien# o' speci'ici#y ran.ed 'rom 7DU-7DGD &n indi/idual animals #he an#ibody #i#er #o 43D 9:1 remained a# a rela#i/ely hi.h le/el un#il JU days and on a modera#e le/el c wi#h C4 o' a# leas# 7 c 'or up #o 7 years Y#he dura#ion o' #he obser/a#ion periodZD &# should be no#ed #ha#E alon. wi#h 43D 9:1 and #he an#ibodies #o i#E #he presence o' 43D 9:7D 43D 9:8E and 43D 9:9 and #he correspondin. an#ibodies was de#ec#ed in all animal speciesE bu# in si.ni'ican#ly lower #i#ers YC4 \ 1DG c 1DKZD

1[

"able 9C !ime to develop ma:imum level of S%D as a function of type and severity of acute radiation sicBness "ype and se/eri#y o' "ime in#er/al a'#er irradia#ion YhoursZ #o de/elop maximum 43D !34F 43D le/elE animal species and irradia#ion subMec# 3abbi#s lymph "ypical mildE 43D 9:1 8- K Modera#e 43D 9:7 4e/ere 43D 9:8 <x#remely se/ereE 43D 9:9 8-1U 8-79 blood J-1U J-1U J-79 Do.s lymph G- K 8-1U 1-[7 blood 7U-9U 1G-GU K-[7 lymph 8-17 8-19 8-79 Ca##le blood J-1G K-1G J-[7

1-79

J-7K

1-[7

9-9U

7-7K

8-KU

3esul#s o' reac#ions #o #he presence o' Wuinoid radio#oxins Y@3"Z measured accordin. #o #he me#hod described in H99L were ne.a#i/e bo#h in lymph and in bloodD Nowe/erE in cases o' modera#ely se/ere !3C #he 'orma#ion o' a posi#i/e le/el o' @3" and maximum 43D in lymph preceded such le/els in blood by an a/era.e o' 7-G hours and was obser/ed in experimen#al animalsE re.ardless o' speciesE durin. #he period 8-1U hours a'#er irradia#ionD "he reliabili#y o' #he aniline reac#ion #o @3"E as assessed on #he basis o' op#ical parame#ers o' lymph samples s#udiedE di''ered si.ni'ican#ly 'rom #he reliabili#y o' resul#s o' <0&4! analysis 'or #he presence o' 43D 9:1 YpgUDUG and pgUDUU1ZE and 'ur#hermoreE a'#er 79 hoursE reac#ions #o Wuinoid produc#s became ne.a#i/eD "husE alon. wi#h #heore#ical indica#ionsE a whole series o' experimen#al 'ac#ors indica#e #ha# #he reac#ion o' 43D and an#i-43D bindin. is an immunochemical reac#ion be#ween an an#i.en and an an#ibodyD !# #he same #imeE #he presence o' @3" is based on a physicochemical reac#ionE which unambi.uously ar.ues #ha# a speci'ic immunochemical reac#ion underlies #he mechanism #hrou.h which #he radiobiolo.ical e''ec# is reali=edD &# is no# precludedE howe/erE #ha# #he use o' conMu.a#esE in which a Wuinoid produc# ser/es as #he hap#ene could produce a posi#i/e resul#D "he weak reac#ion #o #he Wuinoid produc# when #he la##er is measured in 43D prepara#ions usin. !DMD Tu=in_s me#hods ar.ues in 'a/or o' #hisD "he coe''icien# o' speci'ici#y resul#in. 'rom #he <0&4! #es# o' 43D 9:1 in lymph was 7DU-7D1E while #ha# 'or 43D 9:7 was 7DG-7DJD "hese resul#s demons#ra#e #ha# speci'ic

1K

immunochemical reac#ions in animals wi#h modera#e le/els o' !34 .enera#e less 43D 9:1 and more 43D 9:7E which is also demons#ra#ed by #he #i#er o' an#ibodies rela#i/e #o #he de#erminan#E #he le/el o' whichE like #he le/el o' correspondin. an#ibody #i#ers showed more indi/idual #o indi/idual #han species #o species /aria#ionD "husE #he blood serum o' cer#ain rabbi#s #ha# showed an earlier reac#ion o' ac#i/e an#ibody de/elopmen# con#ained hi.h #i#ers o' an#ibodies #o 43D 9:1 and 43D 9:7E which con#inued #o be obser/ed 'or #he en#ire 7-mon#h obser/a#ion periodD "he blood serum o' ca##le exhibi#ed hi.h an#ibody #i#ers only #o 43D 9:7 Y'rom 7 weeks #o 7 mon#hsZ and some do.sE durin. #he ini#ial periodE exhibi#ed hi.h an#ibody #i#ers YC4\7D9Z #o 43D 9:1E and subseWuen#ly #o 43D 9:7 Ylikewise persis#in. un#il mon#h 7ZD &# should also be no#ed #ha# animals whose blood and lymph con#ained hi.h #i#ers o' 43D 9:7 and 9:1 a'#er irradia#ion showedE as well as hi.h an#ibody #i#ers #o #hese subs#ances Y1C7UUK- 1C79KKZE did no# exhibi# ei#her o' #hese charac#eris#ics Yan#ibody #i#ers were 1CKU-1C1UUZ be'ore irradia#ionD "he leadin. de#erminan# in #he .as#roin#es#inal syndrome o' !34E as re/ealed by #he <0&4! me#hodE was 43D 8E wi#h C4\7DJ-8D1D Nowe/erE seroposi#i/e resul#s were also ob#ained #o #he de#erminan#s charac#eris#ic o' se/ere and ex#remely se/ere le/els o' !34E iDeDE 43D 9:8 and 43D 9:9 YC4 ran.ed 'rom 7D7-7DGZD Shile in #he #ypical 'orm o' !34 #he resul#s o' #he <0&4! me#hod 'or 43D 7 were serone.a#i/eE in #he .as#roin#es#inal !34 syndrome #he C4 'or 43D 7 was 7D7-7DG which is charac#eris#ic o' #he #oxic syndrome o' !34D &rradia#ion o' animals in doses inducin. #he #oxic 'orm o' !34 led #o #he 'orma#ion o' 43D 7 de#erminan#s wi#h C4 o' 7DG-8D1E wi#h seroposi#i/e resul#s #o #he presence o' 43D 8E 43D 9:9E and 43D 9:8E bu# lower speci'ici#y YC4 was 7D1-7D9ZD &n #he cerebral !34 syndromeE #he leadin. de#erminan# was 43D 1E wi#h seroposi#i/e resul#s #o #he presence o' 43D 7E 43D 8E 43D 9:9 and 43D 9:8D &# should be poin#ed ou# #ha# irradia#ion in #he dose ran.e e/okin. !34 'rom #he ex#remely se/ere de.ree #o #he cerebral syndrome did no# lead #o 'orma#ion o' #he de#erminan#s charac#eris#ic o' #he mild and modera#e 'orms o' !34E iDeD 43D 9:1 and 9:7 Y"able 8ZD &n our in/es#i.a#ion o' mi.ra#ion o' 43D 9:9 usin. #he me#hod o' ex#racorporeal di/ersion o' lymph Y<D0Z in rabbi#sE do.sE and ca##le su''erin. 'rom ex#remely se/ere !34 we es#ablished #ha# #he le/el o' 43D 9:9 in blood a'#er lymph di/ersion 'alls by an a/era.e o' 8U-GU^ YpgUDUUGZE and #he peak o' maximum sa#ura#ion is displaced #o #he ri.h# alon. #he abscissa YPi.ure 1UZD

1I

& I 973 nm.

)hours)

#igure 08. 1hanges over time in level of S%D 9)9 in the blood of dogs .J,K/ and cattle .", L/ 2ith e:tremely severe A%S J, L 2ithout &DL and K, " 2ith &DL. Shen we compared #he #ime course o' 43D 9:9 'orma#ion in blood and lymph wi#h #he #ime course o' lymph 'low in #he #horacic duc# we no#ed a correla#ion be#ween #hese processesD "husE subs#an#ial shi'#s in lymph #ranspor# were no#ed 9K-[7 hours a'#er irradia#ion and #he maximum concen#ra#ions o' 43D 9:9 in lymph and blood were no#ed a# 8-1U and 8-1I hoursE respec#i/elyD "hus #he ra#e o' 43D 9:9 inpu# in#o blood #hrou.h lymph 'low durin. #he 'irs# 'ew hours a'#er irradia#ion is /ery si.ni'ican#D !s #he lymphos#a#ic process induced by radia#ion exposure e/ol/esE #here is a decrease in #he le/el o' 43D 9:9 in bloodD &n se/ere and ex#remely se/ere !34E in addi#ion #o 43D 9:8 and 9:9 and #he correspondin. an#ibodiesE we always 'ound 43D 9:1E 43D 9:1 and #heir speci'ic an#ibodies in #he animal_s lymph and blood D Nowe/er #heir coe''icien#s o' speci'ici#y ran.ed 'rom 1DJ-7DUD &# should be specially no#ed #ha# 43D 8 was presen# in #he lymph and blood o' animals who ul#ima#ely died YT%]D%Z and absen# in #hose who sur/i/ed Y"able 8ZD "husE #he hi.h le/els o' C4 alon. #he Vdia.onalX in "able 8E which s#ar#s wi#h a mild de.ree o' !34 Yminimal doses o' radia#ion were used in #hese experimen#sZ and ends wi#h #he cerebral !34 syndrome a##es#s #o a immunochemical reac#ion speci'ic #o #he radiobiolo.ical e''ec# in #he ran.e o' !34 'orms s#udiedD

7U

"hese resul#s show #ha# #he radiosensi#i/i#y o' animals is de#ermined by #he na#ure o' #he speci'ic immunochemical reac#ion #o exposure #o radia#ionD "husE in animals irradia#ed in doses #ha# are le#hal #o #hemE 43D 1E 7E and 8 predomina#eE bu# #he absorbed o' radia#ion is no# impor#an#D &' <0&4! analysis shows a C4 o' more #han 7DU Y'or #hese de#erminan#sZ #he dea#h o' #he irradia#ed animal is ine/i#ableD "husE #he iden#ical dose o' radia#ion leads #o #he 'orma#ion o' speci'ic de#erminan#s wi#h di''eren# epi#opes #ha# de#ermine #he ou#come o' acu#e radia#ion sickness no# only in #he bodies o' animals o' di''eren# speciesE bu# in indi/iduals o' #he same speciesD Prom #his s#andpoin# i# is easy #o explain why #here are such si.ni'ican# di''erences in #he biolo.ical e''ec#s and ou#comes o' radia#ion inMury #o animals .i/en #he concep# o' dose i$ YsicZ GU:8U YGU^ dieE and GU^ sur/i/e a# #he iden#ical radia#ion doseZE iDeDE o' radiosensi#i/i#yD Se per'ormed #wo experimen#s #o s#udy whe#her symp#oms o' acu#e radia#ion sickness could be induced usin. 43D 1 and 43D 9:9 isola#ed 'rom #he lymph o' animals su''erin. 'rom #he cerebral syndrome and ex#remely se/ere 'orm o' !34D "he de#erminan#s were adminis#ered in#ramuscularlyF con#rol animals recei/ed normal saline solu#ion ins#eadD "he radiobiolo.ical e''ec#s induced by 43D 1 and 43D 9:9 were compared #o #he analo.ous e''ec#s induced by irradia#ionD "henE on #he basis o' clinical and au#opsy resul#sE we de#ermined #he 'orm and se/eri#y o' #he illness induced by #he 43DD &nMec#ion o' #he 43D 1 prepara#ion in 8 #es# doses caused #he animals #o die durin. #he 'irs# day a'#er inMec#ionD &n #hese experimen#al animalsE a period o' ex#reme a.i#a#ion was replaced by a deep comaE which ended in dea#hD "he resul#s o' #he au#opsy o' #heir bodies showed #he 'ollowin.C parenchyme or.ans 'illed wi#h bloodE punc#ua#e bleedin. on serous membranesE cerebral s#rokesD Comparin. #he resul#s ob#ained wi#h #hose in #he li#era#ure and 'rom our own researchE we concluded #ha# #he 43D 1 prepara#ionE when adminis#ered paren#erallyE induces radiobiolo.ical e''ec#s in experimen#al animals charac#eris#ic o' #he cerebral !34 syndromeD "he cleares# e''ec#s charac#eris#ic o' radia#ion exposure occurred in animals in experimen# 7E who recei/ed in#ramuscular inMec#ions o' an 43D 9:9 prepara#ionE in #hree #es# dosesD !'#er 1G days had elapsedE all animals o' #his .roup had died on days IE 17E and 1J Ysee Pi.ure 11ZD

71

#igure 00. Sheep (o. $-8904. 00 days after administration of S%D 9)9 preparation. Simulation of the clinical picture of e:tremely severe A%S .sheep died on day 03/. "he clinical physiolo.ical symp#oms here wereC shor#-#erm a.i#a#ion 'or #he 'irs# 7 hours a'#er adminis#ra#ion o' #he prepara#ion accompanied by 'i#s o' arrhy#hmiaE decrease in appe#i#e and increased peris#alsisD 4hor#-#erm leukocy#osis .a/e way #o a pro.ressi/e increase in leukopeniaE mainly a##ribu#able #o decrease in #he absolu#e number o' lymphocy#esE minimal le/els o' which were 'ound be#ween days [ and 1G a'#er inMec#ionD !moun#s were 1D7-1DJ 1UUU:`l and UD9-UDG #housand:`l in sheep and 1DK-7DG #housand:`l and UDG-UD[ #housand:`l in ca##leD 3eco/ery o' absolu#e and rela#i/e le/els o' leukocy#es and lymphocy#es was obser/ed in some animals be#ween days 8U and JUD +la#ele# le/els exhibi#ed #hrombocy#openia accompanied by pro.ressi/e ery#hrocy#openia Yin animals o' #he #hird experimen#al .roupZE which de/eloped in#o anemiaD !n ex#ensi/e blood analysis o' #he peripheral blood o' #he ca##le showed #ha# #he processes induced by #he 43D 9:9 inMec#ion and #he processes occurrin. a'#er irradia#ion were o' #he iden#ical #ype H17E 99LD !nalysis o' #he de/elopmen# o' #he clinical reac#ion #o 43D 9:9 adminis#ra#ionE which was assessed on #he basis o' body #empera#ureE and hear# and respira#ion ra#eE es#ablished #ha# all experimen#al animals showed reac#ions o' #he same #ype 'or all #he #es#ed doses and #ha# sheep and horses were more sensi#i/e #o #he adminis#ered prepara#ionD "husE in sheep in experimen#al .roup #hreeE which recei/ed 43D 9:9 in #he maximum dose #es#edE 1 or 7 days be'ore dea#h body #empera#ure increased by 1DG-7 f CE reachin. 91D7fD "his was accompanied by se/ere #achycardia and #achypneaE which were measured a# 1UG-1UJ and [U-KU per minu#eE respec#i/elyD Shen 43D 9:9 was adminis#ered in #he in#ermedia#e and maximum dose le/elsE sheep and ca##le showed

77

chan.es o' #he same #ype bu# o' lesser de.reeD "he maMori#y o' experimen#al animals had reco/ered be#ween days 8U-JU a'#er inMec#ion o' #he prepara#ionD !u#opsy on #he animals #ha# died showed chan.es charac#eris#ic o' acu#e radia#ion sickness accompanied by marked hemorrha.iaD 4ome sheep showed areas o' epila#ion on #he back and abdomenD Comparison o' #he clinical pic#ure and #he chan.es re/ealed by au#opsy and considera#ion o' #he #ime course o' animal dea#hs induced by 43D wi#h /arious epi#opes and in /arious dosesE led us #o conclude #ha# isola#ed de#erminan#s 'rom cen#ral lymph o' animals irradia#ed in a broad ran.e o' doses mani'es# marked radiomime#ic e''ec#s charac#eris#ic o' #he symp#oms o' acu#e radia#ion sickness o' one or ano#her 'ormD "husE #he research per'ormed su..es#s #ha# when animals are irradia#ed in doses inducin. de/elopmen# o' cerebral Y1ZE #oxic Y7ZE .as#roin#es#inal Y8Z and #ypical Y9Z 'orms o' acu#e radia#ion sicknessE #he radiobiolo.ical e''ec#s are o' #he na#ure o' a speci'ic immunochemical reac#ionE which #akes #he 'orm o' .enera#ion in #he lymphoid sys#em o' a hi.h molecular mass MM 7UU-7GU kD! .lycopro#ein Y43DZ wi#h a speci'ic epi#ope 'or each 'orm o' radia#ion inMury Y43D 1F 7F 8F 9ZD 4pecies and indi/idual radiosensi#i/i#y o' animals is de#ermined by #he indi/idual_s capaci#y #o respond #o radia#ion exposure by #he 'orma#ion o' #he dominan# speci'ic de#erminan# o' #he correspondin. epi#opeD &n o#her wordsE #he se/eri#y and 'inal resul# o' #he radiobiolo.ical e''ec# induced by #he iden#ical radia#ion dose in indi/iduals o' #he same or di''eren# animal species depends on which o' #he .lycopro#ein epi#opes 'orms in #he body o' #he irradia#ed animalD Despi#e #he 'ac# #ha# #he pa#ho.enic mechanism underlyin. 'orma#ion o' #he radiobiolo.ical e''ec# has been considered in #he li#era#ure since 1KIJE #he immunochemical speci'ici#y o' #his mechanism has no# been in/es#i.a#ed un#il nowD "he experimen#al con'irma#ion o' #his hypo#hesis o' #he speci'ici#y o' #he immunochemical reac#ions underlyin. #he radiobiolo.ical e''ec#s supplied by #he presen# s#udies has pro/ided us wi#h #he know how #o crea#e experimen#al samples o' an#iradia#ion /accines and speci'ic means o' #herapy and dia.nosis o' radia#ion inMuriesD <xperimen#s #o de#ermine op#imal dose and adminis#ra#ion #imes o' an an#iradia#ion /accine c 4!1 Yspeci'ic an#iradia#ion /accine Z-- in order #o e/alua#e i#s radiopro#ec#i/e proper#ies were per'ormed on Sis#ar line ra#s a.ed 8-9 mon#hs wi#h li/e wei.h# o' 1KU-7UU k.E Chinchilla line rabbi#sE 11-18 mon#hs old and li/e wei.h# o' 8DG8D[F and mixed breed do.sE 8 #o 9 years oldE wi#h li/e wei.h# o' J-JDG k.D "here were 7U experimen#al and 7U con#rol animals in each s#udyD "he experimen# was replica#ed #hree #imesD "he exposure dose ra#e ran.ed 'rom 8 !:k. #o 7I !:k.D Pi/eE 1UE 1GE 8UD JU and IU days be'ore irradia#ionE #he animals recei/ed subcu#aneous inMec#ions o' an an#iradia#ion /accine in doses o' GE 1UE 1GE or 7U `.:k. li/e massD "he con#rol animals were inMec#ed wi#h 1DG ml o' normal saline solu#ionD "he cri#erion 'or /accine e''icacy was sur/i/al o' #he animals 8U days a'#er irradia#ionC 'or ra#s a# a dose o' 1UDU QyE rabbi#s a# a dose o' IDG QyE and do.s a# dose o' JDG QyD "he resul#s are presen#ed in "able GD

78

"able GD
oD Qroup j species 3a#s 4!1 dose Y`.:k.Z G 1U 1G 7U -G 1U 1G 7U -G 1U 1G 7U -7U 7U 7U 7U 7U 7U 7U 7U 7U 7U 7U 7U 7U 7U 7U &rradD dose YQyZ 1UDU 1UDU 1UDU 1UDU 1UDU IDG IDG IDG IDG IDG JDG JDG JDG JDG JDG umber o' days be'ore irradia#ion 4!1 adminis#ered: +ercen# animals sur/i/in. 'or 8U days G 1U 1G 8U JU KG IU IU IU KG KG 1UU 1UU IG 1UU IU IG 1UU 1UU 1UU IU IG IU IG IU U KU IU KG KG U [U [U JG [U U U KU 1UU IU K YsicDZ U [G IG IG KU U U KU IG 1UU IU U KU IG 1UU [G U U [G 1UU 1UU KG U [U 1UU IG KU U U [U 1UU 1UU KU U [U IG 1UU IU U

Con#rol Yradia#ion onlyZ 3abbi#s

Con#rol Yradia#ion onlyZ Do.s

Con#rol Yradia#ion onlyZ

&# 'ollows 'rom "able G #ha# #he maximum radiopro#ec#i/e e''ec# o' 4!1 occurs a# doses o' 1U-1G `.:k. and adminis#ra#ion #imes 1U-JU days be'ore exposure #o le#hal doses o' radia#ionE ensurin. sur/i/al ra#es o' IG-1UU^ in experimen#al animalsD &# should be no#ed #ha# doses o' G or 7U `.:k. adminis#ered 1U-JU days be'ore irradia#ion also had a hi.h radiopro#ec#i/e e''ec# Y[U-IU^ZE and adminis#ra#ion o' 4!1 G and IU days be'ore irradia#ion ensured sur/i/al ra#es o' 9U YsicD no such number in #ablek"ransDZ #o IU^D Pa#ali#y ra#e was 1UU^ in #he con#rol condi#ionsD <x#ensi/e in'orma#ion abou# #he e''icacy o' #he an#iradia#ion /accine is pro/ided in "able JD "able J !able @. !he effect of antiradiation vaccine .after e:posure to lethal doses of up to 08 5y/
oDl !nimalsC Qroup <xposure DoseE Qy [DU [DU 4ur/i/al ra#e in ^ a'#er exposure period o' YdaysZ 8U days 1 Mice <xperimen#al Con#rol KG-1UU U JU days [G-1UU U 1KU days 8JU days -

79

3abbi#sC

<xperimen#al Con#rol

IDG IDG JDG JDG [DG [DG JDG JDG JDG JDG ID7 ID7

KU-1UU U IG-1UU U IG-1UU U KU-1UU U [G-1UU U IG-1UU U

[U-1UU U KG-1UU U IU-1UU U [G-1UU U [U-1UU U IU-1UU U

[U-1UU U KU-1UU U [U-1UU U [U-1UU U KU-1UU U

: [U-1UU U KU-1UU U [U-1UU U [U-1UU U [G-1UU U

Do.sC

<xperimen#al Con#rol <xperimen#al Con#rol <xperimen#al Con#rol <xperimen#al Con#rol <xperimen#al Con#rol

+i.sC

4heep

NorsesC

Ca##leC

3a#sC

<xperimen#al Con#rol

KDG KDG

IU-1UU U

KG-1UU U

"he resul#s ob#ained may be applied ex#ensi/ely #o #echnolo.ies in radio biolo.y and #he produc#ion o' dru.s 'or use durin. space 'li.h#s o' humans and animals under condi#ions o' ele/a#ed radia#ion risk and #ra/el #o areas close #o hi.h radia#ion sourcesE #o increase #he immuno.enic proper#ies o' /accines Y'oo# and mou#h diseaseE para#yphoidE and an#hraxZ and sera used 'or dia.nosin. parasi#ic diseases Yin par#icular we ha/e de/eloped a me#hod 'or e''ec#i/e dia.nosis o' echinococcosis usin. 43D as #he adMu/an#ZE and in oncolo.y Y#o increase #he radioresis#ance o' #he #issues surroundin. a #umorZD "he maMor uses o' radia#ion an#i.enC c manu'ac#ure o' an#iradia#ion /accines #o pre/en# and pro#ec# animals and #he public 'rom #he conseWuences o' radia#ion exposure Y'or exampleE #he Chernobyl ++ disas#erZF c #o decrease side e''ec#s o' radia#ion #herapy 'or cancerF c #o moni#or pa#ien#_s condi#ion durin. radia#ion #herapyD "he resul#s o' #he research ha/e been used #o de/elop ma#erial 'or pa#en#in. as 'ollowsC c a subs#ance c a Vradia#ionX an#i.enF

7G

c c c

a me#hod o' isola#in. #he Vradia#ionX an#i.enF a me#hod o' usin. #he Vradia#ionX an#i.enF possession o' know-how #o isola#e #he Vradia#ionX an#i.enD

Se ha/e prepared #he 'irs# /olume o' a #hree-/olume mono.raph on V"he 3adiobiolo.y o' MammalsDX Se ha/e de/eloped procedural recommenda#ions on #he V<0&4!-based dia.nosis o' radia#ion inMuriesX usin. #he radia#ion an#i.enE which makes i# possible #o unambi.uously iden#i'y radia#ion inMuryD !# #he presen# #ime we are considerin. proposals and issues rela#ed #o #he pa#en# de'ense o' research ma#erials and #heir more e''ec#i/e use in /arious areas o' human li'eD A1E(",L&D5M&(!S "he au#hors wish #o express #heir sincere .ra#i#ude 'or help in per'ormin. #he experimen#al s#udies #oC 1D4D !doyuno/E DThD i.ma#ullinE D!D o/iko/E 4D1D TirshinE !D D "i#E 3D4D "yumenE 1D!D 4o'ono/aE 1D1D !khrimenkoE +D D "sy./in#se/E 4DTD Pirsako/aE 3DMD !leksakhinE +D!D 4erbunE 3DQD &lyso/E MD $er.erE Doc#or "oy and many o#hersD %&#&%&(1&S 1D !ksyan#se/E MD D e# alD Compensa#ory capaci#ies o' #he lympha#ic sys#em in acu#e radia#ion sicknessE Med' radiologiyaE 1IJ9E GE ppD 8I-99D 7D !lekseye/E !D!DE +e#ukho/E meD$DE MusinE &D&D e# alD "he composi#ion o' lymph a'#er puri'ica#ion usin. sorp#ion and ul#ra'il#ra#ion #echniWues, ;et' khirDE 1IKGE oD JE pD 118D 8D $ardyche/E MD4DE "sybE !DPD 3ocal <adiation 4n=uryD MoscowC Medi#sinaE 1IKGE 79U pa.esD 9D $elouso/aE %D&DE Pedo#o/aE MD&D @uan#i#a#i/e assessmen# o' early reac#ions o' lymphoid #issue #o irradia#ion in a broad dose ran.esD &nC 4ssues of <adiobiologyD "omskC 1IJKE 98 pa.esD GD $ere=o/E "D"D Toro/kinE $DPD >iological ChemistryD MoscowC Medi#sinaE 1IK7E [GU pa.esD JD $i=hokasE 1D!D Maliye/E 1DMD "he radia#ion an#i.en and speci'ic me#hods o' dia.nosis and #herapyD RD ;eterinaria ir ?ootechnika,@ YsicDZE 18n8GoD 7UU1E ppD G-1UD [D 1ladimiro/E 1DQD e# alD %n low molecular mass D ! in #he blood plasma o' irradia#ed animalsD Mat' radiobiol' syeAdaE Tie/E 1II8E 1E pD 1KJD KD Qru=de/E QD+D 1he 6cute <adiation >one Marrow 5yndromeD MoscowC Medi#sinaE 1IKKE 198 pa.esD ID ]hdano/E DD!D Beneral 6natomy and Physiology of the 3ymphatic 5ystemD 0enin.radC 1IJ7E 88J pa.esD 1UD ]hi/o#o/skiyE $DDD Molecular mechanisms o' irradia#ion-induced pro.rammed cell dea#hD !u#hors abs#rac# o' disser#a#ion 'or doc#ora#e in biolo.ical sciencesD 0enin.radE 1IKKE 97 pa.esD

7J

11D &/ano/E !DmeDE Turshako/aE D D 4hikhodyro/E 1D1D Pathological 6natomy in <adiation 5icknessD MoscowC Medi#sinaE 1IK1E 1J[ pa.esD 17D TirshinE 1D!D $elo/E !DDDE $udarko/E 1D!D ;eterinary <adiobiologyD MoscowC !.rompromi=da#E 1IKJE 1[9 pa.esD 18D TirshinE 1D!DE $urdako/E 1D!D ;eterinary 6ntiradiation ProtectionD MoscowC !.rompromi=da#E 1IKIE 7UG pa.esD 19D Tlimo/E &D!D Mi.ra#ional ac#i/i#y o' microcy#es and #he s#a#e o' pro#eolysis in pa#ien#s wi#h mali.nan# lymphomas durin. radia#ion de#oxi'ica#ion #herapyE 3aboratornoye deloE 1II[E oD 7E ppD 97-99D 1GD Traye/skiyE D!D 2ssays on Pathological 6natomy in <adiation 5icknessD MoscowC 1IG[E IU pa.esD 1JD Tudryasho/E muD $D E $eren'eldE $D4D Principles of <adiation >iophysicsD MoscowC MQOE 1IK7E 8U7 pa.esD 1[D Tu=inE !DMD "he role o' #oxins in radia#ion dama.e #o cellsD &n& Pathogenesis, 2xperimental Prophylaxis, and 1herapy for <adiation 4n=uriesD MoscowC Medi#sinaE 1IJ9E ppD 181-19[D 1KD Tu=inE !DMD Molecular <adiobiology of the Cell NucleusD MoscowC !#omi=da#E 1I[8E 7UK pa.esD 1ID Tu=inE !DMD 3adio#oxins as re.ula#ors o' .row#h and de/elopmen#D &nC muD$D Tudryasho/a Yed') <adiation 4n=uriesD MoscowC MQOE 1IK[E ppD 118-177D 7UD Tu=inE !DMDE Topylo/E 1D!D <adiotoxinsD MoscowC aukaE 1[8 pa.esD 71D Tu=ne#so/aE 4DPD "he e''ec# o' myelopep#ides on pos#radia#ion reco/ery o' #he #hymusE <adiobiologiyaE 1IKIE 7UY8ZE ppD 87J-88UD 77D Tupryano/E 1D1D e# alD MicrolymphologyD MoscowC Medi#sinaE 1IK8E 7KK pa.esD 78D Topylo/E 1D!D 3e'inE !DPD %n #he si.ni'icance o' remo/in. Wuinoid radio#oxins 'rom #he irradia#ed body in #he e''icacy o' hemosorp#ion in #rea#in. radia#ion sicknessE <adiobiologiyaE 1II7E 87 Y7ZE ppD 7[J-7[ID 79D 0ebede/E TD!D Morpholo.ical aspec#s o' #he e''ec# o' ioni=in. radia#ion on an#ibody 'orma#ionD Cspekhi so%ermennoy biologiiE 1IJIE J[Y1ZE ppD JK-[KD 7GD 0e/inE muDMD Principles of 1herapeutic 3ymphologyD MoscowE Medi#sinaE 1IKJE 7KK pa.esD 7JD 0ekon#E ]hD 4nfrared <adiationD MoscowC 4#a#e +ublishin. Nouse 'or +hysD Ma#hD 0i#era#ureE 1IGKE GKJ pa.esD 7[D Maliye/E 1DMD <arly morpholo.ical chan.es in #he #hymus o' ra#s in acu#e radia#ion sicknessD &nC Morphology of 6gricultural 6nimalsD 0enin.radC 01&E 1IK9E ppD 8[-8ID 7KD Maliye/E 1DMD e# alD 3eac#i/e chan.es in #he or.ans o' ra#s_ immune sys#em in acu#e radia#ion sicknessD !bs#rac# o' presen#a#ion a# #he 4econd !ll-union Con'erence on !.ricul#ural 3adiobiolo.yE %bninskE 1IK9E /olume 7E ppD J7-J8D 7ID Maliye/E 1DMDE &kaye/E 4DThD e# alD 23456 6nalysis in *iagnosing <adiation 5ickness in 6nimalsD 1ladika/ka=C QO1 O443 Minis#ry o' !.ricul#ureE 1II1E II pa.esD 8UD Maliye/E 1DMDE +opo/E DD D e# alD Principles and Methods for 5tudying the 5tructural and Physiological Constants of 3ymph and 3ymph DormationD MoscowC 1IKIE 7[ pa.esD 81D Maliye/E 1DMD Mechanisms o' 'orma#ion o' radio.enic lymphopa#holo.yD 3epor#s o' 3ussian !cademy o' !.ricul#ural 4cienceD oD I-1UE 1II7E ppD 71E 7GD

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81Da Maliye/E 1DMDE +opo/E DD DE TirshinE 1D!D e# alD 1he Phenomenon of 5pecific 4mmunochemical <eaction in the <adiobiological 2ffect in 6nimalsD +ushinoE 1II[E J1 pa.esD 87D +anchenko/E 3D"DE 1yrenko/E muD4DE maremaE &D1D Or#aye/E $DMD 3ymphosorptionD MoscowC Medi#sinaE 1I[UE G97 pa.esD 88D +a#en# 3P oD 719GK[[D 89D +e#ro/E 3D1D ]are#skaE muD MD <adiation 4mmunology and 1ransplantationD MoscowC !#omi=da#E 1I[UE G97 pa.esD 8GD +o=harisskayaE "DDDE 0a/ren#ye/aE &D&D 3eco/ery o' cellular composi#ion o' /arious =ones o' #he cor#ex o' a lymph node a'#er whole body and par#ial body irradia#ionD <adiobiologiyaE 1I[KE p1&&&Y1ZE ppD G1-GGD 8JD ;accine and 5era EandbookD +D+E $ur.aso/ YedDZ MoscowC Medi#sinaE 1I[KE 7UI ppD 8[D 4orokinaE D+D "ri..er mechanisms 'or #he radia#ion dea#h o' lymphoid cellsD !u#hor_s abs#rac# o' disser#a#ion o' de.ree o' doc#or o' biolo.ical sciencesD %bninskE 1IKKE 87 pa.esD 8KD "roi#skiyE 1D0DE TaulenE DD3DE "umanyanE MD!DE Pridensh#eynE !DmaDE Chakho/aE %D1D <adiation 4mmunologyD MoscowC Medi#sinaE 1IJGE 8[G pa.esD 8ID "ori#skiyE "D0DE "umanyanE MD!D 1he 2ffect of 4oniAing <adiation on 4mmunityD MoscowE 1IGKE 1IK pa.esD 9UD ThansonE TD+DE ]hi/o#o/skiyE $DDDE me/#ushenkoE 1D&DE ]/onare/aE D$D 4eyliey/E $oriso/aE meD!DE Conclusions and prospec#s in s#udy o' #he mechanism underlyin. in#erphase dea#h o' irradia#ed cellsE <adiobiologiya, 4nfectsionnyy byulletenE 1IKKE 87E pD 8[D 91D marilinE !D!D Disrup#ion and reco/ery o' #he bonds be#ween popula#ions o' lymphocy#es a'#er irradia#ionD !u#hor_s abs#rac# o' a disser#a#ion 'or de.ree o' doc#or o' medical sciencesD 0enin.radC 1IK1E 8G pa.esD 97D marmonenkoE 4D+D <adiobiology of Eumans and 6nimalsD MoscowC 1yschaya shkolaE 1IK9E 8[G pa.esD 98D !nderson 3D<DE 4hren# RD E Miller 4DPD!D+D 3adiosensi#i/i#y o' "- and $lymphocy#esD 1D <''ec# o' irradia#ion an cell mi.ra#ion :: 2ur'F'4mmunolD- 1I[9D/D9D- pD 1II-7U8D 99D $ondE 1DE Plinder ?DE !rshamboE DD <adiation *eath of MammalsD MoscowE #omi=da#E 1I[1E 81[ pa.esD 9GD Dubois MD e# alD Colorime#ric me#hod 'or de#ermina#ion o' su.ars and rela#ed subs#ancesD- 6nalytical ChemistryD- 1IGJD- /D7KD- JD-pD8GU-8GJD 9JD 0owry %DND e# alD +ro#ein measuremen# wi#h #he 'olin-phenol rea.en#:: 1he Fournal of >iological ChemistryD-1IG1D- 1D 1I8D- 1D- pD 7JG-7[GD 9[D %kadaE 4hD 3adia#ion >iology of CellsD MoscowC MirE 1I[GE 8IJ pa.esD 9KD +e#ers DDE NayesE DDE Ni'#yE QD Chemical 5eparation and MeasurementD MoscowC ThimiyaE 1I[KE /olume 7E K1J pa.esD 9ID 3usnyakE &E PeldyE MD 4aboE DD Physiology and Pathology of 3ymph DormationD Nun.arian !cademy o' 4cienceE 1IG[E KGJ pa.esD

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