M<tho<lof", A..... 'o. It.. Elf"" .. orCb=hIo onR..,rod-run,; F. """"""£<1,,«1b)' V. a.Vook .'" P, J Slo«,," nC 19lIl$COP~

Methods fo r Evaluat ing and PredictingHuman Fetal Development

J E. Jnu.su

I. I:-;TROD UCfION

Whenewr a pregnant wOman i' e~posed to infecliou, "gents. ionizing rad iationor chemical'. including dru g,. lhere i' alwa)\ a possibiluy that sucb e,posu r...may b,,-. ad"erse effects on lhe de'-elopment of the fetus, A tberapy lhal may heof greal benefit to lhe sic~ mother . may be al lhe same time hazardou' \Q lhefetus,

A terat ogen may be defined as a phYSIcal. chemICal Or biological agemprod ucing ' truc tural or funclional abnor malilies in an organi, m e, posed to itbefore biMh, The sensilivit)' of lhe conceptu, to an e,o genously ind""ed damagevaries widelydepending on lhe 'la ge of developmem (Wilson. 1972).At lhe earl)', tage of onlogenw" berore blastogenesisi, completed (in human' during lhe firstlhree weeksafter conception) tbeconceptu" ife, posed 10a no.i ouug ent such a,ionizing radiahon . lends to ha'e an "a ll-or-none" response (Ru, sel and RUSst l.1954). The embryo survives withoul an)' analomlcol damage. or dies and i.abon ed. During organogenesi,. approximatel)' 4- 12 weeks after conceplion. the",n,iti" ity of embr)'o 10 morpho lollic malformation' is hIgh, the peak ofsensilivily beinll durinll the fifth and si~lh wee~ aher conception_The p"riod ofp"a~ sensitl"lty '0 teratogen, includes tbe conslitu\lon of embr)"ocborioniccirculation and early differenliation of maoy orgao,. There are "c,;tiota]periods"m morphogenesis of all ' trUCture. (Schwalbe, 1906: Sne n and Rapo la, 1%9),Ad' er.. influences at critical p"nods rna)' invoh-e not only morpholo gicmalformal ions. bUIrna)' resull in ad'er .. changes of postoa tal functions as ,,'ell

Possible teratollens are lested usinll .. "eral species of an,mal•. In ,pite ofs,m,la"l ie, in lhe de' elopmemal pall ern, the~uliarities of some ,~ie, ma~ethe compariso ns e' lremely difficult, For inSlan« , io"ersion of lhe germ-laye.. inm,ce and "'IS and tbe resultinll temporary yolk sac placenta does nOIoccur inothermammal~. AIle>the metabolism of chemical, in the maternal organi,m maydiffer 'ignifican tl}' belween .~ies,

~n bk 1 CoInpu>ou "" c...... IIea,"'"of pI'C1Ultol d, "lot.... ", of Q.... "' pcd ~""

eH .... n M""~.y ~, -- ,

t('01"1""' ''' •• ,.ge< l.<:n. 'h " l-""8th " ...." I' h " Len". " .-••• ", mu, m d4 }'1 In mm '" dol'" on",m in do ys m ",no In d.,.. •Um<cllul", "' u a ." 1,II ... ..,...., ", Cl6 10_ , ,

"' " O. IS ,• ,-, , , •<...._. , .. .. u -o.J " ~, .'"..

lliw. ....~ au.,. ,•

--, plio.. • •., ... ., lOll . , , .."~...- •, ,

==do')' yoIL.., , 0 2 ..~• •

T riIr._ ... bryo .....,i_~ pnmI ..... ....... k " U 10 l ~- n ., 11 .9 .-. ,

w" h . """'X1oonIa1pru<cM " t 0 2.0 ,,~

" tl .. rl1 ...... " .. ...p)<

co nlpl<' dy ope n ,",,,' .' ."~".. " 1 S 2.0 ." '.' 1 1 1~ ., • •,D<\l",I'n"" dOli" .. !both tn<li opco ~, I S 4 0

" U •• " U - Hl 1(>-10 J 4

0"" or both "oaml'<""'"

Io~ "' r s "< 4.0 6,0 14 26 1 4.1 " r s , 'JS' . II' of limh <I",dopmenl

bud of pro" "," 1"" "",, " y , , .. 2& 32 sc " '" " ," .'W ", ,

•b oo. of pm, m.. l. oJ 0

d lS.. 1e"rom" j.. , z ,-. n " s.o " H IU " '",,.

p ' o, ;,,, . l o, ' ,,,," ,' y •',,"oscg""'n" , , ,

'" " " SS& " " •1" """ ", I . oJ <1;".1 e"" " ,,, ;'y ~'''-0 "<8"'""" " " u J7 42 " ~ 9. ~ IJI,I 'g". 1' . y' . [0'>1pb' <> , ,

'"" " " rc " '" " 1I. ~ "~.

" I,

d ,gu . 1' aocrde . ,. l J 21 « " H < u.s " " id ,WlS. loe luocre les " 19-24 " " " " " " " >

La« "mbryon.1 "' l!<, ..<I,'l<re" h. I",1 "''''m,' ",,, •, " " sa • " « " 171 " " " ~

19:,

" " n8 eyel.. , .-, -n " .W J9 44 sa " " " " ~a" W ~. ,,- a

1'",.1 p<tiod • ' 00 1&2 + M ,m as 19 2 1 n 25 " '"W, '00l'orinal . 1 r><'iod to '00 ,~. 1'"",. 1'"",. I'os, -

oa, . 1 nalol nalal

"

302 M~11ttHh1MA~Jsi1lJ ,Iv Eff r<:/J of Clvmi€au "" k pro<JoM!ri'rt FrmcrjqN

The a.. 1of leraloaen"my 1'"'1' .. 10detmnine Ihe doK ,,'hich .. embr)Olo .ie

in animals ~ausin. intrautmne dealh Or 1TO"'tJ,r.urd ation. Of .. hic:h is1.....I"l!en"'. P,odUClIlI -.nalomal malfOnTlauons..lbe mechalUsm of Ibma~ "ftOloflCnin~aupled General m«IlaBlsm5 of mlerferm "'th prenalal I'mullaDd d,ll"cr-muauoa mr:l..... mhlbtuon of ~U prOW"..... ,1Ollrd.,fd 10 .lln-fd.ynih=J of protClas.. carbob)'dral" and tipids. or 10 limlled CDCl"I)' .uppl);mhibiuon of pl:lalllallnilSfn- .nd pto .. m O)"ftUlnu, ,nlerf~ "'"h ftlal andfetopl:locmtal ~lJOfl; and ahcred Of mluhued formalJOfl ()( _mOt}',,,,,,,,,,,-

lbe I}-P<"of .... lformauon .. <daled 10lbe otqe II ...hJdl nnbt)'O" e~ 10a , ..... ,osa> For mtUlntt" CoocepIUa ahOf.. d dUM . ,tard FalaUOlllImouth aI\cr up""'~ 10 (a folic tcid ............ ) """"' ·ed uopho.bWtic 1XICI'OIi.. illrombolosof lilt inlft'YilJouoopaooand damqcd (etal bema­

IopooaIS ""11Lul 1M b'n- (Sha",. 1'1"'2,. If lbe (etus "'''''ed. a Il~1

amn'opla'lfl J)'Ddromo ...". eharxlenled b;. mall"onnalJOn' ,n 1M c",,,ralI>C'I\ ....... }~nn. CfllUof_1 d)"19Ia.<d..dublw>ds and duhfm .• nd menulm .r<1auon("Ttuef1ch. 19~2, H"", vd and Rood. 19n).

lbe d1"eeu of Ihc chcmlcals ...hieh ha'"e been lesled may be claaslfied asexpeeled aDd UIlCXpecled The expeclcd effects are ....oss mall'ormallOo,.allOruon and cld4 .......11'0.from 'ohlbtlion of grO\nh obs<n«l.On- .doun'J­lrali oo of .ouoop,mn. lnlSlllf.n. chloram hucil. C)d ol'hospllaoude .nd 10100·""mlll ,o 1M finl Inmeiln- orp<ClJIIlIC}" (Scbill"Cl. 1..1970). Gr o" 'lb rctardauonrna} ", oull from 1M lrea lmen! " 'Ih ccmsone (Wa ITeI and Ta )'lor, 19( 8).Mascuhnll.ll !lon of female feluses i. ao expecled conscqu~ncc oradm,nll'r. ' ionof llIdroaenic subst.nces luch IlICSIOSlerone and ;l>der;"al;" ,", (G lUmbae b andDoc har me. 1%0). The ,nlerf.....,nce of dielh) l1lilbclIfOI(Herbal el al . 1971)wilhnorma l 'CS.......ion of panllllClOnepbric ep ilheliu m ,..ilh,n Ihe "all,na l pLOIeleadioilio ' ·allina l .<lenom Ibould be con~idercd al a predic lable e/fecl S,nce""JIM i•• kno"' n larS"1 of cslrOilen•. Telracyc hn il kno" °10form eM Llies " ilhcabum. aod afler applicalJon 10prep nl ,, 'omen il i. deposiled in Ihe fCial baMnuocnll and deciduO\Isleeth (Tno/Jand Ra ,.,d. 19( 8), A Ins expeclfd effecl'S 1MdeposHlon oflelr&C)'Clme,nlO 1Mleo ... lcadrna 10Clla,-.:u( Harley CIII,. 196<1)LDdcn landa hk .. 1M C'lvuh n..,...-edama ,e rd :ll od 10 MOIll)'CUI. lIrq>lom)'Cina nd qUI",,'" and Ihe formallon O((CUI,OH..,. Ifln- "dmuu.strallon of anu·th) 'TOlddlUp du rinJ prqTWlO)

L;Mxp«ted 1.....1OJmIC ctrncuare 1MdI"<CI." hJdlarcdll6culllo predlCl ~'~if lbe metahobm orlbe cMmocal ISk_ n. Fo.- ,mtaDl%. lilt pheDoc:op) o(!be

RIppled eprp!l)_ 'lndrome (0.- cItondJ'odl'PlnIa l'~U) ""h"'b de\eloplIf"" adm.oo<lrallOflof dlClllllll'Olloticw.alllan .. (mch III ...vfaml ) dUJ'UII11wliN lluu monlhs" dlfticub 10 pre<hct(Shall! aDd Han. 19n ), Slmllarl). lbepropcrucsof thalidon>odedo not r"",,, "Illl ibc cbara<ur orthe _ ...'Cll-k_ 'nS)ndrome t:""xp«tN elfeeu ma)"be so-..ma dercet od in I.lUmalupmmn>lI .H.... "''CI.il should a/'tt.'I)' be unc\cnl:oodthaI alrapolalion of .uumal e<pen'""", u ,0 IDIl:I 1:1, .. lId ... 1) 10 a hmlled dcptt

M"'Wlb!Qr F;,'alualmg Il1IdPredkling Human Fetal De.. lopmem 303

T~rat<>lo&"'s and exptllmenw, 1 emb!)'ologisls ha"e pro'ided much infor·malion on the develop menta l mechanisms and their d,sturbances in diff~.. ntverteb ra'es . To make lhe interprelation of "SUllS ea,ier , we ha, -e proposed acomparal i"e c1assificalion of pronalal developmen, of quand",ped. in whichcorre.pond ing stages a re ""'rked by lhe same s}'stem of numbers. The prena ,aldevelopment of man. macaceus. ra, and mou... " compared in Table I.

2 MOTH ER· FETUS 1.1'0 Fl:T U5-M OTH ER TRA1'o'SFl:R ;THE PLACE"'T AL BARRIER

The transfer of substances from the maternal in,o ,he fetal compartment takespla"" either across 'he placenta Or across fela l membranes . Only undertherapeut ical Of e, perimenta l condi'ions ,ub stances aT<adminislered dire<;tlyin,o 'he amn ion. or into 'he peritonea l cavity of 'he felUs (in,rau tenne fetaltrans fusion ), Transpo n mechanism' are related '0 the anawmy of 'he biologicalbarrier. t<>lhe uterochOfionic and uteropla""n,a l (materna l) circula,ion and tothe embr;o-chorionic or feto_placental (fe'al) circula lion ,

The 10lalmalerna l ulerine blood flowisconsidero;l to be 94-1 n ml mmute kgat 10-28 gesta,iona l weeks a nd approxlma,ely 150ml minute kg at the term(Assah et a1.. 1%0 ). The blood flow enter ing in'en'illous space ISintermitlen t.and 'he blood pressure in the terminal uterine arteries is 70-80 mmHg (Alvare:>:and Cald";ro-Bareia. 1950). The blood pressure In lhc int.".il lous space i'approxima tely lOmmHg. and increa... s duri ng uterine conlra C"on, up to 30-50mmHg, The pressure in 'he maternal veIn, leaving 'he inters'inous space .. about8 mmHg. The blood pressure "ithin 'he in,ervillous space i' lo"er than thai"i'h in the chorionic (fetal) vessels.The blood pressure in the umb>hcalaner ies i.approxImate ly 48-5OmmH g and in Ihe "ein 24 mmHg (J>largolis and Orcu tl.1960), The.. cond itions faVOffe'o_materna l tran. port ,

The paraplacental transfer is effec'ed by a Iranspon from decidual ""lis across'he chorion into Ihe amnion For in"anco. prolac tine roleased from decidua lcoUscrosses thec horion and amnion and dIffusesinto 'he amnionic fluid (Riddick andMasler. 19&1).The ,ran,pen of decidual pro lactine requi .... act ive pro,einsynthesis and an intact microlUbular sY"em. The permeab ,llty of amniochorioncan be ea, il; siudied in vilro (Seed, el aI., 1980),

The placenlal transfer tak .. place by diffu,ion , b; facihtaled diffusion, byaCll"e transport (against 'he grad ient). by endoc)"oSl'. Or b}' other poorlyunders'ood mechanisms. and depends on 'he ph)" icochemical properties ofchemicals. such a. molecular w.. ght. ioniza'ion, hpld SOlubility and prot.. nbinding (Mir kin. 1976)

Compounds wilh low molecular "eillh' diffuse more rapidly, If the molecularweigh, exceed. I000u. 'he passage a'"os, 'he placen,a i. usuallyslow, Mo leculesin non-ionized sta,e eros. 'h e placental barrier mote rapid ly 'han in ionized Slate.li pid solubility facilities the'ransper! . There i, a substantial difference be,weenprotein b>nding in 'he ma,erna l and fetal blood plasma, Thi' differencernaye.ert

304 M n llodsj or Asstssing IIu>Effects of Chem ical.<<m Rqmdwcli'"I' FW1C1iorts

a signIficant mOuenceon lhe transfer of chemical•.especially if the fr"" subslancecrOS5.eSthe placenlal ba,, ';el slowly. There are $Ornespecific placentaltran.rer,} stems invol" ng ,pec ificconjugation of tho suMtanc ocr""ing tho plaeonta to acomplex which i' cl<:and befon: lhe substance is reka>e<lafter the transportacr"" lhe trophobla,t.

The placenta . .. pecially lhe trophob lasl. conta in. ellZyme, metabo lizingendogenous a, " 'onas xmob iotic ,u b'trat .. (Hagerma n. 19(9). Somoof them areof fundamenta l ph}',iological impo n ance. for e"" mpl<:.tbe monoamin e oxida ..(De Maria . 19(3) wh'ch deaminatesa number of ,uMtrates,uch as epmephrlne.norepinephrine , dopa mine... rOlonine and some amphelamines : diamine oxi_dase (hi'lamin a .. ) (Weingold and Southern. 19(8); catecho l-O­methyllran,f era"" which lnact,,-at« epmephn ne and norepInephri ne; """e", lper tlda .. ,. 'ucb a, cystine aminopepl ida.. (ox}'tocin.... ) (Mathur and Walker.19N1: choline acelytran, fera.", (Bull "' at.. 1%1); 3/i.bydroxy"","oid dohy_drogena... 11/l-h}'droxYSleroiddehyd rogenase. a very potont steroid arylat ings}' tom. and othe" .

The elfe<:t' of cbomlCal' on the placontal "ascu la,uro can bc SludlM 10vi"o inptlfu sion experimem (Mancini and Gaut ie" 1%4 ). Serotonin. norepinephrine.LSD and some oth er ballucinogen,. morphine and codeine are poten, con_strictors of placenla l ,·«",,1,. at lea' t 10 ' -'lro (Ganl and Dyer. 1971),Prostagland in F, . enhances the effect of oxytocin (Jungmannova er al,. 1975).

It i' evident tha t placOIllalon,ymatlc S}'St,mHan metaboli' e a large numbcr ofendoge nous as well a, e"' geno,," substanc... Hyd rolyzable ,ubstrat .. areu,ually hydro lyzed. and ,,,soa cti, .. ,uMtances and hormonal pept ides arometabolized 10 maCti' " SUM'anc<:$, Placenta l conjugations. wilh a po" ibleexcepuon of acetylation. seem not to occur,

3 PRENATAL DETECT ION OF FETAL MALFORMATl O,""SA" DCHRO;\10SO :\-IALAND GENETIC DISEASES

Te<:hnique. uSC<!for prena,a l detoxuon of fetal disorders can be c1a"i fied a,invam 'e and non-in".,i' -e In, ,, , ive technique•. , uch a, amnloc,nthem andfet<»copy. include p"' etrat ion onto 'he amnlo, ic cavity. Non-,nyasiw te<:hniqu..are based On X_ra, or ul'raso und procedure, . or on different kond' of felalmonltoring.

3.1. !",,,,,""urO'SBa"'" "" Amni""emh....

AmniocOIlthe'i' is u,ua ll}'performed at 16 18 w""ks of g.. tation . The 'o lume ofamnio' ic ftuid at thi' . tage is appro, imatel; 120-3lXlmi. from " hich 5-20 ml aro" ithdra wn and used for ana ly'is . The risk of COmpliCation, " laled to amniocen­the,i, i. Ie.. that 0.5 \ (S,rnpSon and Mart in. 1976). Pot, nllal materna lcomplication, Include Intra -abdominal bleeding. puncture of the bladder or

Method ./or £.'oluolingand PrMicring HumtJ!I FeIOIDe>'ewpmenl '"ont~'t ine and Rh·immunl2al ion. Pos, ible f~lal compl ication, are injune , 10 'hefelu. b) lhe nffi:lle, bleeding from placen'al ves,. l•. InIt.ovular ,nfecuon andabon ion, Leakage of amnIotICfluId ha. also been repon ed,Th~ amnim ic flu,d sample is c~ntrifuged, and lhe celh are used for cylologic

e~amona ,ion. CUllivaIion and ,ubsequ~nt chromo'o mal and melabo lic analy.i "The fluid can ~ examined by biochemical melhod"

3 ,/./ Cyl olog)' 0/ Amnioli c Cel/,

Centr ifuged cells from ammol ic fluid are spr~ad on a microscopic slide. fixed informatin in "apor and, if an open neural l u~ defect i' ' U$p<Cted , th~ ,mear ,.checked for lhe presence of glial elemems (Brock. 1978). The glial elemenl' arel:" pola r. elongaled and Mtectabl~ by a glial protein S-loo e~h; bil ing a sp<Clfkimmunofluorescence (Sarkar cr al.. 1980). Ana lysis for X< hromatm and y.ChrOmal in in uncultivaled amn;ol ic c~ll, can ~ ea,i ly perforrne<l;however. ther.. uh' are far Ie" informaliye than karY01YPing

3.1.1 Amniotic Cell, Ti5sue CullUre.

Cen,,, fuged cell' from amniolic fluid areculliy'aled for apprmtimalely 2-4 weeksin a nUlrient medium con,aln ing calf serum and anub,olIC•. Tbe cul,;,·allon,.. u,,"es, ful in 70--99';. of cases, 1fkarY01yping i' required. colchicine i. added 10the growin g cullure (mlloses are a,re ,led al metapha,.). the medium of theculture i, h)'po tonized and acellc acld·m<'1hanol fixaliveadded: cells ar~ ,preadon microscopic slid.. and 'l ained b) different banding techniques, Using th ismelhod a ll numerical chromosomal abnormallues Can be detecled. Th~ mostcommon l\ lhe mono,om y for lhe X..chromosome (45.X). Th~ Olher qu;t~

frequently occurring palho loglc hr;o types are 47.XXY: 47.XXX: 47. + 21:47. + 18, 47. _ 13,

The dete.;uon of , truCIUr. 1chromO>Qmal anomalies depend. on Ihe extent ofchromosomal leSIOns,MInor abnormalities may remam undetected. Strucluralchromosoma l abnormalit l<:<lo , ol , ~ de lelions. lran, locallon, and rearran ge·menls r.. ulting in lhe forma lion of iwchromo' ome' . Incomplete chromosomes.fr~gmenl' , n ng<hromo,ome , . "'c. Chromosomal break ' are o~r",,<1 afler X­ra} irradiation.

The mo,t impo rtant indicau on for prenala l karyoty pinll of the fem, arechromow mal abnormalilies (such as ba lanced lran slocal ion Or im'ersio" ) ineither p;lrent , advanced materna l age or existence of indi,'idua ls affected bychro mosomal aneuploid;. or anOlher chromosomal abnormali ty in llie famil},

Cultur.. from amniolic fluid of fem, .. Wilh 0P"" neura llu\>e defects comainglial c~ll, " hich are elonga ,ed. b;·polar and exceptionall} adh.. ; , ~ (rapidadher ent (}f RA cells). and comain a specific gha l pro le;n (Sarkar et al .. 1980) ,

Culture s of amniotic cell, can also ~ u<ed10detecl man; metabolic diseasesw;lh known biochemical basis. The biochemical defecl i, usua lly idenlified b)'

306 Mtl /todsj <>rA3HJJUtsI1wEjJ«u "jC!wmkal> .... R~p'<><hKllw Fu>lCI......

em) ......a..,..ys ,n cullum! ullt. Abnut 73 dJlfcrnll mrubo(;c dlSOl"dfrl(such asmucopolysa<bandoses. 11~05eS. ~podmes. ammOK>dopatlua. etc ) aonnO'" be diaJllosed pmlataJIy (Golbu$ ~'"L 1976; Rill".,. 1976).

J.IJ 1lIQcloDorinoJA"')~" <>fA_ir Flvid

(a ] A/pIIt>-fr'..,..nM AF' AFP II lyutbcsizcd '" fcul cododn"mal.u,ICtUfnlOChas the ),,11<Me. hn alMlCU" AF P \11ll1llDi<Micl!IOO"'"IU"'ta pIOl:Iablyfrom r~l u.ntIC. and ,I' allllllOUC~UJ,lIOCI <lcaa .... "'tb ad'"lIl1CU1J~tionaI. (Broc k aDd Sutduft"c, 1972) EIc>"lItcd a:w;:ultnollOllS 0( AFP .,.,mtOUnlercd in opm """,*1 tube M<xu (..... lXephaly. opeD spmabltlda j aDdba'''' also been IepCI<ttd'" MeIOCO<X)P teraloma. ompbalncdo aDdcoapmwneplu""",, aDdID ptep'aTlCM'$ .."II rnal death>: elc>lItcd AFP IS somet>_~t abo m fet....... ,Ib amsaas '" dJF'U'" lUbe. polycyst", bd.Df)'$. UUlulalpancreu. h)-.lrocepbalus. fallot·.teuaJoc aDd <:<>n&=1a1....'D defocu . ~ormal

<:onCeIlUJ,uons0( amDIOllC AFP dunn l ~[lOl>al ..«k . 16-1&are bet .. '","" 20aDd 7DLmL In f.. uses ",Ih opm neul"llilube defeeu. the AFP 'lIl\1f$ eXffd25l1J. rea<:tulll in some cases «Xl-~nl.ml (Brock. 197&). ContamlnatlQll 01"tht amni otic lIwd ..,,11 fnal blond is tbe most common $Oultt oI"fabe poIll""i:['filiiI'.

(b) HOI'_ ' in "",n"'li<" jfuid. F... I K~ CIInbe pm!"' tcd from tbe ammOI'"CC<lCefImlli oo of tnlOStcron e. FS H and LH. In male fetuses. tntOllcron eoo~nlra\lon ,$ much h,. ber. • nd FSH In d LH COllC01llra lion 10,,"'''' IMn inr.male,

The dilferenc .. m amnio lic concentra lions of testOllOro ne. FSH I nd LHd unn g gesta lional .... k l 16- 18 a", , ho" n in Ta ble 2

EStlmal ion of ammollC concentratio n of FSH and 3. 3'. SAri odothyron,ne(RT. ) could be useful for prenatal dla", ,,, .. or fetal hypotbe yroldism, .'Ih .rcon goni,. 1or induced by .~posu'" of pl'eJU nl "'OIDOll to I lrumi .. n$ (Chopraand Cl"lIOOall, 1975). EIo' '&led amnIot ic 17-hydro~)'prOilnl"'OIle "' &I observedin f011LSC1alf.." ed by .drmal (or tlCal hyporpla.i. duo 10 C-2l Slnoid hy·dro~yw.. dooo ency (PooIII01al.. 1980).

Tablo 2 DIlI"om><n III boo_ "",,, 10 in tbe fCl.ll ._ ..... .,ld "' ........ aDd,- -

2O-JO",,'"O.l---.(l.7 .9-U 1DI

__ lAo

_dunq Ie-IS_~-.... M.

T l ao-- I:~C~"'~"'----c",,,,-,,------FSH OOS-O,12 .......LH :1--1.......

.v~,ltoJsjQ, £".,/lIa' I1I6"" PMik tu., H_ F~lIl1lNti~ID"mn" )07

).2 FcI OlCOp)'

Duo;t ",... 1~lXIIl of the f~lus ISpnfonroed b) a fet"'""P" . a speaa lliberopl><:eodOlCOpt. Ahhou&bSInlul~ 'UI.lalin."""ol !he ... un fC'lU$ISl>l>t poosiblt.lSOlaled portiofts oltbe fC'lut sudI all OClIJp. e,.-cs.1ipa.fiD&en.lacs.ulmlal Jftlltalaa aDd tho anI....,. bod y .... n.mclucha,!he lDXt"tioa oflllllbllg lcord. may M chlo;tly CDmu>«l III_ ~!he " ... IizaUOll ol. SlnIC1un:."boK Illalom y 1$ 10 M cum" .. d lilly M ''ftY dJffic:uItbccaUIC ol !heCOllt.lllUllllXlll ollmm<IUClIuideitber b) lllO::OO'wtl Of byblood . l a ow 1KIlp1l1fC'lOKop)' ISpnfonroedl11ldtt grnln l anesltwsr.a. Tbc \lo'olf fC'lOlCOpt() . 2 mmdiamn ....' ISInocned inlo lhe:ul..,.". und...-,,""I COIltrOIafttt a .mallla""rOlom)has bea1 perfonroed; a .utu n: i. It' led Ir cuDd the fC'l""""PC iaxnlOCl TbcIocalizauOClof tM pla«n ta is al }'S ckl....mlned ulu-l.SonnUy .Dd IhtInlnsplaCClltalroute of f..."""opt lnwrllon i. stnctly avoided, Th~ nsl of felaldeath and .bon lOClafter feloocopy i. approxtn\lt~ly Zlr" ,Abortion. art n:1ated10 Ieax"it of . mniotic lIuid and .ubSl'qu ~n t . mnioxhorion lti<.-r.r~ly 10 Intra·amniotICblecdini. Chronic Ie.kail~ of . mnlOlle fluid durini the:third trimtst~r

requi res I Ioni· t....m hOSpita lization in I ppr oxlmattl ) 3'r" of patltntS.F...os<:"PY.110..... prtna ta l dll iOOSIS of IU major u, ema l morpholoaiQIlmalformaliOCluuch IS facillcJeft •.deformed u tenll ln .... defo;t. of CNS.1fOSSmalfOrnllllOCl'of hmbs (phocomelsa.an~). oliaodactybes . pot)"dac.I)'~cs. . }udal:lyMa, clef..,.. ofantmOf body ... U ....d SO"'l' malfOfmlltlOCl'of""tml l sm>lIha.

FC'loocopy " lIidnted '" .}"DdrollJCl.. hodl do DOlexhibit cllromoloml l Of

biocMml.boorma~l>CSmbcnted~ 10MCftdeIwo Io....'$. arid"'iucb an:eharaclcriml by comtIa'lllalom l malfOl'1l\lUOOl

DiIJDo:>RICfetotCOJlyhas lxnI pnfOfllWd In O\lf bosplll l by Z..1qft" In 41cases . 2Oofthem ..'"" tbmr.peutiICIlly abort ed Malformations ..ut found '" 18easel . In 1"'0 cascs '" .. lucb 1M fel\UOeS..ut dl&poscd fOlOlCopa l!) IS

malfOl'D>ed. 1M<hIiO........... not confirmedafter aboru"'" t:" ......ted abomonoccurr<d '" four pallallS "" tb lI<:aJ'hy fC'luSts. Pte-ttrm <k~,~ OIXUfTedIIItbJcc cases. 1"'0 Jlfe-ltnII ,,", 'born. dted, T..-enly plluen.. de~,'Cfed b<:allbycluldrm II fuJl·I£nO. leabgt ol.mnlOlIC ftuKl~ in six ol them . US'n,fetoscopc " 'e d~tecl ed follo..in i defec,- : fanubll elefl.ltp I nd palate (6 caSts).Treach..,. Colhn.· Iyndrome (2 caws). anencephaly (pn:>-;ous!y dll in OSl'<!uhrason iQIily. 2 cases). ectrodactyly (I case). Sald,no-Noonan .yndrom~ (Icaw). MaJe" ',x l,yn drome(I caw). I}'ndac,yly Iype V( 1case). Robert' .yndrome(I CI.Ie). an hr08ryposi, (I cast). Aperl Iyndrome (l ea<el, Lip piH:len ltpI)ndrome ( I casel,

&mpl"" offrl~1 ,u.s..,""tk,ftl~~i(: roniTo/ Tbe n«dJe.cope . .. hlChhallralher bnuted VISualkid for. gtn .... 1fetOKOpy;, a ioodtool fOfVlSualiutionof cbononic ,-eon' In tM placen1l1 plate , Ann . ,uillblt "em is >UWl1~ . •hcpannt«d ~It " dlt'CC<edinlOthe lumen ol the ''tUd, 8 ) du....... bod 1"''''

308 Melhods! or A",essing lhe £f!el:IH,!Chemicol, on Reproducliw Fun"'io'"

fetal blood can be obtatned a nd used for prenata l diagnosis of hemoglobindisord ers (Mac Kenzie and Macit an. 1980),

Felo scopy contro lled intrau terine sbn and muscle biop, ies are posllble andpromising for dlagn o>el of disordm which are nN detectable from amnioticfibro blam . Fe,oscope can be also used for 'he contr ol of intrauterinetransfus.ion.

3.3 "' ....· In...s.. . ~ I.lh""s in Prenata l o;.gnosis

1.3./ Ullrosonograp/!y

Using real.t ,me ultra"'n ic ,"stru ment> rhe concep'us can be ,'i,u.lized during5,h or 6th geSlauonal ,,<:ok>; and its growth. po'i'ion and localiza'ion of theplacenta can be monitored (Jou pilla and Piiroinen. 1975), The iner. a.. in thebiparietal diameter is normally' used for general gro" 'lh evaluallon (Queehan (I

al .. 1976). £ "im auon$offetal bod y and h.a d wlumolare perfor med ,o d,agno"intrauter ine growth retardation (Jordan and Clar k. 1980).

The fetal bea .. rate and the ~t of umbIlical arteri ... can be also visualizedultra sonically, The follo"'ing malformatlonl can be dia gnoled prena tally byultra sound procedure" anencephalI at ,he beginning of the second trimeste r:mle lomeninlloceles. but diagno sis mal be difficult (Campbe lL 1977) : hy­drocephalus and microcephalI in the 'h"d tnmester. inienceph. ly, sacrococ.cygeal teralOmas . mullicyst'" or po lycl'tic b dneyl or hl dronephrosis andomp haloceles "i'ualized as masses in a chara ctcri' ti< location (Bartlel et al ..1977). In some case'. the ,hape of external getnta l,a allo"" the diagnos is of se•. Inthe laSlt"me 't er, [he heart ,,,IHI and ca,'i,ie, can bevisualizedand the dlagno<isof gross congenita l heart anoma lies i. pos",b le (Patel and Goldberll. 1976).

3.3.2 RlUiiogr"!'h"

The u.. of dL,",,' radiograp hy for diagno' i' of skeletal defects i' Iimiled to thetrurd l" moster , During second trime"e t even llIos, .n oma li", such as achon ­dropla. ia arc undetectable (Golbus an d Hall. 1974). Prenatal radIographICdiallflo'is (after JOIh gestat ional w..,k) of anencepha ly. hy'drocephal;. m;croc·ephaly and of mOStof thellI o" skeletal anoma lies has been reponed , Contrastradioirap hy with a water·,ol uble radio-cool ral! d;~1 hOIbeen used in the 'hlrdtnm "' ter fot vi,ua liut ;on of ab$cnt fc'a l "wallowing in Case<of ga't roint""t inalaU""ta

4 CO:->CLt:SIO:\'S

The elucida tion of lleneti<.biocheml,a l a nd ph}'>iological mcchan ilm. in'ol' "edio prenatal morphogcnes is . nd embryon ic funcuon il a nec",,$ary prerequisite for

MellwdJ for na lualing and Prroicling Human Ftrallk> 'elopmelll 309

atlemp, ini ,he prediclio n of adye,... etf""ts of chemica l agents On the exposedmother and fetu •. To pmlic t 'he elf""t. of xenob io'ic subs,ances on prenatald,, 'elopmen t, 'he " ai e of pregnane; a, "hic h 'h e fetus was exposed and theeXl'O' ure coneen,ra,io n need to be known. Predic' ion' based on animalembryo,oxic ity and ,era,oienic i,y are, however, of limited "a lue.

For detection of inadequa,e fetal grow,h, chromO$Omal and me,abo licdisease<and e"er nal ana'omica l malformations. ,he folio" 109methods can beused :<;1oIOiYof cell. exfol iated into ' he am niotic fluid, direc' biochemic.1 testsof .mn ioloc fluid, kal;-ot; 'ping of amniotic ,·ell. grown in ,i'sue eulture' ,biochemICal ana l;sis of amnlo, ic cells grown in ,issue cultures . fd oSCOp; .analy. is of fda l hlood wi'hdr awn uooer fetoscopica l contlol, ult raoonographyand rad iograp hy of ,h. fetus

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A, .. " . 1\ S.. RauI.mo. L.. and p, l,onen . T , ( 1%0), Jf""""",,"' 0/ ",m~ hloodjlo~'

and "",i"""," abolism. A""". 1. Obm l . Gy" ecN .. 79. &(;_98ll . rt ),,)' J. A .. Golbu,. M, S__FIlly R. A.. and Ha ll. B. 1).1 1977). Prenat a l diasn <>Si' of

dy, p","" ktdney disease CIt". Ge",,' .. 11. 375-3 J%.Broc• . D. J. H, 1 '97~) . Alpna-felOpr",ei n in the prenata lJi .~nn>i, nf n",,,,1 tube J . fecl>

'n l ittlefiekl J. W.. J . Grouch). J... nd Ehhn~ 1. J. 8. (Ed• .). Bi"h DejeclJ. P'o< . 5thIn« , n. tI,," al Conf .... nc• . Excerpt a ),led . Foond., Amsterdam and Oxford

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Bu'l . Ii . Hebb . C . and R. t!;o,·... C (l% n Cho lIne a,;et) la.. in tbe h"ll1an pla"n ta atdiFferent ,,~~es of d<,.. lop ment. .\ "111'" ,Lan duil I, 19\1. 1202

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310 Mellwdsfor AS~SJing Ine Effects oj CMrnical. on Rep r'JdUC/i"e FrmClimu

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