Ii—Afl AO h&l 332 NAVAL MEDICAL RESrARCH TNST PETHESDA MD F/O 6,1

COMPARISON OF PLASMA RILIRUBTN TURNOVER IN MAN WITH CARROt . ‘40N0——ryccu)1977 F L R OD 4(EY. POBERK

UNCLASSIFIED P4.

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IENDDAn

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T : ; ’ I I ~~~ T FIF :I~, E C U R IT CL A S C ~ C A T I O N It T H I S P A ~ ,E (Wh.fl 1)51. Fnl,rt.d I ‘

/ REPORT DOCUMENTATION PAGE BEFORE COMPI FTING FORMI O R T NUMBER 12 GOVT CESSION NO.

)~._.R~~C I P I EN T ~ S C A T A L O G N U M B E R

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COMPARISON OF PLASMA BILIRUBIN TURNOVER IN MAN MEDICAL RESEARCH PROGRESS~~~~ WITH CARBON MONOXIDE PRODUCTION ESTIMATED

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SIMULTANEOUSLY BY BLOOD AND GAS MEASUREMENTS . P .,FIM1N~~~~ W~~~~~ EPOR T NU M B E R

____________ - . ~~~~CONT RA CT OR G RA N T N U M a E R J

(~ ~~~~ I~’ . Lee/Rodkey Paul D./Berk \ ~~~i~~ ’/Li~j

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. // //9 P E R F O R M I N G O R G A N I Z A T I O N N A M E A ND A D D R E S S t O P RO G R A M E L E M E N T. P ROJECT . T A S K

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A R E A A W OR P~)JNIT NU M 8 ER S

Naval Medical Research Institute ( ,‘ , Resea ~~~~ No.Bethesda, Maryland 20014 \L.~/~j ~~~~~~~~~~~~~~~~~~~~ ~~

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Naval Medical Research and Development Command // l9~~~Bethesda , Maryland 20014 NUMB~E R O F P A G E S

14 M O N I T O R I N G A G E N C Y N A M E & A D D R E S S ( I f di f fe ren t front Control l ing O f f i c e) t5~ SECURITY CLASS. (of thi. report)

Bureau of Medicine & Surgery -

Department of the Navy f, J~’,J. UNCLASSIFIEDWashington , D. C. 20372 - I I5a , D E C L A S S I F I C A T I O N DOWNGRADING

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L19. SUPPLEMEN TARY NOTES

*published in Fogarty International Center Proe.~a.dings #35Chemistry and Physiology of Bile Pigments, ed. Paul D. Berk andNathaniel I. Berlin, National Institutes of Health,

>. Bethesda , Maryland (1977)0... II K E Y WORD S (Conlinu. on r.r.r.. side if necessa ry aid id.ntlf y by block nimib.r)

• ~~~ Carbon monoxide, heme metabolism, bile pigments hemolysis

U... 20 I~B STRAC T (Continue ott r.v .r.. side if necess ary aid ld.ntffy by block nus,b.t)Carbon monoxide production in man was measured by both gas phase and by blood

—

~~~~~~~~ analysis. Both measurements correlated well with the simultaneous measurement____ of plasma bilirubin turnover , but the blood phase measurement is preferred

since it is subject to less error. Correspondence of endogenous CO productionand plasma bilirubin turnover indicate that heme distrlbtion is the majorsource of endogenous carbon monoxide in man. ,

DD ~~~~~~~ 1473 EDiTION 0 1 NOV 11 IS OSIOLETE UNCLASSIFIED ~~L/ 7’ ~~~~~~ —

~,_ .. / I 2 / I SICU~~IYY CLASSI FICAtION OF tI4IS FA GS (591ai Del. t.ti r.d)

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Chapter 11

COMPARISON OF PLASMA BILIRUBINTURNOVER IN MAN WITH CARBON

MONOXIDE PRODUCTION ESTIMATEDSIMULTANEOUSLY BY BLOOD AND

GAS MEASUR EMENTS ’

F. 1h;~: RonK~ ’~ AN!) PAUl. I). BERK 2

Q

u ant it ~it iv e nieasurenient of en— i i i t he 1)1k W i t hoti t passing th roug hdogenous carbon monoxide (U)) t lie p lasnia . Measurements of B R U

production ifl man was f i r s t sa t i s f ac — Wou l( I I hu s he eXl)ected to under est i —i ( )ri l’~ (lone l)\ (;oburii et ;iI. ( I 963) . It mate slig ht IV to ta l b ili r L l l ) i I l pr odu —has i~~ )W IX ’CIl esta blishe(l t h a t the in t ion

/~~() breakdown of heme results in Procedures for measur emet it of ( ( )e(Jtlifl l oIal’ produc t ion of (.() a iI ( l pr o(lu c t ion involve })l~c1l~~ t h e sub jecthili rub in ( Landaw et a t . , 1 970). Si m i — in a cli)sed t ebr eat hing sy stem to pre—tar conversion of heme to (A) and vent ( ( ) excretion. The rate ofhi l i rubin Was demor ~st rated in I i ssU e (h ai lge of ( ( ) in th e to t a l s~ ste m isprepa rat ions on t a in in g liver ni ic r ) — est imated f r o m observe(l changes insomes in ‘i/To (T enhunen et al. 1 968 L 1)100(1 ( arhox% henu)g l( ) l ) in sat urat lollProcedures were devised to est imate ( ;OHh) ~ ~~~~~~ et at . . I 963: Berk ciI lie (Lii l~ p lasni a b i l i ruhi n t urnover at. . 1974: Beiisinger et al. I 97 IB RT) in man ( Berk et al . , I 969). 1 .v nc h and Moede , I 972) or f rom

Simul taneous est imations of plasma & hat iges of ( ( ) ( oii ei i t rat ion i i i t heR R l a nd Co pr oduction in man have gas 1)11a5(’ ( 1.ogtie t’t at.. 197 1) .shown a hi g h degree of correlation ~I lieor et i(aIlv 1)0th 1)100(1 and ga s—(r=0 .99) : however , the CC) product ion 1)hase analy ses shot ild kad to simil araveraged 114 per cent of B k] fo r (X )—produ tion rat es if su fl iden t lv ac—r norm al sub ject s and for sub j ect s w i t h curate data are obtained. This paperIn ( ieased henie tu rnov er due to de— presents (lata for ( () p roduction oh—

iease( 1 t e(1 cell surv iva l ( Berk et al., t am ed hs si t i t ul t a i te ot i s measurement1970 . 1 his d i f ference is pa rtiall y at— of 1)100(1 and ga s— p hase changes.

t r ibu ted to the possibilit y tha t a small I hese (lata are compared wi th BR]’pa rt of heme breakdown in the liver measurements per formed at the samel I I ~I~ lea(l to (Ii i cc! bi l i rul ) in excretion I if l)C ,

siI p f J4J i ie ( I I ,~ t h y I5ur e.iu of \ l i~ I ,, m r .iioI Smaigr i ’, . ~~~~ i) r Ih i i t t I w I i i . kr’.r..ii Ii I .isk St Ru t I . UI . 01 0103 149k 1. .UI .ISs4 I II4IIIS Uuii .IiiIV (I I Uj € ~uiI .nr IIIII’~C UI t h e .i t iihiin ~ .111(1 . I IV Iii,, f)V & OI)~ II mii’ (I .iS Of f i t i.tI iii

, r t ie i h u g thiu ’ ‘,ir~.s ii! t hr \,i~~m i)rp.ii 1 1 1 1 ( 1 1 1 III t h e ti.i~~.,i s e t ’. ur .11 Loge

124

A _________ _________________ ________________

(:() /~,od ,,, bon; B/on ,! /‘ !,ii ‘V S h i t /U I —

(log e l i t tus ~ :( 10 odtt lioti i t t not nial tilat i ‘.~ ig. . t t i ( t S~ Is.iitn I ¶I 4m ~ I l i e 1)I~(1t U —

i)~ 1)l)t’l14)I).iti)it.ll .111(1 (Ilpl1(’l1~ IIi~ ( ta t ) — lioti of t d t boti t i t o t o m~ t tIe f t o tt t heniog lo—

10111 \~~‘. Ft t ~ I \teiI 25~ : l 2 iI~ htti in m H ’ o J ( l t t t Invest 16:3It ~~~~~

( :oi tut i , RI’ I~.4 I ( 4 I ~. t ltt ’ at ion t tiIOIiO\Id(’ ( olinia,i , (1, f t . ( ,~S1 l)tldle% 111. and SJ)s i t 4 1 ( 5 . \ I l t l \ ‘i .\t .ttl St t I I . I I I .t’~~t’i ( ‘ I I . J i I 91)9. M e a s u t c u i e t i t ~)f ~~~

22 t logetonis ( , t t I ) o t l i1 lOi iOXi ( l&’ j ) iO ( f t I& t i o t i

( ulnir i t . RF. \ %S Ulakeriuue . art l RI’. h rsier t o detei ini t ir the rf t t ’ I of high .~ ( .. a( —

I ~u 3 F.t ldogciulus & a i iX) t l t lO ) IIOX1( I(’ (CI(’t . tt iOi l 4 ) 11 lIlt ’ ( I e s t t l I (t i O t I I . l tC ( If t t ’ ( I

} ) t I,t IIIt I tOt i It ) 111,11). ( liii I t t~ rsl elk At’ rospat e Med I() : 1 27—4i1 I12: I I 72 1 1 7M I)cli%ol ta—i ’ a1 tdo})toIIus . M. RI ( olnit ii .

(;~~loit t i , RI- . (l.~ E) att ielsot i . \SS P,Iakeiiiote. RF~ I-ut stel 1971. ( ‘ , lit ‘ .at t , i t i o h t of ta le.411 ( 1 RI-i Fui siet I’.H I.t ( :.iii xo fliOt)OXi(ie (If ( ~t t I H ) ! 1 IT I ( ) t iOX i ( l ( ’ j )tO( I t I( 11011 t O 0 (41 —t t i I) l i f tot: .ni,iI’.tu .tI nteihod .40(1 S0(I1t(’s tti ,tI s’.oiiiet i. I A ppI I’hs 51(11 10: 19 ~ Iof et t o t J ~j )}) l I’hvsDI I 19: ’ I u ~ I I .~~da’.’ .. s \ . F1\V (;.ill.uhati . J t . ato t K 4

4( Iit iiitl( .ohtttt i , RI . and RI’. F() rstt ’t I ¶H ~~ Ph’. siolo g— 1974 ) . ( aiaholisnt t~l hetite in n o . ( 0111—

It .41 ~ at .lt lI(’tt’t s iii.tt (It ’ t t’l Illi t lt’ blood 1), I i I SOt l of tilt’ s t i i t t i l ta t i t ’ot is 1)10(1(0 I t o l t

.11IX )X ’ , il eIllOg IOhit l ( l I l t e r i l I atton”. Ii) ( I t I~ilit I l l) t t l .111(1 ( .411 ) 4 ) 11 t l lOtO)XitIt ’ ( liiith an J (:h~ li t ~esi 11 1 5 9 9 1 1 1 ) . I t t ’ , t s i 199 1 1 ¶12 )

(:ohtit il. RI’ . I’ ( .ontttit ’ , F .~h1xnjd . .uitl L~ [atin ili. I~. M B JollahIssot i . ( Mett ke . .tit d I•Ph egtllakt’I s I ‘)~l. M%o(aI ( 1i.tl tii’.oglo— ( a’ .all i t i—Siali l I ¶4 72. F:iiliaru etlielit of1)1 11 \ \ g ( J ) i t ’ I I s J Ot l . ,5.tt i t ’ i J I’Ii’. 514 ) 1 (a T 1)011 t l) t )I t t)\ I ( l r [)t ()(l1I(liOil I)’. .tlot o22 l :M7 1) — $71’. le s t i t e lion iii t t ’ , a i i . St .it id J (:lit i I~tI) I t i—

( .Ohtl t t i . RI’ . SB k.t l i t i . .ttid W J %Sillianis ‘ .est 29: ( stipp i I 24I ¶400 ( arhon niotioxidt’ piodu&tioii iti I . & i u t n a i i m a k i . K. aitd RI’ ( oI)lI1Il I 909. F f -

~).t t iet 1l.. ‘.stti) IIt’lIo ls lit ,Itl( ’lIl i.i. j (.Ifl) f t .’( I 4 ) 1 111(’l,hIft)lI5I)1 ,ttl(l ( I iStitI)L l l IOhl (IfI ti’. est 15. I 414 4—468 . ( . 4 , 1) 4 lIt 1110110X1(Ie’ (HI 1)100(1 .111(1 1)4 ol’.

( .ol)(I1 ti . R I . .4(1(1 1. \1.t ’.t ’t ~. I ¶ 4 7 1. 1)ctet iin t ia— s lot es . At iiet J Ph~ siol 2 1 7: 1St — :91110)1) 4) 1 111’. ()g lOl )It l 4 ) 5 % gei~ leIlslo t l f t uni I ‘ i t t It , SR. .uld A t. Mt’edr I ¶ 4 72. “. . i t t a t t u t i i t t

at box’. rnoglobin f~~1 t e l l ! sat U ,a t i UI 111 t he r.iie of eIld Igelo 015 taIb 01 00 ItO xukskeleta l t111l5 ( It’ . AITIcI J l’li’.siol 220:66— Pt0 (tLfl 1 1 ( 4 1 1 III ttot tii.il lntt t iat i Ilett igs I7;. 1 ab ( lit i Mtd 79:85 95.

(~ ‘.hut, i . RI’ , II WalJ ,tt t , ,iiitl K .-~hbotitI 19, 1. I t ’ t i l iri it& ’t i , R I . ItS ~‘.1at’.t’t, at l( I R St Itt itid

RCIISII h I ) ( I t IO t I (It 1)0( 1% (.0 1)4)11 tlI()t)OXi(It’ I ¶4119 . ~I i . t o s Itlt,II IletlIe ( IX ’ . gU t l . i s t ’ .

folk)’.’. it ig IlehiltIl rhage Attit ’t J I~ l’.siol ( ~~~ .l( let iiaiitot of the en /s it o . J Iliol

22 (4 :565 $ 7 1 ( hetii 21 I i ~:~ss 01~t I(:ohtirti , RI’ . %% J %%j IIi ,tn ’ ,s, .iti(t RFI For slel \VaIIa e, I I , RI; )I)lIt t i . F I l,hI)I)OIt’sIle, %V I-

I ¶H14b. Fil et t of ci ’ . thux ‘. rc des irboliont Blakeniore , .4,1( 1 ( 1- She~,at (I I ¶4 7 144)11 ( .itix ,, * IIIt )IIOXIdt’ }JT o(I(I(tIOII in m an. Nict haitoa l I’. tt l( llt ( (‘(1 liii t d ’ . ast U lat .ltl(l

J ( ;h~ In’.~~t 13: 1 09$ — Il 03. esi I a’. .is oUt iit’iiiol’, sis iii dogs ( i t t Rt’s(o hurr., RI’ . WJ Wilh anI s , P W hite, ( laid- 211:34 7—361 ) .

[~~~~: ‘ 2~ ¶~ t ‘~~

-

T.i_j

(: ( ) I ’, odui lion: ~~~~~~~~~~~~ 0/ B/ nod onil (,o ’. J) aio 125

MATERIALS AND METHODS

Patients Studied labeled u uconjug ated hili rub in ( Rerkci a I., 1969). .\ II si udi( ’s were ca rr ie(l

Si i i i t i l t a i i t ’o us ill( ’.i s t i l ( ’ i l I ( ’i i l s of Oi l ! to t 3(1 h o ot s .111 ( 1 l ( ’S i i I tS cx—BR I . tt i d of ( ( ) p 1’0(hui ( t I ( I i i f i o n i 1)t( ’Ss(’( l as ~L iiiok’s/ k g bods wei g ht pciI ) l( 14 )d a 11( 1 gas—p h ase data weic j w I- — (Li’.t i l l nn’ (I i i i 12 ivid ivi&lua ls: 3 normal‘.1) 11111 ten s, 6 patie nt S W i t h (o l ige l l i ta I Carbon Monoxide Productioncs tosis . I pat len t ‘.‘.II Ii sick Ic cel lrlist’ .isc , I pat i( ’tl I W i t ii p

~ t ( IX s srn a l . - - .I he rat (‘ of ( .() t ) 1od t i ( t lOl l Wasi io . t u i n a l Iiemog lobi i i ur ia , and I Pa’— -

- . - ‘ . iiie.tsti red i i i t lie ( los(’d l e l) l c at hi ngI i ent w i t Ii ‘. i t t ’ . I ( h lor i(I e (‘X f) OSti IC .111(1 . . . - . -- . . . ( i l ( t l l t shown in hgt t re I 1 — 1 . 1 hi ssi i l ) s e ( lUCi i t l l ( ’p.i t lt f i I ) I o s l s . BR I aiid

procedure is essetitiall s th at ofl )hn ) ( l— 1) I Iase ( .( ) I1i ( ’, i sUi t ’i l I ( ’ i l I s , w i t h — , .( .obu in (( . ohu tn et ~t l. 1963) refined( l i l t ga . })11~Is t (t a t a, were p ei f orifled- — - . to ur oVI( le au accu ratel y coflt rOlle( 1

l i i .411 a ( i ( l l t i o l iaI 3 1 i t i ( I i S i d u a l s , as ‘. . , . . -P4 i i i t h e closed SS Stem. \ a l’iat ion of

) i ( ’V i( ) t l s I% le})( )r te (I (Be i L ci a t . , 2197 -I ) . I n f o i med ( ot isen t ‘.% ~~~ ( )~ ) P~ wit h i t i t he chamber of less than

t . i i t l t ’( i f i ( )ili ea ch i i i ( I i v i ( lu a I. ~ 2 mm 1-1 g was achieved. Carbondi oxid e of t he insp i red air in the

Plasma Bilirubj n Turnover ClOse(I system (lid not eX( eed 0. 10percent . A connection at the exi t of

1)ail ’. B R I was .il ( u l a ind f rom t 1w the sp irometer was used to removep la5 ma d is . Ipp ear atke urve of r adio— samp les of t he gas phase for .inals sis

~P R OM~~Y F P

~~—.--n

~~~~~~~~~~~~44,

~~~ ~~ t-

L~~~~~LiFigutt’ II — I. S( hefl)~iti( diagram (If (IOSt’ (l 1-es puration s ’ . s t t ’ t l i fo r nileasni Chilt’Ilt( if ( .i t b()I1 IIIOIloXidV Ptt~~

tI(lb0l) 11w ( ontitied gas, about 20 liters, was (li awlIf t (In) I lit’ 11(44 )41 hs I Itt’ 1)111111), I).l55(’(l (I’ . (‘ t lilt’ t aI b( )t I tlioxidt’ ,il)S0i ht’l .itidox’. gt’ti 5rt154 )i 1)t’fOI (‘ (‘111cm it lg t t it’ %-arial)Ie sOI LI hIli ’ spironlelel~ Ihe (4 ) 1(1 trap,~,. ked III 14 (‘ , %(‘ l ‘. t’(1 tO (( 44 ) 1 tilt ’ g~t s and lt ’ hll t ) ’ . (‘ 5% . i t ( ’ t ( ..is sa m ples for anal’. ses

‘.~ ~q e i1t ,i~~ i i f t 1)111 .4 1)4 )1 1 4 ) 11 h it’ spiiu111t’tt’t

____________ _______

I 211 Rni’Ikei and Berk

as R’~~L It t t ’d .t fl( l to add kllOwIl small iiilw coiil;iiuiitig ( I t s (liso-.1111011 in s of ( ( ) fo r t h e (Ii I t i t iOu IUC.ls— r l iu i I l l t’t 11’. lelle (tianhiitel (‘t I a.t (nt I ( , h ( ido rement . 1 l ie effi i t e ‘. ) lume of t he (E li l .~ ) , leaving t hi t ’ u h~Pt’1 ~i uindose(l s\ stei n was n~~’asti red ( w i t I lout h n u i t ii ( I f t iw t t i lw f i l l e d ‘. ‘. iI h ai i I liet he subj ect I I I die 1)00(1) .111(1 Was 17 . : ) t 1II)( ’ was StOl)p el ( ’(t a tul gen iI ’. in —li ters ( act LI~L I ‘.o l t tt ne ) i l l a ( tdi t iOn 1( 1 serted ~~ (t i s soiSt ’ ~hn ’ a nt n ) a gul IaI i tt h e ( a l i h r a t e ( I ‘. a t i ab l e so l t i me (I f t he a ti d fo r m u l i x i ug pu u posns hi’ 14 ) l ( ’ t lieS)) l it ) IUC It’i . 1:4) 1- P 111P°54- ’5 ( I f ( 1( l l L i — .iii.i l’. t i ( pI0((’tlUi(’5 ;~ ( l ( I I I i O I I , t I ~, l 1 l i —

i( 111 . t 1W tota l ~~~I S sot LIIIW Was t aken 1 ( 1 ~lt’5 4 ) 1 5 ( ‘ i lOl is 1) 1( 11 Id .i tid of I f in gaslw th us iiit ’asti red \OIt if lR ’ ptus LIW vol— phase w em e t ak e i t at 2 i ) — l n i l I t i I e h u n t -

t ime (o ntaine (I in the sp ii-ometet bell , v ats o’.t’ i thit ’ next 2 hio i i , s I. - s t ia ll s 5l\xr ps, 3 and corrected to S F PD~

1 I h e sa t i i p k’s Wt’l(’ iis( ’( l. I hit ’ t otal-hod ’.vo lume of gas disp Jaced b~ th e s il l ) — ( :0—hi l i ( I i l ig ( a it ’ . ( ( ( B( : ‘.‘.

jecls head was assumed to bc eq ii i s a — I hen (It’tnlIllitin d h’. 2 (1 ( 0 3( 1 n i l ~ ( ( )

lent to the added lung sOItlille. (dej)t’Ii(Iilig 1 l J ) O I l t in ’ S l Ih j (’( I s hod ’.Oxsgen tension in the dosc(L s’. stem wt ) being .t(l(ie ( I 14) I In ’ gas 1)hast - h it ’

was main ta in ed near 150 mm Hg (:( ) ‘.‘. as added 114 ) 11 .111 ~ ( ( I l l ,itt’I’.F102 = 0.2 1 ).

~~ \ a ltles of the ‘‘ t ’f it ’t — ca lil ) 1 aln ( l ( 14 nuh le—si i ) f )( ~ k g lass u

tive alveolar P~42 ’’ were calculated by l lonwtt ’ u f i l led ‘.51111 ( ;( )

the method of’ Ri les et .iI. ( 1916) a s— Mat 1insoii ( u ., 1’,tst Ri o h ei b i d , \ ) . )

s t i m in g arterial P4 ‘~~2 = 40 mni H g and at t h i n ktio ’.’.ri 100111 t e t u u p e t a l u i c and

respira tors (Ju otieult ( R .Q .) = (1 .85. a tmosp f t e u i ( pt essi i i ‘ . . ~~ g . i s — t i g l i tI ’ se of 21 p CF ( e I i t oxygen i i i t h C s’. r ouge 5% aS used 1 ( 1 iepeaR ’(I1~ f l u s h i. hosed ss st t ’iii pr ov ided ( ‘sti flbl t& ’d the a i i t roni t he t i )s( ’(l s’. s l e t t i

. i i ierial P values f rom 95 to I Of ) hr oiig li I hi ’ 11)1)1 )l)i(’It’l ,uid l) .I ( k i l i l i I02 the ~~‘ sIe l)I . \ t ’l)()U s l) I ( )O(l s .itui p k’smm , neal- the s~t lue observed in ma ti

‘. ‘.eie Ohia i u l e(I 15 and 6( 1 n ij u l u l t ’sbreath ing normal air by Clark and af ter a ( I ( I i t ioi l ~ ( ( ) WIi ( ’ l l S i ) Lil a1Lamhet -ts et i ( 197 1) . Each t a n k of OX S —

gen u sed was analv ied to determine equi l ib r ium between the gas .111(1 thehod’. had been reestablished ,the con t an ) m a t loll wit Ii GO, met bane , -

and nitrogen.The stu d~ was begun bs the subject Anal ytica l Procedures

being placed in the hood and the OX’.—gen content ad ju st ed to 2 1 percent. All measurements were made in1 t ) i t i a l samp les of the gas phase and dtip licate . Blood was analvied forblood were obtained 30 minutes abici total hemog lobin content b~- thet he oxs geti concet it i-atton was ~td c ’.anmethemoglobin procedure (Z i j l—j usted. Blood s~iti i p les were dr awn stra and Van Kampen , 1960; Vanf rom a periphet .iI vein b~- use of ~i Kampen and Zij lstra . 1961). A reac—small needle and a dry syringe. The don t ime of 2 hours was used to in—blood W.iS immediately placed in a sure comp lete conversion of’ COHb to

cvanrnethemog lobin ( Rodkey . 1967).5 I PS. g.is %,,Iun)r’. .41 .,rnI )w ,l1 t rt f l pe’ratu nt’ 41t (1 P’ ” ' Blood CO content was determined by5411 e ‘. ~II~ T sdh or att’d: S I Pt), g~ s ‘ .olunws ted ut ed in I

7I ~ ) 111111 V I V~ the method of’ Collision et al. ( 1968) as1 ( 1 , 1111 1 1,0,. ‘.oiume t rd ( lion (II inpired gil’. w h ith modified b Rodkey and Collisionit )~ .ti ni,,riositfr ifl t ix ’ .grr i

( ( ) Pu,duci,on: ( wflpi,,i~~ n o/ B/no d and (;ac I)ala 127

I ¶1 7 (6 Pnm’nt sat tirat ion of hemo— gas phase from endogenotis CO p ro—g lo l) iii wit h i carbon monoxide (C( )H h) (Iti Ctiot ) ( lur ing the ree quil ibrat ionW,IS ta ken as 10( 1 x blood ( ( ) content period. ‘I’he CO production was then( IIV1 ( led I) ’. I lit ’ to t a l hit ’n iog lobi i i GllCt ilate ( 1 fro m the equat ion( :( ) B( : . i .~~~~., I 39 x g p e l t e n t h i b . ‘I h it’ ( :o }) 10~~~1t h ull I mllhr) = I l lgas j ) hi . ls ( ’ w a s anal ’ . .‘ed 101 ( ( ) bs gas

~ ()~, x( 1 1 l ( ) i l i . i I ( )g 1.4 1) 1) ’. ( R )( 1 kt ’’. 1 ¶ 17( 1) .it i ( 1tot O\\ get ) at i ( l cali)oti ( I i OXi ( lt ’ 1)5

Results wet -c then expressed asst ,i i i ( l a 1 (1 P 10t LI

~ molesIkg bod y wei ght per day .lot al (:OBC was calculated f rom

Calculations the gas data by use ( If the Haldaneeqt lat ion to obt ait i the change in

To calculate CC) production from COlib saturation caused by C01) . hethe 1)100(1 data , the rate of’ COHb Haldane equation expresses the

hange was determined after the ~r~ — equilibrium relationt ia l equilibration phase os-er about a (X)Ht)

- 12 12-hour period. The method of least 02 111) ( 4 ,Psquares was used to establish the slopeUCOHh~~) and intercept of this where (X)Hb and O2 Hb are the frac-linear rate. A kn own STPD volume ~f t ional saturat ions of hemog lobin wit hCO (C01)) was then added to the gas CC) and 02, respe ctively , and K is thephase to determine the total ( OBC. relat ive aff inity constant of hemog lo—The increment in COl-I b caused by bin for CO and 02 ( Rodkev et al , ,(:01) was determined 45 and 60 m m - 1 969). ‘The e( Iu iI ih ) r ium values , P ( ( )utes after the injection of CO. Cot-rec— •iiic l ‘02’ at -c essent ia l l y those of ar te—dons were made for the anmunt of ria l blood. In the closed rebreat hingGO1) remaining in the gas phase and sy stem alveolar , insp ired , arteri al , andfor the endogenous change of COHh eqt i i l ibrium P4 0 are identical , i.e.,dur ing the ree qui l ibration period. CC) i~4 co = (P 14.1r 47 ) Ft (X) . 5 E( 1Ll iIihri I in iproduction in mI/hr was calculated as p is estimated as arterial Pa°2 (Clark~ COHb~~/ IO O t imes COBC. Results and Lambert sen , 1971; Riley et at. ,were then converted to Mmoles/kg 1946; Rodkey et al , , 1974), i.e., Pa°2bod y weight per day . (P Bar 47)’F’102 55 hi -eathing air.To calculate CO production from ‘I he value of’ O2 Hb may he expressedthe gas data , the change of total (:0 ~ as 1 -(:OHh when the insp ired oxs-the gas p h ase (gas vo lt in ie X f ra ction gt’ti is 21 percent 01’ greater. WithCO) was determined over the 2-hour t lwse assumptions and approxima—period . The method of least squares t ions , (:0111) ‘.s-as calculated from thewas used to evaluate the slope Hakiane equation in the following~~C0~ ’. expressed in mI/hr. Follow- form:ing reequi libration after addition ofthe known amount of (:0 (COn), theincrement of total (:0 in the gas phase

1 K.~ h,i,,,m~it 44 essut t - . I’~

( ( ) .i,u f p.43,, partial(~~CO1)) In ml was calculated , taking ilessuir of . i t Ix ~ i tttut ,i,xuk and ox ’ .gr n ii , the .n-teriai111 (4) account the amount added to the L~~1

128 Rudke’. and Be,k

22)) P~, ( :() l ’~I s ’ .h ie i e (( fl 11 — ~~( ~~~~ ~~~~~~~~~~~~~~~~~~~~~~~~ t ile

1’ 0. 220 P~4 (:( ) ( ( ) aI)so rlx’d by I lie StlI)je ( t and- .~( ( )1 lb is the ( .Ll(ULLtC (t change inI h e to t a l (X)B( : Was then cdkLi la t e ( t C( ) F ib (aused bs ( l ie ( ( ) absorbed.

( ( l i l t ) ( X ~ D

— ~~ ~ 1t ))

..~( ( ) l I I I

RESULTS

Variabilit y of CO -Production Estima- In view of the vt’rs small CV oftion From Blood Data (X)BC measured in a given subject

(vide in/ r u ) and the very small C~ s of\ ar m abi lit s of the ( :0—pi-od oct tot i the chem ical methods em ploy ed . t he( ‘sI imat ion is a i-esult of 1)0th biologic ana lvt i t - unce r t a in t y of t lie CC)—S ar i at ion an(l .ii iah t ic un c er t a i n t~- in )1oduc ti on est imation m a y best henieasu m e m e n t of the COB(. and of t1~t expressed by the stat is t ical el-i-or inrate of COHb change. I en measure— the measure~1 slope of COHb change.nients of (:0 l) 1o(I tlction and of This error , expressed as the coeff i—(:oB(: were made on a si n gle normal cient of variat ion of the calculatedsubject 0% (‘I �i 3—ve ai period. I he slope , has been estimated in 59 tIe-meai i salue for COBC was 11 .9 ± 0. 1(1 terminat ions on 32 individuals to beS~~ I i i i l /k g. (ot -r espotiding to a t oe— 8.0 ± 0.61 percent. ‘Ih e same cr1-o r inf f i c i ent of a rm a t i on (C\ ’ ) of 2.3 p er— slope is observed with male and(‘li t . lie -orresponding mean value female subjects wi th 1)0(11 normal andfor (:0 production was -1.9 ± 0.28 markedly elevated rates of (:0 pm-

~.tni ol esIkg per (lay , C\’ = 18 percent , duction. A constant CV for wideo’.ei t ile same period. ‘I his part icular changes in slope is proh~I 1)lv due tosubje ct has a CO production lower the constant CV’s for Ij oth totalt han 111(1st no rmal i i i chi v m du als wi th a henmg lohin and blood (:0 content es—m a n ge ( I F values Fro m 3.0 to 6.:’ tin iation (Van Kampen and Zijl str a ,i~moles/k g pei (las over the 3—year 196 1; Collison e .tl. , I 96S). The dataperiod. In a group of 6 male and 5 suggest that at least half of the oh—f e t ~ia1e normal volunteers (Blaschke served uncer ta int y is due to biologicet al. , 1971) GO production was 8.9 ± ‘.~1ri~Lt ioti .0.6 ~.tmo1es/kg Pet’ day wi t h a range1 roni 6.3 to 12. 1 ILmoles/k g per day Equilibration of Added COcorresponding to a 22 percent ( \ ‘ .1hi us the da y —to—da y measurement of Subjects placed on the closed i’e—(:0 production for a give!) normal in— 1)reathing system equilibrate the gast l iv idua l has a coefficient of vari ation phase to the equilibrium P(. () of thesimilar to t hat observed at iiong a blood in less t h a n 20 minutes . %~‘hengt oup of normal it id ivid u als. co is added to the gas phase , how-

ever , the reequi libt ’ation requires aStand.ittl C It I P 1 I I ) il t e longer period. ‘I ’his is illustrated in

Li) I ’tmliu izo,, : (M m ~art ~on ol f l /nod and Ca I) aia 129

I able I I — 1 w h e r e a subj ect ‘.v as l)Ia ’~~1 CO pro du (-tion were calculated fronioii th e close d s’. steili am id ‘. a lues of the data obta ine(l on the 1)100(1 antih k I~~d ( :oi lb and gas-p hase CO were f rom gas— p hase anal ysi s . The data arenie a s t i i e ( l . ( it’arl ’. , t he f raction of the pt ’setited in l ahle 1 1—2 for sub j ectsto ta l ( :o in t h u e gas phase a~)proaches w i t h an(l wit hoot ( ‘vi (Ietice ofhe t h e o r e t i c a l s a t m i e ( I .3-i 9~ f in - t his hemo lssis .

sI I l ( 1 \ I w i t h i t i t he f i r s t 2( 1 m i n u t e s (10 Clearly ( OBC cati be e s t i tn a te ( lhe (lose(i ss s t e i n . ‘I ’hi e same e ( 1u i l ib— c ( lt i a l lv ‘.‘.cll f to t )) cit her blood or gas

I 01111 t l is t rml )tltu)n is aChie ’.e( i a f t e i data. The ra t i o of C()BC ca lculated,I ( l ( l i t i o l i ~~ (~0 to the gas h ) hiase but at f rom the gas data to tha t calculatedleast -13 miii n ti t es are i-equired. This from the blood data was 0,968 ± 0.01leprest l i t s the t ime fo r absorption for these 12 measurements. This110) 111 I lie gas and d is t r ibu t ion I h i-oug h value i s not s igni f icant ly d i f f e r en tall ( ( ) binding comp ar tments . For from I .00 (P >0 .2) anti the 95 per(-entthe se reasons we have chosen to allow confidence l imi ts are 0.90— 1 .04.15 i 11 i ) l l i t ( ’ s foi i-ee( 1tii l ibration a f t e r There was get iera l ls good agi- ee—a( I ( l i t ion ~ ( () and h ave used data mei it between the I 2 paiie (i iii eas—i a k c n at i) 0 th ) IS atld 6() m i n u t e s of u r en i en t s of ( ( ) p I o(I t1(t ion l i i bloodtee quil ihi - at ion fo r ( lup li cate e s t ima tes and gas p hases (r = 0.95 , P << 0.() 1) .~~ C:OBC. fo r these stu ( iies both gas— and

blood-p ha se mea suremen ts of COComparison of CO Production Cal- I) t ’oduction correlated hi ghl y with

culated From Blood and Gas Data BR ’F (r = 0.97 and r = 0.96 , respec-BR ’! ati ( 1 (:0 production were t ive lv) . Flie r at io of ( ( ) product ion:

miieasured simultaneously . \alties of BR’f was I . 12 ± 0. 14 for the 11 cases in

IABLE l i —I

COMPARISON OF ( .AS CO (A)NITN I AXE ) I1IFLO RE I 1CAL. E Q U I L I B R I U M CO(:ON 1 EN I st BjE(;I MAt E. 586 kg, P 102 = I 72 mm Hg, Pitar = 762 mm H g,

= 18.68 1. sr I’I) , COBC = 9 14 ~~( (1 in I~IIS

I,,,,, ~ I’ , ) , VI I I If

‘I - ( ~ ~~ ( 0 / l b (th’ . ( i/~ t ( (I in

/14 ,~ , 1~1~~

( I,t,,,I —

L~!I V

I , 0 1 ii 4 ) • i . i e ~ I ((2 i )1

I ( “ I ( I i , i 4 2 % 15 4I (7 (I 4- MS 1 (13 14 $

2 ( $4 ( .0 ) added = 16 .72 ml 51 I ’ll2 27 2 I I ‘~~~~ i M S 4462 ‘2 2 )I ( _‘ i . ) i $9 i .732 77 2 ‘i 24) 2 19 5 1590 4)2 2 ‘p4 205 0)4 i 58

Ser te x t at nh II ‘.i Ii a it iii ile ’. to t cx l~1~ t it ii ni of ~ibhre’.-tat Kifl S

j it ti l.itrd II’. Ct ~ iI at ii iii I Wit Ii ,~S%tl 1111)111111’ . .4 ~~iVC’tl III t CX iau uI ..tril Ii ti nt ubse t ~et1 V . . ( 4 ) in gas , t( )t~( ~. and COHh

- —- - - - . ._A_ . t t., t _ _ _ _, ,”, - ,. _ * _ _ _ ’,. ”~~~~~~~~_ _ _ _ _ _ _ — — —

I :ft) RO(I k( ’\ ~~~ B~~k

1 ABI. F ; 1 1 - 2

( O M P . -~RISO\ 01’ B I t , t R t ’ B l N t U R N O V E R WITH CARBON M ONO X II )EPR( ) L ) L ( :- I ’l ( )N

/J/i,ui/ /),j/ij (,u s( ( 4 ( ()

S / ( , /41? ! /V I u l / I l 4 (lul l / it~~ / i ~

I - 4 jL7Hii/I S I ( . l ) /0 ( . 4Lrn ( lb ’l / (;OR(: ~j y,,,/, /I ‘V V I (n I a,’ ? l ’ ~ ’ I s i / i l ), A

~’ ,Iii1 i ’,i! k~’’ A’~’ -I/Ill I I in! -k ~ i A L’ ’! ’- ,

1 ) 1 1 . 51 \ i 41 ) 11 n P ) l l l t i t ( ( t 27 3 1 ( 1 12. )) i)) I Ii 251 ~s , i tual sol t t t i t t - t t . . 5 $ ¶~~ 4 5 4 , 7 :1

).~ ~t I \ iuro, i l l i i I i j , j n - 4 - z - 2 h 1) ~. 7 7 .3 0 . 0 —,

J II I- I I i g * I I i I .il

np l . i i K s n i n l * ) 4 . ..I

tumn 27 3 5 I J i . 5 7.3 1 ) 4 .2 ‘

SI I I St 4 i i igeii i r , iI2 1 u 4 0 1 3 4 ) i 3 0 ) 12 5

51 .11 I— ( i I l 4 ~( ’ l i l ) . l t

23 3 lu . ; In 3 I . )) 4 I I ) I

t I ll F (; i.iigt ’ t i t~t l( it ‘.t O ’.l’. I ~ S I ~ 4 5 17 1 (4 3 I I I,

( : 11 SI ( t igeiiu.t I2 7 . i 14 $ :1 ( 42 I o 2 0 2

0 II 51 ( ) t i g ( t l l I .iI

.j )I)t’ t ( ~~ % l oSt ’ , poSt4 ( 4 ) 4 4 5 - . 5 . 5 l i t I I I 3 13$ ft

J II 51 5 ti n I Itlot nIt- In i

- 7 ) I-l i 5 4 15 . 2 ii 7S b . 51 S u k l t i t ’ l l i I i ~r ,ise . 3 1 1 7 l 2 1 51, 3 1 2 1F I F 1 . , i i n s n ‘ 1)1 ,4 ! t i in t i i t n~il

l)t’ nt i(g I((t )t t i i I rta . 2~) .t ¶1 .3 2 5 0 ¶4 .1 2(i 7

5 t - u-~~ ,ito l u’~~t k w t t n,tt s lot ‘s ~~ l . i I i i l i t t i of .ihl,tt ’vt.,t t i,tts

w h ich hot h i b i l i r u h i i i ~1Ii (h ga s— ~iui~~’ lat ion sI udied , gas — and h loo d— })h asc

data we lt ’ avai la b le and was I . 1 8 ± niet 110(15 gave eq t i i v a kn t (l ata f ’oi’ ( ( )IS tot ( h ue I S studie s Ii - otii winch pi’odti ct lOl l . the statist ka l ui i t ’ert a i t i t s

hot h i bi~i i’uhi n a iu d blood—p hase CO of ’ ans si ogle st ri d s was Il iUch hi g lici(Lita w e t e av~i i l~i h) 1e. hi I 550 pat ie li t S It) for t lie gas— p hiast ’ dat a. 1’lic un c e i —I lit ’ ~) leSet i t series , t h e rat io )f CO taint R’s of t he tw o est m n i a t es of ’ C( )pro dtK-t iot l : B R’F was greater than 2 p i’odU(ti oii are best t’xplCsse (l by th efor 1)lood—p ha se Iii&’ astl i- t ’m eiit s. ‘i hi s coelfic iei it s of va i- iat ion i i i d c t e r n l i l i a —i (sulte(l 110 1 1) .111 ( ‘XI i e l i ) t ’ iV to )%% ‘ aiid t 1( 111 o f t he i-ale ( I f t-liaii g e (

biolog ic ally u i t l i ke l s s aloe ( I f BR I i l l ~ CC)Hb~ in blood or of .~CO~~S instihjt’c t 1’. R . , ai id an ext i’enwls hig h t he gas p hase . ‘I lie coef f ic ien t ( I f s - ai’ i—‘ aloe o~f ( ( ) produc t loll I i i subj ect at ion iii t he slopes f i ’on i i these I 2M. i i . \%t , i i i t i O t ( ‘X i) lai li cit iit’i o f stU ( l l ( ’S was 8.7 ± 1. 07 l) eiteIi t for t i l t ’hese aberrant value s : in WeS ci . hot hi blood dat a and 26.9 ± 5.6 pt’i ceiit Ioi

are cleat -l~ i i i ( ’ ( I n s i s l ( ’ l) ( wi hi a ricil la is the gas (lila. This si gn i f I cant differ—observations. Alt 11(10g b foi the 1)0)1)11— cu te . I ’< 0.0 1 . is exp lained hs the cx—

!‘P~((( ll4 ( /101) Comf iai’oon o/ Blood and Ca i) a (a I ~ I

t 1 e I I i ( ’ l s sin ,tll t a R ’ of d ia iuge i i i gas t i’ol 0 ) 1 i i l O ( l i f \ t h e ( l i t ’t . I t is k n u w i i( .( ) 0 o I 1 ( e I i I b . ~’lo I I , i i i o l t h e tact t h a t t h u a t f a s t i n g ( ffloo iiic i ci al . . 19 7 1) and

98 pen cot of t h e ( ( ) s tas s in the s~ ’~~ei , t ’ ( a l O I ’i ( 1 t ’ s I t i ( t i u i l ( 1.011 ( 111 etsr ihj eo . in o l (toes i i o ) I ~t~)j ) ( ’ai’ in t he ii.. 1972 : Rt ’i i s i t i g c i et a !., 1973) i l l —t i l t L i I . % 1 i 1 l 1 4 ~~~~~ s ( I i I i I l I e . . -~~t i ( I I l i e 1 ( O i l — ( lease s c i t i t i i h i l i t 011111 ( UtI( ( ’ t i t f a t i o I lI i I l I m u t 111 1 4 f .o l i i i I s t h a t s t i i a l l t l i . I I i g ( ’ s ~i i i o t , 1 ( 1 a sIIiillt’l ex t eil t , ~ ~ ~~~~~~

—

I I I 14.4 ’s S ( I1l n b ’ f t ( l i i i “ .1 l i i l ) l i u ig 4 1 I i ’oiii I H I t ) . \ ~~l ( 1 1 1 1 ) 1 1 1 I 1 5 4 . ’ 1 ) 0 I i i i 1 ~ f I \ ( I I ( l t i —

leaks ( . 0 1 1 1 4 11 1w (l et ( ’o te(l in t h e (01-J h n’ci s sv ;is used t O ) ( ) I ) se i5 ( .’ h it ’ \ , I I i , I t l o l l

o’S t i l i i , f t i O i i . h ) t t t ~s i l l 1R’ ( l i i t ’t ’t ls i’ ( ’hd t c ( I i l l (:( ) l ) i o ( l l i ( ’t i o i i ss h i t ’ i i a ( O l i s t a l i t

to I lie a lcula t e ( 1 ( ( ) i l l t lie gas ph ase , diet was used. ‘ I lie (h i ( ’ t W a s o u t s ( 0 ) 1 1 —

~I i i ( 1 h i e ine ssI l l h ia t ig e (A) p roduct l o l l ) t l’Olle (l 10) t h l ( ’ e x t e t i t t h a t a~it ’~i t i a i t ’.is a k u l a t et i f Foil) the gas data . ‘lh e a l o l ics \ V C I ’e ~~I 1 )vi(Ie(l aii (l I lie sub j ectt - t’ l i a h I i l i t s ( I f 14a5 (Li la i l i ( ’Ft ’iSCS fo t J ) a — OI ) s l l I f l t ’ ( I i l l i ( I e l ) t i ( a l til(’tltl on t h et i e n t s w i t h e levated ( ( ) pr oduc t ion . ( lay j ) l ’ iOl ’ 1 ( 1 and ( I i i t h e ( La s ( I f (‘it ’ ll ofl i i t i lt ’ sevei~ sIll)j ect s with BR’T ’ < 10 a iwo ( I I ’ l i i ( I I ’C i l i c a s L l i ’e t i i c l i t s ( I f ( ( )(X of 36.3 ± 7.8 was o)h)served, while l)Fo(ttI(’t iou . Results given I l l 1 .LI ) lethe five subjects wi th BRT > 10 had 1 1— 3 5110w t h u ( o ) i i s t a u i c v ( I f a (li d1 3.7 ± 2.9 , a value much closet- to the coti t a i u i i u i g a( 1C9 oat e (~ Ih ) I ’i C s 15 11 ( 11 a

t o)t’fhc iell t 0)1 var ia t ion obse t-ved from l i i aj ( I r I actol - i i i eli n l i i i a t i l i g s a l i a t ions1)100(1 (t aLl . ~ii (A ) product ion. It app eals t h a t t h e

o)l)5ei’\’C (l t ipp e r ati ~1 lowe r ext rUffl esEffects of Constant Diet ~ ~~~~~~~~ j ) I ’o (L t l t t i o l i i l l l i o i ’ l i i a l st t h )je t ’t S .

~1osc ( :o—p I (I ducmio fI ( S t I l u t d t C s 5% ’CUC 6— 1 1 /2n101es/k g ~~C1’ tia s . in.is t ’qu al l st i i a de af ’te r the subject h lad eaten well he ohser~cd i l l i given i t i ( l i \ i ( l t t a Ih) l’t’ ak f i s t , hut W i t hi 1) 0 ) at t e l i l }) t to c O I l ) — ( I l l I h t ’ (‘( I l l s t a i i I diet ( P R ) or w I l e l i t h e

1 .- \R t I’l I I :~1- t- l - F ( : I Of I f l F l .-~R\ ( :O\ I ROt . ON

RF PI I ( .\ I I E S I I M.-~ I t ( )N OF ( \ R B ( ) N Sl( )N ( ) \ I l ) I ’ I’R( ) 1) t ’ ( I I ONIN NORMAl . V( ) t , t ‘N I t ’ . f R S

(4 / l?uI/I)l !IVIPI ( ( I I ? ( .‘i l l / . I 5 , 5 1),, t~irn , , ,n i , ,f I ~_i,,, ,/, - A~ (IllS I (ill! j,

5 K SI \ , 1 1 , 3 3 )5 K it’s ( , ( I . ’ ( I

5 K \ t ~~ ( ( ( I S T

Win SI \, 12 5 I t I’I l l

K \ , - ~ I I ‘ I I l i S

RI I St 5,.,, I I is I ) , I )

g i l ‘O- s I i ’ s I.” 5PR St 7 0 I )~ 7PR \ i ~~ I I h . 0

i’R I 1 . 1 t i Ii)( . St \ - ~~

i_ I_ It rs ‘s

1)0 • s t’s ‘4 . I it It

1)0 . \ ,~ l i i I” I

_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

I 12 /? io l l l 5 m u ! B ’,k

( 11 ( 1 t~ lu’s’s ( t t I I l o ) l h o ’ ( l I ~t K I . I ho ’ l i i i — t l ( ’ S t ’ t I l u I I ’ e t h i ti l~ V’1”’°t aii (f ( i i i —

( e l t , l l t l I \ I I ) rst 1 l l I . i i i i O ) i j l I h i t ’ 1 1 ( 1 1 t \ J ) I ~1i I i I hi t ’ I .111 ) 4 4. ’ of ~ ~ ~ l~~~’° 1(1 t I ( —

.M ( )l Ih ~ t h i t t i l t In st st ih j t ’t I s 5 5 . 1 5 I l o u 0 I I ) s t ’ l S (‘(1 i i i I hies - Iss t~ i t o l i s I h i a l s ,

DISCUSSION

I l i i ’ s 5 ( 11 ( 15 ( ( 1 1 ) 1 1 1 i i i ’ s t h a t i i l t ’ . I s I i I ( ’ — i’epoi ’tcd ( B ( ’t k et al . , 197 1) . \ I t h u I t I g hiI 1 l ( ’ l i t ’ s ( if ( ~( ) ~I t ( l ( h I l ( 0 ( 111 01.0 14. ’ I l l i h i t ’ i t i s W i ( l ( ’lV i ( ( (‘J)t(’(I thai hil ii i i h i i i , i i id1 4. I S phi .ise a l e . on a sl at ls li t .ul basis, (( ) a t e t h e j)u’o (ltl ( I s ( I f hi eii it’I i i g h ls s i u ) l i l . l l 1 ( 1 those 0 ) 1 ) 1 a i iu ’ d I I ’ ( I i ) i (‘a t a l ) oh i s t i i i l l (‘(i t l i t l H I l a l ’ i u i i o u i i l s a t i dt h e h)luod p h ase I hie ( ( )—jn’ u dI l ( 1 1 ( 1 1 ) t h a t i i e i t h i e i il’is( ’’s I i’ou ii 01 bet’ 5 ( 1 1 1 1 ( ( ‘ 5 .

( 1, 11.1 I i ~~i 11 t h u tw o i i i e t h u n t s t’ ui’i’c ha t t ’ u i i ( l l e re ( e i i t sunlies sh ow t h l ; l t ( ( )h i ig hi l s sn ’ i t h i e,i~ hi ( ) I h i O ’ l ( 1 = 0 .°, ) ) , i t io l u i i a s In ’ Pl’~ 0 lti t ( ’d hi o lo ) g i ( a i l s f i o t i i1 ) 0 ) 1 1 1 ( ( ) — } I l ’ O ( I I l t 1 10 1) ( ‘ s h i I ) i , l t ( ’ s ~~~4 i (‘ l io t i h i e l l i t ’ 50111 ( ( ‘ 5 st ichi i’s clihialo gt’ —l ,ue h i i g h hs rs i tlb r ibe , i s tn ’ ( ’ I lben I s of t )alt ’( I int ’l b an e ( K u h : 1 j ( et al . . I 97—1 )R R ’l - .-~ l t h i o u g hi I lie tw o ( .( ) inch hio (I5 ~uid l I o I V l ) h ie l lo l i c ‘o t i i p o l t i t l ( l s 111 lu ( h —, t l &‘ s t a t j s t i t i l l s sitni lal . t h e g , I s — I I h i ast ’ i t ig ca techo l at i i i i i c s ( \ l i s a i i a t ’ .i andl i leasti r eui ie nt is te (’hll iica llv ni uch mOre Takaha ski . 1 97 1 ) . Eu i’t h ie r t i io i ’e .

demandin g a nd it id iv id ua l sI t1(11e5 ( I t her St t l ( l iCS ( Ei gel et a1 .. 1 972) raisehs th i s tec h t i i ( I t l e have a hig her iIill e i— t h i t ’ j ) Ossi h ) i l i t \ I h at a stliall I i-at -I i o I i i ( If

ent st a i st ical i t l i cer t a int s especialls t lie ( ( ) i t u t ’xp i) c( I i i i u i u a s ai-ise I u ’ oti i

at normal rates of (A) production . hiuit ’~iiJ fl)eI abO lJ SJi ) 11) ihi t ’ gi st l O l l ) —

I t is 0) 1 i i i t ( ’ l ( ’ s t t h a t ( :() j ) F ( l ( I i i t ’ t i ( I l i - t e s t i n a l t i s u i . 1)esp it e these p o t e n t i a lcx ceds B R ! h) s a I ) 1 ) t ( I X i l i l a t C l s 12 - - IS soul-ct ’s ( I t ‘‘ext es’s ( 0,’ t he l i t i(I ( I f

h ) t I ( ’ t ’ t ) t s she th e t t l i t ’ d s l i l ( ’ O I i l l bI Iuo l ( ( ) p io d u c t i o u i : P1 R I oh’sei Ve( l in ou r4 ) 1 )4d5 j ) h i , l5 t ’ . A sii i~i ll pa i l ( I f t h is ( l i ’ s — s i t i o h i e s suggt ’s( ’s th la t i i i l ) i d I i thi t ’ cx—

t ’t’j )~l l) ( S l i i i 5 l e s t l l t 1 1 ( 1 1 1 ) t h e f a c t t h a t t t ’u i t ( I f e x t ~a t i eot i s ( :( ) Pl ’~~0 l t l c t i 0 i i hsB R I sli g h i t l s u u i d t ’i e s t i n i a t t ’ s t ( I t a l Si l t hi 1) iot ’t’sse’s 110151 1)4. ’ sh u tt 0 ) 1 i l e g —hi h i t u h i r i 1) 1(4 ( 1 (1( 1 tOl l , iS h ) l’ t’~ ~t i sls ligi hle .

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