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The Natural History of Atherosclerosis


From the Centre d 'hude des Maladies des Arteres
Coronaires, Department of Cardiology and Department of Clinical Pathology, Hopital d 'Ixelles,
Brussels, Belgium.


term when alluding exclusively to one or another of the various lesions of this condition.
Such habits have made literature more and
more confusing.
As a matter of fact, the only valuable way
of defining each kind of arterial degenerative
lesion is based on histologic criteria. This is
why, referring to etymology, and according
to the recent nomenclature of the World
Health Organization,3 we shall confine ourselves in this paper to the following definitions:
1. Intimal sclerosis: diffuse fibrous thickening of the intima, generally associated with
degeneration of the intimal elastic membrane
and with sclerosis of the mnedia. Grossly, diffuse intimal sclerosis gives rise to some dilatation and to lengthening (unrolling) of the
involved vessels, but the lumen is not impaired.
2. Atheromna: strictly focal lesion (e'tVa' =
tumor ), characterized by the presence of
lipids (a'Thj'py = mush), conferring to the
plaque a grossly yellowish color.
3. Atherosclerosis: more recent word introduced by Marchand4 and referring to a lesion
intermediate between intimal selerosis and
atheroma. Usua.lly focal, atherosclerosis has
an important fibrous component and constantly shows lipid deposits. In their advanced stages, atheroma and atherosclerosis
grossly impair the lumen of the vessel.
As a rule, atherosclerosis is less localized
in animals than in man. Especially in birds,
stenosing atheroselerosis is much more diffuse
than usually observed in human arteries
(fig. 1).
The diffuseness of spontaneous atherosclerosis in animals often makes it very difficult
to establish a clear-cut difference between
mere intimal sclerosis and the first stages of
atherosclerosis. This explains why it is very

A LTHOUGH unceasing attempts have been
j L made to increase the knowledge of human and experimental atherosclerosis, the
pathogenesis of this disease is still far from
being thoroughly understood. The uncertainty
of present conceptions concerning atherogenesis is emphasized by the rapid succession
during the last 20 years of several theories
allotting a predominant role to different substances such as cholesterol, animal fats, saturated fatty acids, and nonessential fatty
acids, and the series has probably not yet
come to an end.
Hitherto, epidemiologic investigations have
been quite anthropocentric: although the incidence of clinical atheroselerosis among various human populations of the world is relatively well known, the frequency if its
spontaneous occurrence in the entire group of
animals having a specialized circulatory system is almost completely unknown. In other
words, most of the natural history of atherosclerosis is still to be discovered. The purpose
of this paper is to sum up the fragmentary
knowledge in this field, and to pose some
problems the solution of which might be of
interest for the etiology of atherosclerosis.
In order to avoid confusion, it may be opportune to define once more arteriosclerosis
and atherosclerosis. As, proposed by Lobstein'
arteriosclerosis is a generic term, designating
several arterial degenerative processes giving
rise not only to a thickening, but also to a
loss of elasticity of the vessel wall (diffuse
intimal sclerosis, Mdnckeberg 's2 medial sclerosis, focal atheroselerosis, etc.). Unfortunately, too many workers have discarded the
generic sense of arteriosclerosis and used this

Circulation, Volume XXVI, November



jturv at the Zoo of Philadelphia has built mtiost of our present klnowledge of spontaneouLs atheroselerosis in wil(d aniimals. alike i(ihiekeli ill)it.lilso of the Very obliging hell) of 1m1aty11y oigaiizit-. (ouit --11111g'.on.5'6 was a *Tlese stuSdies couu(II T C8 ( cir 1 I idout only bec.tioalls iln differenit coiiitries.11' 12 and pigeoli1. 1lir(ll aild ni. Coneentrie thickmug of the itirln : the 1(9 1 stiocytes wuithi hinun riletr cytoplasm sh/io the atheroselerotie natun'C cygnus ofthi's lesion.1" So. The Commilission of Coopa-trative Alnltomy (Kaiisas. dese Ins in iny species of domnestie ail. Thie capturing camp of Einpulti (Conlgo). X >2t7. Pavine ni tier). spoiiau leons Itt inal selerosis or athlcrosrlIteosis lhas beent.1)ELCU(R)IlZ 1.nesis of altelial de (rei('Lerativte ee esses es a mid of eni ntIatull listolry TeMaiJi Ob'senrilet. (legelereati_V(e arte-rial li1siols. th10 wock of' Fox. Tue Scienitific Peseairebl ilnstitulte of Pilill.sels 'University (Prof. Prof.f ( den Bergl aInld Pi-of.SALENE. Fox. icnireasi ig Iecuei(eyll(v of steioEsiIg eorounrx lesimns Hi tlie Ihomel(othernic anIimal's of the Philadelhlia 1950. Jim ov s)oiltxtleousl. In1.fmoals.oiiits ieeessary forL the coiupllet'e 11iderstaialidl1 (of thle re. to dissociate the natur al history of atherosclerosis from the nataural history of dliffuise intiinial selerosis. MAr. Wtv of (ald-iolog) (I)r-s.iegiaj (celer fror theC Studly ()f (`oronary.ia fhil)-potauitt l). Dr. B.itz (Fiancec). Eeei itlY. Morhimatl-ins). if niot im-possitAble.d orl reduced to Car) Stdorla .iiiiiials. toio e(aS( lrO-flnrt jioallxt.v eI. Volumc XX V. einax oiisiilde it fils1111v established thfiat mI(ost sp)cies o fiomeotl)hrlni' animals that: ni has doinest:hafite.1 d{log. The MArlrine Biology Stationl of l?oscotfi (France) . d(o AMediiia.eltire ma-ifestat iolXs ii the anlinial. Tf e Aiitwerp) Zoo (Mr. USA).ry School of Utrieclit s Uniiversit3. Another section.\Vith]54. The Avor k carried out for niearlyA half cen.the oi-ller hanilld . killn-dom hans b:ceen uniIdertakecn for a fe xvv e ari-s. tuinde:r thie mIpi(s s of tilew il.iolall Soce. A[aizl<e.. V. Distiai aniid Dr. Baltt(iffe 1 ( 0rmihi ) showed that tle ineideliee (. and(i Aw-itlh the str'esses t-o wheli tI-le animials arie Su1)4j et(ed. The Laboratory of Paithology of the lrnw. PIcrcier. Novmnber 1.Yerasi II idc1(7 ani-d lE'lliot t' iinsisted upIol I lie.itl tile rate of' over 1l)opllation of zoos.962 . the same ness Gou A.Xot iaInIIIa1IIIils a 11(I ibirds livilig i i cat it v t i viii c1ila d I t.x (lexeh)l) 50m111 towin of arCCi 1)8(1 serss Coroner! artery of javaniicus Figure 1 (otl jeeon tee duck (Dendro Ahor. Bari6ty and AMr. in erterilq shoo-ingfJ importance of the lipid deposits.v xv. Aee wish to thank sotouig otlcrs: The Intrn 1a telt. Zw art. Circulation.ioll. Mr.ieatvx' aterial na phleC tllo seevs. The med(ical Laboratory of Stanfle) v lle (CoII- go).V4 YASTE.42) thIeI filst to shio 1ht.Artery I)iseasc i svstemiatic researelh jilto) the arter ial degeeIra. Ealct -lifit'c'-. Thie ''l Justiituluto Latinio Aincrica:o de \Arttomia patologi.]).t]phv ZV0O since lat. inlvoking agai(n aIs Iet l Cause [avorllou the vxol iti loll of t hese elsioiis I lie 'V lug(Ieist o ot tne ej ivet animal] plop(. White no]l Kat.jc II of Me'eico (PIr of Costero). I I viewv tidt (liffntst- bleeak B X27 difficult. Thle ArAsterdani Zoo Toil Veteriina.

activity of the clearing factor. and the aortas of 72 of them (table 1). We occasionallv prepared frozen sections in order to stain lipids with Sudan black B. ineludiiig represenitatives of Cheloniidae (10) Crocodylidae (4) Varanidae (3) Lacertidae (4) Iguanidae (1) Serpenits (16) 2. A second group of imaterial includes the heart anid coronary arteries of 144 vertebrates living in freedom and killed in their natural habitat. The partially dehydrated aortas were stained in a saturated solution of Sudan IV in 70 per cent ethyl alcohol for 10 minutes. in order to remove excess stain. Bir ds: 36 specimens. Very often. and with Weigert's resorein-fuehsin for the staining of elastic tissue. Volume XXVI.l4 Mfethods were described in a previous paper. in 88 of the captive anim-lals and in 51 of the free ones. and the following determinations were made: lipidograim.ated by postmiortem arteriography. In the involved arteries. iincidiig representatives of Marsupialia (3) Carnivores (28) Pholidota (6) Pininipedia (8) Perissodactyla (8) Suidae (6) Ruminants (75) Rodents (10) Proboscidae (3) Primates (95) Table 2 Vertebrates in Freedom 1. This was especially the case in the coronary arteries of somne fishes (bluefin tuns: Thunnus Thynnius L. total blood cholesterol. Fishes: 40 specimens. Gabe's method (aldehydefuchsin) was used for the simultaneous staining of elastic and connective tissue. and then differentiated in 70 per cent ethyl alcohol for 24 hours. however. and. Frozen sections were made in the stained areas. November 1962 characterized lesions were observed. AMaminals: 68 specimens. Paraffin sections were prepared and stained with Masson's triehronme stain (iron bematoxylin-ponceau-light green). TIn both groups of animilals. Reptiles: 38 specimens. that died in the Zoo of Antwerp or at the capturing camiip of Epulu. elastosis was an isolated process. ineludinig representatives of Spheniscif ormes (29) Ciconiidae (2) Coraciif ormes (5) 3. cholesterol/phospholipid ratio. Material and Methods The gathered imaterial consists first of the hearts aind coronarv arteries of 403 captive wild vertebrates. however. these investig-ations are still in their first stages. includinig representatives of Carnivores (3) Suidae (35) Ruminants (9) Primates (21) Animals Elastosis of the initernal elastic membrane of the coronary arteries was observed in some sharks (I surus Nasus Donnaterre). Among bony fishes. The anatomy as well as the pateney of the coronary arterial tree of most of these anitoals was investig.15 Results Degenerative Arterial Processes in Poikilothermic 843 Table 1 Captire Vertebrates 1.). and the specimens hitherto examnined of invertebrates are too few to fuirnish valuable data.also examined (table 2). includinig represenitatives of Stuthionifornies (2) Galliformes (15) Lariformes (3) Balearicidae (8) Sphenisciformes (2) Anseriformes (27) Cicoiiidae (30) Falconidae (6) Coraciiformes (20) Passeriformes (10) 3. the proximal segments of the main coronary trunks were removed and fixed in formalin. The first changes eonsisted in circular or semiicircular nodules of hyperplastic intimiial tissue projecting into the vessel s luien . in many cases. The aortas of 62 of them were . as determnined by Hahn's technic. Birds: 123 specimeins. cholesterol fraction bound to the beta-lipoproteins. much more Circulation. Mammals: 242 specimens.NATURAL HISTORY OF ATHEROSCLEROSIS of the huge field to be explored. Among vertebrates. iineludinig represenitatives of Selachae (1) Anguillidae (3) Cyprinidae (20) Thunnidae (10) Istiophorus (6) 2. blood samples were obtained by venous or by cardiac puncture. withlout any intimal sclerosis. a few points worthv of initerest alreadv have been recorded. Thirdly.

1/te rujtt c4't44t4r4 otertt1( {//' I t ttl}l av t1iif)4 1 (. I leifA Iltsupc 'tX I. 2A ) Figure 2A Fat l intin4a/l 'cItctoau itt.s so0li44mes rllptulle( aiid its frarinent s were dlisJ)erse(l ill the hvyLerlplastie. )ElCOfIT()UTRT 8H44 Figure 2C Abundi anlt lipid deposits in the thi kerned mail coronary t? n/k of a immature Thnn?ns.'4 WClre0 found ill till of the' 10 illVtstlgated tlnlnielS rPlwey were xoun c.s marinaus L.I/i. d1e'posits (fig. Sudan b1)/c/1 B X (fig. (Certai llial)iallianu reptiles.5. 2C). iII (raptix'ity at letlst. intiluiilt lure speleliliells abmit :3 feet lo(. eatught in thle l)b-v of B'iscaIV (Fainee) The main eoronary rtlerv of two of thiem shiowe(d wilespreadl atherosclerosis witlh abundant 4xtrial'T4ll1flar ljpid. Trplse (ealnge0s. Wexlexr. the peseis(iiee of iinuerous histioeytes witlh tlearoxytopla4S1in -very often eonferre( to tihle lesiou-S (lefillite tendencvy towatid atherosclerosis (fi'. elastic( 111(lineb)ranc t1igure zn FJ"ca/ i14ti4441/l st'l'tr'osis i?/ tltt r'igh/4t c'r)4't0()ry'. alill Miller. arte'- xvitlh slight fibr ous.) A verp Zoo. largte ainaeoida (Eutoec. Nov mbe r 19432 . ios °!/ (4 1tioin/ fish. elosel) reSelhl)liillg thliose reee)itly (dmlesribed inI spawvning' Paciffic Sablions bV Robertso. Volum e X\X VI. Altflhou(g. X ej. tlhickeninlg of the iltimna. intinia. Tllw internal elatst i' mIlcbllltl)'al) mi(lIerlleathi the inltial proliei-ations wa.n.VA4 VrAESAEGER. llt itJct-t X t). fibroblasts ta-nd elolagyenois suhstaiiee wxxeve the mi-laiin constituents of tle iollvlules. Focal initiimial thickenlieng witlh ruiptured interutal elastic inenibrane was also fotunid in a main coronary trunk of a 9-foot crocodile (Cr'oco(dilats iloticts) living for mtiairany mnonithis Circulation. slowevd fra"th rli A that died if mneltation of thie intern'al. eal also (develop dlifflse ilntilall selerosis. This is tlhe (ease x ith eeritaiu opliidiains of the typ)llo)s giolup: tlle coronary arteries of a.7. 2). 213).

and(1 of a youmig mnale ebiinpanzee (Pan Sch/iuvc furtl/hii [Cigi]) Of the wild birds shot in freedom.alia Canilivores Perissodactyla.5. Ini th1e lesiolns. B. ani adult feimale showed more ICIispiciaius faitty.) sihot ill theilr nIaltnlll'- habitat. Table 3 4 ortic .ortie Atheoscletieosis Siidanophilic fatty streaks of atlerosclrot' nature are relativelv ommll11on0 ill thIe aorta of wvild mraImlInals ail(1 birds. the lipid (Iel)Osits were very strik6ing ii thle aorta of a rlhea (Rhcau IRotschild Brab. Frequency of Atherosclerosis among WarmBlooded Vertebrates . Of 1three' (leer. Nouvnilber 1 Free specimens Normal Atherosclerotic Total 9 4 - 14 16 0 2 14 6 28 7 7 1 36 28 1 8 48 1962 13 24 10 47 15 38 13*4 . gross sills of coroiary s-clerosis ws.8 NATURAL HISTORY 1F ArlAHEROSCLEROSIS 0 in captivity at the Epulu camiip (Congcro). and 13.). Reptiles 3 2. of aii oldI baboon (Pupio Paplo Desmarest). Focal intiwald sc lcrit in the caranarq airteries of at. 3A aiid B). 1-ii this animiiial. nlolne( shiowedl any sio'n of aortic atherosclertosis. Suidae Rumninriants Rodents Primates 4 Total Cireulation. X 100.). Thme 1??nt r0/Ifs hitioc tqtcs With a ct atl? cyjotplasmnsShow cc dcj1Jit anned Wheo /h e lerrosis. of ani ai1dult biiffalo (Bubalus b)ubalis L. abmndant listiocytes witli elear cytoplasmn also sugg(-est a defilite ten(ledilr towarl atherosc lerosis (fi(g. Aimioi cai)tive aniiiimals. i. 6-foot cap/iet ecrocodul e. il t e/ X 2. For fishes as well as for reptiles.). Nlanimials Marsupi. of a l)eliean (1P%icU/ins Speick).).) of a yak (Roephagrus grunWiens L.ere see-n. Birds 3.4 therosclerosis Captive specimens Normal Atherosclerotic 1. and were preseInt in nerl1y 30 per cent of tfle exallled sIvecinleiis (table 3). of two wild boar (Su?s Scrafu L. Verv slihlt lipid (leposits wvere observed i1n thle aorta of eight of 35 wild boar (Sus Scnaofui Bf Figure 3 A.streaks. Volume AX VI. of a 30-year-old eanmel (Ca}me/tus bactrianus LI. WolViqed A. (dissemimlite(l ill its thoracic aild abdomfinal aorta. the nmim h)er of investigated specimelnes is too snall fotr ain estimnationi of thle freqneiiev of arteriloslero(sis inl thlese (grolupsll of -verl tebrylates.

anid it was rather commonier to find fatty patches disseiminated along the deseending portion of the thoracic aorta and stretching along the abdominal portion. at least. foam eells and cholesterol crvstals were present in the thickened intimta.. C. about 2 per oeut of the examinedi speeimeiis.]. or by thronmbosis. Of the five examined ehinmpanzees shot in freedom. either miaminlals (niioufflon. C. in lnonie of the fi-ve and four Galago Demidovi [Fischer]) was ani sudaniophilic spot to be fo-und. they were quite differenit: despite their diffuseness! and the absence of atheroma. in ungulata. tapir) or birds (crane. Balearica pavoninta (LA). C. and an old female of Mangabey (Cercocebus aterrim us [Oudemnans]) showed widespread fatty streaks in the thoracic aorta and less important ones in the abdominal aorta (table 4). C. I 4 9 4 ) 8 Genus C. in spite of the fact that the serumii of one of the galagYos was turbid. Dend-rocygn?a javanica [Horf. More widespread in the aniterior deseending braneh and in the eircumiflex braneh of the left coronary artery. three were omnivorous mammrnals (two chimnpanizees. and three tree ducks. were the lesions of the tapir: they eonsisted not onily of lipid deposits in the thickened fibrous intimna but also of raised areas forminig real atheromatous plaques (fig. it was found in onlv nine of the 484 Nvarin-blooded vertebrates. Coronary Atherosclerosis Coronary atheroselerosis is rare in our material. All of the aiffected indiviiduals were captive: two were Total 8 3 23 39 65 923_ 515 401 125 vegetarian mammin-als (a mouffion. and a tapir. miiore comnion finding thani atheroseler osis in the coronarv arteries we investigated: it was ob'served in more than 25 per cent of the eaptivet Circulation.e.17 In this case. Lervia [Pallas]. two adult females of Colobus (Colobus auigolensis). In the little panida. Diffuse intimnal sclerosis is a much. Of the 11 African monhkeys. The muore conspicuous lesions of the chimupanzees were hunman-like atheromatous plaques with foam cells and cholesterol crystals. i. 4). panida). Ammotragus. by imitramural hemorrhage. Cholesterol crvstals were fouind onilv in the coronary lesions of the omnivorous animals (chimpanzees.846VATSESAE%JER. wTere quiite free froni this sterol. also. DELCOJR T 846 Table 5 Inatimeal Sclerosis in Primates Table 4 (o.. Volumte XXVI. The lesion-is of the vegetarian vertebrates. C. C. the thoracic aorta seemed to be affected sooner thain the abdomuinal portion. The aoirtie arch was rarelv affected alone. an oeclusive thromnbosis occurred in the anterior descending branieh and killed the animal. Ailurus fudgen1s [Cuv. Badius Ba dius Badius Badius Angolensis Angolensis Anigolensis Aiigoleiisis Sex Aortic atherosclerosis F F + F F Ml ++- Amoong free primates. free (lucks).o bus MonAeks No. a youngomale of Aseanius (Cercopithlascanius Lemuridae Callitrichidae aind Cebidae Cereopithecidne Pongidae A:t lIemnuridae (one Perodicticus Potto [Gmiiell cts With intimal sclerosis [Audebert]).]) the other three were vegetarian birds (a crownied crane. two had localized atherosclerotic lesions in the thoracie aorta: the lesions were larger anid more important in a young adult female than in a somewhat older adult muale. Such a dispersion of the lesions was observed in birds as well as in maiymals. November 1962 . Atherosclerosis of the nmouffloi 's coroniary tree was very slight anid consisted exclusively of snall sudanophilic spots in the in-tima s diffuse thickeninig. Pani Sch weitfurthii [Gigl]. however. Tapirus Inlldictus Desmarest). C. Neither in eaptive nor in free aniiials were aortie lesions complicated by caleification. anid one little panda.

wlL.962 Figure n Coronary thrombosis lin a 7-month-old :wild.50 to 0. whole mnateriail. amiong captilVe orl amllloln free specimnes (old )looCled.70 to 1. X . boar. very difficlLt to estimiiat e because the vari.. Triel. fibrolls thickeIing the iuitina in their eoroniary arteries. B. w-hiel appqwarently j-intensifiedI wvith age.Velopled on pire initimyial sclerosis. hinalll]Wds 1(i nearix 1 p rent ofdthfe bir'1ds.. Volatnoc XXVI. X 2.1- I - I. of coroarvy atherosclerosis couldl l)e diseoTered ini annualmIs shot in -freedoi.h. 1eigefrt: X 10. 5) :in thlis animial.e. (fig. the less vu-lnierable to itimntal sclerosis wvere ltlelenorous iidae. howvever. per ceniit for.t5. 0. Figure 4 NItherloserotbe plaque in th e ri ht Ioronaryr(Jrer/f of a tapir. WcVigert: A.60( vegetariani ande onnfiv-or(Ins. [The thr-ombuzs deIclopedi in the righlt coronary! ar-terq in spite of the absentce of atherosclerosis in the coronitary arilter-ial tree a1wl( in1 tlhe aorta. but also idividnal. il the ecaptiVe ver. The range is comiiprised betw-eeni 7.NATI'RIAL 11FTOIIY ( ti' AIIElIiOSQLEIlO8ITS 8 47 A . anmd chimpanzees often shiowved diffuLse intimlal selerosis.10. u ild boar. t lie clholesterol /plospliolipid iratio. to a less degree.teltrates of our11 innWterial.primates. anid the mo-s Auileraievwere thwe polgidae (table 5). 52) per ecuit of whici slhowv 1 Of t1mo0re Or less 1i1mportnIt.ations ini the vulnierabilitv to intimal sclerosis are niot only specific.vorouIs andlherbi-vorous aininals than a. This is. 0. 0. throimibosis de1.mong' earnruvannua. Biochemical Results Total Cholesterol anid Phospholipids Thle level of totail cholesterol is by nio neanis echaracteristie of the species. the coronarr-v arteries of deer.5 aIndi 250 nig.. I . roebuck. in spite of' the v\lerified absence of any lipid deposit eith-elr in the aorta or in t1e coroniary arterial tree. Of iiteiest iv as the d iseoverv of ani organizingo thiroimibus in the right eoronary artery of a 7-month-old boar that was hunted and((irculation. Norcmber Although1) nio sign. the freqnevey ()of liffinse intiual sclelSis se(1ems to be hlig. slolt. mnellys. carntivorIols.1 IT.r amlong01' omlnil. Lipid infiltration owas pres(-nt in the other coronary trunks of this animal. Amnong.:-N .9.56 to 0. orag-utans) are witlh mi-an the onily species of thie atfnim-l kiigrdomi in wic-leih dliftnse intfimal selerosis is a genleralize(d featuire antIi begins as ear ly as the first weeks of life.12). ehimpaizees alI grorillas (andl. indilviduil variationis maxlie great (table 6).genierie differences in. TI every case. .e are nio strikingy n.

0 ci m CAm c00i x 0 m cc a: ti. I I t ||||9 |~0)0) M . 0) S .00t r-.o o to - 4 - 01:. c7- C. 0) ct cc 0) C)tC Ci) CC (a)Ci Wc Ci) a)i Ci) a) 0) . 7z 9 C) a) I-c ° Mc F-° .H P C) ° (. o±++ I .0 CA cc 'c~ oo 0 0 06 05 0) to li-.0 M0) a) 0 Ci0) S-S 0 ).ci00 C Iccic c. m C t- (M Cl to :. t c 0 0 00 CIA '0 1. 1._ .tc0 c 'IO '4 00 CA cc l 0 cc in 00)5 0 ci d Ct c Ci 1.+ 0q C5 rY.H e. o- It 1o !I C) 1= V.44 0--) Ci-) 0 0 0) 0 0 *c. H C o )~~~~ C.L 1. C.vcci 00 00 00 Ci00 TiC cc 1:. Circulation.. 001..C ci ci cc r -uccicc 4) Q E-4 00ci V c%7 4- 4.o CA O Ii bo 1000 C) ' 10cc = m OCi t 0 1 ~co00 :. .- 0 ct ..0 '111 cc ci II II <=:s ci 0 10 + 010 cc in~ o d4 c9 cc ci co m tfO II ) MclL-C0 C 00 0In_ I( V II 10~ teo0 I.._14 0 I-1 0) t X Ct~ ~ ~ 0 0)U '0 t[ 0 ° 0) on 0) 0)S c-I Mc 0) P0 0a) 0 0 CH e-_ 1-c 0)q . PL t. November 1962 .r-.P: POrk "00) a)i5 t t .H .VASTESAEGER. 0 .-t lo 00000 f cc to -.(- P.P 0 Oi) 0 OO00 OOOO° 0 V 0) 0) 0 0 Ci 8 0 0) 50 . 1-i s z Ni t . Ci dl :) 0a) 0) 0)~~~~~~) 4 cc 0) 0) 002C 0 9. m +~o O 00 9 Pi. r-- (.l rH H r-- tO r-- t :-Cto 010 r 0 1.0 ! Ot- 0 0 * - )0)0 1 Cc O II C c3 !-4 OIcI Q cn 0 I i C~ I -t n CJ °i rl.. H p t Ic * P0)M~~~ c ct P pH ? c-i .Mc:.- 44 P.1:(M "i O C: It m o: C1 ~C r-.1.00'0 cc.0).0 It 1:- tc> cli m c-i .- OpwQCi c-i ci cA Q r-~- -- O c-.0. DELCOIRT 848 a) I.t.0 tQ 1:- 1 o Io0 ci t in.0 0. Volume XXVI. 1>C1 .8 hz )0SW X 4e 4 el zoP0.H P P P P.Cc. * t . toc cc ic to cim53) to to0 (Z00)000C Hcc C.4 A Co CA 0 ci r-i I o- 00 1:c. .- 0 0) I- 0) C2 7 0e 0 ^4-2 Cs 0)-4 0 o '0) C) 0) '.cli ci 1:ci m O~ 0 cli to ri0 c1-ti *H ] ccl 1Hr e 0 Oc ci r--r -- ccciIc000-CI0i. C) c...¢ 7- H. ct PL a) .H . m CQo ci = 0~ -)tz.C9 100C i It M N c1c 1t0 C11 CACA II Ii 001 0 0 H- cc cc M 00 00 q in.~40) .00Gc 1:r-- QQ.0- 1 0C.0 t. c 0 0) v: 00 LO O= O to ci ci. C) c c 00 0c: ci cl ci ci CJ tZ H r' cc CA CAi -4r~r....= + :--- 0)0) c 0= + = °°oo II Ca 0 0 C.) 0) a C) c) U Ci .C mf CA In C CA t~.m ~rCA q 1t 000 cli i" "r- cc 00 000 0 1:- ci 't oo 0) 0c iOO (= r 1.ccciq ci L10- cc iC C3lV 0O\l 101:-1.4 to I "t C itO~ Oi- 0o c O~ CA . rH ci ol C'0 00 Io IC -ItI I -ec M H= r- r-~rqC C'I cinao toIt. D.Mv *..

- CO +1 00 O -I -.L4X 4-. 0 4 0 V W W a ) O) a) PP.~~~~~~~~~~~:scd m 4 m.I- 00 0H CA 00 HC0 m t CO t CO to t- + tO o t O 00 t--00 O-- C C 100 C'1 0 ° C A t- r C 00C C0 OC] O ]0 x 00 +1 Lo00 to 'IC 0c t 00 00 A 00 O c1 c t1 00 C 00e CA C1 10 C O0C e s CA ]I CQ cJ00 to Uc I 00 ?- M) I 1o li +l 0 10 'OI O0: 03 C\ n H la I Cx1 CQ t- 0 t- to +1 +1 +1 C'1 CA 0- C\l .tH t V V ct a-.C 4 t6 C O~~~~~~~~~.: 0q U U ap U IZg a .~~~ *~~~~ o o " _t ~~~ () +1 *t t0 +1 0 0) tli CO CA $4.Cq -It t- -I C\] di t- v t- L- +1 +1 Cl 00 a) a..- +1 CO Co +1 c U +l t-l 00 41 + 00 +1 +1 a4~ 0) V CO tl C]) +1 10 +1 C] 0-q C] %0 o + C] 0 Cl +1 +1 es +1 10 +1 C occ a O~~~~~~~~~~~~~~~~~L Cc Cc cc cd cdc$c c. aa4 a O CO CO to to 0 O O E-4 0 0-f 00 *-) 1t M * * M1-* -a. 0d Circulation. 0Z CZ 3 c c O O *~~~~~~~~~~~~~~.I . a c.8-49 NATURAL HISTORY OF ATHEROSCLEROSIS Ct I to tl 0o° 1xp0.0 I I III I V I 0 0 +1 +1 C] +1 tl r- Co I + IC OR CO O O10 00 L- r- (O O 11I O= r-- r-- to m 00 C] O mCD t-1s x OX OOq b C) in CD 00 10 CO +1 cli +1 +1 vli C] 00 C] C] 1~0 O0 PC L 100C- O L. November 1962 .t. Volume XXVI.C) U U U U U U U U o V U 0V . .5 a 0 00 0d oa 4n a) C]i ¢ --i a) . 00 e ~~ ~ ~ ~ c ~ -l oq ~Jo CO In r-H " a cl r-- o mcs-q r-"s C0 r-- Co 0 +1 Hf +l +1 100 C] Co +1 to eC C> CO C\l 00 o00o 0 L- -1 t0 D CL tB s °° r r--i r-- r-- r-- 00 LO tL: t i C\] 10 00 C] s m CD O H 000 CCO C 00 CO u C] 1i co r_ 1 r- cll' es 00 01 -t 00 C] C) M 00 oo 0 - 0 "I C) 00 0 CCO 10 M CA CD C] L- t- 1 cl C> C] to1 t.. 0) C] C] 0*o-~j~j~ 0) cQQ Cd Q a) cq Cd~~~~~~~~~C ro a~~~~~~~~~~~~~~~~4 L qPiA 0 7 CO~~~~~~~Iz CO . a a aP.

Sakagami and Zilversmit. in captivity. (bottom) pholidota. t I CAAN. the beta peak becomes very prominent. (center) carnivora. In our experience. I. Arrow indicates: Left. Beta-lipoproteins are by this way selectively precipitated.L 'a. Using antihuman beta-lipoprotein serum.. DELCOURT -W C i ccct. the lipidogram seems to be related to evolution: the more evoluted the species.9 have shown that dextran sulfate is a suitable reagent to separate quantitatively the betalipoproteins of the dog serum. But even in human serum. Isolation and Characterization of Beta-Lipoproteins Burstein"5 described a method suitable for quantitative separation of beta-lipoproteins. Dextran sulfate of high molecular weight and a molar solution of caleium chloride are added to the serum. as is the level of cholesterol bound to beta-lipoproteins (fig. in freedom (straight line). there is a rather strict dependence between the precipitation of beta-lipoproteins and the molecular weight of dextran sulf ate. II. In lipidograms. the more the alpha-lipoprotein fraction becomes prominent (fig. when one tries to study lipoproteins in animals other than man. the total cholesterol is much higher. 6). Volume XXVI. As a rule. in freedom. November 1962 . The changes Circulation. as in human serum. Whole Lipoproteins The lipidogram pattern is more characteristic of the species. (bottonm) potto (lorisidae. Liptidographic pattern in primates.). Center.VOautL PRI tATE4 Figure 6 Evolution of the lipidogram in different groups of mammals. that captivity has a deleterious effect on the beta-lipoprotein levels. But absolute levels were quite dispersed. however. in cold-blooded vertebrates as well as in mammals. either by lipidograms or by chemical determinations. Generally speaking. Figure 7 Lipidograms of a wild boar: I.18 The procedure of Burstein remains questionable. the percentage of cholesterol bound to beta-lipoproteins in wild animals was not very different from the percentage of this fraction as demonstrated by lipidogram. Lipidographic pattern in baboons. table 7). (center) galago (ga'lagidae%. Lipoproteins and Captivity It can be seen. However. 7. Lipidographic pattern in (top) ungulates. the precipitation is not linked with any antigenic property of lipoproteins. Lipidograms confirm that the beta-lipoproteins are correctly separated by this method. Extensive studies should be done on many individuals in each species before one may even attempt to systematize the biochemical results and the specific trend toward atherosclerosis (table 7). we found fractions having a common antigenicity with human fraction only in monkeys and in some antelopes (Cephalophus monticola [Thunb] ). in captivity (dotted line). Right.85Q'' VASTESAEGER. (top) cercopithecidae.

there are still wild specimens living in places almost inaccessible to man and having only brief and incidental contacts with him. the lesions we observed in . As far as warm-blooded animals are concerned.6 Freedom Captivity. as well as with those of Van Zyl and Kerrick' captive specimens of Papio ursinus.62 Captivity. in the phospholipids. captive and in their wild state. that have kept to strictly natural foodstuffs and to a natural way of life. Europe 127 204 211 52 75 78 125 92 119 60 61 57 160 118 42 256 73 189 202 73 169 . either by domestication or by captivity. and stage of development are concerned. Europe 71 51 114 104 193 139 20 10 54 28 9 27 176 156 144 84 119 100 92 37 44 4. Of course the domestic ones have been deeply modified in their morphology as well as in their physiology by the artificial selection that man has brought to bear on them.95 2. sex.08 175 26 149 85 214 100 114 Captivity. Discussion Until now.77 . Total a 18 13 Cholesterol Freedom Captivity. If such conditions still exist. Nevertheless.08 . and also of the multiplicity and power of the commanding means of man over nature.62 . Europe Captivity. are less pronounced. Congo 145 110 35 220 97 133 24 60 256 218 243 160 38 83 1. In order to establish the influence of captivity on the genesis and evolution of arteriosclerosis. it is difficult to conceive that there could still Circulation. they are realized in the sea: the discovery of intimal sclerosis and of atherosclerosis in tunny fishes caught in their natural habitat brings proof that the two processes are not a consequence of captivity in the case of coldblooded vertebrates. our figures are in agreement with those of McGill and found in Kenya baboons living in freedom.81 . most investigations concerning spontaneous degenerative processes have been carried out either on domestic animals or on captive wild animals. Volume XXVI. In order to answer this question.8 Phospholipids %. Europe Captivity. Congo 144 Freedom 4 Freedom 157 58 99 63 212 118 94 170 113 57 33 5 Freedom 272 146 126 70 55 16 27 6 160 73 87 Freedom *Tn spite of slight specific differences. one should be able to compare under rigorous conditions the arterial state of animals of the same species.46 1.7 . Congo 142 20 122 87 158 82 76 Captivity. First of all.56 1. sometimes during thousands of years. All these conditions are far from being fulfilled in our material: this is why we cannot draw any firm conclusion concerning the influence of captivity on the incidence and evolution of intimal sclerosis and atherosclerosis in birds and mammals. there still remains the difficulty of procuring individuals of the same race that are comparable as far as age. has favored the birth and the development of the arterial degenerative processes observed in the examined animals. either total or bound to beta-lipoproteins.2 5.4 co-workers'0 obtained in exist animals that entirely escape human influence.851 NATURAL HISTORY OF ATHEROSCLEROSIS Table 7 Cholesterol and Phospholipid Levels and Ratios Status Species Wild boar (Sus searofa) I1 2 3 Pangolins (Manis tricuspis) 1 2 Mangabeys (Cereocebus ateriimus) 1 2 3 Baboons* (Papio doguera) 1 2 3 Total Cholesterol a 18 Phospholipids . Congo Capitivity. One must then ask if it is not human influence that. November 1962 3. in view of the density of human population and its dispersion over all the lands of the globe.

the criteria of these authors are quite different from ours: their practice has been "to ignore isolated plaques of the aorta and larger branches. that the lesions are mainly distributed in the aorta. the animals remain in their natural climate. our observations confirm those of Fox dealing with the widely separated varieties of the affected vertebrates. at first made by Fox. and diet. our biochemical results bring accumulated evidence that captivity may significantly alter some aspects of the lipidic metabolism in animals. In the first case their natural feeding grounds are little disturbed. Two kinds of captivity are to be considered: in the first one. it was among primates that we found the highest percentage of aortic lesions. nearly 30 per cent of the examined aortas were affected. per cent. Even in freedom. In any case. Captive animals are not normal. however. an animal presumed normal is not necessarily normal. Volume XXVI. sex.22 Table 8 summarizes the results of this study. Such a discrepancy can of course be explained by the. From the comparison of sections made in the same coronary trunk of young captive chimpanzees and of free chimpanzees nearly of the same age. November 196. the following points should be emphasized. between 1931 and 1956. Ratcliffe and Cronin7 observed an inereasing frequency of arteriosclerosis in captive wild animals. This was observed not only in European suidae and carnivora. Our results disagree. They also confirm the statement. in colobi. neither intimal sclerosis nor atherosclerosis is a mere consequence of captivity.t . not far away from their original habitat. independent of age." in ours. we got the impression that the importance of individual variations in the stage of intimal sclerosis in this species may occasionally surpass the influence of captivity on this process. Different local conditions of diet are probably not the only factors responsible for the discrepaney between Fox's figures and ours: in the same Zoo of Philadelphia. 1. Specimens shot or trapped in their habitat are usually disabled or ill. or length of captivity could be established. but also in African ungulates and primates. the animals are sent to zoos in far distant countries. the great majority of the involved specimens showing this distribution alone. Our biochemical results are also rather disappointing for the study of the natural history of atherosclerosis. 2. the mean value being 250 mg. The total blood cholesterol ranged between 120 and 470 mg. with those of Fox concerning frequency and specific incidence of atherosclerosis: in Fox 's material "lipidic deposits were exceedingly rare. No relationship between cholesterol level and age. and in chimpanzees living in freedom bring evidence that.852 deer. kidney or Circulation. unless they were accompanied by vascular disease of the heart. at least in ruminants and in primates. On the other hand. in Fox's material. difference of conditions of captivity in the zoos of Antwerp and Philadelphia. This should be kept in mind when one seeks suitable material for experimental atherosclerosis. As compared with the values recorded in freedom. In all specimens the cholesterol VASTESAEGER. In any case. As far as comparative arterial pathology is concerned. both types of captivity significantly alter lipoproteins as well as cholesterol levels. As a matter of fact. primates were less vulnerable than carnivora and ungulates to degenerative arterial disease: on the contrary. The results of our investigations are very difficult to compare with the figures published by Rateliffe and Cronin. in the second one. DELCOURT fraction bound to the beta-lipoproteins was quite large. This deleterious effect of captivity is impressively confirmed by a previously published study of 60 young chimpanzees living for many months at the medical Laboratory of Stanleyville. per cent. unfortunately the influence of parasites on the lipoproteins pattern is still unknown. we had the opportunity to see how severely tropical animals are infested with parasites (table 6). but in the second case diet and environmental conditions are quite different. the healthy and fit ones escaping more easily. Before the results are discussed.

bat thte jatiima is moloeiEately thickened. 8. Volutme XXVI. btut it is firllliy established tihat atherosclerosis may . atically importanit fibrous changtes of the coronary iintima before puberty. 8. tlhey imla-de 110 elihe(late effort to separate atherosclerosis from otler formus of arterioselerosis. which died of old age (fig.her in the prima-tes thani in the ungrulates. in its hIman incidence diffuise intinial selerosis appears as a generalize(1 process. Obviously. AAWhereas intimal seletosis is foullnd.g cenera like cercopithecidae anid al)es. tley are qutiite sufficienit to establishl that not onilv the ineidenee of diffuse intimal sclerosis but also the age at which it appears vary eonisiderablv from one speeies to another. But7 as far as the incidence of spontaneous atheroselerosis is concerned. show hardly ml-ore advaniced lesions than those of a 5-year-old gorilla (fig. -Although the facts brought about by our11 iinvYesticgations are very fragmentary. The frequeney of atheroselerosis is hig. To what extent alterations of lipidi netabolism facilitate spontaneous atherosilerosis ini thie fibirous initiina is still unikniowin.' Furithermiore.853 NATURAL IIISTORtY OF ATIJEROSULEROSIS sptleen. bottom) proves undeniably the existence of specific differences in the precocity of the fibrous thickeniing of the coroaiavy intima. especially if the variable life span of the different species is taken inito account. w-e have been struck by its greater frequlency amiong certamn species stueh as tree duekls (three of five aninnials had eoronary atherosclerosis) and amnono. spontaneous atherosecierosis always develops in arteries alreadly impaired by iintimal selerosis. We could multiply suci examiples Cand show that of all mrlammals the anthropoid apes and mrlanl are the only ones to present systemnCirculation. beginnling as soonl as the first Aveeks of life in white as wcll as in Bantu people?3 Such universal preeoeity among h-i 111811 beiniis gave manv autlhors the imnpressioni that diffuise intinual selerosis could be a physiologic proeess. Bottoml. Tfhe internal elastic membrane is as thick as that in the wart-hog. too small to allowv even a ronghi estimiiate of the frequenev of both intiinal sclerosis and atheroselerosis in eaeli speeies of vertebrates. The larger number of primnates we could investigfate gave us the impression that in this or(ler at least the frequency and the precocity of initiial fibrosis increase with the degree of evolution (table 5). in all voungr human aduilts. Anterior desceniding branch of a yloun ignInmature fecna7 gorilla. In ouir experienee. and hiigher in the ungulates than in the carnivora. of course. In otlher vwords. top). Nevertheless. November 1962 Figure 8 '1'oj. iintimal sclerosis appears as a basic prelimilmary lCondition for spontanieous atherogenesis. The numiber of our specimenes is. the faet that the coronary arteries of a captive wild pig. On tile othe-r lhand. this is far fi-oun the ease in other species. Anterior descending branch of an o1(1 cO]pti wart-hog. the fact that we cannot know the real agfe of captive animals makes any kind of precision difficult. growingf progressively with advancingf agre. lVeigert X 40. The internal elastic ne mbran 1 iv stretched and ruptured.

in freedom as well as in captivity (deer. total cholesterol. 1958. LOBSTEIN. beta-cholesterol and cholesterol/phospholipid ratio were also determined. (T. DAUBER. camel. 10. tapir). H. Circulation 21: 730. Arch. eamel. LINDSAY. November 196.. in captive birds whose dietary fats are mostly unsaturated (crane. 1960. D. pelican) .: Arteriosclerosis. in captive birds whose dietary fats are mostly unsaturated (crane.: Changes in the character and location of arterial lesions in mammals and birds in the Philadelphia Zoological Garden. reinforced by specific and individual differences in enzymatic activity of lipid metabolism.: In Cowdry. E. 3. 1. physical activity. 6. 153. World Health Organization: Rapport d 'un groupe d 'etude: Classification des lesions d 'atheroselerose. Philadelphia. BRAGDON. 1904. AND CRONIN. References 1. and intimal sclerosis seems to be a basic condition for spontaneous atherogenesis.: Spontaneous atherosclerosis in the rabbit. In this material. Summary In order to complete the fragmentary knowledge of the natural history of atherosclerosis. J. G. 53: 281. 1903.e. 9. p. J. path. S. 7. Med.: Disease in Captive Wild Mammals and Birds. Volume XXVI. Lippincott Company. H. YERASIMIDEs. H. 21. Circulation.. New York. Circulation 18: 41. inn. lipidogram. I. C..: Arteriosclerosis in the dog: Spontaneous lesions in the aorta and the coronary arteries. and especially the beta-lipoprotein levels as well as the beta-cholesterol fraction. 11. Spontaneous atherosclerosis may develop in fishes living in their natural habitat. Kong. 1958. Levrault. Paris.2 . The Macmillan Company. It was observed in any case that captivity significantly alters some aspects of the lipidic metabolismi in animals. The discovery of coronary atherosclerosis in free tunny fishes. AND GILMORE. H. H. Unfortunately. f. Fox.: Traite d 'anatomie pathologique. 5. F. L. AND ELLIOTT..: uber Arterioskierose (AtheroSklerose). These facts indicate that there is no absolute and uniform relationship-i. CHAIKoFF. 1833. brings proof that atheroselerosis is neither VASTESAEGER. 1933. 1944. In 88 of the captive animals and in 51 of the free individuals. Virchows Arch. 1923. in wild mammals whose diet is free of animal fats. V. F. A. 38: 46. M6NCKEBERG. Captivity or way of life (diet. RATCLIFFE. G. tree duck. MARCHAND. applicable to every vertebrate species and to every individual in a given species between the origin or the chemical nature of dietary fats and the development of atherosclerosis. Circulation 5: 641. the authors examined the coronary arteries of 403 captive and 146 wild vertebrates living in freedom. 1952. F. Path. as well as the aortas of 134 of them. RATCLIFFE. 1952. DELCOURT the privilege of warm-blooded vertebrates nor a consequence of captivity. Arch. and stress) are further variable elements in the problem of atherogenesis in wild animals. caught in the Bay of Biscay. the material collected does nlot permit even a rough estimate of the influence of captivity on the development of atherosclerosis. Verhandl.: Changing frequency of arteriosclerosis in mammals and birds at the Philadelphia Zoological Garden. L. M..854 spontaneously develop in animals having quite different diets. Path. Geneve. a Survey of the Problem. J. pelican) . L..ber die reine Mediaverkalkung der Extreiitiitenarterien urnd ihr Verhalten zur Arteriosklerose. tree duck. L. G. 8. 21: 23. 171: 141. T. G. spontaileous atherosclerosis always developed in arteries already impaired by intimal selerosis. Lipid deposits were found in the intima of 29 per cent of the examined aortas. B. and especially in fishes feeding on a diet rich in unsaturated fats. J. but coronary atherosclerosis was present in only 2 per cent of the captive specimens. J. Palais des Nations. feeding on a diet rich in unsaturated fats. tapir). yield a better explanation of the great variability in the incidence of atheroselerosis in different species and in different individuals of the same species.: Spontaneous arteriosclerosis in chickens. T. Fox. and in wild mammals whose diet is free of animal fats. in freedom as well as in captivity (deer. Inbred specific and individual differences in vulnerability to intimal sclerosis. 2. V. Anat. 4.: t.

21. they must have ideas which they submit to the control of facts. 18.: Degenerative changes in the cardiovascular system of the spawning Pacific salmon (Oncorhynchus tshawytscha). Science 98: 19. 22. P. GILLOT. DELCOURT.: A biochemical study of spontaneous atherosclerosis in pigeons. R. Men of science who mean to embrace the principles of the experimental method as a whole. P. roy. AND MILLER. CHAIKOFP. 17.. 1955. 14. H.. R. South African J. R. 20: 97. 1961. M. G. dextran sulfate precipitation and paper electrophoresis. they must be at once observers and experimenters. Circukation. must fulfill two classes of conditions and must possess two qualities of mind which are indispensable if they are to reach their goal and succeed in the discovery of truth. H. 1960. I. L..: Serum lipids and age in the baboon (Papio ursinus). HAHN. AND RASSON. 23. November 1962 . Zool. Anvers 14: 47. H. P. 21. AND DELCOURT.: Abolishment of alimentary lipemia following injection of heparin. Circulation Research 8: 670. J.: Separation of dog serum lipoproteins by ultracentrifugation. Path.. Bull. STRONG. WEXLER. M. GILLOT. LINDSAY. I. C. P.-CLJAUDE BERNARD. H. VASTESAEGER.: Les extrapolations cliniques des statistiques biologiques. 2eme Colloque International sur la morbidite coronarienne dans le monde: Rome. E. 1943. 40: 29. F. Arch. LOPLAND.. M. J. ZANCHI. 1960. Soc. et biol. Resultats d'une enquete sur la cholesterolemie.. 1927. but at thie same time they must make sure that the facts which serve as starting point or as control for the idea are correct and well established... T. 16. VASTESAEGER. T. T. J. F. Path. 15: 26. AND KERRICK. B. p.: Spontaneous atherosclerosis and diet in captive animals.: Sur une nouvelle m6thode de dosage des lipoprotienes du serum. SAKAGAMI. B. MCGILL. AND GILMORE. J. 13. 1961. A. S.: Arterial lesions in Kenya baboon. AND WERTHESSEN.: Thrombose coronarienne ayant entraine la mort chez un panda (Ailurus fulgens). HOLMAN. First. 19. Acta dlin. The Macmillan Company.855 NATURAL HISTORY OF ATHEROSCLEROSIS 12. belg. 1958. D.. B. AND MORTELMANS. Volume XXVI. B. Sc. AND ZILVERSMIT.. Nutritio et dieta 3: 174. D. 1959. 0. Lipid Research 2: 271. 1962.. H. ROBERTSON. 20. An Introduction to the Study of Experimental Medicine.. M. L. VAN ZYL. New York. L. 6: 541.. AND CLARKSON. BURSTEIN. 1959. N. AI. 1960. 15. 1955.. Circulation 7: 234. Circulation Research 9: 826.: Morphologie des arteres coronaires dans la premiere enfance chez le blanc et le bantou. 1961. Cardiologia Pratica 13: 147.. C.: Arteriosclerosis in the cat: Naturally occurring lesions in the aorta and the coronary arteries. B.