Talk:Paper - On the time of the post-natal obliteration of the fetal blood-passages

From Embryology

On The Time Of The Post-Natal Obliteration Of The Fetal Blood-Passages (Foramen Ovale, Ductus Arteriosus, Ductus Venosus)

Richard E. Scammon And Edgar H. Norris Institute of Anatomy, University Of Minnesota

Three Plates

It is gcnorally rocognized that two separate processes are involved in the occlusion of the fetal blood passages after birth. The first is the simple functional closure which takes place, in the great majority of cases, at or immediately following birth; the second is the permanent anatomic obliterati(jn which occurs at a later period. The mechanics and histology of the latter process are generally discussed in our larger treatises on obstetrics and pediatrics, and in our major an.atoTnical texts; but the thiie of postnatal obliteration is often unmentioned, although in a number the statement is made that the obliteration takes place in the first few days or, at most, weeks after birth.

The origin of the current concepts as expressed in these larger texts can be traced, we think, to the first statistical study on this subject, which was pubUshed ninety years ago by the French clinician Billard. This investigator (H)Ilected data on the obliteration of the ductus veno.sus, ductus arteriosus, and foramen ovale in a series of one hundred and twenty-eight children who died in the first eight days of life. He found instances of the obliteration of ail these pas.sjiges on the first tlay after birth. The foramen ovale and ductus arteriosus were closed in over fifty per cent of his cases on the eighth day while the ductus venosus was clo.sed in a still larger* immber. He therefore concluded that the obliteration of the fetal blood-passages proce<'ded very rapid!}- in the first few days of life- an opinion in accord with that held by a niimbor of wTiters in the eighteenth century. The n^sults of Billard's study were published in his 'Traits des maladies des enfants nouveau-n6s' in 1S2S. This work was extremely popular in its time; it passed through a number of French editions, wa.s translated into Italian and German and twice app<>jired in .Vmerican editions. In several publications of the middl.' of the last century Billard's figures are cited and his name is quoted in connection with them. In later works, howevei", the same opinion has been repeatedly expressed but its sourc(^ apparently has been forgotten.

A large amount of data concerning the chronology of the postnatal obliteration of the fetal blood-passages has accumulated since the time of Billard. This material is scattered through the periodical literature of legal medicine, obstetrics, and pediatrics and is also uieluded in a number of rather inaccessible brochures. In the following pages we have collected as much as possible of these scattered data and have arranged them in tabular and graphic fonii. In so doing we have confined oiu-selves to those records in which series of consecutive cases have been assembled. These records include ob.serA^ations on children who were born at term, and also a few cases of children who were prematiu-ely born. In most instances, however, investigators have failed to separate these two groups. A comparison of the records of the few known cases of prematurity with those of children known to be born at term shows no appreciable difference in the time of obliteration of the fetal blood-passages. We hav(> therefore combined them in our tables. In all cases the patency of the fetal passages was te.sted either by injection or by probing, excepting those of Faber ('09) which were examined micro-scopically.

The results made apparent by this combined series of observations are at variance with the current concepts on the subject as expressed in most of our larger texts, and also with the results of Billard. Billard's observations were confirmed by Bernutz ('65) who found the ductus arteriosus clo.sed in fourteen cases in a series of twenty-one children who died between the tenth and twentieth daj's, and in thirty-six out of thirty-eight children dying between the twentieth and sixtieth days. Since this time


OBLITERATION OF FETAL BLOOD-PASSAGES 167

no observer has substunfiatod these findings. .-ilthouRh a number of series niueli larger than those of Billard and Bernutz' have been collected. Thus Elsasser ('52), in a series of nearly three hundred observations upon children of the first month, found obliteration of the ductus arteriosus in about two per cent and of the foramen ovale in about three per cent of his cases; and Alvan^nga ('69) found practical!}' no instances of obliteration of the foramen ovale or ductus art^-riosus before sixty days. The findings of later obseivers (Alexeieff ('00), Theremin ('87-'95), KuchefT ('01) and others) agree essentially with those of Elsasser and Aharenga although they have noted some instances of earlier obliteration of the pas.-^ages.^

THE FOR.\MEN OVALE

The compiled data on the obliteration of the foramen ovale are shown in extenso in table 1. Table 2 is a summary of these data giving bj^ periods the total number of observations and the number and per cent of cases obliterated. Graphic representations of these data are shown in figure 1 curve A. and in figure 2. In the summary and curve the data of Billard are omitted because his findings are so directly opposed to those of all other investigators that we conclude that either his method of investigation was defective or that his definition of obliteration was entirely different.

As will be seen in table 2. less than one per cent of the foramina are completely closed in the first week of life, and less than two and one-half per cent in the second week. In the latter part of the first month the figures indicate that the obUterative process takes place more rapidly as the opening is impervious in about one-eighth of all cases of this period. The rapiditj' of this process mcreases during the second month, for the interatrial

' The cases reported by Bernutz were not examined by him personally but were oollected iit his instance by the interne of a colleague in the Hospice des EnfantsTrouv<5s.

' Haberda ('96) studied the obliteration of the ductus venosus and ductus arteriosus in a considerable series of infants and children. As his data are not given in numerical form we are unable to include them in our summary. The general statements of this writer indicate that his findings were somewhat similar, as regards these two vessels, to our own.


168


RICHARD E. SCAMMON AND EDGAR U. NORRIS


TABLE 1

Data on the obliteration of the Foramen ovale. Numerals enclosed in parentheses indicate number of obliterated cases


ornanTU AXD

DAT*


r

i


< a


5

o


i


9


1!

•4 O


5

a


1


a


O

at


o a


5


3



S


3:



o


s


f*


i<


i!


H


H


i


O


8


S


g


i!


^s




«o

7


6


11


21


25


46


68


15


3


2


1


1



Alexeieff. '00.


3









(1)


(3)


(6)


(3)



(2)


(1)


(1)



Alvarenga,
















'67


44


19


17


33


28


12


23

(2)


18

(1)


1


4


12

(4)



2

(1)



Billard, '28...


118 (17)


20

(11)














British Anat.
















Soc, '98...












23

(11)


9

(7)


30

(23)


286 (217)


Bitot, '37...



34


63














(23)


(49)


EUasser, '52.


150

(1)


63 (4)


62 (9)


23

(2)


3

(1)











Fawcett














6

(3)


209

(207)


Letourneau,
















'58


8

(1)


1














Ogle, '57















62 (49)


Theremin '87


70


76


63


49


55


45


81


65


11


7







18


5


(10) 4


(13) 6


(17)


(18) 4


(47) 8


(51) 6


(9)

1


(7)


4





Theremin '95


6







(1)



(2)


(3)


(6)


(6)


(4)





Wallraann,
















'59












2


o


4


291 (170)


Zahn


293


171


152


121


107


86


158


157


27


20


43


12


42


711



(552)


Totals


1712



(2)


(4)


(19)


(16)


(18)


(21)


(55)


(64)


(12)


(11)


(21)


(8)


(28)


(1228)


• 1 to 15 years t 7 to 10 months.


OBLITERATION OF FETAL BLOOD-PASSAGES


169


TABLE 2 Obliteration oj the Foramen ovale (SB58 cases)


Birth to 8 days...

8 to I.'} (lays.. . .

15 to 32 days

32 to 46 days

46 to 61 days

61 to 91 days

3 to 6 months.. 6 to 9 months. .

9 to 12 months. . 1 to 5 years . . . . 1 fo 15 years. . . . 5 to 10 years ....

10 to 20 years .... 15 years and over 20 years and over


TOTAL


HOHBIB or


FEB CZNT or


HmiBBB or


CABCa


CAIIU


ctam


OBUTBRATED


OBLTTBnATBD


293


2


0.7


171


4


2.3


273


35


12.8


107


18


16.8


86


21


24.8


158


55


34.8


157


64


40.7


27


12


44.4


20


11


55.0


43


21


50.0


93


52


55.9


12


8


66.6


42


28


66.6


1840


1320


71.7


1712


1228


71.7


comnumication is oblitoratcd in approxiinatelj^ one-.sixth of all cases in the first half of this poriod and in about one-fourth in the latter half. During the third month somewhat less than ten per cent of the cases are obliterated so that by the end of the first trimester the foramen or {do is finally closed in about onethird of all cases. After the end of the third month the process again goes on more slowly and the average of obliteration in the second trimester is about forty per cent, that in the third trimester about forty-five per cent and that in the fourth trimester about fifty-five per cent. Our figures for the period between one and five years show an average obliteration of fiftj-five per cent, which is five per cent less than that of the last trimester of the first year. This difference is due. no doubt, to the small number of cases tabulated in these two periods, and does not represent a real increase in the number of patent foramina. The figures of the .second five year interval indicate that twothirds of the foramina are clo.sed: but the small amount of data for the period between the first and tenth years makes it impossible to say with certainty just when this increase in obliteration


170 RICHARD E. 8CAMMON AND EDGAR H. NORRIS

is bntuphf about. In the srcond drrado the percontago of open foramina is the same as in the period l)et\veen five and ten years and tlie foramen ovale is found to be obliterated in about seventytwo per cent of individuals of twenty years and over.

Curve A of hgure 1 expresses graph ieallj' the frequency of obliteration of the foramen ovale during different periods in the first year of life. This curve is readily di\'isible into three parts. Tlie first portion, which extends from birth to the middle of the


" ' >• Ol »3 W go 120 IBO JTO 360.1^

Fig. 1 Three curves representing the average percentages of obliterated fetal blood-passages at different periods in the first year of life. A, dotted line, foramen ovale, B, solid line, ductus arteriosus; C, broken line, ductus venosus. These curves are based upon the material summarized in tables 2, 4 and 5.

second week, is a short segment expressing the obliteration of a Uttle over two per cent of the ca.ses. It is followed by a longer segment rising abruptly and terminating in about the middle of the third month. Nearly half of the ca.ses which are finally closed are obliterated in the period represented by this segment (sixtj- days), l^he third segment, extending from about the middle of the third month to the end of the J'ear, shows a very slow but continuous rise and expresses the obhteration of about ten per cent of the total number of cases.


OBLITERATION OF FETAL BLOOD-PASSAGES 171

Figure 2 is a curve illustrating the froquoncy of tho oblitfration of the foraiiu'n ovale throughout life. The details of the obliteration during the first year, which have just been described, are masked in this figure by the diminution of the time unit. Here again three periods can be recognized which correspond roughly to infancy, childhood, and adolescence and maturity.





TO

%




60


^^^■-^ lS-80y«»rsTl.T


SO


-r^'^



40




SO




n




10





I.I 1 1 1 1



1 i to LSyur*

Fig. 2 .\ curve showing the average percentages of cases of oblit«rated foramen ovale at different periods throughout life. Based upon the material summarizcd in table 2.


During the first period the curve rises with extreme abruptness to a point at which fifty per cent of the cases are obliterated. The ri.-^e is continued but is much less rapid during the second period which extends from infancy well into childhood. In the third period, which extends to extreme old age, the curve rises very slowly to about seventj'-two per cent. In all probability this final percentage of obliteration is reached in early maturity although the character of our data does not permit a graphic representation of this point.


172


RICHARD K. SCAMMON AND EDGAR H. NORRIS


THE DUCTUS ARTERIOSUS

Tabic 3 shows tho compiled ihxta. upon the post-uutal obliteration of the ductus arteriosus, and table 4 summarizes these data by periods. The graphic presentation of this material is shown

TABLE 3

Data on the frequency of post-nalal oblileralion of the Ductus arteriosus. NumeraU enclosed in parentheses indicate number of obliterated cases


OMBBTBB AKC DAT!


Alverenga, '69 .

Bernutz, '65 — Billard, '28... Elaaaser, '52....

Faber, '12

Gerard, '00

Kucheff, '01...

Letourneau, '58 Theremin, '87..

Theremin '9.t Totals


19


118

(16)

150

(1)


52


12 67

II


429 (17)


11


« 


5


P


<


<



a



fi











O


!•


H


t*


(O


o


« 



N


CO


15


28

(1)


17



1

66


168

(14)


2

(2)


61

(6) 4 (2)


143 (18)


8 (1)


23

(1)


56

(8) 6 (3)


117 (13)


38 (36)


2

(2)


2

(2)


40

(18)


50 (26)


75

(28)


43

(23) 4 (1)


57 (27)


14

(5)


9

(3)


15

(14) 2 (2)


68

(55) 8 (8)


92 (70)


53

(48)

1

(1)


63

(52)


6

(6)

16

(12)


54 (54)

12 (12)


89 (84)


7

(7)


23

(18)

13

(13)


4

(4)


47 (33)


• 10 to 20 days.

•• 21 to 60 days.

t 2nd month (data for other periods not available).

■' to 30 days.


OBLITERATION OF FETAL BLOOD-PASSAGES


173


TABLE i Obliteration of the Ductus arteriosus (1095 cases)


Birth to 8 days 8 to 15 days. .. 15 to 22 days... 22 to 32 days... 32 to 46 days... 46 to 61 days... 61 to 91 days... 91 to 120 days.. 120 to 365 days.


10TAI.

KUHBIB or

CUM


311

148 143 117 75 57 92 63


HUHBIB or

CAIES OBLtTSBATBO


1 3

16 13 28 27 70 52 84


ram csirr or

CABIS OBLITUATSO


0.3 2.0 11.2 11.1 37.3 47.4 76.0 82.5 94.5


in figuro 1, curve B. As in the ca.se of (he foramen ovale, and for the same reasons, we have omitted Billard's data and also those of Bernutz from the summarized table and from the curve.

During the first week of life the percentage of obliteration of the ductus arterio.sus is even less than that of the foramen ovale (three-tenths of one per cent). In the second week the percentage rises to an average of two and in the third and fourth weeks to an average of a little over eleven. From this tune on to the end of the third month the process of obliteration is e.\tremely rapid; in the first part of the second month it averages over thirty-seven per cent, in the latter part over forty-seven, and in the third month seventy-six per cent. During the fourth month the average obliteration is eighty-two per cent. Thereafter the percentage increases quite slowly until the end of the year. The a\erage percentage of obliteration in the last three-quarters of the first year is nearly ninetj'-five.

The data a\ailable for the period after one year are small in amount and, with the exception of certain instances in Faber's series, all cases of this period were obliterated. It is quite possible that Faber's material included several specimens which, while containing remnants of the original lumen, were obliterated at other points. His method of examination might easily classify such cases iis patent. It is probable, therefore, that table 3 shows a much lower per cent of obliteration for this period than is actually the case.


174


RICHARD E. SCAMMON AND EDGAR H. XORRIS


Curve B of figure 1 is the graphic presentation of the data summarizes above. Like the curve for the foramen ovale already doserihod and repr(>s('ntrd in the .same figure, thi.s graph is reaciily divisible into three portion,s. The first segment extends from birth to the middle of the second week and shows a temunal obliteration of two per cent of the cases. The second .segment ri.ses abruptly, cros,ses that of the foramen ovale, and teniiiimted at about .seventy-five per cent in the middle of the third month. The third segment, which extends from the middle of the third month to the end of the first year, shows a continuously decreasing rate of obliteration and terminates at nearly one hundred per cent. Probably all normal cases are closed shortly after the first year, although there are numerous records of individual cases of the anomalous persistence of the lumen of the ductus arteriosus in later life.'

THE DUCTUS VENOSUS


The material compiled upon the obliteration of the ductus venosus is shown in table 5 and is represented graphically in curve A of figure 1.


TABLE S


Obliteration of the Ducltis venosus (,76i


cases)



Aoa


TOTAL IfDHBBB OF

oiaas


KDUBBB or

cASn

OBLRBRATBD


FIB CENT OF

CASM OBUTBKATBD


Birth to 8 dara


211

150

169

103

37

20

41

19

12


6

27 64 78 36 20 41 19 12


2.3


8 to 15 dava


18.0


IS to 22 dayK


37.5


22 to 32 days


75.7


Second month


97.3


Third month


100.0


Fourth, tifth, and sixth months


100.0 100.0



100.0




• For the literature on the subject of persistent patency of the ductus arteriosus the reader is referred to the papers of Poynter ('16), Wells ('08) and Gerard ('00').


OBLITERATION OF FETAL BLOOD-PASSAGES 175

The procrss of oblilcnitidn is much more rapid in the ductus venosus than in the other fetal passiiges. During the first week the average is two and three-tenths per cent, in the second week it is eighteen per cent, in the third thirty-seven and one-half per cent, and in the last ten days of the first month about seventysix per cent: During the second month the percentage rises to nearly one hundred and thereafter all cases are obliterated.

The curve shown in figure I, while nuich more abrupt than that of the foramen ovale and th(> ductus arteriosus, is of the same general character and shows three segments. The first segment corresponds to the first few days after birth, and terminates between two and three per cent. The second segment rises with extreme abruptness to ninety-seven per cent in the middle of the second month. The third .segment, which is very short, rises gradually to a full hundred per cent by the end of this month.

THE HATE OF THE OBLITERATIVE PROCESS IN THE SEVER.\L

PASSAGES

In t)rder to study the actinty of the obliterative process in the various fetal blood-passages we have calculated from the graphs shown in figure 1 the average daily rate of obUteration for a series of periods in earlj' life. This was done by determining from the graphs the initial and terminal percentages of obliteration for each given period. The initial percentage was then subtracted from the terminal one and the figure thus obtained divided by the number of dajs in the period. For example, in the case of the ductus arteriosus the percentage of obliteration at the beginning of the second month was fift>--seven and at the close wjvs seventy-nine and one-half. Thus twenty-two and one-half per cent of all cases were obUterated in this period of thirtA' days and the average daily rate of obliteration was seventy-five hundredths per cent. Table (5 shows the results of these calculations for the three passages and the curves in figure 3 express them graphically.

It will be seen by the examination of these curves that they have certain characters in common. Each starts with a low rate


176


RICHARD E. 8CAMMON AND EDGAR H. NORRIS


TABLE t


Approximate rate of daily and f


obliteration of the Ductus venosut, Ductus arteriosus, 'oramen ovale in early childhood


FSRIOD


ODCTDB VEHOSnS


DUCTCS

ARTiBioem


roBAuaN

OTALI



0.33

3.00

0.60






0.04 0.92 1.00 0.75 0.07 0.04


0.10


1 week to 1 month


0.60



0.45



0.40


3 to 6 months


0.07


6 to 12 months


0.04


1 to 5 vears


0.005




of obliteration, rises rapidly to a peak or niaxinium, and then declines to the base-line which represents the cessation of obliterative activity. In all cases the portion of the curve representing the decline in activity is less abrupt than is the initial rise.

Considering the curves individually, it will be noted that in the case of the ductus venosus both the initial and maximal rate of obUteration is much greater than that of the other two passages and that consequently the entire obliterative process is completed much sooner. The curve expressing the rate of obliteration of the ductus arteriosus is very similjir to that of the ductus venosus although the initial and maximal rates are lower and the apex of the curve falls at a later period. While the curve of the rate of obUteration of the foramen ovale shows the three conmion characters indicated in the preceding paragraph it is markedl}' different from the curves of the \cs.sol.s. It ri.scs less abruptly to a lower maximum rate of obliteration which is maintained with but little loss for a much longer period, so that the curve presents a plateau which is entirely absent from the curves of the vessels. Following this plateau, the curve falls at first rapidly and then gradually over a long interval to the base line.

The results of this study may be sunmiarized as follows:


OBLITERATION OF FETAL BLOOD-PASSAGES


177



IstwV.


lukrl mo.


2nd mo.


3rd mo.


15-6 mos. 6-12mo».


l-5yrs


Fig. 3 Three curves showing the approximate average daily rate of obliteration of the fetal blood-passnges in early life. A, dotted line, foramen ovale, B, solid line, ductus arteriosus; C, broken line, ductus venosus. Based upon the data summarized in table 6.


17S RICHARD E. 8CAMMON AND EDGAR H. NORRIS

SUMMARY

The tinio of oblitoration of the three fetal blood-passages (the ductus venosiis, the ductus arteriosus, and the foramen ovale) is distinctly hittr than is cojnnionly assumed. The prnocss of oblitvniliim in each of these passages shows three fairly- distinct periods: an initial period with a low rate of obliteration, a middle period in which the rate of obliteration rises and the majority of cases iire closed, ami a tenninal period in which the rate of obliteration is again slower.

ObUteration is most rapid in the ductus venosus. Although slow in the first week, the process reaches its maxiniuni before the end of the first month and in the third month and thereafter all cases are closed.*

The ductus arteriosus closes more slowly. The obliterative proce-s-s, which is very slow during the first two weeks of life, does not n>ach its maxiiimm until the second month. Threefourths of all cases arc closed at the end of the first trimester and over ninety-five per cent by the end of the first j^ear.

The period of the obliterative process of the foramen ovale is a matter of years rather than months. Beginning ver)' slowly the process reaches its maximum activity near the close of the first nxmth and continues with a slightly diminishing rate during the remainder of the first trunester. At the end of this time approximately one-third of all cases are closed. During the second trimester the rate of obliteration declines rapidl}' and thereafter decreases verj- slowly for an indefinite period — probably until early maturity, although few cases are closed after childliood. At the end of the first year about one-half of all cases are closed, ill th«' second deccnnium about two-thirds, and in maturity about seventy-two per cent.

^ * It has been shown by the studies of Wertheimer ('86), Nitdtin COl), Fontan ClI) and others that the vein which sometimes occupies the center of the ligament of the ductus venoaus in older children and adults is a new vessel developed after the obliteration of the ductus venosus and is not derived from the remains of this trunk.


OULITERATION OF FETAL HLOOD-PASSAGES 179

POSTSCRIPT

Aft or this pii,pc>r was in press we secured :i summiiry of onrhundn (1 ;ind eighty-*,'Veii obs Tvations by Parrot on the obliteration of the ductus jirteriosus. Parrot's findings are in general agroeint nt with thos.^ of other observers which we have sununarized above. Fnfortunatt'ly his cases under one year are grouped in such a way that wc? eaiuujt include them in our table 4, but if this were possible they would evidently affect our averages very little. Parrot found the ductus arteriosus patent in four ca.ses in thirty-three of one yejvr and in one in fifty-four of two years. In seventeen crises of tliree y<>ars and over the the <luctus wa.s always obliterated.

BIULIOOHAPHY

Alexeieff 1900 The foramen ovale in tlie child. Diss. St. Petersburg. Alvarenga, p. F. DaC. 1869 Considi^rations et observations sur IV'poque de

I'occlusion du trou ovale et du canal arti^riel. Lisbon. Ber.ndtz 1865 [Quoted from Gc/rard, (1. ('00«).l BiLLARD, C. M. 1828 Traitd des maladies des enfants nouveau-n^s et k la

mamelle, fondi^ sur de nouvelles observations cliniques et d'anatomie

pathologique, faites & I'hopital des enfants trouvos de Paris. Paris. BizoT, J. 1837 Hecherches sur le coeur et le systtoe art(?riel chez I'homme.

Mem. .Soc. Med. d'Observation, T. I. EL6A8SER 1852 t'ber den Zustand der Fotuskreislaufwcge bei neugeborenen

Kindern. Zeitschr. f. Staatsarzneik., Bd. LXIV. Faber, \. 1912 Die anatomischen und physikalischen Verhiiltnisse des Ductus

Botalli. .Vrch. f. .\nat. u. Ent. Fawcett E. AND Blachford, J. V. 1900 The frequency of an opening between

the right and left auricles at the seat of the foetal foramen ovale.

Journ. .\nat. and Physiol., vol. XXXV. FoNTAN, C. 1911 Lc canal d'Arantius (i^tude anatomique). Th^sc, Lille. G£rard, G. 1900 Lc canal artrriel. Etude anatomique. Journ. de I'Anat.,

T. XXXVL

1900' De I'oblitt^ration du canal art^'riel (les th^-ories et les faits).

Journ. de I'Anat., T. XXXVL Uauerda. A. 1896 Die fotalen Krcislaufswege des Neugeborenen und ihre

Vcriinderungen nach der Geburt. Wien. Klobb 1859 (Amtl. Ber. XXXIII Versamml. deutsch. Xaturf. u. .\rzle zu

Bonn, 1857.)

• Depaul, Diet. Encyclo|x5d. d. So. M6d., 2. a6i., T. XIII.


ISO RICHARD E. SCAMMON AND EDGAR H. NORRIS

KucHEFF, N. E. 1901 (The ductus Botftlli in cliildren.) Diss. St. Petersburg

(Quoti'd from summnrips in Jnhrcsl)cr. f. Annt. u. Entwickl., Bd. VII

(N.F.), and from Ciundohin: Die Besonderheiten des Ivindcsalters,

1912.) LETorRNTiAU 1858 Quelques observations sur le nouveau-n<5. ThJse, Paris. Parsons, F. G. and Keith, A. 1898 Seventh report of the committee of collective investi(?iilion of the .\nntomicaI Societj- of Great Britain and

Ireland, for the year 1896-97. Journ. of Anat. and Physiol., vol.

XXXII. NiDTiN, A. A. 1901 The duct of Arantius in childhood. Diss. St. Petere burif. Ogle, J. 1857 On certain cases in which the foramen ovale was still patent in

the adult. British Med. Journ. PoYSTEB, C. \\. M. 1916 .\rterial anomalies pertaining to the aortic arches

and the branches arising from them. University Studies, (U. of

Nebraska) vol. .XVI. Quincke, H. 1885 Ueber die Entstehung der Gelbsucht Neugeborener. .A.rch.

f. exp. Pathol, u Pharmakol., Bd. XIX. THfREMiN, E. 1887 Xote sur I'involution des voies foetales. Rev. d. Mai. d.

I'Enfance, T. V.

1895 £tudes sur les afTectioos congenitales du coeur. Paris. Wallma.vn, H. 1859 Ueber das Offenbleiben des Foramen ovale cordis bei

Erwachsenen. Vierteljalirschr. f. d. prakt. Heilk., Bd. LXII. Wells, H. G. 1908 Persistent patency of the ductus arteriosus. Amer. Journ.

Med. Sc. N.S., vol. CXXXVI. Wertheimer, E. 1886 Recherches sur la veine ombilcale. Journ. de I'Anat.,

T. XXII. Zahn, (Cited by .\bbot, M. E. in: Osier and McCrae, Modern Medicine, vol.

IV, 1908.)


ltTIU>H»' AH.-»TH\CT fV Tilt* P*PI:K I^hUCD KV TIIK blULIuaHAPIIIC AKflVICC, OCTOUKK It


iiii; i;Ai;i.\ AiTKAirwch; of iiii-; aslma.s oi' iiii-: I'AUs nuKHAi.is IN iiii: iivpophvsis of i'iif-:

ClIKK

UAYNK J. AfWKLF- AND IDA SITLKU

Dr.pnrliiiriil •>) Aiiiilninii, Meilinil Sihiiiil nf the I'niverxili/ nf Miih!ijiin

FIVK PICIHKS

[t is now well n'('()>riuz((l lli:it tlic cpithcliiil portion of tlip liyiJophysis consists of three ilistinct parts. The pars anterior propria is the principal cpitheHal lobe and constitutes the main l)iilk of the ulaml, the i);'.rs int(>nii( dia is a thin lavt'r. epithehal in iiatun". which becomes iiitiniately associated with the neural lobe. 'rh(» nicst recenth" rec(jgnizeil epithelial lobe is the pars tul)eralis — so named by Tilney ('13) on account of its close relation to the tuber cinereum. It extends forwaril from the junction of the pars intermedia anil the pars anterior propria, surrounding the infundil)ular stalk and spreading out for .some di.staiice under the brain floor.

The jnirs tubcralis has been .sometimes confu.sed with the pars intermedia- Lothringer ('86) and Herring ('08) — but recent studies have .shown conclusively that these two parts are different both in adult structure and in developmental historj-. Tilney, in summarizing the develo])ment of tlie pars tuberalis in the chick and the cat, states:

In atUlition to (In- histological ditTorcnccs between these two parts, llic ontogenesis of tlie organ as observed in the cat and the fowl still furtluT einpiiasizes the fact .that the pars tubcralis and the pars infmnlibularis (pars inlernuMlia) are morphologically distinct elements. Till- |)ars infiiiidibularis makes its appearance inniiediately after the aiiianc of the buccal portion of the liypopliysis is fornietl. The pars tubcralis ai'ises as a relatively late stniciiirc. It has its origin in two sccond.aiy ilivcrliciila or sprouts from the ImkIv of the pituitary sac. These sprouts, the tubcral processes, ultimately fuse with each other across the median line, displace the body of the pituitary sac vcntrad and thus secondarily a.ssume their juxta-iiein-al position.

181

TUB ANATOMICAL RI.CiiKD, Vol. 15. SO 4


IS2 WAYNK J. ATWKM- WD IDA SITLKR

Out" (if lis Al\V(>ll CIS) — ill ;i recent study of tlie tlevelopiiii'iit of tlie liypopliysis in the nil)l)if has obtainod somewhat (lifTereiit results. Wliile afireeiiifi with Tilney as to the distinctness of the pars tul)erahs and the pars intermedia, and also conHrminK the statement tliat the pars tulieralis is late in acquiring its adult relationship with the tuber cinereum, it has been found thai in the rabbit the anlagen of the pars tuberalis may 1)1' discerned \erv early. They were found to precede* the definite pars intermedia by a considerable period of time.

It was with the lio|)e of throwiiifi some light upon this point that the j)resent study was imdertaken. Accordingly we have been led to construct a luimber of wax-plate models of the epillielial hypophysis from chick emiiryos, l)eginning with .stages in whicii the tuberal processes might be recognized easily and then j)roceeding to succes.sively younger (>mbryos in an effort to determine the earliest appearance of th<' anlagcu.

Tlie literature relating to the lateral lobes and the pai's tuberalis in the hypophysis of the chick is not extensive.

Rossi ('{)()) speaks of a median and two lateral parts in the early hypophj'sis of the chick embryo, .\ccording to Ro.ssi the lateral lobes are secondary structures.

iM'onomo ('!)!)) observed a pair of "Seitenspro.ssen' in the hypophysis of the dove and of the domestic fowl. In dove embryos the sprouts appear b(>tween the fourth and seventh days of incubation. \o definite statement is made concerning the first ai)pearance of the sprouts in the chick.

Tilney ('13) first ob.served the 'tuberal processes' in a chick embryo of o days and 20 hours of incubation. From this stage the jiroces.ses were traced to the formation of the pars tuberalis of the adult fowl, .\lthough a reconstruction was prepared from an embryo of fourda3's of incubation the tul)eral proces-ses were not seen in this stage. Embryos younger than four days were not studied.

Woerdeman ('14) notes that the lateral lobes are forming in a chick embryo of about 72 hours of de\-elopment. The thickened epithelium which lies in front of Hathke's pocket is constricted off from the mouth cavity by two lateral folds. Woerdeman con


PARS TUUER.\US IN HYPOPHYSIS OF CHICK 1S3

siders that the lateral lobes so formed arise independently of Rathko's pocket.

Bruni ('15) observes the presence of two 'lobi laterali' in the chick at 82 hours of incubation. He also figures and describes the latrral lobes in older embryos but does not trace them into the formation of the pars luberalis.

OBSERVATIONS

We have prcpan-d wax-plate reconstructions of the epithelial portion of the hypophj'sis from cliick embryos of 48, 59, 67, 72, 96, 120 and 144 hours of incubation. A relatively high magnification was chosen for the construction of the models in order that all details of structure might be shown as accurately as possible. For all younger stages, including the 72 hour embryo, the magnification was 300 diameters. For the older embryos the magnification was reduced to 200 diameters.

Chick embryo, 4^ hours of incubation {21-2 pairs of primitive segments). Fig. 1. The hj^jophyseal pouch is well formed but opens widely into the mouth invagination. There is no indication of the lateral lobes. The anterior end of the fore-gut, which will later form Seessel's pouch, extends farther cranially than does the hj-pophyseal pouch. At this time the oral membrane is intact.

Chick embryo, 59 hours of incubation (30 pairs of segments). The hypophyseal pouch (Rathke's pocket) has deepened and now exhibits two lateral enlargements near its attachment to the oral epithelium. As later models show, these are theanlagen of the lat«'ral lobes from which the tuberal processes develop. .As may be seen from figure 2, Rathke's pocket is shghtly constricted just above the lateral lobes. The lateral lobes have the fomi of blunt ridges which protrude laterally and also somewhat nas^iUy. Their long axes lie parallel with the long axis of the entire h.vpophyseal pouch. This embryo shows one small perforation in the oral membrane.

Chick embryo, 67 hours of incubation. The lateral lobes are more prominent at this stage due to the fact that the hypophyseal pouch is beginning to be constricted somewhat from the


184


WAYNE J. ATWELL ANU IDA SITLER



_l.p.


PARS TUBERALIS IN HVPOPHYSIS OF CHICK 185

oral ojivity. The constriction of Rathkc's pocket dorsal to the lateral lobes is also more distinct than previously. Each lateral lobe contains a lumen communicating with the cavity of the main h}7iophyseal sac. Seessel's pouch is in contact with the dorsal wall of Hathke's pocket for a considerable area. This is the eeto-entodennal fusion which has been recorded by numerous observers.

Chick embryo, 72 hours of incubnlion, figure 3. The hypophysis anlage is closely appUed to the brain wall, causing the nasal surface of the pouch to be sharply concave. The lateral lobes are more prominent than in the preceding .stage. The lumen of the pouch extends well into each lateral lobe. One striking feature is the extensive degree of communication between the cavity of Seessel's pouch and the hypophyseal sac. The two open into each other almost to the summit of the ecto-entodemial fusion. This cau-ses the opening of the hypophyseal sac into the oro-phuryn.\ to be relatively larger than in pre\-ious stages. From an examination of embryos of this age alone the impression might be gained that the lateral lobes are being added to Rathke's pocket. A critical comparison of this and younger stages, however, indiates strongly that the lateral lobes of the chick do not arise independently of Rathke's pouch, but that they are formed from it. In this we support the observations of Rossi.

Chick cinhryo, 96 hours of incubaUon. The principal feature of interest in this stage is the beginning recession of Seessel's pouch and its separation from the hj'pophyseal .sac. The lateral lobes are more sharply marked off from the superior part of the hypophysis, but otherwise this stage does not exhibit any striking differences from the preceding. .

.\11 figures represent wux-pliite recon-structions of the epithelial hypophysis as viewed from in front and from the left side. S, Seessel's pouch, R, Rathke's pouch, /./., lateral lobes, t.p., tuberal processes, st., hypophyseal stalk.

Fig. 1. Hypophysis region from chick embryo of 21-2 pairs of primitive segments (end of second day of incubation). X 100.

Fig. 2. Hypophysis from chick embryo of 30 pairs of primitive segments (59 hours of incubation). X 100.

Fig. 3. Hypophysis from chick embryo of 72 hours of incubation. X 100.

Fig. 4. Hypophysis from chick embryo, o days (120 hours) of incubation. X 75.

Fig. 5. Hypophysis from chick embryo, 6 days (144 hours) of incubation. X 75.


186 WAYNE J. ATWELL AND IDA SITLKK

Chick embryo. 5 days (ISD hoiirn) of incubation. By tliis lime a definite h yp< )pliysoal stalk has been fonned. It is hollow and affords a coniniunioation between the lumen of the hypophysis and the oral cavity. The lateral lobes have increased in size so that the transverse diameter of the gland, measured between the lateral extremities of the two lobes, is almost twice the transverse diameter of the superior part of Rathke's pocket. The lateral lobes ar<^ united by a prominent ridge around the inferior and nasal enil of the hj-jiophysis. This solid median protuberance doubtless corresponds to a vestigial '\'orraum' or 'corpus I«)buli bifiu-cati' of other vertebrates as described by Woerdeman. The lateral lobes are beginning to be solid, also. At this stage they sometimes contain lumina, which, however, no longer conmiunicate clearlj- with the main h3^pophyseal cavity.

Seessel's pouch is represented by a solid bud of epithelial cells just dorsal to the hypophyseal stalk (S, fig. 4). Curiously enough Economo labels this bud the remains of Rathke's pocket.

Chid: embryo. 6 days (144 hours) of incubation. The hypophyseal stalk is much elongated and has become solid. Near its connection with the oral epithelium may be seen the bud-like remains of Seessel's pouch. The superior, or distal, half of the hjT)ophysisis is bent dorsalh" and forms an angle of about ninety degrees with the inferior or proximal half of the gland. The constriction near the middle of the gland is pronounced. Distinct 'tuberal processes' have formed from the lateral lobes. Instead of projecting so much laterally-, thej' are now directed toward the brain wall. The tub(>ral proc(\sses are not located at the extreme nasal end of the gland but are seen to protrude from about the middle of the inferior half (fig. 5).

SUMMARY

The lateral lobes, from which the tuberal processes arise, may be distinguished in a chick embryo having 30 pairs of primitive segjiients. From a careful study of stages preceding and following the rupture of the oral membrane it is evident that the lateral lobes are not fonned independently of Rathke's pocket


PABS TUBERALIS IN HYPOPHYSIS OF CHICK 187

and later added to it, but are rather fonried secondarily from the nasjil wall of the early hypophyseal anlaK*'.

The lateral lobes, in all foniis studied, appear earlj- in devel«»pment. This would indicate that they and their derivative in higher vertebrates, the pars tuberahs, are of fundamental phylf>genetie importance. Thus gwat interest is attached to the broad homologies drawn by Woerdeman ('14).

LITERATURE

Atwell, Wayne J. 1918 The development of the hypophysis cerebri of the rabbit (Lepus cuniculus L.). Amer. Jour. Anat., vol. 24, p. 271.

Brum, A. C. 1915 Sullo sviluppo del lobo ghiandolare dell' ipofisi negli Amnioti. Internat. Monutschr. f. Anat. u. Physiol., Bd. 31, S. 129.

EcoNOMO, C. J. 1899 Zur Entwicklung der Vogelhj-pophyse. Sitzber. d. kais. Akad. d. Wiss., math.-naturw. Classe, Bd. 108, Abth. 3, S. 381.

Herring, P. T. 1908 The histological appearances of the mammalian pituitary body. Quar. Jour. Ex. Physiol., vol. 1, p. 121.

LoTBRiNGER, S. 1886 Untersuchungcn an der hypophyse einiger Saugetbiere und des Menschen. Arch. f. mikr. Anat., Bd. 28, S. 257.

Rossi, U. 1896 Sui lobi latcrale della Ipsofisi. Monit. Zool. itul., 7, p. 240.

TiiJJET, Frederick 1913 An analysis of the ju.xtra-neural epithelial portion of the hypophysis cerebri, with an embryological and histological account of an hitherto undescribed part of the organ. Internat. Monatschr. f. Anat. u. Physiol.. Bd 30. S. 258.

Woerdeman, Martin W. 1914 Vergleichende Ontogenie der Hypophysis. Arch. f. mikr. Anat., Bd. 86, ,S. 108.


THE IDENTIFICATION OF ENDOTHELIAL LEUCOCYTES IN HUMAN TISSUE

THIRD REPORT OF STUDIES ON THE MONONTJCLEAR CELLS OF THE

BLOOD

F. A. McJUNKIN Deparlmenl of Pathology, Marquette Univerrily School of Medicine

TWO FI0UTIE8

In an earlier report by the writer ('18) it was shown that the phagocytic mononuclear cells present in the peripheral blood arise by mitosis from the endothelium of the blood vessels. The method that was devised for this purpose consists of the intravenous injection of lampblack suspensions and is not, therefore, applicable to human tissues. The tis.sues of animals injected with carbon suspensions in which the endothelial leucocj-tes and cells are characterized bj' carbon particles ingested by phagocj-tosis are, however, well adapted for testing the action of various stains on these cells. It has been found that the staining method u.sed by Craham ('16) colors these leucocytes in a characteristic way in both animal and human tissue. Since paraffin or celloidin sections caiuiot be used, Graham emploj-ed frozen sections for his stain but owing to their thickness, they are not suitable for accurate cell identification. The purpose of this paper is to record a new method of tissue imbedding for obtaining thin sections to which the stain is applicable.

The .staining method of (iraliam depenils on the action of solutions of alphanaphthol on parts of the cytoplasm of cells. It was shown by O. Witt ('82) that a blue dye (indophenol blue) is formed by the oxidation in dilute alkaUne or acid solution of alphanaphthol and dimethyl-para-phenylenediamine. Winkler ('07) and others found that myeloblastic cells react in a characteristic way in tissues treated with these two substances and the

189


190 F. A. MrJUNKIN

grunulcs of the reacting rolls wore said to contain an oxidizing frrniont (oxydase or peroxidase) that oxidized (he two conii)ounds and caused them to unite with the production of a blue color in the neutrophilic and eosinophil granules. Later Loele ('14) found that the treatment of mjeloblastic cells with alphanaphthol solutions alone produced the same blue color in the granules. The part played bj- the cell granules has usually been regarded as an oxidizing one ('oxydase or peroxydase reaction). As applied by Loele the phenomenon consists of a pur])lish or bluish color in the cytoplasmic granules of the cell produced by treatment with old alphanaphthol solutions. If a dj'ostuff is produced by alphanaphthol alone, and this seems likely, an aromatic compound or compounds must be supplied by the leucocytic granules. If such is the case "indophenol reaction" is a better term than "oxydase reaction."

Leucocytic granule stain (Ciraham). Since hydrogen peroxide is added to the alphanaphthol solution to make it immediately activ(> and the swollen granules are heavily and permanently stained by treating the preparations with an aniline dye, the method of Graham is better than the other indophenol staining methods devised. To applj' this method, remove thin sections of formalin-fixed tissue attached to slides from the distilled water and stain in dilute Q-5) hematoxylin (Dclafield) for five minutes; wash them in distilled water, place in a saturated solution of lithium carbonate for five minutes, wash in distilled water for two minutes, and stain for ten minutes in 10 cc. alphanaphthol .solution to which 10 drops of 1 per cent pyronin (Griibler) have been added immediately before placing the preparations in it. Prevent evaporation by covering the dish. Wash the sections in distilled water, place in saturated aqueous solution of lithium carbonate for from five to ten minutes, wash in water and differentiate and dehydrate in 95 per cent alcohol for one-half minute. Complete the dehydration by inniiersing the slides in xylol and raising them above the surface of the liquid two or three times and blotting with smooth, soft filter paper. A flat oblong .staining di.sh, the .size and width of a slide, is used for the staining and differentiation of the slides. Mount in


ENDOTHEUAL LEUCOCYTES IN HUMAN TISSUE 191

coloplioniuni-xylol or acid-fr(M> balsam. Tho tinu' that tlio prpparations remain in tin* saturatod lithium carbonate aftor troatmont with alphanaphthol is important because this removes the excess of pyronin. With some tissue better results are obtained by staininp only five minutes in the alphanaphthol-pjTonin solution and differentiating for a shorter time in the lithium carbonate after this stain. If the one per cent pyronin solution is to be kept for some time, sufficient formalin should be added to make it 10 per cent formalin.

The alphanaphthol solution is made by dissolving 1 gm. alphanaphthol (Merck Reagent) in 100 cc. 40 per cent ethyl alcohol (made from absolute alcohol) at a temperature of 50°C. and adding 0.2 cc. hydrogen peroxide. Commercial hydrogen peroxide containing approximately 3 per cent hydrogen peroxide, as detennined by titration with decinomial potassium permanganate, should be used.

Soap method of imbedding. 200 grams transparent glycerine toilet soap are placed in a 5(X)-cc. Erlenmeyer or Florence flask, that contains 200 cc. distilled water. The soap must be so hard that it is brittle and cracks apart when cut with a knife, otherwise the soap solution will not be of the proper consistency. The flask is placed in the paraffin oven at 52°C. overnight in order to dis,solve the soap. Remove it from the oven and place for three hours in an incubator at 37.o°C. The contents should be a syrupy liquid and should solidify when a small amount is poured into a paper boat and allowed to stand at room temperature for one-half hour. If solidification does not take place, 20 grams of soap should be added to the flask, and the contents again melted in the paraffin oven. After the correct consistency has been obtained the soap solution is placed in 100-cc. wide-mouth bottles with cork or glass stoppers, with about 50 cc. to a bottle.

To imbed the tissue small pieces are taken from 10 per cent fonnalin and dropped into the melted soap contained in one of the bottles which is placed at 37.5°r. two to three hours and occasionally shaken. The liquid soap in the bottles usually become solid jifter remaining at 37.5°C. for a day or more. To melt the solidified soap, the bottles are placed in the paraffin oven for


192 F. A. McJUNKIN

an hour, the s(jap coolctl to A't'^C, the tissue added, and the Inittles rephic«'d in the incubator at 37.5°C. The soap solution with the tissue in it is eniplitd into a box of suitable size made from paper as in paraffin imIjediLing and the tissue arranged on the bott«.>m of the box with forceps. The box should be made from paraffined paper or the paper may be coated by pouring melted paraffin into it. At the end of about one hour the paper is removed, the soUd soap trinuned with a knife to thi; desired size about the tissue and the blocks attached to a heated metal disk just as paraffin blocks are attached. The blocks after about one hour are dropped into a siiturated solution of sodium chioriile in a pint Mason jar, and the jar placed hi the incubator at 37.5°C. overnight.

With forceps remove the block from the saturated salt solution, attach to rotarj- microtome and cut away the block until the tissue is reached. Carefullj' trim the block and allow to dry for from three to six hours until a ribbon 6 to 8 microns thick cuts perfectly. The di.sks maybe detached from the microtome and after the proper drj-ing again attached, so that the ribbon comes from the very surface of the block. The ribbon is placed in distilled water in a fiat dish more than 6 inches in diameter, and sections floated on shdes on which there is a thin coating of fixative made by adding 4 cc. of a very thick .syrupy celloidin to 16 cc. oil of cloves. The preparations after being pressed out and carefully blotted with filter-paper are placed in the paraffin oven for fifteen minutes, when they are removed, washed in 95 per cent alcohol for thirty seconds, and placed in distilled water where they remain less than five minutes. It)nization in the large volume of water in which the soap sections Jire first placed develops only a shght alkalinity, and in the thick soap solution ionization is practically absent. The sjiturated salt solution hardens the blocks since it prevents hydrolysis by mass action. If the tissue is quite fragile the ribbon may be placed in sjiturated salt .solution instead of distilled water.

Reaction of sectio7i.s obtained by the soap method of imbeddimj to the stain. The nuclei are blue and the granules of myeloblastic cells, eudothehal cells and endotlielial leucocytes arc red. (iraham


ENDOTHELIAL LEUCOCYTES IN HUMAN TISSUE 193

notrd fh<' red granuli-s in both ciidothclijil cells and ondothcliiil leucocytes and ixpluincd their presence there as the result of ingestion by phagocytosis of niyeloblastic ceils or the cytoplasmic granules of these cells. He does not make it clear whether the grannies were found in all endothelial cells and leucocytes.

In the thin .soap sections it is evident that many cells of endothelial origin contain the granules and that they are not present here as the result of an accidental phagocytic phenomenon is shown by the small size of the granules and their even distribution in the cell cjioplasm. The granules difTer from the myelobla.stic granules .seen in neutrophiles and eosinophiles in being f(>wer in number, snudler in size and more di.scretely distributed. In ti-ssues in which only mature ipol\Tnorphonuclear) niyeloblastic cells are present a casual glance is sufficient to distinguish the endothelial leucocytes containing the red granules since they are mononuclear. In ti.ssue containing myeloc.vtes, the heavier staining and greater number of granules of the neutrophilic and eosinophihc myelocytes separate them from the endothelial leucoc\-tes. The differentiation between myelocytes and endothelial leucocytes is well shown in sections of bone-maiTow

(fig. 2).

The characteristic action of the stain is best .seen in sections of the liver or other organ of an animal that has received intravenous injwtions of a lampblack suspen.sion according to a method devised by the wTiter ('18). Endothelial cells containing carbon and definiieiy lining the sinusoids have nnl granules scattered in their cytoplasm (fig. 1). Likewise carbon-containing endotheUal leucocytes in the vessels show the discrete red granules. The neutrophilic and eosinophilic leucocytes are more conspicuous than the cells of endoihdial origin owing to the greater number and larger size of their granules. There are a certain number of neutrophiles, eosinphiles, and endothelial leucocytes that do not take the stain; anil it is only in the endotheUal cells with a distinct amount of visible cytoplasm that the granules may be distinguished. The failure of some C(>lls to stain appears to be due to an error in technic but all attempts to correct this have failed.


194 F. A. McJUNKIN

BIBLIOGRAPHY

Witt, O. 1882 J. Soc. Chem. Ind., p. 255.