The Pathogenesis of Deficiency Disease. IV. The Influence of a Scorbutic Diet on the Adrenal Glands

THE PA1H.OGENESIS OF DEFICIENCY DISEASE. No. IV. THE IMU + NCE OF A SCORBUTIC DIET ON THE A.DREI~IAL GLANDS. I3Y B )?nVKTL1c UTWNANT - 00L0XEL ROBERT AUCARRISON, ALD., D.SC ., F,RAP ., [Pasteur Institute of Southern India, Coonoor'.) [Received for publication , June 12, 1919.] TH E influence of a scorbutic diet of crushed oats and autoclaved milk on the adrenal glands was studied in guinea-pigs. The animals were isolated in separate cages. The experiment was continued until death occurred . Autopsies were performed with the attention to detail previously described .( ) Aerobic cultures of the heart's blood at autopsy gave negative results in ionr cases, positive results in one ( No. 5, Table 1). In this case the organism isolated , failed to kill guinea - pigs on subcutaneous injection ; in large dosage. MO RBI D Axaoxy. Increase in size of the adrenal glands together with greater or lesser degrees of congestion were the main naked - eye changes observed. The yellow colour of the organs was usually found to have given place to a reddish-yellow tinge. The superficial vessels presented varying degrees Of engorgement . Minute hmmorrhagic effusions under the serous coat were frequently encountered. The weight of the adrenals of guinea - pigs dying in consequence of the scorbutic diet was approximately double that of health ( Table I). Whereas the gloss weight of both organs in healthy guinea-pigs ranged between 0 . 390 and 0'520, with an average weight of 0' 467 grams, in subito e.V. licerfsed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) I Q Ir O WEIGHT OF ADRENAL$ PER KILO or 013-W `r s - W TOTAL g  ADRENALIN Q  PER KILO OF TOTAL. ADRE NAWN PER GRAM or GLAND Os•W Fs.W 0.00080 0.00055 0.00050 0'00045 0'00040 0.00055 0.00030 0.00025 0'00020 0-00015 0.00010 0.00005 3.0 2?.S t ?'S 2.25 2.0 . Robert McCarrzson.  189 guinea-pigs fed on the scorbutic dietary the weight of both glands ranged between 0.850 and P 150 grams, with an average weight of 0 . 955 grams. The increase in, weight due to. the scorbutic diet was even more marked when the weight. of the glands was calQulated per kilo of original and of final body-weight (Chart I). CHART I. Char I. l;howing weight of adrenals and total adrenalin per kilo of original (OBW) and of final (FB-W) body-weight, also total adrenalin per ;ram of gland in (a) healthy guinea-pigs (black columns) and in (R) guinea-pigs fed on a scorbutic diet (shaded columns). ADRENALIN-CONTENT OF THE ENLARGED GLANDS. The adrenalin-content tivas estimated by the method of Folin, Cannon and Dennis.( 3 ) In each case one gland was - used for adrenalin subito e.V. licensed customer copy supplied and printed for Helsinl<i University Main Library (SL103X00759E) 190  The Pathogenesis of Deficiency Disease. estimation, the other for histological study. On the presumption that both organs were affected in like degree, the total adrenalin-content of both adrenals was calculated from the results obtained with one gland. In some instances the right *gland was used for adrenalin-estimation; in others the left. The results of these estimations are shown in, the following table (Table I), and are illustrated graphically in Chart T. TAnLr 1. Showing the adrenalin-content of the adrenal glands, the total adrenalin per kilo of body:weight, etc., in (a) five guinea-pigs fed on a scorbutic diet, and (b) in four healthy guinea-pigs fed on normal food. Original h unbar  weight of  of guinea•  guinea• pig.  pig in grams. Final weight of guineapig in grams, Weight adrenals in g rams; Weight of adrenals per kilo °f original body" weight in grams, Weight of per Ii IO renal s of final bodyweight b in grams. Total adrenalin in both adrenal , lands in b grams. Total a pex kilo in of original bodyweight g in grams. Total sdrenalin eer kilo of final body-' weight;' g in grams. Total ad a alit per gram of gland. Experimental or control animal. 1  2 3 4 6 6 7 8 9 10 11 1  550 2  520 3  500 4  550 5  620 390 400 320 320 390 -850' 1.029' 850. • 900 1.150 1.545 1.978 1.700 1636 2.211 2.179 2572 2.656 2.812 2.948 • 000113 • 000143 • 000096 • 000116 • 000086 ^ •000205 • 000275 • 000192 • 000210 • 000165 000289  •000132 • 000357  •000139 • 000300  •000112 • 000362  •000128 • 000220  •000074 Experimental. Do. Do. Do. Do. 528 364 '955, 1.814 2.633 • 000110 - 000209 • 000305  -000117 Averages. 000443 • 000354 • 000414 • 000347  7  570  8  610  9  610  10  670 620 610 610 670 • 390 • 520 • 460 • 500 • 684 • 852 • 754 • 746 • 629 • 852 • 754 • 746 000253 • 000216 '•000253 • 000233 • 000408 • 000354 • 000414 • 000347 • 000648 • 000415 • 000550 • 000466 Control. Do. Do. Do. 615 627 • 467  •759 • 745 000238 • 000389 , •000380 • 000519 Averages. NOTE ;—I am indebted to Colonel Cornwall, r.,ftA, for the figures relating to guinea-pigs Nos. 8, 9 and 10. Owing to difficulty in obtaining supplies of pure anhydrous phosphoric acid and of sodium-tungstate I was unable to estimate the adrenalin -content of the adrenal glands in my controls to this experiment (with the exception of Pig No. 7). The figures given for cases Nos. 8 1 9 and 10 are the results of estimations made by. Colonel Cornwall for lriother purpose in three healthy guinea-pigs from laboratory stock, subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) Robert McCarrzson.  191 ANALYSIS of TABLE I. (1) The total quantity of adrenalin in both glands in guinea-pigs fed on a scorbutic diet was less than half that present in healthy 'guineapigs (Table I, col. 7) although the weight of the ' organs was more than twice as great in the former as in, the latter. (Table I, cols. 4, 6, 6.) (2) Similarly the total adrenalin per kilo of original body-weight (i.e., the weight of the animal before the experiment commenced) was little more than, half that of health. (Table I, col. 8.) (3) When calculated against the final body-weight (Table I, col. 9) of the guinea-pig, - the total adrenalin per kilo was found to be less markedly reduced ; that is to say, the total adrenalin-production does not diminish in proportion to the diminution of body-weight. (4) The total adrenalin per gram of gland was less than one-fourth of that found in health. (Table I, col. 10.) (5) It will be noted that in guinea-pig No. b, in, which a coliform organism was cultured from the heart's blood at autopsy, the adrenalincontent was considerably less than in the other four animals in which no infection was detected by aerobic methods of culture. A pronounced reduction in the amount of adrenalin in the suprarenal glands was thus found to result in guinea-pigs from a scorbutic dietary of crushed oats and autoclaved milk. In conn~xion with these findings it is of interest to direct attention to the fact that the total adrenalin per gram of gland in healthy pigeons is teA times greater ( - 0023 grams) than in healthy guinea-pigs ( - 00023 grams). This greater proportion of adrenalin in birds may bear some relationship to the fact that they excrete uric acid as such. The work of Bauer,( 5 ) which leads him to believe that 'the function of the adrenal cortex is the making of pigment from the superfluous, uric acid in the blood,' is of interest in this connexion. Ile considers that the pigment, which he believes to be made by the cortex, is transformed by the medulla into adrenalin. A further point of interest in connexion with these findings is the contrast they afford to the great increase in the adrenalin,-content of the adrenal glands which occurs in pigeons wholly deprived of accessory food factors of all classes. In another place( 4 ), I have shown that the adrenalin-content of these organs in birds is largely dependent on the class of accessory food factor which is absent from the dietary, subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) 192  The Pathogenesis of Deficiency Disease, H.I8TO-PATFIOI , 0GX. The histological changes in the adrenal glands which result from a scorbutic dietary comprise :-- (1) Haomorrhagio infiltration, and (2) Degenerative changes in the cellular elements of , the medulla and cortex. H(vfnorrhggic infiltration. There are two features of importance i n connexion with the haemorrhagic infiltrations :— (1) Their cire imscribed character and situation in the adrenal cortex ; and (2) the fact that they may occur before any clinical evidences of scurvy are observed, and even before any notable scorbutic changes are to be found at autopsy between cartilage and rib. The circumscribed character of the hmmorrhagic infiltrations and their situation in the adrenal cortex . are shown in Fig. 1.. A higher magnification of a similar se.ctim is . shown in Fig. 3. Through the kindness of Colonel Cornwall, I.M,S,, x am enabled to show photomicxographs of sections of the adrenal from a caseof street-virus rabies in . a guinea-pig. These (Figs. 8 and 9) illustrate the radiating distribution of the capillary spaces of the adrenal cortex in guinea-pigs, They provide. also an instructive contrast to the sections of adrenals from guinea-pigs fed on a scoxbutie diet. it will be noted that in the latter the congestive process is iwt confined to engorgemeAt of the capillary spaces of the cortex but results in more or less circumsoribed haemorxhagic infiltration of scattered areas of the cortex. These hmmorrhagic areas axe of varying sizes ; their more or less uniform distribution around, the periphery of the cortex is their characteristic feature. I have not observed this curious distribution of haemorrhages in the adrenal glands of guinea-pigs dying of infectious diseases in which both cortex azad medulla axe often extensively congested. The heemorrhagic areas were distributed more or less uniformly around the pexiphery.of the cortex in three cases in the present series (T'igs. f and 3). They were present on only one side of the adrenal cortex in one case. Tn, the fifth case (Fig. 5) tbey were absent, a moderate degree of gene,alized haemo _ rrhagic infiltration of the cortex alone baing present. The changes ire the cortical cells consist in (a) loss of their tessellated appearance, (b) , vacuolation and disintegration , of cells, . and subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) FIG.  Pbotograpb of section of adrenal gland from a guiztea-pig fed on, a scorbutic diet of crushed oats and autoclaved milk. Note: fourteen oircumscribed areas of baemmorrbage distributed tbrottgbout the cortex. Some of these are indicated by the letter' H'; the others are unmarked. The pbotograpb was taken by interposing a Ions (X 15) between the bellows of the caiuera and the object, The m.a,;i_tifica.tion is rongbly 20 to .30 diameters, subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) PLATD S.I. Fib 1. ROBERT KOCA RRISpv,— .Patilo;;enesii of Deficiency Disease, No, IV, subito e.V, licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) Pf,ATS X11. I 1 Fw. 2. Section of adrenal gland of beahby guinea-pig, X 55. Note ; normtal appearance of healthy tortes (C) and medulla (11). „ .i. Section of-adrenal gland of guinea-pig wliicb died ii), cousequeuce of a scorbutic diet of crugbed oats alid autoclaved milk, X 55. Note : circumscribe(; areas' of bw orrbage .(.fi), disintegration of cellular elemwits of cortex (C), and of medulla (M). Compare w b Fig. 1, subito e.V, licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) PLATS X11. Fib;. ?. Fig. 3. )t01WICT RICC AItIt(SON,—P.tt,l1n ^cYe1wwiy of De-ivil-ney Wso:rr, No. 1V. subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) `Robert 'MeCarrisgm..  1:9.3 (e) disappearance or loss of staining reactions of their nuclei. These changes are well seen in the photomicrographs which illustrate "three different cases ; they call for :no - farther description. The medulla is much disorganized and , shows cellular disintegration with scatterad. blood cells lying throughout its ;substance. Usually it is possible to detect in healthy glands the chrorne-staining granules of the medullary cells. In the case of guinea-pigs fed on the scorbutic dietary these granules are few or altogether wanting,.. I have been much impressed, with the fact that the histo-pathological changes I have described were .present in the adrenals of guinea-pigs, fed on the scorbutic dietary, which exhibited during lifeno clinical evidences of scurvy. Depression of adrenal function may, therefore, be regarded as a pre-scorbutic manifestation of a dietary deficient -in acerssory food factors of the ` C' class. SUMMARY. A scorbutic diet gives rise in guinea-pigs to-- (l) an increase in size and in weight of the adrenal ;lands ; (2) a marked diminution in the adrenalin-content of these organs; (3) hemorrhagic infiltration of the adrenal glands, usually circum. scribed in extent and situated around the peripl;ery of the adrenal cortex; and to (4) degenerative changes in the cellular elements of the adrenal cortex and medulla. CONCLUSIONS. 1. A scorbutic diet causes pronounced depreciation in functional capacity of the adrenal glands in guinea-pigs. 2. The impairment of adrenal function occurs before clinical evidences of scurvy manifest themselves. REFERENCES. (1) MCCA1tRZSON, R.  The Pathogenesis, of Deficiency Disease,' I.—Indian Journal of Xedieal ,Research, 1919, VI, No. 3, pp. 275-355. , (2) Do.  The Pathogenesis of Deficiency Disease, ' II.—Indian Journal of 11Tedical Kesearch, 1919, VI, No. 4, April, pp. 550--556. j g R  13 subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E) 191  The Pathogenesis of Deficiency Disease. (3) 1'oLIN, 0.; CANNON, V. B. ; and DrwNis, W.   Journal of Biological Chemistry, 1912-13, RIII, pp. 477, el seq. (4) MCCARMSON, R.   .. ` The Genesis of CCdema in Beriberi,' communicated to the Royal Society, 1919, 27th March. (u) DAUER  , .  .. ` Funktion der Nebernieren.' Virchow's Archid, (Berlin), 1918, 2251, 1, subito e.V. licensed customer copy supplied and printed for Helsinki University Main Library (SL103X00759E)