Ondm·stepoOtt J manal of F etetinm'y Science ancl Animal I nd1tStTy, F olum e I, ~Yum lwr 1, J 1tly, 1936.
Sulphur Metabolism.
II. The Distribution of Sulphur in the Tissues the of Rat s fed Rations with and w ithout addition of Elementary Sulphur.
By -T. H. KELLBR ~IAXX, Sedion of Biochemistry and ~ u tri tion, Onder::;tepoort.
THE injedion ::wd oral rHlmin ish:1tion ·of elementary sulphur as a prophylactic in YaJ·ious di:;eases has hee11 practised for mauy :vear,;
and the popularity of this drng- in medieine seems to iucrf'ase of late rather than clenease. Yet, c:uri.ously euough, little IY<lti b1011·n about the extent to "hic:h this element is ab~orbe<l and stored iu the animal bod~-- HoweYer, a fe\\· iuYestigator:> haYe stucli.e\1 the storage of some uf its metaholie products (sulphide and sulphate) lmt, un- fortunately, their results do not giYe a true picture of the metabolism of elementary sulphur as such. K eYertheless, their recml!;:; h:we some bearing on those to be presented in this article and are, therefore, reYie"·ed.
It •nts fmm(l at the lo\\·a Experimental Station (1924) that sulphur, when fe(l to e"·es in claily doses of 0 · 5 oz., was retaine(l in the animal body to a c:onsiclerable extent, though there was eYi(lence that some had been o:s.i(lisecl to sulphate:; ancl excreted in the urine. Support seems to he lent to this point of Yiew by the autlwr·s IY·ork (19=35) on the nbsorption t1ll(1 exeretiou of sulphur in rats aJl(l sheep.
Denis tm(l Hee(l (1927b) analysed the blood of (logs that had been fe(l a stan(lanl ration to "·hich had been ad(le(l po1nlered sulphur to the extent of 0·5 gm. per kilo. per day and found in some cases an increase in the non-protein 1mlphur oompounlls ancl in others a marked (lecrease. Tasaka and Xakar.lnYa (1981) ohsen ecl that the chang-e in blood picture after su l1cutaneous inj eetion of colloi rlal sulphur is similar to that after suhcutaneous injection of insulin,
onl~- less marke(l. A(;c;onling- to the work of Kuho (1931) the feeding of sulphur ancl injediou of insulin clecidedly i11 creasecl both the tot a 1 and neutral sulphur c·on tents of most organs, espe(~i ally those -of the suprarenal capsules "·here the sulphate eontent wa,; also raise(l. )feyer-Biseh and Techner (19::l]) found no chang-es in the hydro- lyr.ahle and total sulphm contents of the liYer in rabbits that receiYe(l sulphur parenterally.
Denis and Le(·he (192=::ib) in a series -of experiments in which hypertonic- solutions of ii-oclium sulphate \Yere administered to dogs by intraYenous injections fm.mcl by analysis of the blood, muscles, and viscera that there was little absorption of the sulphate iron by
SULl'Il"Cll )IETAUOLISJ\I II.
the tissues, although eYen t\YO hours after injection the fmlphate L:onteut of the bloo1l might still he ten times its initial Yalue. Arnoldi, Liss and Hosmn (19=~2) fed mineral water containing Jleutral sul- phates to oue ancl alkaline sulphate:; to another group of rats and fmuul that the aYerage Yalues for totnl sulphur in the liYers of the ::miHt;ll,.; were higher for both the groups gi,·en sulphate \Yaters thau that of the control animab.
Seither Tjehmann (1~92) nor }[e~·er (1898) coul<l filHl any hydrogen :;ulphide in the blood o± clog;; that had inhaled this ga;; OYer a long periofl of time. Denis anrl lleecl (1927a) analysefl the blood of clogs th:ll barl received sodium sulphicle by intruYeuous inje<"i ion, or hy injrd ion iJJto the intestine oml ohsrned that thr1·r ,,·as soJlle- times a 1·isr in the inorgani<· sulphate hut no inn<>ase in thr ueutnl fntlp hur fradion. These inneasecl Yalues \Yere n otr<l particularly in the ca,;p of animab in \Yhom the ];icllleys harl been tied off.
Taken a;; ;) "·hole the literahll'e gi,·en aboYe serms to shmY thai sulphur is stored to a cedain extent in the animal bocly an<l it \\';ts,
therefore, the objec·t of this investig-ation to stntly morp fully ibe extent to which .and the form and tissues in which sulphnr j, :;tm·P<l in the organism, which information, it '"as thought, might suh- sequentl)· th1·o"· ;.;ome light upon the <·a:nse of the toxic effect of tlte oral .administration -of flo\\'ers of sulphur as ohservetl by f-)tP\'11 (1931), Le,,·is anrl Le,Yis (1927) and Lawson, Hedfield an<l Bo~:1.e (Hl34).
Exl'EIUMEKTAL.
"\Vhite 1·at,; \\'el·e 11secl in these expel'iments. 'l'he young rats usecl in the first experiment were the same :~nimals that "·ere emplo:ved in anothe1· inYestigation in '"hich the gr·o1dh of rats fed a mo<lifiecl Osborne-Mendel diet '"ith :~ncl withont arl<le<l sulphur '"'as Rhulie<l. 'l'he c-omposihon of the rations onrl the experiment-al conditions are given in the thinl paper (Kellermann, 193G), of this series and \Yill, therefore, not be describe<! in detail again. For i.hP
;;ake of d:nity, ho\1-eYer, it c;m he mentioned ~-h:~t three gro11ps of
~-oung rat; '"ere fe<l a modifietl Osborne and .Menclel low protf)iu ration with :nul without the a<l<lition of 0·8 per ('ent. of elementary Rulphur. Groups II an<l III receiYecl i.he sulphur ;;upplf)mented rntion. In ncldition the :~nimals receiYe<l distilled water except thm;e of g-r·oup ITT that were g-iYen fresh orange juiee rul libitum instPad of water. On the last cla~r of the experiment referred to nhoYe i11e rats \\·ere kille<l anrl the tissues removed for annlysis. '1\Yo groups of G4 anfl GK a<lult rats of about nine t-o ten months of age \Yere use<l in the seconcl expe1·iment. They were raised on the st·ock ration use<l in this laboratory (Kellermann, 1934) and the only differ- ence in the treatment of these groups >H1S the substituti-on -of three parts by weig-ht of fiowers of sulphur f-or the same amount of yellow maize meal in the ration of one of the groups fm the lust twenty-one days.
The 1·ats \\'ere p11t un<ler ether anaesthesia ancl then bled to tleath after ,;eYering one or both c;Jmti<l arterie,;. The blood "·afi collectetl in a beaker that contained lithium citrate as anticoagulant.
After the pooled bloo<l \Yas strained thr-ough muslin in order to
J. H. Ii:ELLERMANN.
remoYe any hairs, it wa:s allmYerl to cool dmYn to room temperature.
The brain, femoral muscles and thoracic and abdominal viscera "·ere remoYed, and great care was taken not to contaminate them 11·ith any hairs. 'l'he tissues to he analysed were then po-oled an<l finely ground up.
Only the total sulphur determinations were made on the tissues of the rats (young) in the first experiment. They were made on the dry materirtls, after the tissues had been dried in an electric oven. 'l'he tissues of the rats (aclult) in the second experiment were analysed for total sulphur, t·otal sulphate and volatile sulphide. All the determinations were made on the fresh tiRsues, but for the sake of better comparison the results are exp1·essell in milligrams sulphur per 100 gm. dry material. The moisture cleterminatiom; 11·ere carried out i.n 35 c.c. weighing bottles kept for about hYeh e to fifteen hours in an electric oYen which \Yas run at 98° C. The total sulphur determinations 11·ere made according to the method of Feigl antl Schorr (1923). vVhen the determination \\"aS made on the fresh mate- rial the g-round up tissue (2-0 gm.) was thoroughly mixed 11·ith the Na2C03-K:M:n04 mixture and then <lriecl over a steam bath before the rest of the method ,,·as proceeded \Yith. 'rhe total sulphate sulphur determinations ''"ere made aeoorrling to a modified method of Deni~ and Leche (1925a). \VheneYer enough material was avail- able, 20 gm., instead of 10 gm., of fresh material \vere autoclaverl, as described by Denis and Leche, with 200 c.c. of 20 per cent. Hcl (i.e. 1 vol. of coneentratecl Hcl made up to 4 Yols. with distilled
"·ater) in a 400 c.e. Jena beaker. The contents of the beaker were then transferrerl quantitatiYely t·o a 500 e.c. Yolmnetric :flask by warm distilled \Yater, cooled and the flask filled to the mark. 'l'he liquid, after thorough mixing 11·as filtered t.hrough ~ o. 588 of Carl Sehleicher and Schull's filter paper. The sulphate in an a.liquot (250 c.c.) ·of the filtrate \HIS then precipitated by the addition of Bac12 solution and determined further a<·eording to the graYimetrie method as BaSo4 • The Yolatile sulphide wa;; also <let.ermined gr::n·i- metricall.r as BaS01 aR clescribecl by Heffter ancl Hausmann (1904).
The results obtained with the y·oung rats are p:iven in Tables I, II and III.
T'AlJLE I.
Distribution of Total Sulplttt7• in TissllPS of
r
OU71g Rats ferl Jfodi- fiecl Osborne-!.l!Pndel Ration for Si.rty Days.(Five animals, three months of age when killerl, were used.)
Tissue.
Brain ............................ . Lung ................................ . Liver ................................... . Spleen ................................... . Heart ................................ . Kidney .......................... . Muscle .......................... . Blood ......................................... .
191
Moisture.
Per Cent. 77·9 77·5 71·6 75·1 77·3 74·7 72·3 81·6
Rnlphnr in Dry :\fatter.
)fgm. Per Cent.
521·1 593·6 582·2 647·1 856·5 726·4 680·1 878·0
SrLPIIUR METABOLIS11 Il.
TABLE II.
Distribution of ,Sulphttr -in Tissves of Young Rats Receiring J!)leml'?dm·y Sulz7ll1!r -in Food. (Llforli.fied Osl;orne-Mendel Ration Contai'lling 0 · 8 per cent. Flowm·s of SulzJhur for Last Si.dy Days.
(FiYe animals, three months of age when kille(l, were usell.)
Tissue.
Brain ............................. . J_..ung .. ...................... . Liver. ............................. . Spleen ........................ . Heart ............................. . Kidney .......................... . Muscle .......................... . Blood ................................ .
rr ABLE III.
Moisture.
Per Cent.
77·5 7fi·9 69·6 n ·9 76·6 74·3 7:3·3 80·7
I
in Dry Total Sulphur :Matter.:lfgm. Per Cent.
532·6 763·8 563·\l 796·9
\)2;'5·5
7()7·() 673·\l 887·0
!Jistn:bution of S~tlzJlua 1.n Tissues of
r
oung Rats Rece1>uing Elementary Snlphur 1·n Food. (Jl odified Os/;orne-ilf endel Ration Containing 0 · 8 pe1· cent. Flowe1·s of SulzJhuT Flv.s Oran,r;e jw'ce adlibitum .fo1· last Si:dy DaJ;s.)
(Five animals, three months of age "·hen killed, \\·ere used.)
Tissue.
Brain ................................ . Lung ............................ . Liver .......................... . Spleen .................................... . Hc>wt ........................ . Kidney ................... . Muscle ........................... . Blood ....................................... .
:;\loisture.
l'er Cent.
77·5 73·6 72·3 63·5 72·7 70·8 71·0 81· :3
Total Sulphur in Dry Matter.
:IIgm. Per(Cent.
525·9 644·2 551·6 747·3 832·4 748·7 666·6 804·0
It ,,·ill he seen that in the first gtoup "·ithout the addition of elementary sulphur the blood ana heart containe(l the lal'gest quantity of sulphur. Next in cmler came the kidney, then the muscle aTJ(l spleen, and lastly the lung, liYer and hraw. Comparison ·of the data in Tables II and III with those in Table I shmn that sulphur feeding increased especially the total sulphur content of the lung and spleen, and also to a certain extent that of the heart and kidney in group II. The lungs ancl spleen of the snlphur fed animals in group II contained respediYely 170·2 and 14.9·8 mgm. aml in gT·oup III 50· G and 100 · 2 mgm. mOl'e sulphm per 100 gm. dry material
J. II. ImLLERMANK.
than the same tissues taken hom animals fed the basal ration alone. The hearts and kidneys of the animab in group II only contained G9·0 ancl 71·;) mgm. per cent. mm:e sulphur than the respectiYe mgant:i of the eontrol gr·oup. On the -other hand the livers and muo;de of the sulphur fed animals eoubined less sulplnu than the same tit:isues from the eontrol animals. HO\vever, these reduced values are so small th;lt they are of no significance. Furthermore, it shoulrl be mentionecl that on the whole, less sulphur 'vas stored
in the tis~ues of group III than in those of gl'Oup II. It may he
that the extra basic elements introduced \vith the orange juice causerl a faster ::mel more complete excretion of the oxidisecl t:~ulphur in the urine in Yie\v of the fact that Moraczewoki and Sliwinski (1934) have found that the elimination of both thiocyanates and sulphates is rapid on an alkaline diet but slo\v and some"·hat prolonged on an aei<l one. l;nf.ortunately, the datn available are fQO few and the differences in the amounts of sulplnu stored in the Yarious tissues of rats feel sulphur rations with an<l without orange ju.ice are not marked enough to warrant any definite condusions and further work is ne<.;e~~ary in ·order to elucidate this point.
It should also be pointecl out that the accurac)· of the results presenter} in Tables I to III must he accepted \vith wme reservation iu vie11· ·of the fact that the tiRsues \vere first dried hef.ore being analysed ancl it iH possible that during the process of clrying some of the sulphur escaped as Y·olatile sulphide. In order to iest out this point t'vo groups each of fiye ymmg rats were feel the modified Osborne and ?\Ienclellow protein ration, with ancl \vithout the addition of 0·8 per cf'nt. elementary sulphur, over a periocl of GO days after
\l·hich their tissues "·ere analysed for Yolatile sulphide. 'l'he latter 1vas tested for qualitatively on the yarious fresh tissues (poolerl for ea<.;h gToup) by steam c1i::;tillation a::; described by Osborne (1928). Kotwitht>tamling the fact that the animals "·ere in a Yery poor condition, no Y.olatile sulphirle coulcl he detected in any of the tissues by this method except in the liver \Yhere only a trace was present.
Bearing the observations of Osborne in mind, it would seem, there- fme, that fresh rat tissues and beef in poor c.ondition, unlike the
:fl.e~;h of sheep and guinea-pigs in a similar conrlition, do not emit
sulphide on boiling. The liver of the sulphur-fed animals emitterl about three times as much sulphide as that of the c·ontrol group,
1vherea~; ·Only traces of sulphide were found in the kidney and muscle of the sulphur group. Fmthermore, t1veni.y-fmu hours after killing, the muscle of the control grQup did not :vet emit any sulphide where- as that of the sulphur group showecl about tlvice as much sulphide b:v that time as the fresh muscle from the same group. \Vith the pr·obable exreption of the liYer of the sulphur gr·oup, it weulcl seem, tlwrefore, that he sulphide lo::;t through ch~·ing coulcl not have been of such magnitude that it would haYe seriously affected the Yaliclity ()f the conclusions drawn.
As a matter of fact the results pres en ted in Ta hles IV and V sho1v that negligible amounts of H2S were present in the fresh tissues of adult rats that rec-eiYed a stock diet with and without the addition of sulphur.
7 193
SULPH"GR :llETABOLIS:II II.
TABLE IV.
Distribution of Tlm-iovs Forms of Sulphu1' in Tissues of Adult Rats Receivin.g Stock Ration.
(Sixty-fonr animnls, ::1bout nine to ten months of age, "·ere usecl.)
Tissue.
Brain ......... . Lun!! ........ . l.iYcr ............ . Spleen ......... . Heart· ........... . KidnPv .......... . Muse!~ ........... . Blood ........... .
Moistun•.
Per Cent.
77 ·1 75·2 72·0 75·.) 77·2 75·8 70·8 79·2 -····- - - - - -
Tot.al S.
:'ll,!!lll.
Per Cent.
751·5 803·9 895·(i 1110·0 1364·0 1062·0 933·:3 876·8
Sulphate
s.
l\Igm. Per Cent.
77·87 30·4.5
ll·~!l
11·21
!1 '(i:1 30·12 4·2-1 10·64
TAJJLE
v .
Hydro!!en Sulphide
s.
l\[gm. Per -Cent.
1·03 trace 0·68 trace
"l:ndekr·
n1ined ".
s.
::\fp:m. Per Cent.
672·60 773 ·4!i 883 ·1::1 1098·70 1354·37 1031·88
!J:W·Oii 866 ·W
Distribution of Tlario1ts Forms of SulzJlwr in Tissues of Adult Rats Recei1,·ing Jhgh Intake of Elementary Sulz1hw·. (Stork Ratio11 containing 3 per cent. of F'lowe1·s of Sulphur fol' last Tlcrnty-o11e
Days.
(Sixty-eight animals, about nine to ten months of age, "·ere use<l.)
Total Sulph:l.tc H1·dro!!Pn "Undetcr- Tissue. ::\foisturc. :;,, s. Sulphide· 13. minedS. "
-- · - - --- - - -- -· -- ---
:\1gm. ~f!!l1l. :Mgm. :'lf,!!ll1.
Per Cent. Per Cent. Per ·cent. l'er Cent. Per Cent.
Brain ............. 77·4 764·3 81 ·41 0·90 os1 ·OO
Lnn.2: ............. 76·2 861·8 27·:19 0·25 834' l(i
Liver .......... 72·3 925·5 16·7() O·iiO 908·:!4
Spleen .......... 75·7 1171· 0 19·77 tr·ace 1151 ·n
Hearf .......... 77 ·1 1445·0 15·0:? U2!?·98
KidnP\' ........... 75·4 1088·0 50·75 1·36 J035·8U
l\lnsd~ .......... 72·4 95R·J 7·-H 0·30 9ii0·33
Blood ......•..... 78·6 883·9 18·01 0·36 864·63
..
_ _ _
- -- - -- - · - - -·A further study of these tables shows that the heart l'Olltained the largest quantity of total sulphur. Xext in onler came the spleen and kidney, then the musde, liYer, blood an<l lung ''"ith brain the lowest. Exactly the same order also hol<ls true for the sulphur group.
Comparison of the data in 'fable V \Yith those in Table IY shmYs that sulphur feeding increased the total sulphur content of the heart, spleen and lung. These organs of the sulphur fed animals C·ontainecl, respectiYely, 81·0, G1·0 and 57·9 mgm. more sulphur per 100 gm.
d1-y matter than the same organs taken from animals raised on the
,;(ol'k J<ll ion alone. Thi~ ~ub.;Lwtia!es the 1hta obtainerl with young J·a(,; ('l'alJles f Ullll JI) fpcl the lh;lJonJe-}Iell(lel ration \Yith and \\·ith-
Oll( ad<lPd ,;ulphuJ". l-lo1YeYer, it ,;hould be pointed out that the toLd ,;ulphur t;On!ents of the tissues -of the adult rats fetl the stock r;tf ion alone are all, except for th hloo<l, percei.Yabl.v higher than tho.-;e of the ~<Jllle i i;;sues ()l ym1ng 1·:~h fed the basal ()~honte-:Jienrlel ra ( i-011. The granrl aYerage of the tissues -of i he eight organs for ihe adult an<l young rats ,,-en' 87-±·G nnrl G85·(i mgm. per 100 p:m.
dry material, respedi.Yel_,-. It should he bmne in 111inrl that the ymmg r;Jts ,,·pre fe1l a lm1· pr-otein, c~·sti ne defic:ien t di.et that \Y:ts :tlso Yer1· lo,,· in ~ulphur content ,,·herens the <lllult rah were rai~rrl
<Jil a llormal ~tock ration, anrl -onr of ihc contributing far·tors for tl11, lm1· toLtl :>ulphur c-ontent f01· ihe iiHsue,; of iht) young nils }Jrobnl>l_,-,,·a.-; that the ti;-;~ue;-; of tht> h !tPr :mimaL \\"<'H' con~equenih
1·er~- lo11· in l!on-pro!ein sulpln1J".
The 1lab of Kamh:l~·ash i (192l)) sern1 to support this thPOl",\- Jri~ fig11res for ihe toLd sulphur, t>xpres~r'd n.-; a pt>l"<"Plll<lg<' -of ihe hc>sh III:Jteri:Jl. aTr slightly lO\n'r tlwn thosr of the author, calrul:ll<·rl on thr ,;ame lJasi~, for <!1lult J":tl~. rnforiunateh·, he onl:v rlt>irnninerl ihr total sulphur in 1l1t> ft>moral musclt>s al\11 liYN of mnle r:tis wpig-hing· 180 to :lGO g11l. F-or these i issup,; he found 242 :nHl :2:2:2 mg·ll!. prr cent., 1·r~prdiYely, whereas the author's figure~ are 1SS·-1- anrl lfi.-J·3 mgm. for the respectiw iissuPc; of the young anrl :2/:l·.j <lllrl :2.)()·8 1ng-m. for those of n<lult 1·ats. Frmn thPse fig-nrt>~
ii i~ oln·iou;-; ihni his rlata, -ohtaiJlPil h~· means of fusing- the <h~
lll:llt>l·i:!l ''"ith n mishue of I\.:\0" flllll I-T:J'\0" etc., are likewi;;e appre- ciabh- hiuhe1· than ihe results obtained hY ihe m1thor 11·ith the younQ' 1·nb .. ihe ·Pau.-;p -of 11·hich, as llll'ntionerl l;t>fore. shoulrl most pr~hahl~
lw looked f<lr in tlw lo,,· sulpln1r ininke hy thesE' ~lnim:1ls.
\Yith 1·egarrl io the total ~ulphate con(·ent of the tis,;ues in the 0ontrol p:rmql the hrain 1·ontained hy far ihe brgesi am011nt of this
;;alt. ;\ext in -mrlPr came tlw lung- and kidne:--·, \Yith appnlxim<liPl:'i- Pqnal amount;;, theu the liYer, splet>n anrl blorHl abo 11·ith approsi- matel_,- equal nmonnts IYhert>ns the hem·i and musdP contained ihe lt>ast. The high fig-ure for l;irlne:'i· cHn, as meniionerl h:-· Denis flll(l Ler·he ( t92,)h) probably he PxplainPrl on the basis of un:11·-oirb hle inr·lu,;ion of iract>,.; o{ urinr, \l'hile ii is JHJ.-'.-;i.blr ihaiiht> hig-h Yalues for tlw hrain and lung \Yere rlnt> to the clecoll!position of ~omP unsiahle sulpln11· eoni:1ining- lipoi1l. ('omparison of the sulphHte l'onteni -of thP ki(lneY, splt>E'll anrl hloorl. 1'hrse tissuE's of ihP ;-;ulplnn feel
anim:1l.~ con!ainr<l, resprr·tiYrl_,-, 20·(i3, S<),j anrl S·:ll mg-m. more Siilpll:dc' sulphur per l()() g-m. rhy material than the SfllllP -m·g-nns from animals fe1l ihe si-or·k r:dion <!lone.
\\' it·h rt>g-anl lo thr 1listrihutiou of hy<lr-og-rn sulphide, it is r-le<lr ihat iht> fePrling of sulphur lwrl no rfft>d 11pon tht> r·oncentrahon of ihi . ..; fraction in ihP ti:-;si!PS. 'l'his is to he expt>derl in Yir,,- of the fad i li~1t Hng-garrl (1921) hns sbmYn that hlo-od plasllla possesses lhe propt>l"i.1· of osidi>-:inp: b~·1h-ogrn sulphi<le rapirll~- h:v i he withdra.,,·al of oxyg-rn from thr' blood corpuscles. Although he clirl not rletermine thr prorlucts of oxi1lation, results ohiaint>rl h:--· ihr author (1D3G) set>m io ~lHJ\\. thai they are sulpl1:dr~. Thi;: finrling-. tog·eiht>r \l·ith lhe ohsen :dion of Denis anrl Leche (Hl'2.-Jb) that sulpha!r's, injected into
195
SULPII"CR METAUOLIS~I II.
the blood strenm, are only slmdy excreted, expbin ''"hy the hloocl and most tissues of sulphur-fed rats eontain more of this constituent than the tissues from control nnimals.
The " uucle(enniuecl " sulphur values of the tissues ·of hoih the sul11hur and control gToups nm pnrallel, as \Yill be Pxpeded, with the total sulphur contents, and comparis·on of the fanner Y:dues in 'l'ables IV andY shmn; on('e lllOJ.·e that feeding of elemPntary ~nlphur
increased e,;pecially the sulphur content ·Of ihe lung, spleen all(l heart.
Su~DIAltY.
(1) Tbe distribution of sulphm was determinea in the tissues of three groups of young rats. One
''"" s
feel a moclified Oshorne-:Jiendel lO\Y protein basal diet, one the bnsal diet supplemented with 0 · 8 per t;ent. of elementary sulphur, and the remaining onf' the sanw supplemented ration plus ·orange juice arl hbitu m. Sulphur feeding increased especially the total sulph11r coutent of the hwg :1nrl spleen.~ext in order came the hem't and kitlney. 'rhe total sulphur content of the lung of the sulphur group was 170 · 2 an c1 ·of the ,;plePn 149 · 8 mgm. more per 100 gm. dry material than the content of the respec- tive tissues taken from the c·ontrol group. 'Jhe aYerage incrPase in the amount of sulphur in the heart and kidney \Yas about half of the average increase in the lung and spleen. On the "·holP less ,;ulphur \\"US stored in the tissues of the sulphur group fhat receiYecl orange juiee in addition, hut the diffe1·em·es are too Sl\lall to "·anmd any definite conclusions.
(2) The distribution of Yarions forms ·of sulphur in the tissues of t\1·o groups of adult rats, one fetl the stock mtion, the other the stock rati·on supplem<:'ntecl \Yith 3 per cent. elementar·)· sulphur, '-Yas also determined. Sulphur feeding likeKise increa,.:etl the total sulphur content of the heart, spleen and lung but not ,;o 1\luch of the kiclney. 'l'he three former organs contained, respPdiYel:v, 81 ·0. G1·0 and 57·9 mgm. more sulplnu per 100 gm. <lr~· matPrial than the same organ;:; taken from animals raisetl on the stock 1·atiotr alone. (3) Sulphur feeding had little effect upon the sulpha(e content of rnt tissnes, thus substantiating the ohsen ation of Deni~ arul Leche (1925b). The only tissues in ''"hich the sulphate content ,,·as slightly mcreased were the kidney, spleen and h lnocl. Tlw l'Pspecti \·e tissues of the sulphur fetl animals contained 20·()3, 8·55 nn<l R·27 mgm.
more sulpha.te sulphur pe1· 100 gnr. thy material than the ,.;a1rrp tissues taken from the control animals.
( 4) There \Yas found to he no difference in the con centra ti·Oll
·of Yolatile sulphide in the tissnes ·of animals feel rations ''"ith and
\Yithout the a<1tlition of f!owe1·s of sulphm.
BrBLl.OGH.APHY.
AR~OLDf, W., J~TSS, G., A:"<:D ROSAM, L. (1932). lJf'r Sf'lmeff'lgehalt der Lebe1· nach Zufuhr einer Sulfat sowei einer Alkaliquelle. /.tschr. f. cl.
aes. eTp. Jled., Vol. 80, pp. G33-6.'l6.
DE~LS, W., AKD LECHE, S. (l925a). A method for the determination of total sulphates in tissues. J. Hiol. Chem .. , Vol. 65, pp. 561-063.
J. 1-I. Ii:ELLElU1ANN.
DENIS, "'\V., A:-<D LECHE, S. (1925b). On the distribution of injected sulphates in tissues. J. 13ivl. ()hem., Yol. 65, pp. 565-5i0.
DEXIS, W., AND REED, L. (1927a). The action of blood on sulphides. J.
JJiol. Chem., Yo!. 72, pp. 385-394.
DENIS, W., AND REED, L. (1927b). Concerning the effect produced b~· tl1e administration of sulphur on the concentration of certain sulphur compounds in blood and urine. J. Biol. Chem., Yol. 73. pp. ·51-57.
FEJGL, F., AKD SCHORR, R (1923). ZtschT. Aw:Ll. Chem., Vol. 63, pp. 10-29.
J. Chem. Soc. (1923), Vol. 124 A. (ii), p. 784.
H.-\.GGAH.D, H. "'\Y. (1921). The fate of sulphides in the blood. J. Rio~. C!tem., Vol. 49, pp. 519-529.
HEFFTER, A., AXD HXCS"J[A:\K, :\1. (190J). Uher die "'\Yirkung des Sclnl·efels a II f Ei ll·eis7.korper. Reitr. Chem. l'h ysiol. tl ]>at h.' y ol. 5, pp. 213-:233.
IOWA EXPT. STA. REPT. (1924). pp. 38-39. Expt. Sta. Bee. (1925), Yol. 53, p. 76R.
KAMBAYASHI, Y. (1929). Uber den SclnYefelgeha]t YOn l\Iuskel und Leber der Ratte. lliochem, Y,tschr., Yol. 215, pp. 402-,106.
KELLElDIAXN, J. H. (1934). A well-balanced ration for stock rats.
UnclerstepoO?'t .J. l'ct. Sci. and !lnim. Ind?cst .. Vol. 2, pp. 6J9-653.
KELLERl\IAXN, J. H. (1935), Sulphur l\Ietabohsm I. The ahsorption and excretion of flo11·ers of sulphur. Ondcrstepoort ]. l'et. Sci. nnd .lnim. 1 nclust., Vol. 4, pp. 199-228.
KELLERMAXN, J. H. (1936). Sulphur Metabolism III. The effect of flo\\·ers of sulphur on the growth of young rats fed an other11·ise 11·ell-balnnc:ed ration. 'L'his J·onrnal.
KELLERMANN, J. H. (1936). Sulphur }[etabolism JV. The oxidation and reduction of elemenkuy sulphur by animal tissues in 1·itro. 'l'l1is Journal.
KUBO. 8. (19:11). Folia Endon-ino~, .Jupan, Vol. 7, pp. 176-177. ('IIC111. A.bst, (1933), Vol. 27, p. 5823.
LAWSON, G. H., REDFIELD, K. T., ANn BOYCE. 0. D. (193J). Tho toxie effect of sulphnr on guinea pigs and rabbits. .J. LaiJ. ond ('lin. J[ed., Vol. 20, pp. 169-171.
LEH:YIANN, K. B. (1892). r1rcli. f. H.vaiene, Yol. 14, p. 135.
LE\VIS, G. '1'., AND LEWIS, H. B. (1927). 'J'l1P effect of elementary sulphur on the gr01dh of the young 11·hito rat. J. 1/iol. Chem., Yol. IJ. pp. ,)1,5-523. YI:EYER, E. (1898). Arch, J!Jxp. Path. ?tncl l'hcmna/;ol, Yol. 41, p. 325.
}!EYER-BTSCH, H., AXD TECH:\" EH, F. (J 931). Dber clas Schic-ksal paren- teral Yerabfolgten Scln1·efels. Hioche11t. Ztsclt?'., Yol. 231, pp. 110-112.
l\'IORACZE"'\YSKJ,
w .
v., AND SLHVTNSKI,n.
(1934). Dber clas Yerhnlton cler Sulfate im Orgnnismus bei ,·erschiedener Kost. Biochem. Ztsclir., Vol. 272, pp. 269-276.OSBORXE, W. A. (HJ28). A note on the volatile sulphide from muscle.
nwchem . .J., Vol. 22, p. 1312.
STEYX, D. G. (1931). The effects of sulphur on merino sheep. lith Rept.
Diu. Vet. 8er. nnd rl11i/lll. Inclttst., pp. L!81-492.
TASAl~A, S., A:-<n XAKAZAWA, M. (1931). Polia Endocrinol, Japan, Vol. 6, pp. 84-85. Ohem. ,-lbst, (1932), p. 4~77.
