The Eisfnhowfir

8/14/2019 The Eisfnhowfir

1/102

THE EISfNHOWfiR I IHHARY

51 02690 1656

nntinih)Him!iijnMmjtijiuii;Hi)iitiinM)nitiiiinrilinmmm;nfi"Mli"lihiniiriiniH'ntjii)ii'"n!iiiM[[.iin'iii-i,i'.i,i'iiiiMntr]miTJiitjiintnfnitM>j>iîjtiiiniiiHiii)>iincii(miiMiiiH1iO'jriiimiiuMit>i!nMiiiMm)1)H!fnt>nt>Mni((*r)M>'MHiftii!jnJiiiiiViinn!)yfifi)tMi'(Miiinn'MMiMnririijMimnmtirni'iMiiii'inni>ii]initj(iitH!n;t'ii!;nnmii;Mtn)H(n)t>riri'îiirininiiint)iintnO)iiiiinnimttitMinji>iiimf*i!i!

SiiMiVîtlHIinitKiI^minnnflitHtHnKiriHiHniifiminiMMttit'nnjiijtriiiijliMKimidiMtuniMHnpmoiriimrMfiHiiMHut'jnih)iMiMiiiPMHriiHM)M!iifMiHMifijjYmiifnrtmttit'ititiftiiitnijnn'(iiiHMrilintniniH>ilin!iMinrin'm)ii"HYiniit]iiiin!nnnii1miiH!liMi>iiit(rnTMiilniMTMiOinnOiitin!iini!iUiiiunrtini!|H>)!imiint>irrniJHtfii'iHiMiMtniiiiiuiMwrKiiDmniiMiniPiMiiHi'lHIHrr(>rtnMtl|rmMimil(imiir>lt)l!IMMIttlHmiln:r>; i:' "M- !niniHntmiiliiiiftit(mmmiininrtir) '(' '.(I.' iri, 'HiimnnTHiiM)iTinint)Mi'7nniftitiriririiiiHnitii''t'in ^ '";" tM'j^Tlt>iOii)tHViiMimnn!rtiiiinirnti>)iliiiMnrijtt)M>.Mi'ifi'"ft(f'H>'i'!ii

....,,..,,Ji3nifrtnit>>i!it(iîtiiiriiiitii}HiiH)i!ii"iriiinnirMiMiiiirittiiHttMMirThtniKiH'iirtu1nii(nmiiiiMt!hryM)iiifiiniM!iirfii'iit(iin*iiMiMiiiiMiMi>!n;'fntiiiihn|i"iriiiMiinminH>)tiiniitjii!nTntilriitiTTiijfiiiMnri!'i(>u'(n'fiMtnTirilliMiMIitKliniltiri>>l1nlii!iii|il)lH>t['nmiMtlM!ttn'i',>>r'i-i"r'ri'(rf'MiintfHttitrtii|!Mi"f"i'Ji'i''"'*f"''>"*'"rn'Jt(nT)HnriTim"ltt'lt!(MiTii:iti' ,'[-.' i|nini)MH>)iilii)'!nittitnnimiMM-M>"tr mi ,IMlllHifMnMUllllTHIMtHmilMl'!"'' -'.'',!.

niiiiVit'iMnnr!;

/'

n


2/102

in^.ifis


3/102


4/102


5/102


6/102


7/102


8/102


9/102


10/102


11/102

STUDIES OH PBSLLiTUiffiLY SKiaiiJ JUTS

DISS3Baa.5?I0H

Sutirltted to the Board of Univoirsity Litudies ofThe Jolins IJopkins University' in Gonfcrmlty

with the BectdraTients for theiJegree of Doctor of Philosophy

Florence PowderraatorJanxairy 1921


12/102


13/102

It Is vdth maab pleasxire tSiat tiie vTriter talces thisopportunity to express her appreciation and gratitude forthe insijiration and help given to her by Dr, ii. 7. i!oGoll-am,and also to Dr. Williara K, Howell for his rnany kindnesses.


14/102


15/102

Eable of Contents

PagIntrodtic tion ....*. 1Frccediu^ and .'Jethods 5Discussion of i^esults ...... 10studies on the Blood .. 12Summary ... 30Conclusions ....... ......... 31Bibliography 32Biography ..... ....34


16/102


17/102

fart XDITIiODUCTIOK

Uie Department of Chemical Hygiene haa for son years beendeveloping a conrprehonsive plan of investigation, one of the objecteof \*ich is to i5erfeot rrjethoda for the ev-xLiîtion of eîCh of thQssontial dietary factors iRfJiioh is ooncemed with the ntatnltion ofmannalB* At present quality and qiântlty of protein aare rocogr.lzeds i-ni>ortant factors iu deternlning the valtaa of a diet. OtheresBential factors v.Jich ire of practical irnportance are: a properitdjiffltrnent of tlie several linsral sle-nents "hicHi the diet "ust furnish,and the three unidentified food factors 'itJ ich are- respectively resj;>aB-sible for the prevention of serophthalmla, polynextritls, and scurvy.It cannot be said that any one of these factors is of t^reaterinqportanoe th-:m nothar, except that failure v/ill supervene sorufi-whatsooner in one case th.aa in another, althou{î this depends almostentirely on the eztent of the deficiency.

As a piiase of this general plan, a series of e"periput8 wascarried out in 1919-1320 with the view to coraparing with as groataccuracy as possible the biolo.'Tlcal values of a series of proteinsfrwn different sources. In carrying out this study, certain principlesnot hitherto er^ployod were made use of in order to hoigjhtan thesenBitivjnass of the test. It ^rs.B learned in c?arll6r studies that ifthe quality of the protein is excellent, young rats are able to growin a nomal manner îhen the food nixture contains but nine per ceratof this factor, Tb anirnals 'ilso show ligb fertility and fairly good


18/102


19/102

-2-

sucooss In the rearing of youstg* 35ioy are also able to renalnvigorous to an age of ajproxirnateli' ttrenty to tvronty-four inonthabefore exhibiting rmrssd si^ia of deterioration.

If the quality of the yrotein in the diet is not of the be8tand but nine per oent is incl\2iod in the food mixture, tii {^^OAsth naybe nomal, but the aniiails ahov/ lowered capaoity In all the otherfunctions enujTjorated above, ThQ extent to vtloh tbay fail Infertility, or show high infant ;-iortaHty, or ago early, forms a verysensitive method of oo.TTsaring the ViiluaB of proteins froen differentsources when tliose are fed at this pl


20/102


21/102

-B-

and nomality of their tissues, as oell as tlialr jtiyslologloal a0*iB have still other possicllltlas for dotarrainlng the actualjjhysiologioal condition of experlriental ani"nals TSyaa we mayreasonably expect an a^d animal, irresjiootive of TiiOther it has livedlong aad has dcrveloped the signs of ago slowly, or has lived \>\xt ashort tine and aged rapidly, to show certain changes in its tissuesTshieh are conconiitaat with deterioration of the tody structures,if'olln (1) has described data -^^Mch seen to indicate that the efficiencyof the xioneis touds to decrease jarogrescively frtKi yonth to old agStand as a resalt the end-products of protein '.oetabolistn tend to re^rialiiin the blood and lyrsph in greater amounts as age advances* Aztal^siaof the blood for these products, wldoh are usually designatedcollectively as 'aon-protein ultrogea'% should be of value in connectionwith other criteria as to the actual physiolosicj.1 age of the aniral.A certain a-iou-'it of aaerala nay be expected in aniniils v.).ich have becondebilitated to a serious extent. In fact, jtay anilytical data of hiâccuracy on anlnails iftiloh are in a poor jihysical condition is ofdistinct valite ^-hen it can te corapared with sinilar data on youthfuland nor'-.ial ani^ûls*

It was also thou^t Ifcat interesting results migît be obt..i7iedby a study of the calciuT, In tie blood, as it h:id been ol: served thatlaany of the animals cm these diets had brittle teeth and bones*

The present ixiper deals with the chiioal an^l.-sis of theblood i.nd the data collected at autopsy, on aninals representing


22/102


23/102

groxips In a series In which in nost cases the sole fault in thediets was in the jrotein noiety, althoug^x in a fev? of the lots tJiere&B a deficiency of oalciun or fat-soluble A, or hoth. A listof tlie diets is given in the tables in "^Q aipendix.

It was hoped that the data collected on tho anitnals ondiets containing nine per cent protein rai^t contribute sone thingto the controversy aver the valxB of low and hi^ protein dietswhich has been debated for so niany years. ôr^nsrly the data &obtained either by a large nuinber of dietary studies or by nitrogenbalance experiments. Ho account was taten of the quality of theprotein car of the prolonged woll-being of the subject. The earlyworkers in this field, 7oit, liubner, and itvsater (2) from evidenceobtained by the statistical method, recor^raended 118 grns. to 127gns. (about 15 per cent of a 3000 calorie diet) as the projiepallowance for raen doing active v/orlc. Biis tradition was questionedabout 1904 by Chittenden and his co-v/orlcers (3), who advised anintake of 36 g!ns. to 40 gns, of protein (about 7 per cent of thetotal diet) per day for active raen. These figures were based on

experirênts lasting five months, during vMch time the -nan were ingood health and raintained their nitrogen balance. A similar dietis recom'oended by Elndhede (4). On the other hand ItoCay (5), aftera study of the diet and jdiyslcal and riental condition of the rjcesof India, concludes that "the hiêr the level of protein interchangethe more robust and energetic and the more nanly the race."*


24/102


25/102

-6

Ihe nost recent work cm a large scale is that cirrled onby Benedict and bis oo-woriEsrs (6) during 1918 and 1919 on a squad of^'oung men. A typical diet contdueu ai-out 55 ^na. of protein and 1600calories, Benedict stated that there are "definite and significantdistxirbanoes of blood composition, nomal sex expression and neuro-muscular efficiency and mental and jjhysical mirest", and yetthe conclusion was drawi by the writers that 'protein cvartailmentis an assured and physiological ly sotuid procedure." Apparentlythe ill effects of the diet are blamed entirely on the low aaloriecontent, althou^^ theare is no proof that it is doe to this or bohbother factor in the diet such as the low protein content, or toboth. The dxaratlon of the experiment, like that of Caiittonden,is but for a very snjall fracticm of the span of life, and so theevidence of the efficiency of ttieir diets in pranoting norrialnutrition cannot be taken as conclusive, "Joreover, the carefi^lEoeasiirer.ents of Benedict indicate that sorne conditions, at least*were niet which were sinilar to those foxmd on animals kept on dietscontaining 9 per cent protein.

Part IIPBDGliDOHB AMD iETHODS.

The rats for this study, 142 in all, \vere selected whenthey Ti@r: between the ages of 200 and 600 days* The anirnals were


26/102


27/102

-6-

divided into 45 lots, each lot representing a different diet. In13 lots (containing 37 rats) the animals êre on -aintenance dietscontaining various amounts of protein. Sorae were low in Ciloinmand fat-soluble A, (j?or a eorrrrlete record of the diets see thetahles in the appendix). In all the other lots the protein was theQnly deficiency. It was alv.'ays present in an amount equal to 9per cent, and in quality coenplled vdth one Of the three grauies

mentioned in the introduction.2he animals on the diets producing very early aging, and

those representing the second or third generations on defectivediets, were killed vSnen they vêre liot-Teen the ages of 200 and 350days, because they would not have survived Hooh longer. One anirralwas 711 days old. None had reached the end of the nOTTiil span ofa rat's life (about 900 days), and even an the best of these dietsthe ani.'nals viere usually In a poor condition before they reached600 days.

!Che rats were given no food during 8 hours rrior to beingidlled. They were weired, etherized, and saîples of hloed taJcenftro-'n the end of the tail for erythrocyte and leucocyte counts andhamoglcbin determinations. An incision was then :Tiade and blooddravai directly fron the heart vvlth a s. ringe or suction apiartitusmade by uttadiing a hyperdaimic needle to the end of a 10 ocgraduated piriette. Since calciu:n was determined in nost of theea-nples, sodium citrate was used to prevent clotting. A few


28/102


29/102

-7-

pulverized crystals were put Into tlie ejringe or dra\8n up Into thepilêtte.

Observations wero made on tie general condition of theanlnal, and desiîated as '*good'*, "fair", or "poor", on the appearanceof tlie hair and sKin and the brittleness of the teeth. Ilhe amout^tof superficial and bodjr fat, and the condition of the lungs andother organs, was also noted, 1?he lungs ware marked nor-Bul +, ++,

++, ++*+, according to the presence and extant of the lung infectionfrom vdiich nian^- of then suffered. 3316 heart, liver, kidneys andsplaen ware removed, and eaoh nas weighed.

Blood cell counts vyere made in the ustial manner. Theper cent hemoôbin was measured with a iaahli hema^neter having ascale such that an individual having a hemoglobin content of 14.3 gns,per 100 ce. of blood gave a reading of 87 per cent. Correctionswere accordingly nade on each reading so that 14.3 gms. of hoT;ogloblnper 100 CO. of blood corresponded to 100 per cent.

Cheniioal Sxanlnation of the Blood. To precipitate theblood proteins a 10 per cent trichloracetic acid solution sra.s usedaccording to the methods of iTolin and tSta, and others (7, 8, 9).One part of blood -vas precipitated by nine parts of trichloraceticacid. 'Bie precipitates were allowed to stand for at least 6 hours,then filtered and preserved, with two drops of toluene (7). non-protein IJ was deterrnined according to tie .et^-od of ji'olln a;id V/u(7), Practically all the determinations \vere rnade in duplicate.


30/102


31/102

-8-

lat the standard {HH^lgSO^ was decomposed ^dth concentrated liaOHand the HHg aspirated through H2S04 Tiie (HH4)2S04 was precipitatedwith alcohol, filtered, md. tliS cryBt;Als dried. They were piarifiedby dissolving and reprecipitating twice.

Caleivcn was determined in the tricliloracetio acid filtrate,a ooi'ibination of the inethod by Lyman (10) and Halverson and Ber jeim(11), (12) being used. Lyman recOTraends the use of 6.5 jjer centtrichloracetic acid as a precipitant, but equally accvtrate resultswere obtained vdtth the 10 per cent solution (latle l). An a/nountof filtrate conta,ining approxi.mately 2 cc. of blood was n^asured intoa 50 cc. flasi: and t'a.de alkaline -isl th 2 H 2Qi40E, using methyl orangeas an indicator* The solution was neutralized with . .Wj . li HIiOs madsacid \;-ith 1 cc. of 4 per cent oxalic acid and then 1 cc. of 20 percent sodiun acetate v-as added. ^he flask was stoppered, tho contentscooled under the hydrant, shaten until ^ faint precipitate appeared,and ^pt in a refrigerator ovor night. ISie contents of the flaskwere centrifugod in a tube \*ich had been drat-Ti out to a point. Theclear, supernatant liquid ^as drawn off with a capillary pipette.The precipitate isas washed \'dth 5 cc. of ciis tilled HgO, racentri fuiged,and the H2O dra\m off. Tliis v,as rep ated three times. Tiie preciritatewas then dissolved in 5 cc. of 2 per cent }i2S04 diluted to 25 00.and titrated vlth joo ^'*1i*^4* '^-^^ rnothod gave excellent resultswith imovn sraall qu-antities of calcium, aSie trichloracetic aciddid not interfere as is shovji by Table 1.


32/102


33/102

-9-

3


34/102


35/102

-10-

Part IIIDISGUSSIOH Qe' H2SU1TS

Autopsy j?lndii^8Iho ooat of a nar'âl irat Is sleek and well fcept, Vt.e

hairs are of even length, the sidn is clean, and there is aconsiderable amount of body fat. Uoae of the experlnental animalscould be considered in exoellont condition, and only four trere ina good condition. The latter iirere from lots 2188, 2578, 2180 and1895 reêctively, but since otier animals on these sajne diets -.erein a poor condition these four cases out of the 142 animals exa:ninedcnnnot be considered typical, but represant, rather, exceptions tothe rule. It is evident tliat at the level fed the proteins ofthese diets are conducive to a loss of body weight and a poorgeneral condition during the prits of the aninal's life, 'Jhis lossof iwBight is due not only to a decrease in flatt; tissue, but also toa decrease in muscular and organ tissue. The niuscles of the legsbecome stringy. Those betwe-3n the ribs aire often like a very thinsheet, 3ie \rei^ts of the liver, kidneys, spleen and heartrespectively vary greatly in different anirîls, but in general theyar roughly in ji-oportion to the body vî^ît, thôse in poor conuitionhaving a low body v7eisht and small ca-gans, (!I!he vseights of all theanimals and of the organs are givon in the tables in the appendix),

Zhe Genital Oreans. Special mention should be ".ade ofthe condition of the genital organs because of the low fertility ofraany of the aninals. In nany of the males the testes vrere ..larl-OBdly


36/102


37/102

-11-

undersized, and rrany of the fe-iales had a purulent or hydra salpin-gitis, j.niA others, a c:,-8tic degeneration of tho ovaries.

Of the 66 males on \*ich autopsies were performed, 29(44 per cent) had s'.ill testes or seminal vesicles, snd in manycases both had de.-Tenerated, Of the 76 females, 24 (51.5 per oent)had Infections of the ovaries or uteri, or botV, and 4 aninials 1-aduiid rsized but uninfected ovaries and uteri. Ljinoe no histological

studies were rade it is not possible to determine vihether the lowfertility v.as due to a lack of active spernatozoa or to the infectionsin the fenale. It is not unlikely that both factors enter in. Inthose lots sho\rtng a high fertility, only 1 of the 13 nales hadsmall testes, and only 2 of the 11 females had an infection. Inthose lots showing a low fertility 19 of the 36 2L*les (53 per cent)had 8- all genitals and 13 of the 42 females had Infections, aM 2 hadvery srnll ovaries.

Little work has been done on the relation of diet tosterility in rats. Osborne and iiendel (13i report that with a fewexceptions animals wsre sterile when kept on diets in viiioh 30 i>ercent and 40 per cent of yeast respectively supplied the protein. Ihediet was properly supple '-jented in all otiier respects. The grov;thcvave was nomal. They reported autopsies on only 2 ?nales, and tiiesehad testes of from one-quarter to one-third the nor'rial size. ^s Inour anlrials, the protein content was the only doubtful factor in thediet, Dru'iTOond (14) ad i^'unk jnd Douglas (15) have reported thatsterility occurs on a diet defioiait in v.-ater-soluble B. Only one


38/102


39/102

-12-

lot (2286) consisting of 2 males and 3 fenales, were on a diet lowin this factor, and in this lot 1 rnale had only one testis, the otherniile appsaring nor-ral, T/hile the fertility of t}e fenales vvas low,

Lunfr Infections. It has long 'bean. 'Knavm tS-iat rats areparticularly sixsceptifcle to lung infection. Eektoan (16) hasdescribed 2 types of lesions, one being solid arA red, usually withnodular reddish-yellow or yello'.^ areas, and riddled vsith abscessescontaining a thick yellowish jfos. In the second type, the lesionshad sharp but irregular margins containing a gelatinous, translucentgrey, or opaque yellow fluid. !I?he gelatinous aj.pearance correspondsto the appoarance of the exudate in broncho-pne monia. 'Swo typesof lesions were observed in our animals, the on having nodular areasfilled with a hard yellov; or greenish-yellow cheesy vaaBs, the secondbeing similar to the second type observed by Hektooji. IIo reddishnodules isere observed, silthough occasionally a lung otherv-'ise nomalappeared henorrhigic. Almost all the lungs had authracosis. Oftlia total nu"!ber of rats e.-smined, 36 (25 per cent) had nomal lujigs.Apparently the condition of the animal ta-s not the aaly predisposslngfactor because 10 of the 36 animals were in fair condition, ji.nd 2 t erein good condition. There vTere only 5 of the 45 lots in ^. ich all the

rats had normal lungs, and of the five, three contained only on rateach, so no conclusions can be drawi trorfi than*

STUDIES OH rnS BLOOD

Galo ium. An abnorra.1 calcium metabolism is frequently cited


40/102


41/102

-13-

as one of the concoraitants of old age, Tiie so-oalled hardeningof the arteries due to the deposition of craloium is a very coramonoocurrence in the aged, ^cCillum in his Textbook of Pathology- (17)speaks of aprofoxmd difference between the calcium rietabolisrn ofjrottth and of old age. lie states that inore calciua is absorbed fromthe bones than is deposited there afresh - a condition of senileosteoporosis. In oases of osteomalacia the histological picture isthe sane as in riclrats, except that the endochondrlal ossificationis coiiTplete. lioCallun says that tjio catiso is debated, but that itis thoxjght to be due to an adrenal instifficiency or, in woraen, toan oTarian hyperactivity. There is no snggestion that U.e cause orone of the causes nay be a dietary deficiency, as in the case ofrickets.

It was stated in the introdiction on page 3 that msaiy ofthe sni:-Eils stiaaing signs of premature old age had poor teeth, 3oniepulled out easily from the garas. Others were so brittle that Ifgrasi>ed betvjeen the forceps tliey would break in two. SUiis was trueof alîost half (44 per cent) of the animals. In some cases (30 jjercent) the ribs were brittle, in otLers they were as flexible ascartilage. So it seeded probable that tiiere had been some derangenentof the cu.lciujn nietabolism. The occxirrenco of hiî calcium content inthe blood and tiê brittleness of teeth and ribs is shovn in Table II.

Bie norroal calcium content of the rats' blood vsas found tobe about the sa-ie as for hu-an beings - betvreen 7.4 Tgs, and 11.1 Tigs,ealciun per 100 cc. tlood. Halverson and his co-workers (18) give


42/102


43/102

-14-

^able II

Lot I Peroentago ; CharacterIfo. protein : of ^-rotelnM

: Galeiirn in t: the blood t;, ngs./lOU cc :

Conditionof teeth

Condi ti onof ribs

2195219521912191

21882188219521952180

999999999

21802179


44/102


45/102

-15-

Xiable II (oontlnued)

Lot


46/102


47/102

-16-

a content of between 9 mgs. and 11 rags, per 100 oc. blood for nonnalmen and women (30 cases) and Jansen (19) reports a content of 11.5mgs. to 12 mgs, per 100 cc. of blood in middle-aged individuals,DeterTdnations were rnade on the blood of 67 of the rats studied, andof this nttnber 26 were above 11 rngs. and 2 were as high as 22 rngs.

The relation between the calcium content of the blood, t2iebrittleness of the teeth, and tl^e brittleness or extrenie flesibilityof the ribs is not altogether clesur. However, no knlTiil with a highcalciun oon.tent in the blood had noriml teeth and ribs, al thoxi^ soraeanimals with brittle teeth and ribs had a normal amoitit of Cilcium inthe blood.

It seems probable that tiiis abnormal calcium n^tabolisni isliniced with the protein content of the diet. '.Vlth one exceptionthere was no lot (of those in vyhich the diets contained 9 per centprotein) in which all the animals had a normal calcium content in theblood and normal teeth and ribs, (In the exception tJiere xsas only oneaal lal in the lot, so it was not significant). Tl-ieBe abnomiuUtiescould not be due to a lack of calcium In the diet, as the calciu."i ofti:e foodstuffs was supple ;r( ted by the addition of 1/5. CaCOs, talcingthe total calciu.li content of the diets between .6 per cent and .75 jiercent, an a-notmt sufficient to meet the needs of the anln^^ils. It isevident, therefore that 9 per cent of even tl e best conbi nation ofproteins frora the atamdpoint of growth is not sufficient to enable thebody to carry on a nornial calcivr.i metabolism throughout life. :!COollumand Parle and their co-workers (verbal comr-runieation) in their Tvork on


48/102


49/102

-17-

the etiology of rlolcets have jrodoced defective bone formation ondiets having sufficient caloiu'n, but contilning inadeqiiato pi-otein.Ihere is no evidence in this study that the inadequacy of tl e jiroteindirectly affects the bony structures or if it acts by producing changesin the endocrine organs as suggested by LlacCallum.

Seven aniraals were studied 'if^. ioh had been living on dietseontainlng 22 per cent of protein but low in fat-soluble A, and vdthand without an adequate arxjunt of c.ilclun, 2able III shows the effectof such a diet on the calciura context of the blood and the conditionof the toeth and ribs.

Although the nunber of animals studied in this connectionis snail, the results sUf^^est probability tfcat besides the ixotein,fat-soluble A is also concerned in calcium metafcolism, Shis alsois In agreement ith the work of LijCollum and his collaborators oariclaats, in 'Aich they show that a deficiency of fat-soluble A. inthe diet causes faulty bone formation (verbal conr^^amicatian). Ihatthe fat-soluble A, as vjsll as calcium, is concerned is abonxi by theanimals in lot 2405 having a diet in which a deficiency of thisTitainlne is the only fault. 'JIbese animals have an abnomally hl^calciu.-! content in the blood, and both the teeth and ribs are abnormalin textxire A paper by '.ells and Benson (20) is ver;/ suggestive Inthis connection. They studied the ossification of foetal epiphysealcartilage by Implanting it in the ri*ltoneal cavity of rats, and theyconcluded from this \/ork. that the ability of these cells to take '^calciura depends on the presence of sone non-living constituent.


50/102


51/102

-18-


52/102


53/102

-19-

ghe Er.vthrQo:rte Coicit aod I e :0;Tlo'bln Content, Wieerytflirooyte oour:t and tii percentage of hemoelobin serves as anotheriridex of the oonditdon of these animals* The red oou..t of normalstock rats from this laboratory varies between 8,000,000 and 10,000,030,althou^ occasionally an animal will be found with a cotmt as high as12,000,000, Ebe herrjOfdobin content nay be as hi^i as 144 per cent invigorous ioxtig -ales weighing about 350 grams. GZbe average is abotit130 per cent, Donaldscm (21) quoting Eivas gives the eryt^lrocytecount of the norsTal albino ub between 7,400,000 and 9,200,000, and theanoimt of hemoglobin between 85 per cent and 100 i>er cent. It shouldbe noted, 1 owever, that tliese animals had been fed on garbage (personalooranamication) and were of inferior stock, x-'ive rigorous males of tiiissane stoclc reared on a diet toiopm to be adequate gave hemoglobin readingsbet'sseen 123 per cent and 137 per cent.

Of the 123 erythoci^te oouots nade, 74 (61 per cent) were above7,500,000, SoEB of those below this figure were very low; t?;o aniraals,for e:s:a;rrle, having a count of 2,500,000 (Table IV),

It will be seen fran the table that 'nost of the aniiails on tiie9 per cent protein aiets have high counts. The exceptions are lots2379, 2371, 2390, 2388, 2491 and 2494, in all of which a legume seed issupple mai ted vdth either a cereal, dried skim milk or stesik. Sorne ofthe anirnals, however, on diets containing a legiene have normal counts.The lOYsest counts arc those i^iade cm the aniu-nals on maintenance diets,all the very low counts being from these groups. Of the 15 animals onmaintenance diets cmly 2 had counts above 8,000,000, the average of those


54/102


55/102

'20-

Table 17.

Lot 1


56/102


57/102

-21-

Table 17. (continued)

: lot :


58/102


59/102

-22-

Table IV. (continued).

Lot i


60/102


61/102

-23-

below 8,000,000 belne 4,800,000, In the one lot (2760) on araintenance diet in


62/102


63/102

-24-

by different investigators on the valtje of different foms of ironin the diet. (32).

OKB HOH-PHOTEIH K Oi;' 2FB BLOOD

Since i'olin perfected his methods for the detertdnation ofnon-protein IT in blood, the literature has contained many referencesshowing the i^nport-uice of this factor. It la a -leans of detectingrenal instif ficiency in the removal of nitrogenous vBste except in theease of rmxisually r:ipid absorption or of .m abnormally hi^ level ofprotein distinction s-uch as occurs in fever, Tre ar-noimt of non-protein K in the blood of normal individuals is given by varioas writersas betTTeen 20 and 45 rags, par 100 oc. It iray be as hlg^ as 350 ngs.in 100 cc. blood in severe cases of urenia (23). ?olln sad Denis (1)state that In "strictly narnal" itaterial the range Is only between 22and. 26 ngs, Hovsever, a "strictly normal '* individual is a rarity.Persons withoct a specific kidney lesion gave readings between 24 rr^s,and 62 T.gs. per 100 cc. blood, 'with only tv;o speci^iens below 26 rif^2hey (1) also fottid that as individuals grew older the non-protein Hof the blood increases,

for the nomal rat ?olln and Denis (24) give an average of36 ngs. of non-protein K per 100 cc of blood, TThe follo-.ving figureswere obtained frorr. normal riale anirmls about 1 year old in thislaboratory:

S&\>le V.31.4 r^gs. per 1(X) cc. blood35.1 igs. ditto33.5 ings.42.5 mgs. "38.2 rags. *43.6 nigs, *


64/102


65/102

-25-

OJablo 71,

: Lot


66/102


67/102

-26-

lable 71. (continued)

:Lot


68/102


69/102

-27-

Table v " ( CO nt Inued )

:Lct


70/102


71/102

-28-

Duplicate deter inat ions of non-protein H were inade on 103of tte aniTials s tTjdled, Hie amount foimd varied frora 30,5 ngs. to92,0 rags, per 100 oo. with an avera^o of 50,6 rngs. Taicing -itj rags.as the upper level for the norsial aiai-nal, 31 anL-nals (79 per cent) hada non-protein K greater than this a'notmt.

Since the diets of all these ini.Tals wore conducive topremature senility, although in varying degree, we -..lauld expect to finda high level of non-protein K in the blood from the t.-otIe of ifolin (l)quoted above. 2lo one diet appears to have an advantage over any othjsr.Those in which the 9 per cent jrotein content was of tiie best quality,and these Tzlth $i rmich hi^ier content have about tJie sa~e degree ofvariation froci the normal. In general tl ere is a fair agre6:3nt inthe results obtained fron animals on the sa":e diet. Shere are a few

exceptions, as far axanple in lota 2363 and 2070 In which the variationsare fron 41.3 "igs. to 79.4 tt^,, and 42.4 mgs. to 79,2 ings. respectively,Ilovver, a certain arsotmt of Viriation is to be expected, especially amongsulaiornial individuals. It is interesting to note that of the fouranirrals on Hie Kaffir com maintenance diet the three which are in poorcondition have a high content, the Tourth is in good coaditi on and has alow contetit. SliKje the second and third genarations raised on thesediets are frequently very poor, it vaa thought ttat there night be apro-^ressive Increase In the non-j^rotein H In these generations. Thisdid not prove to be the case, as may be seen In Table 7II#


72/102


73/102

^29-

Table 711,

First generation - 52.7 mgs.t>l per 100 co Hood (average of 74 anliiiala)Second 51.6 r.gs. ditto ( " " 24 animals)Third " 47.6 -ngB, " ( " " 4 animls)

There is no apparent rel?.ti on between the nttrriier of days onthe diets and the non-protein 3S of the tlood. ?}iis is aho\m by I^ble 7III.

Table VIII.

200 days on diet - average 70,2 rags, non-frotetn 11 (2 animals^260 t n ' _ n 47,5 r^gg^ ri .f ' 400 ''- ' 55,2 mgs, " ' I n ft500 '- 'I 48,1 n^, " " " " "650 " ' - 50,7 mgs. t " i it600 " >t _ rt 55,4 ngc, " r n 700 'I > - ' 50,0 ngB, "

' " ' "


74/102


75/102

-30-

Part 17.

1. iiats isapt on a diet containln;; 9 per cent proteinvary In soae of their physiologic J. reictiona acoording to the qualityof tlie protein. IThis is âvm by differences in the rate of grov/th,in the ti'ne at vhich old age is apparent, the degree of fertility,the condition of the genital orgîus, the infant i:.orta.lity rate, andto a lesssr degree in the condition of the olood as shown by theeri-throcyte count and the percentage of hent^lobin, HowBver, theanlrtsals showed certain changes irrespective of the t, pe of protein,danonstrating that the anoimt as well as the quality is significant.Almost all were in poor condition, the calcium nstal'dism vvas abnorcâl,and the average of the non-proton nitrojâa of the blood v;as high. Thelimg infection vîich v^ls so co'amon scpxxjared to occur indgpendontly ofthe type of die t.

2 A diet deficient in either fat-soluble A alone or inboth caloitm and fat-soluble A is associated with an abnomial calcicmnetabolism.

3a Animals on high levels of jxotaln inta:;:^ (up to 22 percent) but on diets inadequate in other respects, rsay have a hig^ non-protein nitrogen content in ttie blood, especially v/hen tley are inpoor condition.


76/102


77/102

-31-

Part V.

CONCLUcIOHS.

A diet oontalniag 9 per cent protein v.lll not jierrAt therat to complete a norrrjal life cycle. 31ie degree to vLidi this isaccomplished depends cm the quality of the protein -ixture.

A deficiency in fat-solul)le A. unfavorably inriuances tfcecalcium metabolisn, ij'actors in the diet other than the xrotelncontent also influence the amount of non-protein nitrogen in thehlood.


78/102


79/102

-32-

BIBLIOGBiLPHI

la Polin, 0. and Ianis, W Protein -etabollsra fl-om the otandrointof Blood an^ Tissue Anal:,siB bixth paper. Jotir. Biol.Ohem., 191S, 14, 29,

Z Lusk, a. The Science of Nutrition, Slanders, Phlladelihia, 1917,3, Chittenden, B. H, Physiological Liconoray in Nutrition, .^tolres.Hew York, 1913.4, Hindhede, II. Protein and Nutrition, B. Seyiaottr & Company, London, 1913.6, iJoCay, D, Hie Protein Hlenient in Hutrition, Umold, London, 1912.6, Benedict, IJiles, Roth and S Ith, Hunan Vitality and Efficiency

Tinder Prolonged lies trieted Diet. Cimegie Institutionof Vi'ashington, 1919,

7, ?olln, 0. and iifti, H, A Sjist^a of Blood Analysis, Joxir. Biol,Chaii., 1919, 38, 81.

8, Gireenwald, I. Hie EstiJSite of Hon-porotein iUtrogen in Blood.Jour. Biol.Chern., 1918, 34, 97,

9, Book. The Betination of Amino-acid Nitrogen in Blood, Jour,Biol. Chem., 1916, 28, 367,

10, LjTnan. A 25)id iiethod for tJ-ie Determination of Oalci-um In Blood and Lillk,Jour. Biol. CheJTi., 1917, 29, 169.

11, Kalverson and flerg?i!a. Determination of CalclTan in Blood, Joia*,Biol. Cheni., 1916, 24, Proceedings, p. 22,

12, Halverson and Bergeira, Determination of Snail Amounts of Cilciumin the Blood. Jour. Biol. Ohe-;., 1917, 32, 169.

13, Osborne, T. B, and I^ndel, L. B, Kutritive Value of Yeast Protein.Jour. Biol. Chen., 1919, 38, 223,14, Druranond, J. 0. A Ctudy of tiiO


80/102


81/102

-sa-

ls. Kektoen, L Transactions of the C^ilcago Pathologic il Society,Octcbep, 1916, pp. 105-106,

17. r.lao(iJ.lTm, W. 0. A Text-Look of Pathology, W. B. Saundors,Philaoelphia, IJIS.

18. r.alv0rsQn, lohler and iisreei-p.. Tlie Calcium Content of Uxe BloodSerufi in Certain Pathological Conditions, Jour.Biol. Chera., 1917, 32, 171.

19. Jansen, W, H. The CalcitKi Content of Hur-an Blood, II, Deut.Arch. Klin, Lied., 1913, 125, 168,

20. W^lls, H, Gr* and Benson, B, L, studies on Calcification and0. sification, II. Joxjt, Lied, nQsearch, 1907-03, 17, 15.

21. Donaldson, B. H, The flat, IJeTiolrs of the Wis tar Institute ofAnatoryr, Ilo. 6, Philadelphia, 1916.

22. Sherriian, H, G. Chemistry of i'ood and Nutrition, /Jaomi llan Co.,Hew York, 191S,23. Eawk, ! B, Practical Physiological Gherjistrj^ Blakiston's,

Philadelphia, 1916.24 Folin, 0, and I.Iorris, J. L. She Normal Protein r^^tabolism of

the riat, JoiAT. Biol. Che;-,, 1913, 14, 509.


82/102


83/102

-34-

BIOOBAFET

Die nrlter waa bom in Philadelphia, Pa., on ieoe:)er11, 1894, She attended tie publio schools of Philadelphia andBeading, Pa., and was graduated fron the Oirls ' Hlgji Sohool ofBeading In 1911. In 1915 Bhe received the degree of Bachelor ofScience in Cheniistry fi-om the PennsylTanla State College.

ProiG 1915 to 1917 she VAS eraployed as a cherdst at theBaltimore Olty Health Department, and dijring 1916 and 1917 was also

a gradxiate student at the I'adical oohool, Jorms Hopkins University,In the latter part of 1917 sJie was specialist in food research inthe Onited States Departineat of .Igficult lire, but was transferred tothe United States S'ood Ad"^inletratlon in January, 1910, to serve onthe editorial board of the collegiate section. She nas instr^^3torand fellow dtaring 1918-1919 and 1919-1920 respectively In theDepartiTont of Ghe'doal Hygiene, School of Hygiene and Public Eealth,Johns ropidns University, and a graduate stMsnt at the University ofChioago during the sunner quarter of 1919.

She was granted leave of absence in 1920 to serve asspecialist in nutrition in the Extension Service of the Hew JerseyState College*


84/102


85/102


86/102


87/102

iT p^i^ti^ I I J 1 I f^i. ^ 1 1. 1 i .1 I : U I , : I 1 . . : ' . . . I . . > . . . .

FOLD OUT^/c

_--^4- : /^,9f''/

^/ly I


88/102


89/102

77.: #-J^ 17(5'

/ y///ccy/u ^/t:ur^o/^ ; //^/^

FOLD OUT

/Alb//('i/>

Z^^1o-

_il/::

Z^X

7-

^.^

Z^^

73.6 90-9^f-F g^.^

V. ^ -72^^^R-f^ 7f;:/Xf :^VyB^ ^f?^^^ L^^y.j jj,^^?^7 Jl.o^^/ -^^///^ ^cy.^9 ^^SfVf::2- >.^^/7 ^^z

^^1^/^^?^.J' _^f^y/- ^^77


90/102


91/102


92/102


93/102


94/102


95/102


96/102


97/102


98/102


99/102

i"


100/102

tH


101/102


102/102

Documents

The Eisfnhowfir