The permeability of red corpuscles to various salts of gluconic acid

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Material Information

Title:
The permeability of red corpuscles to various salts of gluconic acid
Physical Description:
v, 181 leaves : ill. ; 29 cm.
Language:
English
Creator:
Ansel, Howard C., 1933-
Publication Date:

Subjects

Subjects / Keywords:
Erythrocytes   ( mesh )
Permeability   ( mesh )
Hemolysis   ( mesh )
Gluconates   ( mesh )
Pharmacy thesis Ph.D   ( mesh )
Dissertations, Academic -- pharmacy -- UF   ( mesh )
Genre:
bibliography   ( marcgt )
non-fiction   ( marcgt )

Notes

Thesis:
Thesis (Ph.D.)--University of Florida, 1959.
Bibliography:
Bibliography: leaves 171-180.
Statement of Responsibility:
by Howard Carl Ansel.
General Note:
Typescript.
General Note:
Vita.

Record Information

Source Institution:
University of Florida
Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 000886140
oclc - 24516715
notis - AEJ4290
System ID:
AA00009135:00001


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THE PERMEABILITY OF RED CORPUSCLES

TO VARIOUS SALTS OF GLUCONIC ACID












By
HOWARD CARL ANSEL


A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF
THE UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE
DEGREE OF DOCTOR OF PHILOSOPHY











UNIVERSITY OF FLORIDA
June, 1959















he writer vabes to eNMress his deep ut p-reeta to
Cbhatl of his Suparvlaory COa ttae, tr. WIUa 1. th under wose

attattie ti~tse this lavmstlstia -us m eo sbt. ULm umber-
staMae ad em a thra mtut hsve ben lavalUambl and ut In-

astrinas
amtitubb as extada d to r. Mar M. I mhwltm* for s

teim at it.oo so mmraoul atvma. Is adsistam Is the qplmatta
of carttsin ~elael **mim bea baon eqpeally helpt al d is
afnwiy -qoaa HtoA.

Ms fataest d bo y 1e other mbers of 0 citwe's

SqiWrmay Coamttee, tes. D Vme M Xlatear, HMo I. XMeKtt* st
Imbl B. tinker, has laded b na slatag.
S writer tois Sbted to both the Qauate Couael mA t*a

Amica Iuadbtlxm for -ba-mutmi Muosttn fot tSbr aeuagmt
mat fmiimaL asidstmes.













TAcx w cOsuW


bhw


L fTAEs .W. .. .. .



Sagrf V S J..SASgu. *,

mU ... ..* ..*..
Wa0msantes . .
telsttiug Aility *

ftrue Sanflt ComBlltAtt99 .*.....
frsee o ..

Cobalt ..* 4 .
eOOmr *..99999.t*C.9


tal tions ad ood .
p esmm ty ot rytroqtas .
aitaAtiasea wit ProWes .

P abmenlity of S Aed CeU IMadane

Effbact a of a en lysa .

BBER'DAis ********* *****



t.fAR. ** 94999999*9 **9
CoIectisn of Blooa *


9999999


9999..,
999999.


Pltep tion of abolutias .w *
Qaw1titattive etmdmit m atia of CP ant leatlysts
fltmztastsia of Oar Ccat umalysis Mbs AMaling

t1mastion of iWm Or Ouse In Plarsmtc of
lotOsle sn na . 9 9 .
35 spewtaets 999499999999*****
teamtntata of Metal*ProteIs Pralpitatsn .
leeuing Poaint btsxmtnaUhas .

iil


* au
S. 30
30




.* 35
99 S
99 1











patsp


bwab
Vtr

result

mmxlt



$0AM2ll
RMULI
hteault
hlMn~
heM~f
-.4ORM

-SSS EO


V asLo.t... Poin .. .. ....... 150
4it of BIda .a .... .** . 15
flueat meef .i* ZOB p tctt O* Biool 193
rEat c*It Iand Zen.stam a Umais 153
a 3 Dm4t Ptala nokttas XEirit *.. 155
Sixth oda sad seata ausasatae 156
s wit Oslain m eat u eatnct ...... 157
a vith cadat(n) a Maiame(zn) mmaatm 199
s wit Zrc(n) giants . 160
Svitt Co (n) lumates e 163
Svith a tuta . 9l

* P O C * 9 171

i M .*.. e.g.... ... .. .. .. *.. 171
LiSm @**********99*****1**












WEX OF TABMS

amles


Ch.dte.1. Uwsd ft Praparg SolnttUna

Nm-lyIs hta wtiaMSag too
sodium cwtom te

Soaluc Calarid


M~amasI Gbmmate
Ratsaln Glumenate
wamnt Glmuqafte
O"sMm WANKatm

Mamamee(ZZ) Wnniast.
MOSMO(U) &==at$
arm(n) sluacate
Coblat(ZZ) GlnUcmts
steR GUManate

Coneti2vA Ibiysis WhLes
hDNtm orafaingto pB

rwesiags tt Ofate

ammuy SALe


1.


6 19


34 *-


6 -79
80 *97
98 105
106 111

112 115
U6 118
139 122a

ins


29




ha 4 59

60 69

70 79
80 .89

90 *99
100 -0 1"


U38 133


136 141
12 *1459
146 -148

149














I* adjustment of solutions Intended for tat s adaistratia
to the propr taaicity I of vital ooane. Te value of eqfomlag the
hebmoal a method In the dete ntmatiao ot values has been poLated out Wr
Ba at co-worters (1, 2, 3, 4, 5, 6, 7T 8). Tar s-tuehmixd att e
omioti efttft of a fiateee a M erthroyte etped n not ahly aM te
monoentvatla o th s bstance but also a wther aor oat ate compom

pmfetabtew the cell m ame or ftets ite proprStie bs otion$
aileutated to be isomotie vith blood according to 'jyt oaah esla dtas
are not necessarily eotante to the red blood cell.
a.ixseir at MaNS (5) reported that lase asetate aWO MilUy
* bonaLytfe & ralums, ah beymad those oalulated ftom nollitin
pr*p ty aftason. uWir wvok w Bmt4 that of SWrtM d IMaI

(O), th foudA e"larly hi hmlytie aL "u for lao aulfate.
bs paurs. of the pramest wrE as to ImaR= o tte S.IMatOs
eloml related to ia cn e poaoic table to deteaans dter or not
their action pwalleled that of mine. ASd interest ia this savestlWtla
Sstimslated by the fact that these elements, i.e., co~e oeotlt, irom,
i a an, a tsa a are so calle "tmce el uto*, eammtal to hus
nutrition. To obtain a eapative %ture of the activities of these five
eatses, they wre all eNayed a salts of SLunale aeida. ats pertaining
to their effects =a a red b]o, ell it oef nmeatus value sisee lsea
MaeaLs ay W oo be accapanmted into vitamin Inajeotables. Boduma,






a



potassim, mieslm, Md oMaila glu ea U te oee also studled NA their
hoIoi values A e tetuland th- eattloi are nacmlly pment In larva
mnm t In thb Ismom body, being hnoMI a major a mlnta,' ad an widly
udea Is m.4icia.












M oit W m LUI.IRAUW


uaslA said to a ntahbiraeaquproite awid, aBm(C )hCows,
ditovered Ia 1870 as a proa&ut oft o4tm at ogluess vioth ehlorIa
by Mtnts at -a hso an (9). utreoum (10), ia 187", ae="a me la.
vwetl o n at laotse sold fteamm tiim, firt lestates .us*1 aeit.
It we not att the 1930's, however, stlt glueaite s lA beeM


At 0A prosmt time, gsUmic aei Is puaeroi cmmerusvae l by
the vtsMotaive d ofd tsm o th ashya de otf glueo (31). Iis
oazdtion NOy lso to ecarri out Is aiullao msat with the utilUatiom
oat husbiattes (12), or %LstarolytlanU y 1o11 aginS am obes Imo iso
the resme of at ter eluble itoftd (13). Althou& 2mL, aeid m ay
be prepM as a wt Ua product (34), th hi* cosw at e oMla procedure
has asseuitated th promtiao of a 50 per cant aq&ioMB wLautio to
represent B asiL at oamere.
h a Wus solution stuemi aei exists In equiltu with its
easnysmaltn vm anad salte-lat-U ess, th eoemostratias ofat vary
Ith orgiaBl glieaai asia os caeanttima mA the tmntwe.tu qo.
aIsfottia of It i aqnmos alutltm, gLuocalo aed pas se Satet amreIMtaly
aartint mgtLusoa]tone (15).
axum (16) fomI that ftree glo0ie olad -M aMU tolerated Asaa
takn Vith te daily fbod ftr yeam 1y hmans an Ilasm protela










aetor Jury nor isturbeanse to the dptlve aorma. OI twssis of

stuAtes oamated vitha 0.25 M. lA soltutitms .bawwterd IAntwmouly,

it we reported tnt m nic aselms wa the last toxi of *6 follwsas
aelds listed ta od. orlf tee easda todaity onlte, m.alot pIashoborWe,
tartrtuo, gnmioBs h, hiiroahlawrlo, lactte, 1orie, proiplrae, suelnia ,

aatie, =A lmaMie (17). BiAts re bMen tratetd dAily with beth sub-
utMmmous mn = tawmnus saJeetU ns oat s5nmane foil wileut Oay aUL-
catio of 11 fAfets &fter ame wek (18). a 19-oo, O&mitt ofat
tods of the AMriom 4Almel A- oelatian prmou.e d purified aIta.
lactM of lacimoe Ael to bo aft or us Isn foods (19). a3n paro.
portions of lusamie asi Ld e t to ftta md oils produelt prolet a i

waMe 1 Ibth to beom ro aid. (So).
Lumamfte acld hM beun tmIstNtetl to be uMft3. la the olwutia

oat existase shohte stams iS the bom a kley (16) u wveU asa th*
pmvtirn of tair fa=mtimn (21). Bth gluesamle asA el A gluouro-
ts-3aotme ahe bern use .fftetlively to a1dify uaris (2, 23).

It ba be n rmprte (a2) *ttt gLum el odv p3m'tmble3 to um.. Ue
acid si est 0t blts to tramtmt plan riti d r-ative bsllU .

aB tm rnate *ffeatts w repwarted to hem b eqally efteotive vitb
nu,p homwmr met pltiants .], neumd uffret4 lese uatrotateti al a

rwal ditreme isn treated vith gLuoade weit thn fea mastllo said
wa used.



3 lme .e gumomate amian is without awhrked ~us.xMo_...

activity wA toae ity, ttallie gluoamfte m ay be ~ s e tive









aoens oat the stimi vthout many caLntias arifla fro the pjeos
of ta glxmSUate mlal (25). Mka metallo salt of CaluoaL eit b*e
te prgIraa .' pe lftlea, soh as the gm L of stlram, Mam,
autt y bull=$ sau. cadmiums, caldim, cobalt, oaOperS Iron, IMM,
mga lm, OMn potasamlo d, om, and slas (96, t, 29 3D).
Alisoui mot of te mlts are not eurratly bto tesma netalto,
the possibility atf utilsing swem of tb for tsae iIemNt astt
ticn bas bam wumsmt. (2a).
samnto ait hasb also ben epsoe is a prpesti of malts
of drus- ibhc are arcade bases. Thae balue brAus Cbolae,
aWAtoalnl, clanI ., v rlxw, pwosaoe oeds, gflnimae, 0gdals ,
stamymop tal, A t str ia. gluaantes (31, 32, 33). A good My ofat
the ma lts hwe bem ftnm otw wy aad ins =u m with rsprts or less
tootaltyr tm A= a orole base wve MLe L at aiM d
with inWarlte assos. Ar Instanse., eerflae smltte =a Ipartet to
be -o irltaia tin apeanra gnwc ws injeted lIt- Iamularly vlt little Irritatioa sM aly aiLA tolte
effects tns stmilAr tajeotlas with psaoota. SBdd Ise usually nr
sulted In bWotaN sB sat a ocaswaal fttllty (3V).
trivlalt soft. antimamy gluonate bw baa ALoed isn ateive
twr.a. -t of wlary .astowdd-ia (35). Presott 6a (u) zwprtd
teat p*esrattve for ham aod wmin blaoesd uemA r t-.ItMlaes bas
beea ju mat a gLucasia acidt o a=antia with tetiary atlfatie


00 Utszmture is replte Ii uses ftm calelm g -osate, nrylag
tfte a eountmestic at emarl at local Miasitivitis to steptaeWda









(36) to es aBamac eltntsm of ,vl ald qal aseo atet with
drug a tiaem (37). o alt hbw be umd by atwuaes fttlatMtam
to Mitoe paMents Ufftag strim cateastial tubntcufla (3). It ba
bea une sa eaStatc with 5I4 muortato I* U*s fatmt oC xae.
toAL wrtitt () a wl as vita a alt oC d-tuboua rsae in t
taatmt ofr qstie dltera (40). oleam gLuo mte as ealasos with
pitoeia h be e & aa sA $ ov w osod auaia t o atbor wit aty amt
efforts (41).
A"thmeu n ta s wl&te of alal=l toeity with sesum
gcan.t (be), s al hbe ben 4 wry U1 te s Mea to
ate. ae levels of ehtortntoyouii wea reported to bwo bew
Inraset two to oan ftlt by staitmas ea tsiatIata or seotm
uaoate (43).
h mStaiLms, poestus gusoaste ta ftmad Umats use Io -a
oa hlasmoaews.da, tan t 2anhMu, Sn -wt MargiatS e qwatI.M ("A).
IZthu alsi be suemfuy a04oyS, anly (4) a" pIafnsury (h6),
ls "W teamat at aflcuW 1rgicJ34 a1 be bm faow to be a"
better talan thm pottesi- a 0hlarl.
mesim & g lant as al3 tolarte by -a se atal 0n *ta&
Sautstred wily pugstmlUy (4T). It hb been aincsthun wlqFe
it Gan at lmmorf lh (48) a to OWetaBa sa ow atpwlrtitam.llty
heWrtofr ubeafta by otber hnmam Va (46).
Ammonm pLamaste be be lajetedt Iatmamsy r Inateo a at
vith s apreulable austatad Injury I dotese .gim tant to a lajeotion
ot 30 ak. Ao a af on the beis of rmass body wf at (149). It w a*
,AoPa ft istur of the lom.rUeg oa tb p3 Twa r O ifw, wvith xalts










inamtting its effoetivemne t 10 Go. doses.

Clating Ability
M term "ahelate" wa proposed Ia 1925 y Magan 1 j. (90)

to de aste those cyclic structures aristing tc the iion of metallic

atoms with orgpainc ad inorgmle wolA.ules. aehl (51), ia 1937,
1ammited opoudm epable atof fonsing chalates with metallie oatioms.

6 4atitngiuhsA ehnemical group that uilte with metals 1 the repl-te-

mnt of a hydrogen atom, as carboay1 ead oale hydroil groups, f *m

those tht chabine with metals by simple coordination or addition without

th Fe r la t of ahydrogn, sa earbotyl ead alachdUei groue.

h ghsmal, the ability of the supr aelds to fam mwter soluble
lea vith metallic las art es fro the capacity of their earbomal

aia hydray1 roaups to bat these oatioas In ri frm by musm of o-ee
ordiamte and comlnt boans (52). 1he oblating ability of thae sur

actds has ben found to be, as the awnag, more proamnced la aieklt.
solution than either neutral or aei media due to increased toisation
of the hydrogs of the hyirAr l groups.

Glualaeo acid ad Ita derivatives are capq l eo ftonmag ater

s*bAlt chelates with certain metallic lton. Although h soMiu gluemate

ad neutralind glucoale seld are, ia thBselves, relatively ineffetive

seqaestramt for alelum, they have been homn to be superior to *seiun
citrate, soditm tetroqbophate ad the tetrasodium salt of ethylm.iin.a

tetaeacetle eold, Am Ima strongly a&aLte solutions. odiim glueonate
me found to be uperlr to the three aftrmetioned segestrate i

eomle nag ire, over a rangs of pH 6 through pa 11. a Sl.uommate mw










inferior, hwmver, I thl ohe lat.tmn aindim, cobalt, imn ., ma
sine (11).
Xemtly, psesok an JWret (53) shdin that ueami aid om
vrY stwr8 rOl m- with WMt of th tramAitis eOnmWnt, 4ontraWy
4 ea0 ier opiala of otrw lareAtipgtori (50). ?mgoh a04 Awat (53, 95)
"tNeia soer le s gsuantA stsme polaruqiaoslrl, meaeobshoto.
uetrissUy, porn imtrially, an deeramtrially to order to postulate

possablte bat riag structures for theW uBterals md to tetmine
eglitbta eoastainta at various tnia strengths. Pmok at and&r

(27v, 2) oedHAoted stailm 9xpelaeWtu fr oadia 4A lrm(MI)


MIior I t MabVe (56, 57) hma aerm that th stblAity of iam
bivlBt metal emwleu dxeaased in lA* follovia g tars palladlua,
oprq malelp, lmd, cobalt sL aAl m, m ma m-
neIim. tl wa e oapmrs co, ble at fofna ta aquae plaar
badss, teamd the troUmgwt wealate. She stabUity of ta oaqL-M

@pnoae to dereamm with laemsaig bsoiolty atof the mwtal, w.mpliftd
b7 the weak abelate fomdt with maestum.

Tem Ewme t Casitdmtios
WrIan Pe past emtur7, a epeaslaUy wvitn te last thirty

yews, a steadily lncrentg auboer of el1mtn hwre bem fuand to be
Inavariably parent In living tissue. Definite biologil aseuations

have bee dleoored for ewratin elimnts preamt tIn vwry iute mmts,
relative to the umtities of the mnt cooutitumts tIn tiasnes.
Me*e elsMents have beem termed "tnoe elmnts ."










buteat d Z Diadm (58), tI 1989, *mne h b a bWood spectro-
grphieally -t reputed that alumdnau, ealai=, iron, mmestni, am-n

pnaesa, lsophu, potasslun a.l]eap eUver, tit lkmia, ad qeo we
presMt to Ll sawmes studied mad that hraioum, oeebalt, g pmim ,
lead, SaIbeO Stratui ial t wm re maiftet In m of thm. Toady,
further imelants a ben addd to the growlag list ud within this

grop awe a I.itd au er of elements for ubidh essential bytaolog1al
raes have been -mfivocally esrtablitaid. bse awe cobalt, eoprw,
ldi4ae, ,rmam e M nN, and Bno. Other elmnts as bartmB, t Lorlme,
.olbdoDm, at strcat1i are gra&aSUlly *aquirag Justifl2stit In_
olusimt i this V of esseatia elments (59).
Senae raiotaotive lotopes of *thse elmats Sm be edlnlstered
In mnal sytiaoloioal doses, mA yet measure aeauately, 1 study at
them ma other admbtas hs beem neatly f tidltted mad st liflt (60).

2mn
Mamwai (61) wus lnftumttl Ia placTin the mdietaml use of

ira a a sound bals through his disa ery Iny, 17146, tat Irm Is a
Lharaterhstio eat esuntial emstitumt of blood. 2S 1886, Zsuottffky

(69) usmt tohat pua crystals atof borse bmgloobn eantalated 0.335 per
cmnt aor irm. MWay tm emtsltaS mmees bwe boew reogmsli, In-
cudla8 two nm re eamtly disouved favoprotein n1s-C, eytom
siuntams (63) mnx amthlA a ois. (64).

ZIrm osaurs I the zd bloot ells as hig*lobln. AML mm.ian
hmolobimas M cop. ose of four ferrous tron porphrim (h m) attftd
to e- globla. siogldbin vrertaibly fose a red mtleamtar oapoMnd










wlth ogn that ealiidts a definite absorpta speetrt IUa between
tme hme me m st Obs.a l au .ues It belU ved to be ooi..pid4 through
eletirostatio sttctimon betimen imiXed proploalc a lA goulp sms
aurftoe basio grois otf bm, ma" coordination ot te Lo vm with a
nit2o.a-ontalaing ro of the aglobn, pem 4e to be th0e bseuol.e

srpm at histialn (65).
IOm Is the s.m done ot *esmrlay oew In the free state, bet
riemar Is oampttsly boua to a spee fic protein, anom as tinsifarri

(66) or sluvrwgqlla (67), the saole function of wheh peaws to be te
trm portation of OIrn. It bu bee suasted (68) that a s L mammt
of Irea eists to the imleto e Io equilibr1f with si oihbilla aet
amcovmts for he ls tr&ms au aes mo mbrmes, vweas same se to

thin thbt a ame cMplesz uepatge fottor lavalvng feritat aeoouts
for te treaftfr (69).
alme thae ltem bladibg property of slider hiiUa 1* speolfle -t
sIame irm camot exist the e te the free state for extended

period of tie, the Oqagaty t of ermu to transport iren ie believed to
be govern by the sitdaupihln eastent. Noasly, about le-thrda of
the sideuhillsn is actively tIampartins Im, with the rom ln
siaeraghill coprlisntg the latet lir-bndlng eosaity of the aesam.
An totaesouMs SaiJeatloa of irem producese as aereae la a proporties
of lrm-eastuated a derophilla, with my amess of tre learTng the
blood stue ad entering the times (65).
owr nmy years investlgtam hwre searched tfr as Im oumA
uitable for paeteral adeirnsetima (70). As early as 1896, ier
ctrate eatA Irs propiesate were Injerted subutmreouly In the










trib'et t of ohlaorosi (71). P rYieimIm mammniWLt ag ra r ta Gmi MA

ira. amod.ate vwe eisemwd with the pan following their Injatioa

(7t, 73). Hmeth !&-. (74) onoducted the first earful study of to
utilatiaL. of am taijeotlia mW comluded that "emapt ia ram
n taecs t Ua sdiainistrtia of ir a by the uyteanI rmote smald be
avo4de4.*
h 1936, Helleyer (75) Iajeoted Iron aseoarbate itIw ously
vith eouI remlts. & the falling ye a, Iroam A4 m ime
aittte (76) a4 ferrous glucumate (77) ro mafAnistwea paueetwealy.
urem gluemuato wv r'orted to be voa of protein pjaIpitatiag
aUti as vill as coming as losl or meral reactim an Injection
Into h m.

& 19"5, the anetin of ta e etliaumI Mulary ComItte (78)
stated that atramulmar lneotao of tron w th usly pwmimulble
par intal route. It wu furthew cantaded that snae as salt Atf Ir
vith protein aroieitating properties sabould be useA, as dhote@ of
Iroa prnsUations we liaited to two rpam -iem mand maltm eitrte
at ftraouM luaacate. h latter, iwbel free tree irrtatlan a Sn.a-
Jeetimon, s matied as the drug fat aboloe.
MaMsive doses of Iroa as loll.a l ferria hArooidemm d as
acnlntal ftrte amXds w ltas aeste intranmmaouly In 1946 vith aod
retieulyte I---veLopmt (79). aI the follawelg ynri, mi.a (60) di-.
comered tat relatively larug m wtas of Msoeoaated ron axide oaul
be talemated Iantravously. Sevensl peare later, bowrer, a fttalIty
ns reported after aa Injetima of abautst irL a O I a (8L).
h 1954, ao irna. etrm oamapomd oamntainig 5 pNr eoat af tIM










I .afed ( s). it wS rWrted to be extremely statb la lar

a a vith low td e 1 oue aty l teUmt4 wil t d
stSantlag tryropoIss2i.

Cobalt
So occurrence of cobalt to Min1 tissue, speillly tm aw-o s,

s dwau mstte oMnTlu (8ely ) 198 (83)-. SI S ly s saw e as
deflalte tmS atiam as to the bIlol20osaa funetict of etbalt. four
yers e Iter, amer, sea ~ uO t at sth sen tattaU of obalt
A^ relativJly large but subletbal dees stgmalateld hitop.4esias -A
afneas poleOthmia ian rats (8t).
S1948, the pure antienatious mmae tetar "a disomInt to
be a otbatt eotalatag ocqoomA (85, 06) soa after 4dsstee as vitmia

Bi ad lAter termed uwaooblnmla. It me isolated ftrm lmSin
Uvr as a aed erystaliae om empouA iaftainng 4.0 per oat eabelt.
s Interest tl cobalt saitde fr01 Its iFmrsm to eyIooamimLn
rests a its vauei as a am.peatiflo e7ythrpoietio ettalatbat to the
teteuM t of variou types of mais (87, M).
Althou I preSee etemdaatims of the cobalt centmnt of to
blood are a xtrmly difficult to pertim, du$ to the vwy lee MasM-
tratimms Gmnly presat, Ieparord (89) bag estimate t eobalt It *e

pam a the petked "ea elns of 16 bma sjesats. 1s fmund the aum
eoAmtratle to range fom 0.28 to 1.T70 og. per 100 AL. of blood with
the peaelt sels ceetalalft fee 0.60 to 2.3s meg. of obalt pew 100 at.
It has bum reported (o90) that cobalt does not atffet te nature of
hNL.Obt a r its oWa7I aerrn capacity, nor does It WprouOe a










oabaMc1n pthaifg metblolowbna otfe worker, bomwer, be observed
the pramoe of mt ie-olobi a MB norm l bfn blood w tlaubsated at
3o trw tmaty-fwur biourl In te p.re.eoe of 0.2 Psr omt obalt
blwde (91).



e pressue otf opp a Imo M tisouse bus been kt m sesm
t0e vk of 4'BmtUa7 (91) in 1833. ?t VM0 not until 1981, howwm ,
to te t"i frt evil e of a uhdolole.l timutim otf t element was
Iatt"itsd tarsa lveetlptcsa. 2h t*t ywr, N3atu*y (93) diesovore
the Wsewoe ot esppe a is M lhm brain and In 195, MBrmU (94)
presmtsd rlenez ofr th coper aomtet of blood WnA tisms, o.oasstsg
tto riudma at lihet a tha elemait.
A 190, be% =A oM6extee (95) S aOM. med taint eopW in
dditim to se is oU amntll a the ftemttin orf I-bms1.in ti 10ts.
Mks diiswry w- oca her.ater f vwriesd by aunm er umaIM (96) Ao
futeer estdbliMed that cooper Is esse trial ftr hur topoiesfs i a vide
MrA of mlimL opeIses. 2 1940j, Sahultse (97) stated tat th evidwee
Iammted that oWaer fotlitates both the mob1mimati of Irom twro the
tissmes d te utili atiam of Umt by te blood fbolag roms.
A number of oppr.-proteta eapou bae been isolated from bot
mtfom mg plit sinues, severl of ieRb were dom to be auses with
onum. tuatklisu. a ently, tse aqm bautyryl CA 'eb; iimgsee vas
identified a a uprofvroprotesa to idtch eoer oaours as part of the
prosathetio group (98).
Soe oopSr sn te blood to geerally *quayr distributed between










the cells ad the plasam (99). 21e co0 er coateat of ery6thruotes h

been dmou to be of the harder at O.0O0075 per omat. Mh oem e wr can t

of the laukocytes ad platelets Is osldaerbly w ss, bout me-fourth
of that the rse blood seU (IoM). h bt blood pal go, 90 per

sent or mo of the cowepr tsts I th ftium of a blue eoger-protela
coontad teHMed fsrulna3den (101). 110 rmanIng plam eOwer is
thought to bto loosely boaA to a proteia slace It to ainaly namitalyn.
abne (Ice).
msw aL nvwtimtias with adlometlTe orner e hwe ben msd of
its sabeoptin, dIstributimon a eamretio far various species of mdama,
all tenaAag to ,!astMate te por absorption of aally aStataAtved

eCoper 0ouread to aInjeted eOqw (10B).



BartaiA am A Iaedil, aon (103), Ia 1913, m mg the earliest
workers to d r that mw e is I comstatly presmat in *t tissus and
mar s of ma am mlial. It has been soem by dalyiLs of livr emt

ki&y hm-ates, tht appre able momts of -ui- we loosely
halA It these issues although s me at It is magr fiamly bomd (M10).

S e amhmtao of attdmat Is belUeved to be pretwrtial afflaity of
nmsm er ft ll U l proteIn. flasently, mmmsne has beern shmm to
be as essemtal oarmt of arstmasse, man m~em of ltver, emosmble
atof sptttng arglaie to fem uris. and owrthine (105).
las uhmutmeous latro&aatIm of nrem s- Matt lnto "bbits
bhe ltA to their comWitoian An an wMs, mdaly the klt&L liver,
bwalp, md a espea y the blood (106). It has bheo da thO mMnnsme

Is moe refAlly abeorbed Am abAlmastered paraterally Vm *m tola









oaily (107, ta).
Uft a fe --m comtent of te Wbood me SMeGOn.y meaer,
e to l diffttrully coo ateed a ti detea m ata at thme low oue-
ttiaUs mam y present. roe mad coo-waers (109) stated tat whole
bhum ble d astats 0.12 to 0.18 mg. of mau per aL*. about
tioftld at of htif p reset ta the corpcles.



t presmacsoft tie as te anal.e ad ltver of m wa tirst
naua&bL d lfa 1877 (nO). Sines the, Sala bee beeI fmA il al oft
the tusmis aed cams of thm body that b e bmee a m e orw it, sme
lavnest tors believing it to sour It all living sells (11).
Bavefri erly attwpt e w e wriedt out without aucsse to dGoa-
stte a essetial foatics of lsoe I m im abuatlog (a1, 14
114). h 1939, asset .ercmaetal evlae hod this eament to be
preset 1In prosthtic roup or the e z que avbaoe ayte (115).
CaatOe osyesr.o eatalyss te reticm, I v + C0o "gOO
therefore plaaig a lmapartet role in the eid-basse equiltbrtis of the
bodo by eastabaysin the conversis ofa caurbso dioxide tram t tUassue to
esantse ad la Id s tde vl ch is sbeequeatly Ian Lam to release
eta atl isa tI e bus= (In)*
Vtlle. ad Gibsoa (a6) studied he aaes ceoott atof b b blood
ad fenud 8.8 2.0 aS. tof sa per lof tal et blood. 31cu1ations
hod that 7 per eat of ta total s in tIe blood aM situated i
at ytaeroeyte sU per Gt lt the ploua ad 3 per eat Ia th
le maytes. he iaSriaual tlukoayte, bowe w, coatamised tin.










mauh am as the sndivythla ryroetm. ater nomI coaditms, the
who1 of the aS a Ia erythrooytes em be amconted ar as aboa
ahys. (117).

e omcetrsatim oat gue In the peridphemral lu oo s at
patiemt with crtaft types or eaRla bas been sh6m to be gremtly
redued below ama levels. ljectimes of iAne glueo be have tailed
to rtise the as" level in uch cases (u18).
aSies oe c arboie asbdrame activity las yet bam sham ar the
pleam the Mh present In the plaa is thouaa t to exit la other
mmwr (119). It semo possible that Asos may oeuw ra 19eb.imtia with
oe or prota i, seone patlmst with albaidauria eraete,
ma the amgp, serm tbme as mao atae ls their wtae daily as do
amml laiyvtldual (M10). It has bees omanwtateA that after atm-
eaoiusly aInjecting a ame salt into a rabbit, the alass oatet of to
erythroeytes risem aly as long as ths plaim level ramanes elevated (121).

anIus ct1s yaA wd looda

Frgn ility of Urythocytes
ae pemmabiUnty of t e red cell aona to vario, eleetrolytes
has been ta roumuiy rw-ewed b previous warkera (li2, 123). a this
seetiam, madtimal iatmatimn w zu the heamlytie ffeeto
i ime penetrating by matallie catimsa will be prseseated.
Sthe earlier literature, the erythrocyte was treated as a
atiea-saeszernb eell (IA). With the advent of the iseotopie tramer
te'aaiqws., the ability of certain oastms to penatmte the red sell am*

bne w- Iastasted (las, ia6, ar7).










Potauism at rubt1aum hlorid 6 (wom sItrtud to be loe
offestive Ot sa a odi hlaride to prwevmtig omotle b~u sols of amt a
Wi od iheress nceim and lithlm dUloriss was aom to be -m
afftetve (i28). Although the re. cell meta s iaBmmmsl to Isum-

oMmr noaml ocoodtins', potausim is able to sloky t sverse Itt.
stuaml a*n* ofm Intraoelular potassilm is alvalmt to aorotd.
astely 1 par ouat pr bour at boWy t-mp tur (109).
BquMem and Croe (130) ound that ltAel earth astUms tn-
hibteld hatolysIs by both Mtatnin a bypotecie solutem a the
ftelMIBg Oa" of Iasamuag etTvity s sImeit, tSaiU stmratisu,
barst Aiamls earth lmes were id to thIeoan lb e7rthote ram*
tbmm by ftenmin Insoluhe mits vlth the VoqshtIte cids priest In
theS eSl al.
Croes (131) later foum that aloll earth eaten IamsMse the
rte of hbmlyass iloa^d by smeatf in t*h f llowAISa orers g myoIs
eoletua > stamtSum >bertsm. Althoui misulm mat emluin InenMse4
the rate oft hmisis aNed bry sae im Onwtyl sul4 te and 923n la,0uy
salttsA, they7 ha so effect as bo alias lams by oeolotrimarthy-
ammino m c r d.
& th preseme atof lA (132), wrythuots lost potaseia. mA
aomme loes resistmat to hmotlyus by sysotlodc soltL*MU a eaffet fa-
rowiing with m Saem.ne. la the aem tati~t of laL. Ih efftoet mas
att butedo to a sactian oI thea sell mlamen. At ocastt lIa eoaem*-
tratiSn, eM set pK ladoated potassMim lss.
)NM s-ttl (133) state th t tVo aotlon of mvtaUe set at
rythrosytes aepmds a both the moaentratana a ativt of teitr









eatieas. B boew tt ew treay mal, amomta of mtaes muffeied to
probene humlye"a us haeytis e ativity increassi is S fleowlag
oed stane< oamalu< irc, oabat < nickel, e tlmtin < < weoger,
erew,, s naiMum< P esm awt etitler (134) aoted hat sodium e hloreb solutions ASL
wame ia Otet with stale bt a hbaoyt efets. Soe taere of
hmSmates deopMt a he tId, fe tOeratur, Se sonentata et
the ret Mood erpasuwI, an ta tm durstlam of eastm bet a n he
frtal a" ti eslutan. sM Iholytie activity eeme to dcsem Sa
te fbonlwlaa oar owner > lead > slvw > iaetl > .ull3b -mB>
natimoW >ta >Ma&l=ma. e hbmlyolsts seme to Iaresse wv the
lutin of the ryteriocyte saupasas ad a Intbitd iftan aim am
sewbituttd fr the amUse s1lutins.
It h a bee dm (135) *at 1in, pepper, l ea, 4a Sae sits
*I affAted hbm4wis, with asI salts dmonat B Wa te bist hem.
lytie activity. rvlaenoe state r at mstal sesemA to be ristUS to
thme wuyte poer of t.s smt, fts lvre u valatnse state te pasrter
Us limelyti pme.
b6 a study ot th boWiatlyt actcla o epsr ite a reabit
U7rUtneure,, nUAs a* Omflrw (136) suXspa" aSdd eNA celIs La
itotalo eUea or mearos salut s. 2hy uW A efl per *etts,
citate, tear&te, bensate, bsaBwrogtate, at s leylbatvate ali
ateS siUtairl teyr hnmlyme the eythca yt. Is caatntratsse aboe
1.3 x 10"5 Uoa/lUterA, msued ealututstia Is eaactRatms Io tlr
.1 -04 to 1 x 10-5 al/Lt Iter m wvre amL-umlyti s co ansnttlimaM
or 6.6 a 1O- mNol/liter or iss. BSAy OaluA tet *s0 aimn -w ast










a t tor ia th effect of cwmw nlt. am wythroc~ts treted la suab
a *mw. Say at reported tt a t e s cometimat1ans lower
than 1.3 xz o mle/Alte bad no efftet t h reisatsm of wd R lood
sell to hkyptmnc wolutwns (137).
BamLyt t ha bmolgtie method for the Nteamaattin of vlies,
wtmaa d MiB (4) mapsted that slac sAulmte wm Ow v ed bHood
eUl mase resiutat to bomalysts It bypotaic saolutims byr stothMnag

t0 ary.eoyt merm. L7t I.a e d ma Ua (5) omatly moteA me-
usmal bdwrlar In hnmalyte sq a Mta lmavalving saa metates lao
Iombntllum ofat tse sIM =mit seem to protest s erythroqtes
apaust oimote bmlylss. Extrfmly hMa L values on obtatud for
both dlas ametate sad Sla sulftte tr t h lutwti at'r.

khtersotims with Protelas

mItallis cattme a saA wsntaimtely associated wita proteias
ethrou ut bi oiAC1ml sPtms. Ourd md Mlo= (13) enu aAud that
,t motum of Tarius a talpprotesa oaqmI difftt" wrstly absdpnas
am the 12-steristies of U two iadIvihMal oimamts. 0 stated
ott protel. ith their my remotive side *.t hare the *poential
oqSpwty to bbed mot metals ery lt*tly, so that ma metaWLs eatim
tht nateri a livla arpam SmPOs fa m prtnt pwrt atts time bmoUd
to potelas.
a metl-pWroten e sqln, sa. asa p ro~u as auzboayL, mid-
amole, w alaftryl are wus1y at a mw e lgartame I* bliMaAn the
metal ims tVm n ae te tuwtl minmo eat cutoxyl proae. Cha es la
0te eatfartim of a speaifc protein may armte mr site, as ta te









- Osf at ~ anu.t W*" W-- a law laese it ts btdalas epmiety
~e Em e A It to w eMd ft W9 to 37o (139).
almlioproteiu ae r...oi4ioa as a stretuMal twpe a tiek a
speufi metal atoem takes a Iantegral port Ian t structure of the pro*-
tea. he metal In such eases eamiot be mwe wiftaot destoOylag thew
proteia etrutae. s0 ature of bmtallfrotuas h remmt.y be. rxe*
viewed 3r T le (11)).
A t- omeal proisa icniplM ths mtal S is bo~ R urwauibly,
In m i f e abai l Weola supMop as se olu6 4MitU t am g E lpft1 m
Ub affect. L te aditim of metal Salts bose motal Sei sateret
Maggpmwaby ith th prote usually loaw twe saolubity ft ths
protein (1)1).
2seM are two ianl types of mtel-potean aCO Ms, simple
jl- Inmta h moh metal m ead o w with a s*a Itlgd gra1p
ad shelfat epl em In bifch Mna metal iea Mombes with *a ar mar
ad groups. Aew tye Sy attest the Molublllty tof h pratetal Ar
emalets, ialeetria hbua hMowLobla at 1K 6.6 a" 00 Is ak lesw
8*.OUe after the additim of SaIe state 0N0 In a s6mlal0r maIIe *Q-
talaag only aoi saIts (l4). the other b00, trea.(r h(),
Sa olate metal proteasu O sM i fee awb So limis V t metal
ftree pretein udSr eortaln caldtioe (Ma).
agme" i t eo M utbitUty atof etlprotea -pms- with o ns
la s* are fll fetn. At 0 i, to t preease tof 0.01 to 0.05 N. oaw
ustantims of l)aA m tlase s ts, it me found that Vi soluMllty of
serum "tua e greatly aees Ovor p 3 manes at 2 to 8 ftr te
maltes of lS sad 5 to 8 ftr thUs sa salts (141). ] th *Ma stWV,










the effeettvesa of cmse trastioal elments In precipitating him

mram albumin In the tI renos of 6 to 7 -w found to be as follows in

doeresag orr of effeetivaessu cpuer, casta, sae, ad nidel.

Ih mual, the aditim of mall proportion of metal .lts to

protein solutius results i the rapid fbmtia of very bul y preepi-

tate (143, 1 .4).

A on and W y (14) have stated that theo bIn Is typical
ccmaolable proteai, the proomessae do mtu t m n lng tho e of

other proteins. hey have MflaS that hmogobi Ist depoalyserid

into hobraon. these woreras stated that te damturatin of
him abin Is a reverable procedure buat tat the floculatia of te
datured protein eaks it appear to be irrerereiLe. They fouis biMn-

globia to be deatured ty est, aeid, anall, lea]lc mhnaleal shaking,

otasaba iodIdeb, potessim thiocyonate, o0IUa eallVate, ad eean-

trated us ealutionas.

Smad co.workers (146) htra me tisto d that rreible m tgl ie.
enmatutian, caled pseudoam$tuxstan, affects Ur proethtic group

oaly ternfar being fianatally differat froa ls datnstia of

other proteinsl. SThey found that hmoglobin deatuwed by flaking could
not be reverted beck to its artigal fxm.
eaiiiaso (147), In 1938, stualed tbhe effect of the ditvalat ias

of copper, siae, ad aercury a the protein of rabbit blod pigumet. be
determined the rma of pi within tich globin at oysbmolaDtiua ew

precipitated by it mounts of inc sulftte, eoggewr sulftte, ead
mourwie seetate. BLobSn wa evaqletely precipitated by sine Im at p1

7.5 to 9.6, by iupric io at pB 7.0 to 8.2, amm by mrcueite is at pR 6,3










to 9.0. Oholobin Wvas moitelry paispitated by 1so t on at p1 7.0
to 8.X, w eqwrc lo at Ip 7.0 to 8.3, and Bmswuri n at a 7.0 to

o0.O. prtpate orf th sainc-m mo6xiO own o ud ms ot a
nesdi color, remae~ nstd b far aths, asd vs redily solAe In

sitMr ilute acetic mi or dilute nmoI&. th protate t Meared to aee
suffered no piament .boM as a reslat of its pspitatioa. It faoWd

the amormi spectrum for oxyamO nA the MsI a es Gol be r-omed by

repeated syu brlatiaa with pure Iydrogemi. I* eavuri e-nsmLobus
compoaA precpltatted vith the minll inmot of muroMuie state e-
maimd garmntly mnohKwam fr ame time ere that peclptated vith

a iargm erse ot the salt omued the pi uat to uaep a *cneMp *aliU=
to the ftorasuta of pSiah-mtI. he aop..r-oji.mlubtda s alnost
i oeattay ohnpmd to a romiAh subwt nmo rw urdleM of a proporti

of tw eoer emalt used to precipltate it. he brooatoi uaotnmm Ia
a.kalln soluti ad a peotrum semmwat asiilar to that of a*llale

bhmtan oad oa rdnetim with sodim hydGallt th Spetruam m that
of amodiamogenm.




th epmembility of the red ell. mbimm to varlm s arpiao utd
smargale mopom bas been reviewed by roernt wmioer. (8, 18, 123).
Te purpose of this seetia is to briefly easidw th* p]amwmlity of the
erythroyte inmbwr to oargae ni sa so that a better udeMrstinalug of
s possible activity of the glamate o myu be reuahed.
Ow red a toi recopawed as being suite pemr nbi to m ter mao
to sOm amians, but only ihouly psoomble to eatimm the permu.abulity of










mthe m iome to aulon is estimated a being about 10 tms that to

potassim (I18). Pbuaer (IM8) o. azlsd that rapid volm eabames, ia-
med by bmgaig t eeamtmtmra a of salts or of amistrolytea In
the supmapin mdim, tad to be brought about by the mmaIt of umter
alami let s, I& eell Utds to behave as an omoa-tsr.
sBber (1A9) recogi-ea the -ameetuinm of our -muntn4las
of ey throeyt perme.bility to arALc easim. IU pWmgated several
eMctrs tOat wat W e cmealdred sa epsnina* omaotie ucmas of
Mel volume sit imolys4 these fta's mea ea a) wdther the oarolo
mels are surfse active, b) lwhtler the orem'o asida falmng the salt.
are strg or wemk, e) whether the orguiae aids ar 1pold soluble, A)
whether th la s an strategy hi$kmted, *) whether te toe bt e b oro
twrce or bW Amlimatita prperties, and f) hbather the es are pomisonous,
-etta either On th surtuas or ei th tire protopast.
anll1a ar a an m (6) recently reported tht the order ia whlch

sow orgaie aicus apeared to protect rabbit erythimy-es nLfast ao
otice molysi m tartrate, sucmiate >oitite > >.miniointse>

aoetate, miea yleate, mlD benmate. flr huma bood, the arr of pro*
tetila appeared to be tartrate > aast p...a.miaalio te+ > p.ema-n
beizoate > s.lUylate ) >bnuate, vtt t eitrte falli betan sueinate
Sod p-mni b deaste, pediAag aw caaoantratlon. VBy noticed tht Us
protective action at the orWp I auimo seized to Inureme with = In-
ease of polar roups presemtj iaeaeased hyatla, with slU r diflaNat
mad imarea.itnLy diff4 elt t wa uag of ibamm parew, ws offered as
a possible epanation. Nfdmtio of the wiaa eouLd. idtndly ia-
eremse the aefttiv ean omatratio of a solute by I'eaesiag the mount










at samilable tatr fIan a particular soalutim. Suah a msoaatin oulan he
Ia Iamreafed t1untWiy.



1emiao s of erythroqytes a hnspotmia mmdia must be dmtnamteas
fa Ut owneu ly wa e a JRe the a eqialbim i a tbood eosetitmats
wtl reject to it. B mettle fmnlityg of Me red blood smls w bet
Uereasead at aolr pembity to Varisa ins altaNed 1th a obamg ts

IB*
tIksm (150) ftouBd tat tim restsos at erytrmoqteeO to 0e8
hyaroa sm varied with the almla species. AS -m VAU eroytqa tes
om ae pretestt resiUteSm, )molftLi at $ 3*7m iamons ythe*
heart fto uwaftmts holdyo at $ 6.0. STa vrlatica t roes"tate
we asmried to tbe btet tht 7rAqtesof various Specte oteatn
nms eomss" at, prostaa vi ditffar tc oet i tic Paoist. *e em
%amcb sae mow prme to aggnstmatta are mIN strmaX erqasbcl aN
aoneyaetfl -o boelesis.
CArstibiers (1.51) elated that as bmoaytUo atst. of mdot sA
baRes SigmaA ae sme eritIAl lmnta e4 subse t ly tbe smlla 4A t0
which ostat aciAd csA la wemamds ls with *a eall proteasu.
AsOerstng to bArris (152), ienom Slsu plned a a mmt eam*
tafa etr miatmass which do not pasetate implSy w a r eaaaes
tate oft electrolf, there isa abmwg la tb. IatMcslualo 5 11tb
a pgatadl dLftnam develinas aronss a* seol Sa0m I eaqaeo-
jtes ra Intaotaed Inato much mda iam r to a mboSwM t iatemal
diwlide .U fsr extfmlt bvywl Ias ma the osf latertar albUnm










amA t extemal solutti altd. So al ianity, if 5afrva9p, am e6a
bmoly4il for this ree cells keep better I& modia sat bufftre
the Ian wtawl or ikalis sluUmias. Am cells beeaa mbemitely
alnkiU., Ote laiimata of etlr bhumagnHob it laresd amnisimltatilf
fewer aSsa to alataea ele tricl mutralty vita- the ael. Also
reulttag S a secrs ia tS Intal o motic M r..u of tuV aUll,
a de rsew las ta ce vaolel, a a increase La t resiatemme ofa tha
cel to lAs by hyrpotcale solauti s (153).
Rtam red blood cells ta smlUns misa omxtaliiu es tte buffers
begsa to rwmte ast p 7.0, a th rIete td ta S We ismemsetric
point of b-mot'ia, p1 6.6 to 6.8, ma ts cretimas pearst u mt
about i 6.0, below Vilah te sells -e aosits mUL they ft~s to
halyse at ,1 5.4 Crmatioa dar these cadittelUs appes to tobe


also morso n aoe p 0 at the eslls assme a prfectly soSb qaeriml
ftcm at p1 9.2 (15l).
It has bea .mstrats (1") that the Vl e f *ot wryooyt
gfafnlay laernas with tie attan of l nLfture sol ntA h bwlyais
results. amoti hbmaolya, bosgebitan ws out of e the ela
teas s amlt lUfeemt hnoLyjts a pmorton o bwe mao. n omuls
fix e in toE stms.
hSaksy (136) oftua E order of sffetimeve. of lacomaic aids
Is podelmsd bheialysi of bimam ad dog erpyhrocyte to betI floqhario,
Wl o, Ihyvalr-ie, o d aitric eael, listed i dr atof 4deenag
efteettrmmes. tomd hIsmun red aopryules to be les resistat ma
semUm ret eesl to hemlyta ceased by acidity. Me oaemtsmtilms










oAeGin i4iA bmolysSi, i.e., VE 1.0 to 2.8, thez a injury to the eUll

milmme, b aumd at which could oewur i owotle bm utels. *e KOa a
that Ia b@ioly~. lwmed b Imaoremie aeidc a reastim betmwm t* celU
cmatltuents mlA ti me a we eaqplet befwe hbmly la iVrt moribed to th mlpd pietratin of the *eat isnto te ll,.
A 1931, Ja&obs aA tWprt (157) mevid that s*nss ito s as
little U O.OL of a wit; mA tametume Taristlan ll as 0.5 ofat a
despee auldd brw a mismubl alefftse a t obsere degr oaf amat lts.
h a later pewr, paerwt f (158) dmmutated but by lowriag te
3p fram 6.79 to 6.59, a salt eaidntmstla t of 0.2 per aet ws neede&
to protest 50 per cent of te suAspndaed erythrocytes ftram bh yimag
wbwuas only 0.500 per eat salt oaeeiatratima WA btoa stead prior to
lowrfat h e n.*
gamIe 7ryetooytes hre be"U domU to beoms psmenbls to lithim
a iA modtm tas at pB 6.67, to potesaim iom at pa 6.8, to nibtMi ieBs
at p1 6.92, to 0s1IU io at 1 T7.06, mad to lUw oNam mtastimn of
amentmiu Lieu at pB 7.48 (159). SWe eagm of abhlride mad ailflt Imea
thram the elU mWem hasb bee aibomm to be lomnd by lomwrtag thO
o f selutim Isa hiMch tW e I a* sw pulnded to batel ]p 7.k (160).
rman Se Ilseur (161) foud that te cat red blood ol hbs a
huap p1f optlms (p1 7.2) far *eouim pmeM 1 a ty te effects otf an
petesbmi peu- iUty w~ere smU la oceariso. ow eat rytheoete
Uamily eaital a Ma mtrtla ot sadum as its mAf Ibtr.aelulaw
metiin In ematrut to the hium ew7thrUoyte whieb estelsa mnaly
potaetLm.






2T



B$ aoaeaantxmis of bzdzoin Ios or bprvexrl io. re
alable of proa&Udng iwrrable abanmws In tbe arythaoyte thmlvesw,

a1thouds over R considaabei part of the VX rmo the affats re

rvrmaoble.



















dmWe m1oils usmd i this Iv stiptmi vwe obtained ftra aem-

mereaal sources. 2e abhlaals are listed In bl* I a long with their

mauress, fonmiA, Sades, and lot tmbers. Th abbwvlatin.s usd and

their meaning are "s ftllomws

A MA Wg a sd Chmieal Co., am.

C chm Po ua m eturs Corp.

M uhlmiackrdft Chuemil Works

M Imerok ~d Co., h2.

P Chas. Pfiser mid Co., ae.

Vethods

Colleetiau of Mood

Oe rabbit blood used mw obtained froa healthy mlie ad fmle

rabbits by heart puactor. A 17-sw m, 2-inak sterile dipoemble needle

atta eed, to a 10-1. sterile syrtna ms mployed to abe a4tznae Into

the heart. eM cardi.e puacture was adl v throui the left later.

oostal eeos as meai to the sterum me possible In order to avoid

lajury to the plural cavity. A rs =tely 10 al. of blood Wa with-

drMm and imeadilately dafibrinated by gatle rotatica with 10 to 15

Lass beads contained l a 50.*. Floarence flask. After suirilag the










U 8 IH
UMD 15 PUFARI SOIMS


Cmlx Souras Punam, oane. zot No.

Calelu. Muecate A Ca(C611O). UMP W 0,1716
Cobalt umuseato a Co(CbOU ),2hago efnined 8380
Omar Oluamte P Cu(C00100 Bltiand 5996
ZIrn mlam te A e(cOguo07)2 o X n53797
abawetim lumcate P Mg(C,)g2*2O %0 flnd 5208
tmameue MWueaste P e(C Oq )3HtO Iefined 48
Potassium Olueamate P t Rtaefined 51368
tiod Carboaate a %tCO3 we t CJ
Sotim aloraides Mw Nam Iedst 51587
bodium 0ueamate P 1x, nBetan l 0a1118
ZaS G beat te G(C6 ac O7)QgO Meflted 7te

a soanim chloride for biological wrtk.

flask far sprodmtely five minute, thea defibriasted blood w ecanated
into a Xo-Wl. rlsmmwr fltak sea malted by entle rotatian of te
flak tar about five minutes.
fte hum blood w obtained chiefly trm the veias of the arms
of a twiety-five year old hiate mae. Several e iMtm ts wea conducted
aqplagys blood damated by mie. volunteers. Ot blood w drman f9* the
am of tbe daar w ith ahypodewmic yiesg and needle and treated s the
am. mener as the rabbit blood.
Presh blood aples wre used througAout this lavestigticn.










Prpratim of Solut1is

8alutiene th micl subs tens wm p srud br vwiSM a

Mft proper awots aof t ehicals Ad placing tIb in vaolmtric flasks
uhiak wae brought to volam with triple dat1l1d water. AU of te

delamesm oaemBtstion wens os tutd In teus of tM omdgLrous salt.

AS OW9ndmte 4dilution oag k a g- teuume for sesh sIlt by
comdotvag a pr3lQalry eprtamt In whihob the eco tBtixt o at of beleal
suffleist to PWre t bhmlysls alnltd m V VSI m m conntratan WMr t

without preventing bmlysia coustltuteA the ls ta of the a asired roua.

Various iStems4mst ate aomtatams vese i n prared "A the qpsllmtnt

earriet out.

2I eerimate .VloyIlg soluticas eoatalalag both sotdius bloarde
man tM Chiol uatr Iave t Aittlca, tUh am proed.re wse followed

quMtatitatve etelunaatico of
Par Cot BIAlysls

se ithod uned to detemiue the & e* of boly7ss saa @essen-

tlly that of uater (162). It Is bseds m the ftnt that the guatity

of wed, FytmfglbIan *eoed f*= the .rythroot7s by ha7pot=ai alum

tim tI a di ect fuawtloi of the proportIca of ira es paseles hmnlys.
s ainat ofr ogqa. ol n llerated an oepltate baum slo, by tie

lahag of mrythroeytes ti 0.1 pr cent oadim earboakste oluttc, repre

seat unity ft preaing the dtarw oa hmolyits ocurig In a given

strength of Msalt solutli. fite amtitative atnmmtie of t Pro.*
psrtia of bweolyas t a salt salutima of owy Stragth la aiateved

by dntriftglag damm ay -utmnlyise ells Just prior to dtsumniag to
about atof oaM iobin preset a the yupmuatat solution by anw oft










a photoelectric colorlamter.
OM method espure a e Ssriaats eamisted of volsatreatm y
t oerftg dplleate 5 al. portions of standard sodium ahlride molutiao,
rsmgIg a oeaseetatioa fro 0.32 to 0.h4 per eat *for ao=u=as blood,
0.94 to 0.U per cent for Ngro blood and 0.32 to 0.58 per ent for rabbit
bloot into colorlster tubes. to the e soluticas, 0.05 Al. of fresh d
fit eatA, amteSd blood was tda at the tubs em crefullyt inverteS
Mnal taime to obtain oalqte mbding. ae blooda-ast m wtures wre
thea paced to a watr both at r for forty-ftive iautem. At the ead of
this pwrtod Mt ixtures weo estrifugset or three aautes at amrcia
lately 2000 r.p.m. A ettamreaa botoelectrte olarmeter with a
No. 5g eam filter w usd to measure e amount oat 1 t absobed by
bm spummawt lItid.
Coaplte hbmoolyis of hbma mt rabbit blood w .ftfetetd by
plains 5 *. of a 0.1 per cant sodiua GeN ate solution ina calortimter
tube, admtla to it 0.05 al* of blood Mat iang. After allUtag th tube
to stand for the pr sted fortyefive aantes at 3*, tem preM astia
wus on itrfted for three falntes at the mount tof lit absoited d-
teaUnedt olorimalrially. ive such tubes were prepat Owr seam blood
aeqis, two being rat at the begiantng atd three at the ad of the ex*
peraistal rn. Se sverae of the five resdingW repreaated se 100 per
aot hmalyats value atd us ssematly emloped In i aleuinatg the
degree of haslyis atof the blood t the various salt -oB-intatios.
A bank vas prepared by playing O.OS Am. of bloodt I 5 al. of
0.89 per aont salsn earidse solutida, allowing to stand hr forty-five
anteis at natdifuging. The aolorlater -ms calibrated to mero by










utItt a g tm ruah s rpaed l .nk.

ow per cet hmilysis oomuring Io dilinete tbe ofat each
atrngth at salt salutim wa calculated by ditflaM the ar rei eAtn
obtalain tr the two readlAa ot each alit 'eNseateatia by t the a.era
rising obtainS tr a the eoplete hmalypis obtained vtT softies ouawhate

e autta. tee ealeulatis m.ltliied by 100 ea rersentd the per
cent ot total bmlysti to seem a strt of msnt molutia.
ItrIatims sto erytbwrsyt aOeotie tfxilty mto solutos of the

m- alt eoim tratis with 4Lff rea t snqts of huma bAd has been
aotea (2). AMy Mr due to this variatian Vas uinmtmd by detawtzfag
th Saigrs at bemlysis for each blodt mple ta sodtim Sloride esluticas
ofat varylg eaemntratL. tem a diaet ocaprisam bet... tb dertee of

bho mOt oceurrtng An a certat. o amentxsta of soim ablata saolutlan
ad san olution of eme other malt was obtaldA.

Deteinatie. of Pr Cent Nmlysia
enb dealing with Caloen St lutime
21 soutams atof weral gluate statust d a t this wk wh r
oClalret. or the qatitative dotxnatolfa the per oeat hamlysts
Ito dealing with .u soluteans, -e of tw mahods aM Gw luGSd do5

peata a uwthtr w or t the color of the solution proetnd a am1tive
eamlrlw=to reading with oqba gl rMo a.s
3h the first imthat a!t=iml* salSutia prqeast by making
erythrotes toI triple stilled water was ad&t to lie amnts to two
colotasirn tuem, a eaitaning 5 Al. of triple distilled water mad

00 other oitaMing 5 at. ot the colored Slumaate olutia. o a third

tube, coatatsin 5 a&. of the colored solutia, a poartiam of triple










distilled nter equal is voles to the oyhgmlobtea slucb planned
eo s ofr St firt to tubee wa ened. as colarsntwr rnela of this

tir t*eA added to the readne of Oe tube oRtalStle mly war and
aqlhLroe. a we. -e .omared to nhe o ft of t taOse eia St g ae

sa a. atS m almed gluoate solution t m tf th eanelaw e4

within es taomtal err w the tw colors wre odenreid to be atitiven
If thie re o md to be te can, olarhetaer readings of gi st.eaa t
sab lutin e uade before the ad tlam of blood ad served as Mb ts.

MBwr vslas WTan btatcted fr the colrtnlastri readlag obtaitnd

after te blood vas added to pezuit a aeurate de te Aation of mo
amuat tof lt being borbeted only by the oq, obtan pwresmt is t

supentffit 11quid.
If S. colors of ;osbamBglobn uad teS guw ste saoltite were not

foud to be adittive, the smood of ta two methods erised a-e lqaoyd.
& V1. .mt aod s per eat bc rlysls as detIeumad by trso"es, that
1s, from he Sauat of o;temglobn ma lait Ins the S .ryt-tr ybes aot
yt bawllyus. Uh stBaderd procedure sloqep Om emisuu with ola-
I" snoution as carried out until t point ftLloalug 4triftgticm of
*S tubees, just prior to tS tis hat S. oolarmlmre rndlag wea to
be takon. autead of -lx the reingsB, th supsatest caution of
tas colored gsbu to d an xtcon frCMa labed ells w asmtienlasly
deasted frtam eah tube, beang careful not to disturt t ut imx lyed
"sl* elitgtg tan"foualy to the bottom. Ohe w hne yd ealls wMre ten
wiMed with 0.85 per ont sodalum aoride saolutioA, r*oeetriftaged at d-
catd, the process betag repeated until S.e mipestant liquid free
of color. he supezatat lipad of the last vahwamg was demted et










0.1 per omst so twe mobomate solutima adde to a ok st ndd ot VW

eLoa to tu me, MaOotngs avO of 5t.0s al. Ia tb was 4ently
1averted to Bnuame emaqpst beaoiLrys, th *ost *oels aMnttfted dom,
mat tas remati eolr read os *a coLmrlter. m timo aeg the
m mmage ria of tuo pintleaw t b* ftre the readtag at 100 per at
bhmilyss, ash coaoratrio value of the I o mol*My ex amd ftre Ob
cells aOrgInally bvolye x wa obtatmed. Prea this, the per omst boMi-
lyis was camulated i the usuAl a r.

alMUleatim r of Vlues forb Vas a pswe tm
of Lootuie Solutlons
& acor to etenuise the vablue ofat the giemastes used Is this
studj, the per oat booml7es of a blood as le Io ryla o m tiM saa
Of odiu hlorid sb 003V ONtoas wVll sf vryla Iaestns of
gLu4mts solution a etesmined. y plottlag thoe pewr seeot bamlyis

asnst the stmtb of t *he olutoa, a sigBoid tMpe of curm iaor1ably
resulted wih soaim eablmde soLutimns, and usmally ftr solutlus of
~thew elt.. "riee thea abobrio the degree of heolysi Sa bet
*t goalm ehiari mad th gLunanate s olutlse, asthe a etatinu lta

Oni/100 M.o at so&=m caLorid md 0s glaiate (emuaiag 9, sO, ead
71 per cent homlymia) ween obtled.
lntag the eoam tratlam of sodn aharsie may other ma-

ponud gibiag es -am bsree of at molys, the bmoljt1 L value fr the
compound balm avestigated way be alculated aeooarttas to i follotag
e9"tions










(1vale on. u ofB stl mwa or or other
Na m A toI &. atr om-u oae a I1 o
( --.algauuw = N> vs1 ..- __.__MaJ2.S2at5
(ak. -nd iar v8Ltt Of seCt) (02. -OU2eauar VOeWt o mpoumnd)

Sh uina th abov *qmutim U s vatu. at L ft~er sm otim d ie me take
a 1.86s, th mompted vai e for 0.9 pe c nt (0.15 X) sotum olrwft
soluta. ni 1 value of *ott= hlarids s w almA1m1te fsm sactivity
oosfearts. s 1 vilue. of t nmateoa uaed ta tbHu eMIm tr
w l ulted ftom onm ane'tma aiulag s50 90 a, 75 paw' emt
bMolySia Of bm MA XsVit Xtaroesytos.
a olaulmtatisa r v fa- or Opna- stes lmtulnSa ea p pe mme
atof 0.2 pw omt, ou chlaride4, 0.2 a. pae 100 Al. of sedi, dlstried
wa wftrtfrme, tha eosatitrau s of egaodm 'hiarid oamd" s, 50,
mat 5 par mat hamNlreA. s waronsa of these three vase me em.
siagmd to be a rpesmtastve value. A mltala of two ah r snp
vabums wm otaidme for bet himn ea nebitt blood for eam enemsa
astemta L



h prelataiwy hmoliias eeimati emplejyla -on l asats,
a dbtmlalg of s ood sw dbMervemd a several osoulam.. It we oug
gsted that me possible esm at ach drtaBna Mas Ow saidity of the
oblutioms bMas tested (163, 16). Oase the atire hbulm I menthod
1e da a1 the 0 omunte toioArtutric Atestauktimn of taberpet oQI-
blomWSAo -7 ete woulW soe aub dbtasmxnitati warthLss.
As qamrimMt ma carried ouut to detenxuniae tha mas asMeiity that a
solutica mwy bmo vlthout alterlag the olar atof the a-h e-mna









1b rated air m holyned wrytroytes.

t volwetrasfly adding equal quantiteS of bealrness d reait
blood to test tube oatainian triple ditilled d mter Ma to aeoter
tube fntatia g a equal voue of aidifned triple di ed water,
ootoarisrst oaarisoms of tSe liberated ovabimo lei w possible.
Svarying t degree of acidity of the ter ise sriee of tubes, te
S t aich the lood e e to produe the m eelarlttot reading
as *m tube contatlfl malted triple ditllled water enaly


2 other eaerimt, the flaet of incree g the hiba gm iso
Sce atratla on ha emlysi wa m dete.u t. Me p mw aoveret we teat
ahlo woult salude the O value oat thn sta ate solutis Cle were
mare aidie ta the retrane eteia chloride solutAis mpLos thi
rap vas I 3.5 to 6.a. 2w method lLod coamnstite ofrc volimetrIay
treastferring tpllate 59 al. partions of standard sodium abladte *sle-
tiaus, rawnI n to an ttMtatm froma 0.38 to 0.58 pP oeUt, na t otolo-
rimeer tebe. B2 pf values of these solutions wre detamlned with a
battery qpoAmte een IN metwer. Zibs portions of modiv dslaride
solutions atof the -m caGOtatians but acidified with eomotrated
hyaroahloari aid to varloua degree ofat aidity re voLvatxlaluly
taeftwd to eoloriMtmr tubea. Ib each of the tubee 0.05 IL. ea fresh
fibrtanated flbit blood wm addd a to tubes carewftay Iavrted to
obtaa oaplete S a. ahe tubes we then placed in a mate bath at

3 r fwrtW y-five amiutes after vbt h they are treated i to e usal
mearn for the detenulnation of per cenot hemolysis of t es blood isa soeab
tube. JB plotting the per cent hmoelysis asust the oamoetratias of









t4e eolutias, lgootd curves resulted for both te date btalae4 tfr

Us aidflfed as vll as the mddifled sodir ohlrwld solutml A

dieet ocopart at ofa tes of the ourves an tber relative po

sitios prodded a meas of attributAng f portion of the hbmolyss oo
eurrftg Sa s bloot-glueamats ulztumws to the hydrog toUs omm-

tnwfca.

Dhtaindmtl of Ietal-4hvotin PrelttSlm

satmre B =att~t w1 wae to deteamine hmla te value, a

pr.amterr test we am to abtemdne iethor or maot pr-elaWtt *oa *e

euma *m a &n lUtMt of wqe mcd*n me ad"t to a solution of te

luaomte. fto per set saolutis of s&i glhamate are sqpst.ly

mixed wit equal portias of oaqlmoxgobin solution prexpue by laklag

wmeabt .r7th0ytMt, allowed to stod in a wtw bath ftr forty-tfive

=ttWts at yy3, omtritapd, mA obaearvM for einme atf p'ssipltatla.

Darflag M couWe of a bumlystlc sa, muh prelpttatiom aouIA mae

the oljarlMtric reaifnp to be ppro .aimmnafly lar tbam tgo Uihh
woulA result tem th prqporticm of nOqhowalm actuaLly reLemsat

fram thM 3.aM erythrosytes.

diitimal *mp'rt1mets ur om rrieid out to btestlae dutesr or

sot sm- protelas are pan4ptattd tr *b* gluemate molutim.* spr

we m.pawted Omr blood e.1a by entritfgttp a t wad eA St lihe
poirtls to 1 per cut malutims of amh aluoansat. S mixture wom
obearvs rmpeatM ly fr fmrty-flve mlutoe. &artig b tim they *W

Sn a tar bath at 37.
nureng each stamainatla of a umlysta valu., tha g auoatoo

blood mixtures wre observ for avidnoe of preoelpittscm.










freslng Point -etminatmas
fte aqpatus used to detaarm ts freesiag paints of saouticas
at various salt ofat global acid me that of Bftal (165) a. mootfied

by ftm and Adm (1). The modfication of the o r nal aopRtus w
merely thea *aatin of the toy uotor employed to run the atlrrzi


ha uwau tus, in pu t, coasisbe of a vaomUa DmRa tUo, aU B.
hi atd 6 mn. Insidea dlter, beld upriAt by a rag ad rltamd.
TSe b*e s fitted vith a nrezr stoner bmrtg a alvp bo*te sn th
Meter mA thee smiler holes weoams the ed. A baMry gLas tet twe,

about 2 an. o=4 Mad 3 Ma. Iastud diamtwr s fitted Into th.e lri

bole -ad passed oIB into the DomR tube. ato two of the *atler hol
wwm fitted aer tubes, ae tesinmtiag jut below t uMder arte

of the stober. S othwiar o wper t* xa to the bottan orf u amr
tube adlMa Itn eoll, the eoa beltg psifsrated with a s mxe of Mull
hoaes. Ihsde me test tbe wa a seemd sgss trbe of allatly miner
dim -im a wad wported by a atof rubber tibsin hpla between he
lis of the two tubs. ere wi a very =U11 ye bete tahe msurftmes
of the two tubs. Al ortnary labomstoy thIammter u snMwt%& into
t fourth boU In the staoer mad ws used to observe t- itp Iture of

the outer bt darlns ftwesiag point etersnatlmcas. A salomn dit.
ftewmtial thmmter with a platim view stirmer looped loosely rouod
Its bulb ws placed In the inemost tabe ad hold by meam of at a rbbwer


Prior to opwstlas m0 agpmuts, twe eUled enoer tdb us eam.
oeated to ta sit lain by of rubber tubing. s at oaqw toe










Mas fitted ith a section of rubbe tubing wbich tweaIted out of doors,

thus rvita s the exit for vaporland ether. About 50 al. of the soLu-

tien to te tested ua played in te Iieimost tube this .aatity es
aftilelat to cower the b of tho Saman th -minm te. A quntity at
ethyl aleo a as ade to the sddle tube unti Its this w rv to
a leel jut above that of the m iutna ln the ta tube. f the asr
tu*e im a libeoal su19 ay o there, its eurtse* qaeoesmeatay lvwal ith
the Oether liquids. Barig th4 dot!iIBtious, sir MS psed trou the

ether casulag it to evonmste at a rate strolled by the Iateinty of
afmatio, s rporu.tima of eter betai th bauis tr the OwooUag We*


9he t-*e ture at ther basth wus lowered to wuo aINmStely
*] bef tha e us evidiae of .armat of the Zri Oy elbmn 1t w

wkn fibE- Mmetor. Jk order to frees tUs solvent porktiU of te test
solvitam, It vw neiesswry to reOSe te tV arttuZ a sther b6th to
aOnAt 4.0o. NY caefutly controlling thIe sat of asta t, I t C oolna

proess s paremt at a lIr stad4 po*e. Sr=ng tie ooAUng apeustioan,

ewb -wIpe ws stirred vigroualy eA ooatumaly bat at so a to
stribe the tmmter Vblb or U earry was air don Iate to UA4d4a
AMB the solvmt beWA to free sad separate fra the slautiaa, the

stirring ats m a eInMreased a ntiaA until a tatiomery trantufre
uws reaeLaL aeuronlIng oceurod In eaeh cases I**., the t-,___tu-e
ftell below te freesing point of the aoluta O th ni roce suely to a
fsend ponlat sad rmiad ther for sevnma se ais before felIling ome
apn.* Te teMeutuiwe of b athis pibItr point isr 1ssorbea as *
freesia point atof the solutiotan.










nve hundred attI11lit of triple distilled wter m ued i
the prwparmti of e46h of te 0.025 #a0 0.010 a03a noltiLonMa to be
teute. a ftemslag points of tea 95 0l. Magmes of 00& malutn we
!.I-% =a rowedm. S VoAt t a R I es or "as emm ait-
feamtila thnam ter wahi r mres ted the ftrefds Pift at 'vter we

obtained frw th0e mw. of A5 prste amtSesmafttlms. Bat t bmanur
waus eu~ntely reat to 0.0L of a depgre sad etlmated to 0.001 at a
degr-. Sa Dwje= temmMur- eu fitted vwti a madrifyl g los to

a1i the a pestar. Iw ftlsag polat deqr"easat of te v sriou .elutlarn
wer askB dtems 2sd by merly MIbtractI a the ftreutas paftt taw=-Mtiwn
of mter ftm te ftreassa point tawwmtume of emeh solution teste.
*0 rrosasi point data obtained were emayed in the -m-leinams
atof L vtalM mm nsg to M tram"la




where AEt is as abg In tM fretreas point of wfter after th elm

ditise of Solaute, K t& e Mal ftweesIa poIat conatit aur Mter (1.86),

Lie tme vw't wJrr ftttor, Ma K the mialfty of the solution.
ats at oA various bmalytic adL ftu*sig point tnalatl
s- prewmted to the fOallowRug tbles.









II







i
ii


M

f~b
I;;


I
^ t


4










At B ^I

i ttI
s eVtlf> t a~ wy


j att 5 I
I
j* IIA 5^5


a & a a ss
ses ^ ^












~I







Il I
" )v) if p| ^| pP p
hu l l^a^Pf(aW
sa % %%









Si
II

i!
Si

i~i


*


as|
'1'
p|

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V.S


I










* *s




^S S S[

Sil i s I
^ $ S 9


9s 5









1*


5





*




I


* *





* *


VA
I *
p



p




II,


p


I




47






y a ^ $




J I s a (a s





I
as- Ok


sess AM







I. o 0 ,'4 ~4i L M o
: A : b U
o w. m .0 *o

e < W M Q 4


I


Si-





I

II

I
'U


BJ




I!
II
Ig

I
2


10
* 4


i404
* .4f tf t oi c
;~ gR W 8 t
*~ N i^ '^
MA *R<1^*a t


: SS t I ta :
S*SS 8 g ag
aR i s g gi a a p

"awman as
Qo o eQ do oo


* di -*

:11

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s
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:~ :Go


S* *
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'ow
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1,!



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l

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M* I- I-


Ss ::1
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m 4 co
aV%

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* *
* 0
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)


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at
0 I
0
1
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At lit -x A X 1












* p.
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* 0
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a

*
a


00
a




4

v~ a
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a:


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* C


* 0











00


00 00


a *O
R : cs a





g a u*?





g aS a *a


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'4. 04:
































II


o t~ ~
* 6f*
* art tw, 0
a a a I

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o
a
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p9 S

on.

g gu1


mmO 4
0 8 0



R 5'








8 5 I

aA f

IS i

as 8
die j


m










I
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0
4

5


a
s


V% W .4


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d o oo 6 dd


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0
0
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a

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*
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Iv, 0
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ti U%
p.
fv~*
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Sat

* a
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: M

St ^

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o wt en us f
a ti: JS


SR ^^ ^


S MS :

f0 t- NO q
*O F

r4 d % V%\0 Q
M@4MS10#


4i

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1I


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1 0 i


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* 9
* 9


0000 *OO~OOdO a


,.* a
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8 R:

3 4
at*


t &


U


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Iv'

t- 0
'0 p4

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lA


4 9 A 0: 1 9s t 5







0'O0 0


* *
1^ Go 0A


t co
* 4 4
v0 0
000


I


I



ii

ii
I
I
g
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ji







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s : s
9 9 t

Ia A 'A
J a *I


000
dd f


a 5 ^
3 S $ Y)


1X :


0 f~b
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% '*4
'4
ft .
as
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0 0

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o.
0

I
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a
0




0
a

*.
0


3 41 3
C 0 S












o l .. A 0 N .% 0
'0 ;* 05 w a

r : : 0 :




|: + .. |

, + : : A Ma a i s ie i
S S 5 5 % I 5 .9












*

00000000000 ,+: m
















0 O~
0 0


* .-











W :e





0



0 *
m *


o
0
0


'0


COO 000









0* '0 E
I^ 4 M

S 01 0





* 9 9

^ 9 9

01 @h W1
* 9 *


Ii
J!













I



I


* *~ *
\ MW 40
VmW ,40

oQ o o o







0 0 o 0 0
w *
* *0




00 O *
e- so fr- r U

^ | ^ 40I

000001 <( P

101


00~ 0~ 00 00


* 4


5 8 s


9 5 9lga
0 96



i : ^ ys
4 6


4.

II


aa *

* l- e


J? |
0 6 o


m X
O 0*









% t V%. 'O % 0 v M










o o
&i : : "< 4 4

fl I
j 4 O Qi a O QQ
8 j i a a '1















Is



I1




a

B

s




i'U


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At
a




8




01
st



I

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g

q

q





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t 9
0

1~i4


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^a
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SA


Ao
& 9




%0


^ 0O
9j 9
*m 0


0 a '
* *

o

a a


0 0


- *

a *


0S0
d 6 6










U'

Lw
0
4
0
0
4


'I8


8
Ii














'


0 0


0 0 0


* 4
* 4 ,
* 4


I


I



I


a
!t

J

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1$1 s 1 a A A
0 0 t ; 0 4t 0 0 0 0


I I I I A~ I I I


sA A a I d










uma 6
YSO GO WEAZ3IS OFW RABW ERCEOCI EB1 V 0W e AMMa
Bouswma w VARIOUS cImIF nEL s




s1 amt N Mse I a m a amA 9 1m1l, 1


A. 99.7 100.0
0.90 333 36e
Av. 96.6 100.0
1.00 337 36
A. 96.3 98.6
1.10 a89 290
A8. 2 85.0 90.0


6.6 7o.o
1.30 130 154
V. 36.o 40.4
1.40 a 641
Ar. "T 11.1 3$07
1.50 11.1 u.7

-. 0.8 2.7
1.60 0 0
Ar. T 0.0 0.0










SmU 7
C1-1IASBIM = G.L/AOO KL O ODIM MM AMD X-am UaUm3S
Cm 815, 50, AND 75 TM CaM mUmrI m
OW mIItR I21-OC"IP,


Sol Chlarlde



o.18 o.1,Q

0.391 0.4180
0.o39 O.U02


b1m Wadte



1.36, 1.36

1.g5 1.87

1.16 1.18


xar 8

TAUM OF L MR M r A T zm ArmnTMg A cnmRAM
CAWBM 25, 90, AD 75 PR CME =MOT=B OF
RUM mnclTml


9w Cmt n~l

25

50

75


mAot sMle. 1I

B.17
2.25


2.29


aoo6 nEmte a

2.19




2.28


So
50

75


---- v


I










TABLE 9


o CmR aimozas a cABI sacmxa ranw
0.4 r CMW SO==a Cmrm M VARIOUS M
8083m arUsvwaSe


8aaUX's


mmu Om


tlmaef ChlorAa 3a 3 iiE 4 !M 3 epl. 4


406


400
*r.

3m5




Av*
hr.."t
960


130
1*

48
Ar. "
0
0
AV*


*^
3aOL


360


365


30

a






87

4
Io
O


100.5


96.5


95.5


85.4


3,.7


11.6


0.0


100.0


100.3


100.3


96.4


89.3


67.9


23.9


4.9


0.00


0.50


0.60


0.70


0.00


0.80


0.20


0.90


1.00


1.10


0.80










TABM 10
Ca ATIBs GM WL/iaoo ML. or SODIUMl acmu
CABMS 25,50, AMD 75 Mm CET maUSIB
w main KmanoIIrn


Mlood S1e 3
0.4"99
0.437
0.A17


uooaamou 4
0.183

0.472
o.AW


-muB


SS, 50o AI 5 M cm ma= Sr nun WA 11mr cOTS
u ilplm or 0 o.a mR Cmw amZl m


95
50
7,


Lo.0 s Im* 3


0.848
o.760


am&s al. 4
0.998
0.947
0.878


25
50


mw.!



















-ma la
VAZL W I% SL 'U a GLUMU At 91gA P g
CAaI I, 90, A= 75 NS aE nweamsma F
omia aBImeo aa gw jame
0.2 m am fn CrRAm


Am M aiotda Load amdsLw 3 ,oot NeoLv s

aw 01.94 1.97
50 1.9 1.
m? e2A. .2.. ..01











wE 13


pia Ol aMUNs r a Om I magXcE3 SODU som wmC
10WICB Qy VAIEOB COiMl1R ABM



puer o t __________ m__ m____

_mme _.. -. amene -1, .. l 3. ir Amela M
Soo Wood d Mood Moo

0.80 4 91
AT. W"1 97.6 96.
0.90 478 475
AV. 974" 94.8
1.00 49 69
AV. 88.8 O.4
1.10 0 20
9-.9 39.5
1.90 1417 89
A. 27.7 17-8


AV. 3.7 8.0
1.40 0 0.
Av0 0.0 2.2
1.50 ... 0
Ar ...* 0.0











c MM iOL/100 M.S W SUM rW D saa -A l a GUm==
C-M 50, AND 75 MB C3 ROM==
0w Bma SSSSCSSS


P O.N0 0.1t 1. .f6

*5 o.awel o.,8 lom..us

50 0.34 0.373 1.13 1100
7T 0.371 0.365 1.06 1.05


SMUS 15


VAUtNBs aW L s W a Km a A COma S
cuajm sO50, AD 75 PO CMan msBia w
-m 11 1 I'M=


Per mat samlyts RLoo 3ample 30 meBod MgLe. 31


50 2.36 2.3

75 .13 a.4
kAwm 2.37 2.36










Mnas a

0.1 UR M AIMN= CIMACI AMD VSAMTOA F



Per coat Oat %Mooa!aine
locate a.m. !I. a.! 33 !..vMIq 3 Iu. 33
0.00 0.80 49w6 4.
*.' 100.0 Io0.o
0.0.40 467 399
hr* 0 93-8 93.6
0.50 0.80 o 380


0.A 0.a0 3M6 22
L. 77.6 9.8
O.to 0.20 o 1


0.80 o.ao 6o


1* o1 0.0

.o00 0.80 0 0
O 0.30 .'.... I0.0 0.0











auS 17
Mammm au WM./oo Ms. or am acmanZ
CAom 95m0, 50, 7A5 r am P
p -m Im aroBrsn


Pw Coaftt Rmou is

25
50

75


lood sM~a 3

0.43
0.410o3
0.38t


0oa. a e 33
0.396
0. 3B

0.368


O 50D i m am= 18w am
9, 50, Am m mg cm ]amau s ar umm II.-nn
v us moms (w 0.2 m cm on= On=


1T rcns mmoStd a*ae 3B stM 33

o5 0.757 0.660

50 0.689 0.60,
750.595 .550


b


. ..,. .



















TfAl 19
VAUX= jrI6S R BOMGUMXAT'
QCMBgg %j 50o AN 75 M CMH HMOLUUX Mr
Iwo*~ IBKHINV 1E Ill." plBI jmU m to
0.2 PAR CM BOOM CXamm


Pr oamt smdds Mbaood 004 l32 M.on ameDl 33

85 1.96 2.06
90 2.05 2.013
oa ao3

2.O0 2O0
7, 4 .. .d .
bw...e. a3s .0












PM OW m an OF A a1 1 1Vm VR YOm


1owant5 9 mptls 60 3 flW 6


A.. ,,, 98.0 101.5
1.10 2 295
A. r .o ,9.T
1.20 191 274
Ar. 64.7 83.9
1.30o 2 24
A-. 45. 61.9
1.4o 3 110


1.50 90


1.6o 30 0
Ar. 8.9 0.0
1.7o 9 0
Ar. -0 31o o.0
.80 0 0
Ar. 0.0 0.0










TAaX sM


-m0WtWSiOM


to W./100 ML W soIM C=U0o=R MA=D OSUmU oJMUhAn
CA, MMI 550a, AIM 7" IM CM IMnS
omr RAM sm ivy=


Sodium Chlorie Ptesal GLusat

.ps 5 a~~mt 6 Jal il 6
a5 o.iy 0.412i 1.43 i.ka
50 0.388 0.39 l.aB 1.35

05 0.364 0.386 1.1 1.25









VADM OFr FR PomAsRW nGLUMA3 At c RM B
.i. T25, $0, AM 7 M OWI mIcBIa Or



m oent waaly i leood aMaW 5 mroso asts 6

S5 2.22 8.16
90 2.26 2.21


Avups a.tss 2.22











TARE 23


0* W HM 9 80OMF CAMI A VAR0000 M 3 IM NO
NgBgf aMnfl EYraw
o, M W = M = = T n C -I Iifl m


Pu mit M Omt eolsmE 2amt wP at
potaats1 seuLm UA Moo o0o oo
gueaimte 2awsb Q 7 go 8 pl 7 A B


0.80


0.0O


0.ao0


0.20


0.20


0.-20


4o6
A.*


Ao.




350




AV*
169


79
AV. -


-ov
366





351
351





a98


179


83
-+-


101.0


96.3


a7.6


71.5


100.0


100.6


97.5


97.3


95.S


4e.6


19.6


1.2


48.4


6.0


0.50


0.60


0.70


0.80


0.90


1.00


1.10










waasA
CrormnATuS wLAwO ML. or -mum mrom
CAUBM 25, 50, AN T5 M CBT COT BIS
W RAMR Mr1cmaY


wr Oat Bo.is

a5
50
75


ELood. iLm 7l


0.3T1


mlood sau 8

0.483
0.472
OAWT


as po, 95 m -- Lmjia r sia uase
u -mea n or 0.a om amor soZ mzm


Phr Omet Nmlysls

e
50
75


3Lkd 8sLe 7

0.973
0.883
0*780


aloot Hlto 8t

1.05

0.996
0.9ge


Im


mm


Ill I I II 1 TIllfTI I I1 I



















ms6
VAIa L lF POAUSITm UCOcAS AT c a-l1AIm
CAMu= 25, Af50 lM 7 MI A NOmmUN W
A0.2U H -S M M MOR CSAM
o.a m am SGarm assJA


Or, t Bm ly m ls Bloo S0 pXl T7 Snood Hli 8$

a5 1.96 2.01
50 2.05 2.08

?5 a 2E.0 2.02
Av.gMs 2.03 2.06










um g Nz1t sT
B a aamrUsza o af sRAmocas a oossa
0UfU2MM OF YAlIVUS acipWSlSSSI


otae Nami 35Out __ aMU_ 3_

0.0 449 478
Av. ** 98.0 "06
1.00 439 aRS
A. 95.9 89.5
1.10 418 366
Ar. 91. 777.
1.80 316 315
Ar. 71. 66.8
1.30 158 91
Ar. .3-5 40.1


Av. 9 2.5 1,51
1.50 11 12


1.60 0 0
0 0
-"r" 0.0 0.0











TAE SB28
SSM./100 ML. or 80mM cML3E A=D POmiUSI SGMan
CAUIwM 25, 50, AD 75 PE CWEM"? B
o s-aw aamanas


Sodium Chloride


0.39o.
0.398 0.399
0.35 0.382
0.372 0.363


Camuitvation


1.- 1.36
1.25 1.7
1.19 1.14


VAIMS O 9 MZOkSSIM GLIDMAU ATC AAEB
CA 25, SO, AND 75 IU caM MaaU Or
NMS 11 M-Iovim


Nr Camt alyiadm Mood ~lpl 3om fLtal 35

25 2.23 2.18
50 2.30 02.5


MaVM 2.29 2.27


ur Oaft
BNOMOljn











TA 30

0.2 ER CMT T02A CSS ZMI AMD VAB1OO
MPouuma GEDmMUa"


Am-Gnt -oCmt
lOMM0.00 i
0.00 0.2O


o.Ao


0.50


0.60


0.70


0.80


0.20


0.20




0.20
0.20


1.00


ep as36 ai s37


ad.p a"


AV'*

475
Ar. *

471
AT.*


Ar. *
Atoo
A. W

250
AV. 2W
105


19
AV*

0
Ar. -


CCWIBASM 01


100.0


95.0


96.2






67.2


21.0


m


)Io


292


89



-I-

0
0
-0-
0

-


3.6


0.0


0.0


0.0


;1.2











Wara 31
MrASIs .OL/Loo ML.. of SmO C~azU
C l 25 50, AD 75 PER CMrT Emans
W NmLi z


Emood ample 36

o.41o3

0.303
0.304


mooo ame8la 37

0.396
0.31

0.368


"BMB 3B
Oan mnRAMsB V OM./IO ML.. OF POmSOm GWCA!M CAJM=s
29, 50, AM 75 JIM OW UaraXa W m I 1ss aanevalM
a MB RpmKlCor 0.2 m eam asoam cawmm


. .... .. N. ..O W i


1Mw Coat HMolyia

25

90

7W


SLoo amqle 36

0.783


0.628


n ooa ae.g. 37

0.690
0.637

0.578


har Cent nemoysi

25

50

75


m


-


INIIIoI IIHI[ IN go,



















as 33
VAzm jima NAmaS1B= EMEs M At
CAP 5 30, AND 75 M OffT 3MH W
maW -R 61 inmIs ri aum ini I
0.2 m cm soam UARM


wr Cot BKMImOjSs XLood Smpl 36 Mood e pae ST?

29 2.0o4 .Ia
o0 2.l 2.12

75 .Lf, .22..
Aramgi 2.12 2.14















US 34
V-t c MmJ W PUAM AAIgocim n lA mmm aGZccagMM
SOWMES (X VARICU CCPRIB



PW M .r.
*O" 0.sto Road Mosoood
aqnOmte !m!T! 9 ateule 10o glaw" 9 am^U to
1.00 366 37
A. 100.6 10o.2
1.50 346 3S6
Ar. 95.3 99.7


Av. *.. **** 86.0
2.oo00 193 153
Av. 15 52.9 48.9
2.25 109


2.50 80 15
Ar. 76 IS 0.9 1.7










TAM 35
comcrHAn1 W o./100 ML. or we=M CmmxIB AnD ammmns LWom
Ca-M 25, 50,AID 7 If CT UEMhBIS
OW PARM mRmcs


odio Chloride gMl ua GhueateN
wt Cent Coettmtirattan Stan
aue9 p sne uo apaM 9 emrr t
a5 0.416 o0.l9 2.32 .-14
o 0.399 0.-03 2a.0 1.99
75 0.,0 0.386 1.8.4 1A.







UMS 36
VAUUS Or 1 TO M RflIM O)OCB AT CO2 rMAZDU
CmMn 95, 50, AND 20 =N W l U 3L 5 Or



Ae Cwnt Hsulyruet mood aMRA. 9 m &loo mple 10
a5 2.37 2.58
50 2.61 2.67


.. .. ..ll 2*57 2.67.
2.57sssa 2.67










Mr 37


0.2 ma C2T 0 Csrim AM VARI S caa = OF




W COwt w Cut Clormr eer Ot aL
m-uL te =al lse a.e f t 12 L a 11 p..e 12
0.000 0.20 aT3 299
o..o o.m 27
A. "S 100.0 99.0
0.250 0.20 276 300
Av. -W w 101.5 99.0
0.500 0.20 265 296
Aw. 96.0 98.7
0.750 0.20 217 erT
AV. ;M "0 79.4 93.14
0.87" 0.20 226
..e .226 7..3

1.000 0.20 76 I83
Ar. 17.9 4g.0
1.125 0.20 .. 89
+V *.. ... 29o.0
1.250 0.20 9 T
Ar. 2.6 2.3
1.500 0.20 0 0
0 0
Ar. 0 0 0.0 0.0













mmamsIa v a./ioo -. ow msm =uarm
Caomm 0S, 50, MA 75 M am mm=wa
OW MDIBW


P. comt sIa-'n .


mlood amla 11


0.415

0.39*


0.41,

0.1.8


TAa 39

oM201mas a m./Ioo M.. 0 nasm aimas cam=
25, so# An 75 P- CUTi wamnorIB C9 BA py al It BI
1 umeze CR ( 0.2 E OW SO=1 Camum


ser Cent Iamlys
25
50

75


1.03
0.889

0.775


M am fte" u

1.07
0.975
0.878


-


*



















TAM 10
VATas t KB IMOBmZBm GZUCA3UM AtB
Cmff 25#, 90, MD 75 mO OW BmaXZ a
Raun j1l sCI1 m ul ma wB
0.l2 m B SO3M camum


mr oCat 3aolyes
25
a,
90
75
knaUM


eLOOG mpWl 11
a2.96
3.19


3.15


m3ood fale ]a

3.10
3.19


3.3$
jLuL
3-3










MMES 41
PER CEOT HmmaLISIm of UMM EBRngCES m= YaaS3m G CWM
SOUPS O VARIUS CC MZA S


Per Cat CaLorltan r am.nom Per COt Bmolyws
maealum of Iood no a d at
Qunawate 39 340 3' 9 40
0.50 406 445 47o
kA. "q VS" 100.3 99O- 95.1
0.75 39 435 458
Ar. 1m 1 96.8 97.3 92B3
1.00 307 34o 38
A*. 3% 78.7 77.4 7.7
1.25 192 244 233
A*r. 47.6 54.1 47.a
1.50 133 160 150
A*. "T 4 32.5 36.9 30.8
1.75 65 7 44
Ar. 15.6 16.6 9.1
2.00 0 5 7
A*. 0.0 1.1 1.4
2.15 0 0 0
0 0 0
A". 0-- 0 0.0 0.0 0.0










sm he
9 M./100 w.. w OB=M cZrirm A= mw1 MaMAW
CMAtaJa 25, 50, AND 75 P cUrT HaLIss
0F EAWA ERYROCTE5S


Per Cent
SMax2ys.


0.39

0,38
0.3"


Sodium Chlrid
Ca0Mttmtni
.A Blood sMpt.

0.393 o
L 0.380 0

S 0.363 0


40
.405

.398

.381


Coame*L amate
.pA mBooa smu

1.55 1.63 1.51
.8i 1.30 1.18
1.05 1.05 1.03


AWA 43
VAL=S Wn Lm M WsA M GUlCORMA At ORA s
MOMrs 5, 2 0, AMD 75,MC a OmLlg Omc
IIg arocls


Per Cnt ma7ala1s Xoo ample* 38 Mood Smle 39 Mood amse O40
25 3.36 3.18 3.53
50 3-92 3.86 4.03

75 -T 3.8 4.07
rm 3.97 3.87 4.14


[.cC DHMXB










TAME 44


PER COET BENUMB OF IMIO RmrOCTP 2= S0U1MO3 CW32SI
0.2 M OUT sOM CiIaXC AD VARof1 CnMMam Or
iMnBXM Buemm


wer Cat
mweussa


48S


462
AV*
446


Ar .

398
389
Aw. 393
144


69
A.


&*
T
Arl -j+
0
0
Av. "I


Rin,,


100.0


96.1


0.0 0.8


Im Cut
stcadm
OAXOrld


0.0O


seaOU 42


*e

48

481


0.375


0.500


o.6e5


0.750


0.875


9-.3


0.0lO


0.20


o.ao


0.20


0.20


0.20


0.0o


96.6


73.4


29.7


366


241


79


48.6


1.0


15.3


il


amolm 42


aemas 41
"t^rIMa
e.S l ra


amma. A1











rars 45
oa m MONI T a v /LOO wL. ow oom c=am
CaM 25, 5O, 5VAn 75 P MaiuSa
Wf IF" IsIIFF.Bi


13' ceot NatnsL 0


50

75


Mood afple 41

0.403

0.390

0.378


Blood $Lw4 ae

0.447

0.47
0.4W6


Ws1m 46

2o 50o, amTm m am Cm3ms Hw mm Hs mXul
.. .. 2 a E 0.2 .m m CHT I aTO I R mjuX


25

50

75


blood anpUe 41
0.819

0.750
0.669


*ood *wast te
0.950
0.864

0.750


*


Ow



















UMx 47
VALA OF 4 IM S OWCOM ASt
Caaf 9, 50, ad Tg aR agHf mAa=a7 M O
0.2 MnB CmmT B sm1 CEamiM


tr Cwt ontyas MMBood amplen 4 EM&s ample Am
2 3.46 3.43
0 3.3 3.46
75 3.1 3.60
Auu*sy 3.37 3.50





90




IuR carp muM5ZS MwAXW ms1eMc! s a Cza3m e rAna mCCW
Bou = OF VARIOUS P I




,ci m 9Mo ELoo.o

rw .5 T 100.0


A. 8.6 960
2.00 259 185
A. 8 .6 81.3
2.10 236 156
ir. 7T7. 69.2
2.2D 22k 1SO
Ar. I72.6 54.9
2.30 194 4
A. 6.4 40o.6
2.4o 183 54
4 1 58.8 26.3
2.50 140 52
140 44
k*. 18 45.8 a.4



2.80 63 0
Av. f19.9 0.0












CAM= 25, 50, AND 75 PEI CBr m=Mas
If I I W i I3 ...O. III I IIIII


PMW out
Bilymirs


Soium Chlaride

suspe 13 sftn.U 14
0.466 o.410
O.a5 0o.395
0.3" 0.378


CaLcbm GLuomato



2.7 2a.43

2.16 2.06


TABLE 50
2mu 30
VAsM o i Pa CAZCzW M At C'As ICoaAnT
CAmM 9, 50o, AND 75 MM CMT merB OFW
ABI 'IlROCly S


1r omit siyi.
95
50
75


ood aO pl" 13
2.34
2.38


8.41


mbost MEW* 14
2.31



2.41











wu am51
0.4 Pr cOR 8SUJXM MM= AM TAUDOMO C1MCWIA=MB p
CAtCmJM (BIomAm



Nr Cat Pr Cmt Colaufr fMIs phratt b InuII
ll~dL Bd388


300


ec1


388 aph


340
* *frl
*W


0.20 299 00 o10
,.1- -4-


*49
Ce.


160
IfiD


100.3 100.8 100.0


**... 8.0 87.4


101.4 71.T 83.1


**.. *.,* 63.0


79.3 29.6 46.1


41.o0 10.4 1o.


4 A2
444"" """S"


S144


19.3


3.6


0.8


7.3 e.6 0.0


0.00


0.20


0.20


0.70


o.80


0.90


e...
&* ***
eei
al
tf*e se*e


0.20 S 119 10
**


1.00


1.20


1.40


1.60


0.20


Lv


0.20


0.20


UP 11 WIN










wam 52

CmBo~aSzIx = QLzAi ML. at iPm um
CAM=B 9, 50, As T7 PER CMT MOROMrZ



S mt BeBnoLY ]a ood saaW 15 footd a3.Bl 16 mood saapl 17
25 0.473 o.ea o.0eB
50 0.5o 0.399 0.410
750 .4 0.30 0.39o











ABan 53


a.~ as mL. r "Mm mmm oaozo
MO M CUT.0 M CII IAU I Ch1 1 CAPBI I
50, P375 0. ca W OM8 Vr WIa



Pwr 0Mt SXmtlystu Umo 0 aig 15 loodt atl 16 MoM apto 17I
5 1.32 1.03 1.09
50 1.16 0.905 0.990
75 1.0B 0.785 0.860




















VAWas W j CA7eXM s aBXm3X aS Maumm
atmazalT, So, a0 vE CAN 75 M =


2-8
0.2 uma ca amzamm


S". ..t nain l.s Uoe. a.a 15 I Ioo4 aIt 16 loo i I ..

S2.8 a.95 .87r
$0a.96 3.M 2.91


Avg g."95 3.05 2.97









TABBU c5

SOCTYMBS Or VAKB00 COMH3AEMAZB




tWombft a e 3 3ee MMaple 43 aumm _
1.50 443 4oo
At. 94.0 91.a
1.60 o 372
A. 90" .6 79.T
i.70 4w 320
Ar* 4 .5 70..
1.80 3 ago
-. "3 70.8 &A.4
1-90 300 23
a. i- --] a.o ,.9






2.o oo 74


am30 56 17


2.40 30 0
A, 6.0o o.o
So 103 ,IIII I [ IIIII




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