Download Study on Blood Flow and Pressure in the Cardiovascular System of Animals and more Exams Aviation in PDF only on Docsity!
AMERICAN PHYSIOLOGICAL SOCIETY
PROCEEDINGS
FALL MEETING, AUGUST 23-26, 1960
ABSTRACTS OF PAPERS
Abs
An usten’skfollowilag an athor’s nzamedenotes “by invitation. *=
tracts are arranged in alphabeti‘cd order of fi ‘rst- namedauthors
EFFECTS OFHIWRENAGEONPERIPHSU B~ODFUX!, F,L,Abeland Q. R. hp% l Dept, of wsiol,, Univ. of Wisconsin Med. Sch,, Madison. The Mdtate effects of moderate haorrhage on the tistfibution of blood flow mr8 imrestigatsd. (^) DOgS Were bbd XXpiw all ~OUXt 8qa to 5% of the calculated blood volme (5 II&, per kg.). The bleedings were repeated four tfmes at 3mrkervals of ten dnutes, Blood flow was measured by the electromagnetic flometer Am the reti, superLor mesenteric, ad common Uiac art&es@ Peripheral F8SiSblC8 was calculated for these beds. The r8SdtS show marked constriction sn the Uiac and mesenteric beds foI.lowWg loss of $ and iU$ of the blood volume, but little chmge in resistance of the renal bed. FoIlowhg the loss of 20$ of the bled volume, there was a sharp increase ti resisIxme of the I,lkac and renal beds which was not accompanied w a comespo~%ng increase h resistarrce of the sUp&or mesenteric bed. It is concluded that the rerml flow 9s atabed after sfnau bhod losses at the expense of mustie, skh, and intestine; but that flow to the intasttie rma$ms relatively Ugh fdbwing more severe bleading,
THE PHYSIOLOGIST 5
OXYGEK TENSIOTU’OF IIulUll CEPXBRAL GRAY ANE WHITE NATTEE. John 73. Adams* and John W. Severinahaus, Cardfovascukr Research Institute, IL of Ctiif. Ned. Center, San Francisco. A new polarographic membrane-covered needle oxygen tension electrode
1 mm in diameter (Beckman) appears-- to have less than 5% 8rror in un-
stirred materials. It has been used in 23 neurosurgical procedures to estimate both surface and deep under N20-barbiturate an8 sthesi
tip 3 ,a; some
patients were awake and 20 were were supplemented wi th halothane. pG2 was surprisingly consistent, and not measurably affected by distance from surface vessels, in contrast to results reported with bare platinum electrodes. The mean vtiues and ranges of &2 in mm Hg
were: cerebral cortical surface 13 (&20), cerebral white (depth ca.
5 mm) 30 (11-381, contused cortex 31 (20-58), surface which had been coagulated 54 (2246). Several readings taken within cortical gray agreed with surface readings (2 3 mm Hg). In one subject comparative blood values were: PaG2 139, PaCO2 45; jugular venous ED2 50, Pco2 50; cortical surface vein draining test area po2 60; 3 gyri of frontal
lobe, surface tissue pO2 h-6 mm. During hypothermia (29%) both
carotids and both vertebrals were occluded for 3 minutes. (^) PO on temporal cortfcal surface fell very slowly (from 25 to 15 mm 6 g). We conclude that actively metabolizing neural tissue has a much lower oqgen tension than had been astswned from studies of venous blood, Supported in part by USPHS H-3963 and H-4275,
S. Afonm*, J. F. Herrick, Q, Q. Rowe, and C, W. Crumpton.Dept. of
bd. and Cardio~Imomuy Reasarohlaboratory of the U, o'f Win~onsln
School of Me&&e, Madison6, W%mormin. -
The flmction of the clretmy q&m to distribute tIs heat
gumrated by cellulm activity and to equaliw the body tmperaturr ti
wellkmm, This^ function haa b-n^ studied sxtenalvely^ by^ very^ coqm-
tent lmestigrtoss awes a period of mang years. The lltesatuse, how-
eves, aeema to contain lImItad mperimenrta3td&a on osoUlatory pat-
tesm of the @mamio thtmml variationa sxiating in the venous8yaWm.
Tim thermistor haa proven to be a convenient and reliable device for
8tudyin~ ths 8llUk11 but Swicantc WtitioM fn tsmparatore of the
flowing blood. Thesevarfations^ are^ kewd to cart&in bodily proe4ma08,
one of wMch i8 the breathing fisqueacp, At higher body tvatW98,
the variatton ap&ronfz%d with the bmathing frequ8ncy appear6 to
modubktea slower type of variatim in teqerature which i8 n& 80 svi-
dtbllt~ l'el0t4d t0 P~iOlO~iC~ Vari&tiOll. &9COrd8 of th3W V0Si.0thM
tin be ~easntsd. F-1 conclusion must await further investigations.
THE PHYSIOLOGIST 7
AN IMPROVED METHOD FOR THE BIOASSAY OF ANTI-INFLAMMATORY
ACTIVITY OF ADRENO-CORTICAL STEROIDS IN RATS. Ralph W. Alexander and I ,E, Bush (intr. by Natalie Alexander) l U. &uth. Calif. School of Med. and Radcliffe Infirmary, Oxford, England. In 1950 a cotton pellet assay was introduced for assessing anti- inflammatory activity of cortisone-like steroids in rodents. Although simple to carry out we found the precision of this assay to be rather low due to the shallow slope of the log dose - response plot. VaxrageeninV1, a water soluble polysaccharlde extracted from Irish moss (seaweed) has been shown to cause a striking granulomatous response to guinea pigs. We therefore Incorporated this polysaccbride into cotton pellets and produced large, easily dissectable granulomata in adrenalectomized rats. In several assays using l-6 mgs cortisone acetate and l-4 mgs cortisol acetate we found that the precision using %arrageeninll impregnated cotton pellets was twice that obtained with plain cotton pellets. The reason for the Increased precision was the steeper log dose-response slope obtained. A , the index of precision, ranged between 0.10-O. 17.
RELATIONSHIP BETWEEN CHANGES IN CARDMC OUTPUT AND PERIPHERAL BLOOD FLOW DURING EPINEPHRINE INFUSION IN MAN. M.J, Allwood,* E.W.O. Keck,* R. J, Marshall* and 5. T. Shepherd. Mayo Foundation and Mayo Clinic, Rochester, MGota,
Cardiac output (C.O.) was measured by the indicator-dilution technic in seven healthy men aged 30-35 years, lying supine, Fore- arm blood flow (F .F,) was determined by venous occlusion plethys- mogwaphy using the Whitney strain gauge. Epinephrine bitartrate (lVSuprarenin “-Winthrop)^ was^ infused^ intravenously^ at^ 2.5^ to^ 15~9. per minute. The onset of changes in blood pressure, heart rate (H.R.) and F.F. was not always synchronous. Control values for F.F. were 4.0 (range 2.1-8.7) ml./100 ml./min, andfor C.0, 7, (range 4.9-8.7) L./min. With^ lOBg./min.^ infused^ for^5 min., F.F. initially reached a peak of 15.9 (LO-26.1) ml. and the C.0. was 10.7 (8.8,13,O) L./m& After a transient decrease the F.F. was 6.5 (4.0-9.9) ml. and the C.0. wa8 10~0 (6&-12*3) LJmha With longer infusions (20 min.) in four subjects the F.F. and C,O. showed a further slight increase. In the three subjects with the largest increase in C.0, (70 per cent), oxygen consumption was increased by 11 per cent. Thus the increase in C.O. is not wholly due to increased metabolism. The initial increase in C,O, was largely due to an increase in H.R. Subsequently, increases in H.R. and stroke volume were of about equal importance; this contrasts with a similar increas e in C.O. during ac hiev ,ed mainly by an increase^ in^ H.R.
S (^) upine exercise which was
8 THEPHYSIOL0GIST
A QUANTITATIVE EVALUATION OF NON LACTATE HYDROGEN ACCEPTORS
IN ANOXIC RAT LIVER SLICES, Norman R. Alpert, Richard
Chenoweth+:- and Richard Winzler*. University of-is,
college of Medicine,-"Ehlcago, Illinois
Anaerobic muscle produces large quantities of lactate.
Under these conditfons pyruvate serves as a hydrogen accep-
tor, facilitating the oxidation of DPNH, and thus allowing
glycolyais to continue. Liver, in contrast, produces vsry
little lactate during anoxia. For this tissue, fat forma-
tion has been implicated a8 a possible substitute hydrogen
acceptor system. Experiments were carried out to evaluate
the quantitative significance of this and other non-
pyruvate hydrogen acceptors in anaerobic liva~ slices. It
has been assumed that in liver sl2ces under anaerobic con-
ditions all carbohydrate which traversen the glycolytic
pathway and doea not appear 8s lactate may be involved In
a non-pyruvate hydm gan acceptor transformation. The total
carbohydrate which disappeared anaerobically per gram of
liver slice incubated in a high X medium fur 90 min. and
X20 min. was 21.8 and 38.1 mg while the lactate which ap-
peared was .s and 1.7 mg. Thus a considerable amount of
carbohydrate Is not accounted for as lactate, In addition
anoxic liver uses glgmgen a8 8 substrate in preference to
the exe enous
and 25 P
glucose. A Pasteur effect of 10% at 90 min.
at 180 min. wan observed.
in part by USPHS Grant H2345.
This work was supported
gradad drgraar ef eb8tmmtion of thm main prlmonary rrt8ry by a brlloen rt the tip of 8 orrdllre erthatmr, Sbimf8r eb8emrtiOl8 wara msd8 fa de#r tith intrat errdire 8ept8, Ob8traotimar ef tha mais puboarry artery to l lmvata tha right r@ltrioUlrr Sy8tOli43 prW8or@ to mro than 80 Y. Hg in dog8 with 8trirX roptal defmotr oru8ad right-to-left 8bulat8 throwgh the dsfaot of alp ts 55 par oert of 8y8t*mi@ flow and daorarr8d the magnitado of the left-to-right 8hont8, #o Iylt~m~tfu ehrng88 in maa11 aortio pr888ur+ and inrigaifiorat to rlight imQr8rBa8 ia maan rdght atria1 prmrrare ooeurred. In OontraIt, ia dog8 with introt repta l888er dagreer of ob8truetim predtmiag
Hg were 888o@irtad with hOr888+8 in mman right &trial )rmrru&8 ml rtrikiag door~rr~r itr rnem asrtio prarrurr ud oardire eatpmt. The hmmdymrafe 8t8tu8 ef there no-1 dog8 detwiorrted rapidly ia
oontrrrt to that of the degr -5th rtfirl raptal dafaatr that mra
orprblm af Puimtrinfag right vrotriaohr hypmrtaarion OT+F pariodr lorgor than 30 miattt@8 without rigaifiernt door@888 in 8yrtWi@ rrterbl pFelllr. It 58 oonolrdd that uader therm ucrdftioar tha
10 THE PHYSIOLOGIST
CHOLINESTERASE/hOAMINE OXIDASE RATIOS IN THE AVIAN
CENTRAL NERVOUS SYSTEM. D T.L. Folkerth* and
kc.M. Hanson*. Indiana Univ. Med. Center, Indianapolis, Ind,
Some investigators think that acetylcholine (ACh) and serotonin
(5-HT) act as neurohumors. Cholinesterase (ChE) and monoamine
oxidase (MAO), the enzymes which keep these two important com-
pounds at physiological levels are widely distributed in the central
nervous system (CNS) as well as in other parts of the body. How-
ever, it is extremely interesting that in the CNS, high ChE activf-
ties usually are found where MAO levels are low. Davison (Physiol,
Rev. 38, 729, 1958) summarized some human tissue data and showed
that m/MAO ratios for caudate nucleus and putamen were over 100
whereas the cortex was under 10, These findings suggestthat each
system may be selectively involved in a specific brain part, or may
interact with each other under certain physiological conditions, Sev-
eral investigators have shownthat 5-HT can inhibit ChE in vitro.
Aprison (Fed. Proc. 19, 275, 1960) reported 50%inhibit-a
ChE activity when ACfind 5-HT were present in equal molar concen-
trations (3x10-3M). Further studies in which the brain ChE and MAO
activities are compared in other species seem in order. Since we are
using pigeons in a neurochemical-behavioral project, it was decided
to determine the ChE()IMACh/gm/hr)/MAO(p NH3/gm/hr) ratios in
specific brain parts. The ChE/MAU ratios for the cortex, caudate
nucleus, mid-brain, cerebellum and medulla-pons are 85, 135, 174,
415, and 76 respectively. When compared with a peripheral organ
such as liver (14), the brain ratios are much higher.
DETECTION OF RETROGRADE PASSAGE OF INDICATOR FROM AORTA TO LEFT VENTRICLE IN ZX)GS, Egas Armelin, * Leon Michaels,* H, W, Marshall,* D, E, Donald* and E. H, Wood. Mayo Clinic and Mayo Foundation, Rochester, Minnesota,
Detection of indicator in the left ventricle immediately after injec-
tion into the root of the aorta is proof of retrograde passage across the aortic valve and can be used to assess aortic regurgitation, Dilution curves of indocyanine green were recorded simultaneously from the left ventricle and femoral artery of 26 control dogs and 38 dogs with experimental aortic regurgitation, all with closed chests. The effects^ of^ variation^ in^ sampling^ site^ in^ the^ left^ ventricle^ and injection site in the aorta and time of iajection in the cardiac cycle on the ratio of the areas of the immediately appearing portion of the left ventricular curve to the femoral curve (regurgitant fraction, RF)
were studied. Synchronization of injection period to coincide with
systo lie^ or^ diastole^ caused^ little^ variationrr^ in^ RF.^ However,^ the^ RF values from short duration injections (40 msec,) were time sensi- tive; RF values were minimal with injections in late diastole and early systole and m%ximal, in late systole and early diastole, Physiologic regurgitation was detected in normal dogs during a short, sharply delineated period just at the end of systole apparently related to valve closure. The correlation coefficient between RF
values from nonsynchronized injections extending over several cardiac cycles and percentage regurgitation from back perfusion studies at necropsy was 0.85. This was superior to dilution and pressure pulse methods studied.
other indicator
THE PHYSIOLDGIST 11
A NEWTHEORY OF THE REGULATION OF MEATHIX, B.W,Amstronq, EIJLHurt, 2,s J,M,Worban^ and^ R.W,Blfde.^ U,^ of^ Md.^ School^ of^ Med.,^ BaE Md, (intr. by D,B,DilL, Army Chmfcal Center, Edgewopd Arsenal, Md.) During muscular exercise the Axed venous Pcoz (mco,) and acidity (H%) as well as arterial. acidity &a> are greater than at rest while the P&co2 repins essentially constant, The^ relationship^ between ventilation (V) and each variable, except Pacoz, is statistically sfg- nificant, These cunsiderations suggest that the hyperpnea of muscular exercise can be explained if the classic concept that humural control of breathing is mediated only by receptor mechanisms S;n the arterfal blood stresm (the respiratory center, the carotid and aortPc bodies) is expanded by postulating a chemoreceptoy, probabl in the pulmonary artery, whose activity is related to Ecoa and Ip F. This concept has been tested in dogs by measuring alterations of Pco2 and @ of arterial and mixed venous blood, produced by injections of lactk acid or sodfum bicarbonate solutions, by CO2 inhalation, and by muscular exercise, The resulting hyperpneas cannot be explained on the basis of arterial or mixed venous changes; they^ can^ be^ explained^ if^ both^ arterial^ and mixed venous changes are considered. mrvard Fatigue Laboratory data (J. Physiol. 66:9,1928, 69:18,1930), from norms1 men at rest and exercise, have been usgd to quangitate the cuncept. Some relationships found are: $ = 1.1 H a + 2.3 mcoz - 140. SE: 10.0 L/mi% r t l^ .^ H+a^ l piico = 0.92; P: < 0.001. Ventilatory responses to diabetic acidosis, shock, CO2 inhalation, N&Cl and NaIICO3 administration agree well with the t "predictedvw by t&is equation; it also "predicts" the i of exercising subjects reported by Mitchell et al (J. Clin. Invest, 37:1693,1958). The Glomus pulmonale recently described by V. Krahl (Bull,U, of pld. Sch. of Pied. July 1960) may be part of the postulated mixed venous chemoreceptor mechanism.
A pIJ:mO METHODFOR DETESMINING THE ACID-USE VALUES OF DfiOOD. Astrup (in& b 5, W. Severinghaus). Department of Clinical
Foul
Chemistry, Rigshospitalet, Copenhagen, Denmark. A micro-method, suitable for capillary blood, is described for deter- mining pH, Ro2, standard bicarbonate, base excess and buffer base of blood. The method requires measurement of actual pH and pH after equilibra- tion with carbon dioxide at two known tensions. For this purpose a new micro equipment has been developed, involving a capillary glass electrode (for 25 ~1 samples) and a device for equilibration of micro quantities of blood (30-50&l) with gases. Capillary blood is drawn from ear or finger, For the actual pH, about 20-25~1 is sucked directly into the capillary electrode and the reading is made wediately, or it is sucked from blood drawn anaerobi- cally into a hepariniaed capillary glass tube. (^) About 80-9OM of blood from two capillary glass tubes is divided between two of the chambers of the equilibration apparatus. This is shaken mechanically (2600 RPM) and after 3 minutes the pH values in the two samp3es are measured, By using these two values and the known gas PCO2, the rectilinear pH/log pCC2 line is drawn and the PCQ, standard bicarbonate and base excess can be calculated. (^) The method is highly accurate, as the 3 vtiuss can be found with an error of less than 2 2%.
THE PHYSIOLOGIST 13
ON TEEl?ATUREOF!CHEUNC~G~G DxBMETERINIsoLcLTEDMIcRoscOPIC
VESSEUUNDERPRESSUR,EV~~ON.Silvio~aezand&roldLamoti,
Dept. Anes~esiolo~, N.Y.U. Media Center, NewYork and Dept.
Pfigrsiolog~,k3.e &xiv. %hoolofIkLt NewHaven,Corm.
our studies of tie effects of pressure in isolated micxw3oapic
vessels have disclosed twlo cons=&& characteristics uniqtle to -he
smallest blood vessels; (a) a selective clo8ure of pre&pUlary
sphincters at positive pressures; and (b) an w2mging vessel
dimeter (plateatx) under considerable pressuremriation.
Thenature of the latter v8scxCLarfeature (b) hasbeen further
asdlyzed by the following aperimentel q3pmach: 1. Th0 effect of
staticpressurewas ~stedo~isolafedssterioles,r~gfr~~to
&2 micra in dhumder, using various perf'usion fhtids: rat pIasmsand
semmga$ gehttie, 3%d&mm, 3%albumin mlutions aad kerosene.
fAkthoughdW andgeUt&ne solutions &.mwincreased~~tvis-
cosi~ at lower rangesof pressureas determisedby the OsmXdLimo-
m&er,no sQnificantc%ffer~~e appears~plateaufomationtithalx
5 fluids eaplqd. 2. bHUWb~ S&Of433&WbMRltS, th8 XW3JMBleive-
ness to topicml epinq&e of t;he isolated vessds under deuwnemt
andimmmentof sM&qressurewas detemnined,keeping vessel dfa-
meter constant, The results indicate that the log of the doseof
eptiwe required to pmdwe a gwen ahangein the vessel radhl
isproport5owCL to the 0rIginaI tensionofthevessel. The result
of tiese experimentswarrant, for the present, We following comlu-
sions: (a) t&t^ plateau^ fomEktion^ is^ not^ due^ to^ lack^ of^ pressure
tramnissionbutis inherentintienature of thevessel-; end
(b) that the shorter plateau deserribedfor ir~~ervatedpwtian~
my be related to circulating neuro-hmors or tissue mediators~
SIMULTANEOUST-1824 AND RISA TIMF,-CONCENTRATIONCURVES IN SPLEZVECTO-
MIZED DOGS. Carleton H. Baker. Dept. of Physiology, Med. Coil, of Georgia, Augusta. Time concentration curves of several tag-protein complexes were de- termined over a period of five hours in chronically splenectomiaed dogs anesthetized with morphine and sodium pentobarbital. T-1824 blue dye in distilled water, human albumin solution tagged in vitro with T-1824, or autologous dog !i
lasma tagged in vitro with T-18x were injected at the same time as I 31-tagged hum= e albumin (RISA) or in vitro 11%tagged dog fibrinogen, The T-1824 solutions containedT.5% of dye per ml. of solution. Sufficient dye was injected to have an opti- cal density of about .500 in the dog's plasma and enough radioactive 1131~tagged compounds so there were approximately 3000 counts/min./ml. of plasma at 10 minutes post injection. Arterial^ samples^ were^ taken at 5, 10, 15, 20 and 30 minutes post injection and then at 20 minute intervals thkreafter. It was noted that the disappearance rates of the Il%tagged proteins are the slowest of the various complexes and the semilogarithmic slopes are smooth and linear. However, the disappear- ance rate of the various T-182b solutions is an essentially non-linear one with many abrupt changes in concentration. It also appears that the disappearance rate of the in vitro tagged T-1824 protein complexes was less than that of the distgled water solution of T-1824, but not as low as the 113%.,agged compounds. (Supported by USPHS grant H-4573).
14 THE PHYSIOLOGIn
CELLULAR SITE FOR SYNTHESIS OF HUMAN PRUTHROMIBIN AS RE-
VEALED BY FLUORESCENT ANTIBODY, William J. Baker * and Marion I. Barnhart. Dept. Physiology and Pharmacology, tiyne State University, Detro- it,gan.
The objective of this investigation was to identify the specific cells engaged
in synthesis of prothrombin utilizing immunocytochemical techniques. Purified human prothrombin was used in rabbits to elicit potent antiprothrombin sera. A
single injection of human prothrombin (l-4 mg/KgI wasmixedwith AIWHI3 as
an adjuvant andinjectedIM into a rabbit. Antisera were obtained 10 days later
whichwere of comparable strength to those produced using Freund’s adjuvant
which required considerably greater amounts of antigen, 3 injections and about
5 weeks to harvest. These antisera did not form precipitin bands with purified
human fibrinogen nor exhibit any commonantigenic components with antipro-
thrombin sera to dog or bovine prothrombin. The described immunologic evalua-
tion was madewith the Wilson modification of the Ouchterlony agar gel diffusiun
technique run at several concentrations. Gamma globulin concentrates were pre- pared by Ethodin fractionation of the antiprothrombin serum and conjugated with lissamine rhodamine B 200. Such rhodamine labelled antihuman prothrombin provided the fluorescent tag for the study of human liver imprints. (^) Liver
parenchymal cells containingadequate concentiations of prothrombin attracted
fluorescentantibodyandproducedpatches of brilliantly fluorescing cells. Not
all liver cells react with the specific antiprothrombin. This may meanthat the
liver parenchymal cell is specialized or that there 1s cyclic prothrombin synthesis
(Supported by the Michigan Heart Association).
CHRONIC AWm DOGS WITH TOTAL CARDIAC DENERVATION. Basdeo Balkissoon* and Edward Hawthorne. Howard Univ. School of Medicine Washington, D. C. Total cardiac denervation compatible with chronic survival in dogs was accomplished by a surgical technique involving right thoracotomy and right sided cardiac denervation followed in two weeks by left sided denervation. On both sides all branches of the thoracic vagus above the cardiac apex were sectioned and all thoracic sympathetic ganglia through the eighth were excised. At thoracotomy, mercury in rubber strain gages were placed around the ascending aorta and the middle of the left ventricle for recording aortic circumference (AC) and ventiicular circumference (VC) changes in the awake dogs. Bradycardia was char- acteristically absent following administration of epinephrine (0.05 mg I.V.) in seven chronic, denesvated dogs. This response was unaltered by atropinization (0.02 mg/kg). Awake, normal dogs characteristically showed a bradycardia after epinephrine in this dosage and this effect was blocked by atropinization. Semicontinuous monitaing of AC I VC, and heart rates over 24 hours (two minutes every half hour) revealed a remarkable constancy of heart rate in denervated as compazced to normal dogs. Rapid (one to four minutes) infusions of a blood-de-an mixture into a peripheral vein of standing denervated dogs increased end-dias- tolic VC and stroke volume as indicated by the VC gage. This was ac- companied by an increase in systolic and diastolic AC, Infusions in awake, intact dogs only caused an increase in heart rate, Awake dogs with denervated hearts appear to obey Starling% Law of the heart.
16 THE PH’YSIOLUGIST
PURIFICATION AND PROPERTIES OF MImHUNDRIAL MONOAMINE UXIDASE FRO!l BEEFLIWR. Libero^ Barbato*^ and^ LI^ G.^ Abood.^ Univeroity^ of Illinoie Medical School, Dept. of Psychiatry, Chicago, Xllinoio. Mitochondria from beef liver have been extracted three timea with a 0.5% (final cone) Triton solution, respectively at pH 7.5, 8, 8.5. The last extraction gives a clear oolution, with a S-fold increase in specific activity. A^ final^ twenty-fold^ purification icr obtained by treating the oolution with calcium phosphate gel first at pH 8.5, then at pH 7, and cluting the activity frmn the gel oeveral timeo with 0.2 M K2HPO4, The Michaelia con8tant (h.) of the crude mitochondrfal Triton oolution ia 6.1 x 10M3M and that of the purified entyme 8 x 1Q'k Activity is measured either manonetrically, with tyraaine aa arubstrate, or apectrophotaetrf- tally, with beazylamine a8 8ub8trate. The activity of a crude aitochondrial solution is not linear with time, while the act%vity of the purified enzpe ia linear during the firat two minute@. p-chloromercurobeneoate give& 100% inhibition at a final coneen- tration of 3 x 10 2 x 10-5.
g4, S@6 at 1 x lOa and^ 65% inhibition^ at This inhibition does not seer to be competitive. The activatkng effect of^ KCN and^ the^ inhibitory^ effect^ of^ iproniaeid have been confined using as 8ub8trate kynurmine. The enzyme appears to be al80 activated by incubating in phosphate buffer at pH 8 at 2!5*. The effect of phenantroline ha8 been studied both with bensylmine and with kynuramine.^ With^ benaylamine^ as^ rsub- strate complete inhibition occur8 at 3 x lo'%! and with kynura- mine, 3 x 10% gives only 50% inhibition. The enzyme ohowo an optimum at pH 8.5 an&an inflection point at pH 8, which could not be resolved in another maximtlll. (Supported by ONR)
MODIFICATIONS OF PROTHROMBIN SYNTHESIS IN THE DOG USING
FLUORESCENT ANTIBODY, Marion I, Barnhart and Gordon F. Anderson *. Dept. Physiology and Pharmacology, Wayne State University, Detroit, Mich. Immunochemicai procedures have identified liver parenchymai ceils as sites for the synthesis of prothrombin. The present study is concerned with the dynamic aspects of prothrombin synthesis and makes use of agents known to modify the prothrombin function of the liver. A correlative study was made on circulating levels of various plasma proteins and the cellular state of the liver following coumarin drugs and vitamin Kl therapy. Coumadin (3 mg/Kgl was given 1.v. and 24 hours later the plasma prothrombin was essentially zero while the ci rcuiati ng fibrinogen remained normal. Fluorescent microscgpy and specific fluorescent antiprothrombins were used tn the cellular evaluation of liver function Coumarin derivatives appear to interfere directly in the synthesis of prothrombin rather than promote storage and inhi bit the release of prothrombin from the hepatic cell. Vitamin K1 administration produced renewed synthesis of prothrombin as indicated by brightly colored patches of fluorescent cells. Even though normal circulating plasma levels of prothrombin were reached within 5 hours liver ceils were not as brilliant in their fluorescence as in the normal pre-treatment animal. Also, not ail of the liver parenchymai ceils reacted uniformly with the fiuores- cent antibody. The questions of specialization of function or cyclic production of prothrombin by liver parenchymai ceils still remain to be clarified. (Supported by the Michigan Heart Association)
THE PHYSIOLOGIST 17
SIaW, E.J.Barte1sto and H,EI.W~ (i&r, by S,C.Wq) Ikpt. of Pharma- cology, Colle@ of Physlciam & Surgeons, Colunibia I&ILK, &w York City. It was reported previously that cmtid occlusion mm&ted in an in- crease in venous return of venous origin, By preventtng tMs inc;rease in Venous mtum from reaching the heart the psssor respoxlse to carotid occlusion w&s mmkedly diminished, In the present study f'urtkr Investi- gation of this venous contribution to the cmtid occlusion response was csrried mt. In anest~tized,thoracotornizeU,vaESot~~zed dogs the follow- ing nreasuemwts mm made: vemu8, 8rterial and left intraventricul~ end-diastolic (EDP) pmssusea with Stat- guages; c&&c contractility with a W&tan w, and ctiac ou-@ut with a rotmter. Carotid occlu- sion produced: little cw in central venous pressuse; increase in cardiac contmctility, EDP and mterial pzssure; ma;lntename or in- crease in cardiac output and ConsideHble lncmase in totaS, peripherti resistmce (TPR), When, during subsequent carotid occlusion, the in- cmase in venous =tum of venous origin was aspirated from the inferi- or vena cava, the following chmges ensued: reduction of the increases in contractility and HIP; marked reduction of the arterial pressm rbe, and a p*cfpftous fall in cardiac outputI ?phe immase in WR was x&modified. The decrease in cardiac output was much greater thm that w!xich could be accounted for by tie volume of venous blood aspirated. Re-i-ion of this volum at the "asptiatfon rate", during the B~JIB carotid occlusion mm, re-established typicaL carotid occlu&.oxl IX= sponses. of si@ficance is the fact that mmmJ. of arterial blti, in arz identical mamer, had $51 insignificmt effect on cardiac output, The ability of the co-tent heart to ~spond to carotid occlusion by main- ttining or increasfw cardiac^ output^ is^ dependent^ upon^ reflex^ venocon- striction and the Eaultmt increase in vezmus return of venous origin, (Supported by a grmt from USHB B 31-C)
HYP0cAmT.A Am ERYmRoPoIESIS. R. G, Bartlett, Jr. and N.E.Phillips.* U. S, Naval School of Aviation Medicine, Pensacola, Fla. The unfailing correlation between hypoxia and erythropoiesis ac- companying altitude acclimtization seem to have given rise to an un- questioned cause-effect relationship. Ewever, accompanying a notlce- able hypoxia there is also a hypocaptia produced by the respiratory
loss of carbon dioxide stores. Therefore, by-pocapnia also is posi-
tively correlated with erythropoiesis, and such a correlation raises the obvious question of a possible cause-effect relationship. &peri- merits were designed to ascertain whether or not the hypocapnia ac- companyiw altitude acclfmatization might be a factor in the increased erythropofesis observed, There was a significantly lesser percentage of reticulocytes in the tail blood of mice subJected to a carbon dioxide rich but oxyQen 'poor atmosphere (55 CO ,^ lO$^ 02),^ as^ compared to animals exposed to a similarly low oqgen azmo sphere without added carbon diotide. There was also a lesser weight loss in the animals breathing CO2. During the first few hours of exposure the hypocapnic animals were defititely less active than were^ the^ nommaptic^ animals. From these preliminary observations, then, it is evident that the hype- ca@a accompanying altitude acclimatization may be a contributing factor in the erlytbropoiesfs and other factors observed.
THE PHYSIOLOGIST 19
WE USE OF PERMANENTLYVISUALIZED ORGANSIN STUDYING THE EFFECTS
OF W" FORCES IN DOGS, E, L, Beed$ng+ and F, R, Steggerda, Holloman Air Force &se, New Mexico. Xn 1937, Steggerda and Gianturco demonstiated that the colon of cats can be made permanently opaque to X-rays by injecting Tt-rorium Dioxide (Thorotrast) into the wans of this organ, In the present experiments attempts were made to tisual.izie other organs in the abdominal. cavity and then to record distortions of these organs under varying (^) WI forces in the apparently normaL unanesthetized dog, Organs successfully timalized with this technique ticlude the colon, urinary bladder, kidneys, stomach and diaphragm, The liver and spleen also may be visualized quiti satisfactory by itiecting the opaque material directly into the blood stream, A suspension of finely ground particles of I&SOL (Kicropaque) has in some cases been substituted for morotrast, Ily using a Micronex X-ray machine, the tube of which was mounted over a decelerated sled on which the antial was positioned, X-ray pictures w8re made of the amount of organ displacement in comparison to organ position noted before the stmt of the ex- periment, Adjustments were so made that the X-ray picture was taken at the point of maximum deceleration, The X-ray machine is capable of giving exposure ties as short as l/2000 of a second and thus affords a clear outline of the edges of the organs visualized,
Ablation of the cortical taste area produced transient impairment of taste dlscrimPnation (quinine &drochloride) and coagulation of the thalamic taste nucleus produced permanent impairment of taste diticrim- ination as measured by the traditional preference techntque. dddition- al removal of the olfactory Wbs had no effect on thresholds measured in this fashion, Different results were obtained if thresholds were m8asured by a conditioned avoidance tachnfque using electric &ockr NormI. threbolds were lower than by the preference method, both cor- tical and thalamic removals were followed by only a teqaraw loss of discrimination which returned to norr& levels with further Qaining, and finally, removal of the olfactory bulbs at this stage produced a marked and permnent fncreaae in "tastem thresholds, These reaultt;a suggest that quinine solutiona have an olfactoq component which 1~ not used in the preference situation but which can become a conditioned stimulus in a shock avofdance situation, Thhua if the animal8 are motivated hy ahock they wfll avoid a quinine solutfon on the basis of its olfactory properties at concentrations too weak to taste. (Aided by grant^ s1932,^ mm3,^ mm%)
20 THE PHYSIOLOGIST
HUMANSUBJECTS WITH D-m MENTAIZFY, Heinz W. Berendes, &on Mart+, Robert 5. Ertel, J. A. WCaxtw, John A. Anderson*, and Zkanz Halberg. Cmbridge State School & Hospital, Cambridge, Minn. md Medical School, Utiversity of Minnesota, Mirmeapolfs 14, Minnesota. Circadian (about 24&r.; circa, dies) periodic changes were mapped for 5 ph:nylketonuric patients with verified high blood phenylalanine levels (X=24.9+54 IQ&), 5 mongoloids and 5 medically normal subgects. On the 10 patients, rect (^) #al temperatures were detemined at 6-b. inter- vals for several months. These 1ongitudinaL temperature profiles re- vealed periodic components that were synchronized with the 24-hr. rou- tine of institutional life. Beats on the temperatures record of some cases suggested the i&era&ion of additional free-running though cir- cadian periodicity. Blood profiles wLth 4-hrly. sampling during a 24- hr. period also were obtained for WRC, eosixlophils (chamber oount), cortisol (Mason) and 5-hydroqtryptamine (Davis). Circadian periodic- ity was ascertained in such transversal profiles, with significant phase differences among the variables studied. The blood profiles provide a refined standard of reference, revetiing e.g. increases to 10,000 or more WBC/& at peak-time (z for mongoloids at 17:oO = 11,3oo/nnn3) I Moreover, in both groups of patients blood s-hydrogtzyp- tamine levels were significmtly lower by com~ison to ~~omals, liv- ing on the same diet (P( .05); the me= values were .14&.041)1g/ml for phenylketonurfcs for normals.
,.209&.024 ~&ml for mongoloids 831d .316~016 &ml The finding of lowered blood 5-hydromryptamine levels in these patients with defective mentality is strengthened by the peri- odkity approach which rules out possible differences brought about by sampling in different stages of circadian rhythm.
BEHAVIOR UP TW NON-EMBOLIZEDVESSEIS ACCOMPANYIN GRADED
DIFFUSE PULNONARYEHEWLISM,Theodore Bemthal, A'lan Warres~
and Joseph T,TayXor,III~ Department of Physiology, bdfeal
College of South Carolina, Charleston, S, C.
The vascular bed of the left lower lung lobe In dogs was
embolZzed In situ In steps with 7511glass beads durfng
auto-perfusion with arterial blood at several perfuslola
8 and the effects upon le ft lobar pulmonary artery ,ow measured. changas in sha,pe of the derkved
flow-pressure and reslstame-pressure cum08 and In value
of the ratio
floW &t high PWfUBiOn pI?eSSUlV y8re analyzed
l
flow at low perfusion pressure
Such analyses indicate that, aside from the obstruction of
blood flow occurring in the embolized vessels in the
perfused lobe, the non-embollzed vemel.$ there also offered
progres~lvely lnoreaslng resistance to blood flow as
embolS.zation proceeded and with characteristics suggesting
that this was due to moderate progressively increasiq
vascular tone. (Supported by Grant H-1320, National
Imtitutes of Health, USPHS)
22 THE^ PHYSIOLOGIST
Dssplte reporter to the contrary "denemation ditwemis~ier gene+
tally held to seault from increaaad glomumlar filtration rata (@I%)
in the denemated lddney. *me foUoW studieawxm undertaken to
d-&rate the wrtance of &blea othar than GFR fn pmmotlng
mcsanive acmtion of aodium fmm the danervated kidney. Folla acute, unilafaral, renal denemation in pemtotirbital anesthetized dogs,-clearance s&dies wmru umd to m&ate ren& tiction ti the raparats l!aney8* Ratios of fhnction, daMwv8tad/innemted ahey, during intravenaw $nfb8ions of hype&ode Nacl, ure8 or mamitol indicated that eroCSS8im bxJEF8tiOll of aodium by the denemated kidney waamgp;s=tedbyNaQ inrU8lonor Sldspendently~increaaedGFR ad diminfshed conctitant with incma&q water and solute excretion. Depth of anmthesia wan not a tiabla in tharsa apdmenta.
Iddneymranotdef%ned,butths fai&m of th8 dene&&edkidneyto respond was cle~3.y dknatrabls and attributable, in part, to a defect in 8odfm "reabsorption". InstmanceBktdical I&march Pund.)
(Suppoded by a grant fmm the Life
THE REN IN AND ANG IOTENS I NASE CONTENT OF THE KIDNEY OF NORMAL AND RENAL HYPERTENSIVE RATS. Pedro BlaquieW, David F, Bohr, Alberta C. Taquini,J& and Sibley W. Hoobler, Departments of Physiology and Internal Medicine, University of MTchigan, Medical School, Ann Arbor, Michigan. Renin and angiotensinase concentrations of the kidneys were determined tn the following groups of rats: 1) 7 normal; 2) 12 with renal hypertension produced by unilateral clipping of the renal artery; 3) 11 unilaterally clipped, which failed to develop hyper- tension; 4) 22 with renal hypertension produced by clipping of the renal artery and contralateral nephrectomy; and 5) 7 untlateral cl tpped with contralateral nephrectomy which failed to develop hyper- tension, Kidney extracts were assayed for renin by two different methods: 1) direct intravenous injection fnto a rat sensttIzed by ganglionlc blockade, and 2) the in vitro production of angiotensin which was subsequently assayed In a simrlar manner. Anglotensinase was determined by incubating the kidney extract for two hours at 37O with a known amount of synthettc angiotensin, Renin content was re- duced in kidneys of all groups of rats with renal artery clips, The greatest reduction occured in the rats that failed to develop hyper- tension. The reduction in the untouched kidney was greater than that tn the clipped kfdney, One gram of the normal kidney is capable to inactivate 1 mg. (3000 Dog Units) of synthetic angiotensin under the conditions described above and no change In its concentration was ob- served in the different experimental groups* There was no significant difference between the weights of the kidneys In the normotensIve and hypertensive groups. (Supported by a grant from the Michigan Heart Association,)
THE PHYSIOLOGIST 23
EFFECT OF ZYMOSAN ON PROPERDIN TITERS OF DOGS SUBJECTED TO
HEMORRHAGIC SHOCK, (^) EL Blattberq* and M. N, Levy. St. Vincent Charity Hosp., Cleveland, Ohio. Dogs were injected subcutaneously with 50 mgm. zymosan incorporated in adjuvants, Control animals were also selected but did not receive zymosan, After 2-3 weeks, all animals were subjected t0 oligemia at 35 m-m, Hg arterial pressure fos X50 minutes, then all shed blood was returned to the dog, Periodically, blood samples were drawn and properdin titers were detemined by the phage neutralization assay, The zymosan injected dogs had considerably increased properdin levels at the time of hemorrhage, In all dogs, the titers remained constant during oligemia and fell in the post-transfusion period. Properdin titers fell to a greater extent in the injected dogs than in the control dogs, but rarely fell below the dog's control level,
THEAFFERENTMECHANISMOF THEBAINBRIDGEREFLEX.
Clark M. Blatteis (intr. .by Steven M. Horvath). U. S. Army Medical Research Laboratory, Fort Knox, Kentucky. The effective stimulus for the Bainbridge reflex was studied in 36 morphine-premeditated (0. 5 mg/kg), chloralose-anesthetized ( mg/kg), closed-chest dogs by ivc, right atrium, right and left ven- tricles and pulmonary artery catheterization. Two different stimuli were used: the caval ostia and the right atrium were in the first in- stance perfused with 500 cc/min and 1000 cc/min of whole blood for 15 seconds, in the second instance distended by balloon and prong- catheters for 3 minutes. Heart rate (ECG), blood pressures and oxy- gen consumption were measured continuously; cardiac outputs were determined before and during each stimulus. The observatians were made on 3 preparations: intact, carotid sinus baroreceptors bilater- ally procaine-blocked, and bilaterally vagotomized dogs. The f3ainbridge reflex was not elicited by an increase in the rate of flow or the volume of venous return; in the intact dogs the heart rate was slowed rather than increased during the perfusions, and it was not affected in the denervated preparations. Venous and arterial systolic pressures were increased in all the preparations, but least in the vagotomized dogs. Central venous hypertension alone likewise did not induce reflex cardiac acceleration since localized caval and right atrial distension produced no heart rate and blood pressure changes in any of the preparations used. Cardiac outputs were increased during perfusion, but not changed during distension. The heart ac- commodated to increased filling by increasing systolic discharge,
