sign in sign up
<<
Home , Ackerman (surname), Circulatory system

A r ^ T ri Advisor Department of Approved by BY i960 DISSERTATION Ohio Ohio State University The Ohio State University Kenneth Rae Coburn, B. S. OF CARBON MONOXIDE OF DOGS TO LETHAL resented in Partialegree Fulfillment Doctor of of Philosophythe Requirements in the Graduate for School the of the THE CARDIOVASCULAR AND RESPIRATORY RESPONSES Pi Q ACKNOWLEDGMENTS

I wish, to express my sincere appreciation for the guidance, enthusiasm and invaluable aid of Doctors Fred A0 Hitchcock, Earl T.

Carter and Joseph F„ Tomashefski without whose assistance this study could never have been carried out.

I am grateful to all those of the Cardiopulmonary Laboratory of the Ohio State Tuberculosis Hospital for the aid and assistance which they offered. CONTENTS

Introduction ...... 1

Survey of the Literature

A. Early History of CO ...... 4

B. Absorption and Excretion of C O...... 5

C. , Carbon Monoxide and ...... 8

D. and Circulatory Changes inCO Poisoning...... 15

a. Blood Changes . 15

b. Blood Vessels and CO ...... 16

c. Behaviour of the ...... 16

E. Effect of CO on ...... 18

F. Recent Integrated Studies...... 19

Methods and Procedures

A. and Surgical Preparation...... 21

B. Respiration Measurements . 22

C. Experimental Protocol...... 25

D. Blood Analysis...... 27

E. Statistical Analysis ...... 30

Results

A. Ventilatory Response ...... 32

B. Blood Changes...... 32

C. Cardiovascular Changes ...... 33

Discussion...... 35

iii Summary and Conclusions ...... 43

F ig u r e s ...... - ...... 44-69

T a b le s ...... 70-75

Bibliography ...... 76-81

Autobiography ...... 82

iv LIST OF FIGURES

Page

1. Equilibrium Concentrations of CO with Respect to T im e ...... 44

2. The Haldane Shift ...... 45

3. Representative Data Taken from Swann and Brucer . . . 46

4. Schematic Diagram of Experimental Set-Up...... 47

5. Terminal Yentilogram...... 48

6-26 Individual Data Plots of Experimental Animals.... 49-69

v LIST OF TABLES

Page

1. Cardiovascular Responses to CO ...... 70

2. Blood Gases ...... 71-73

3. Blood Changes ...... 74

4. Ventilatory Response...... 75

vi INTRODUCTION

One of the first of the insidious and deadly by-products of

civilization appears to have been carbon monoxide, CO. As soon as

man became civilized enough to carry fire into a restricted environ­

ment the of CO poisoning became constant companions of man.

While thousands of years were to pass before the mechanism of CO poisoning was discovered and described, it has been associated since

the earliest times with the presence of fire. For thousands of years

the only common source, of CO was from the incomplete combustion of

vegetable fuels, but with the advent of the fifteenth century the increasing

use of coal as a fuel led to a tremendous increase in the incidence of

CO poisoning. With each development of more effective means of

power production the problems grew until, with the invention of the

internal combustion engine, it became one of the major problems not

only of industry but of common usage. In our present industrial age

with natural gas heating, myriads of gasoline engines, jet propulsion

and the widely diversified manufacturing processes required for the maintenance of our standards, we find that CO intoxication

has become one of the most widely distributed and frequently en­

countered types of poisoning.

Carbon monoxide, CO, has a specific gravity of 0.9671 ( air = 1 )

and a density of 1. 2504 grams per liter ( STPD ) with a molecular

of 28.01.

1 It is colorless in all concentrations and is odorless in concentrations up

to about 80 per cent. In higher it has a distinct garlic-

like odor. It melts at -207° Centigrade and boils at -192° Centigrade.

The following list which was compiled from the Bureau of Mines^

shows the approximate proportion of CO in gases to which relatively

large proportions of the population of the United States may at one time

or another be exposed.

Type and Source CO by volume per cent Blast furnace stack gas 28. 0 Bessemer furnace gas 25.0 Producer gas from coke 25.0 TNT blast gas 60.0 Fuel gas 30.0 Gas range using natural gas 0.2 Room heater using natural gas 0. 5 Automobile exhaust gas 7.0 Natural gas heater 1. 0 Arc furnace melting aluminum 3 2, 2

The of CO is due to the fact that it becomes bound strongly

to the hemoglobin of the circulating blood and in this manner interferes

not only with the normal transport of O£ from the to the tissues

but with the reciprocal function of the bLood in respiration, i.e., that

of transporting CO2 from the various tissues of the body to the lungs

for excretion. In this it would seem that CO poisoning is merely

another form of anemic and that in describing the physiological

relationships of one, such as hypochromic anemia, one would in essence be describing the relationships in ail. This is not strictly true inasmuch

2 as CO has a discrete action on the blood vessels of the body which must be taken into the overall picture if a concise knowledge of the mechanisms by which it acts is to be obtained.

In addition, CO is known to combine with cytochrome oxidase in

such a way as to inhibit the action of this enzyme. However, the

affinity of cytochrome oxidase for CO is 1, 000 times Less than its

affinity for O^ and it .is questionable whether this mechanism has a

role in CO poisoning. ®

In view of the fact that the rational treatment of CO poisoning

depends upon a dear knowledge of all the pathophysiological mechanisms involved in the entity of CO intoxication, it was decided

that a rather comprehensive investigation of the cardiopulmonary

responses of to lethal concentrations of CO be undertaken

in an attempt to clarify the relationship between the ventilation effects

and the cardiovascular response to CO.

Of particular interest were certain time relationships existing

between the respiratory center and the vasomotor center throughout

the exposure to a lethal concentration of CO. Is the ultimate cause

of death in the intoxicated animal due prim arily to respiratory

failure or is this merely a manisfestation of a more subtle process. 4

SURVEY OF THE LITERATURE

A. Early History of CO Poisoning

The first case of CO poisoning probably occurred when one of the first to use fire in a cave rolled a rock into the opening of his cave to seal it for the night. By early historical times the effect, if not the cause, of CO poisoning was known.

Sayers and Davenport, ^ from whom the unnumbered references to follow are cited, report that Lewin ^ lists numerous incidents of

CO poisoning which were described in the early Greek and Roman literature. One of the first was reported by Livius during the Second

Pumic War, about 200 B„C0, who wrote

the commanders of the allies and other Roman citizens were seized suddenly and fastened in the public baths for guarding, where the glowing fire and heat took their breath away and they perished in a horrible manner. ^

CO used as a means of self-destruction was first reported by PLutarch who wrote that Catalus kilLed him self by closing him self in a bath with the vapor from many glowing coals. It would appear then that the lethal effect of enclosed fire was well known and documented by the ancients. The nature of its action was, however, to remain obscure for many years. Cassius attributed the ill effects not to fire but to the dry heat it produced. As late as the eighteenth century CO intoxication was still being superstitiously attributed to the work of the devil. However, in 1648 Bacomis de Verulamio noted the fact that the gas was odorless and he referred to this gas as vapor carbonum. Previous to Verulamio the rather vague term fumus had been

applied to the causative agent in CO poisoning. Van Helmont, in 1667, was the first investigator to use the term carbon gas, while Boerhaave

found that all red-hot organic matter released fumes which were lethal 4 to animals.

Pure CO was first prepared in 1796 by F. de Lassone who reduced

zinc oxide with pure carbon. Lavosier knew that CO could be burned

to produce CO^ but was unable to fit it into his general oxidation

theory until, in 1800, Cruikshank showed that CO was what he called the gaseous oxide of carbon. Although the existence of CO was now known, the causative agent of the associated poisoning was still in doubt. As late as 1812 the Dictionnaire des Sciences Medicates du

Paris stated that .it had not definitely established which gas, i.e., CO 4 or H2 S, was responsible for the lethal effect.

B. Absorption and Excretion of CO

The absorption of CO by hemoglobin takes place via the lungs and will therefore be influenced by the degree of physical activity of the subject, hydrogen ion concentration of the blood and any of the other numerous factors which modify respiratory function. ^ Some

CO is absorbed through the skin and mucous membrane, but the amount absorbed in this fashion is so small as to be considered

6 7 negligible. * To illustrate the complexity of the factors

Q governing the absorption of CO, Henderson and Haggard have pointed out that if two individuals of different size are exposed to the same concentration of CO, the smaller and younger one will absorb

the CO more rapidly on account of the more active . The

displacement of O^ in oxyhemoglobin by CO is determined by the ratio

of the respiratory volume to the size of the body and the amount of

blood. The respiratory volume of a person at rest bears a direct

relationship to the surface of the body while the amount of blood has

a direct relationship to the weight of the body. As the relative size

of the surface increases with the inverse ratio of the mass it there­

fore appears that the asthenic person may be more sensitive to CO

than the stout person because the respiratory volume is comparatively 2 greater than the amount of blood.

The excretion of CO from the body is accomplished by the lungs and is the reverse of the absorption process and, as would be ex­

pected from the shape of the curve in Figure 1, the period of time

required to excrete a given quantity of CO is many times as great as

the time required to absorb the same amount. For example, it re- o quires about ten hours, to reduce an initial saturation of 35 per

cent COHb to the 5 per cent level, although sufficient CO may be

absorbed in a matter of a few minutes to raise the saturation from

0 per cent to the 3 5 per cent level. It was also noted that the de-

saturation time required to reduce the COHb Level from 35 per cent

to the 0 per cent level could be decreased by a factor of four by

100 per cent O^.

Figure 2 shows the time characteristics of COHb saturation with varying concentrations of CO. ^

Sayer s and Yant noted that under the same conditions mentioned above the use of a mixture of 92 per cent 0£ and 8 per cent CO2 de­ creased the time required for desaturation by a factor of five or six.

It should be noted that the time course of COHb dissociation follows an exponential curve and, therefore, as the saturation of COHb de­ creases, the time required for the dissociation of the remainder in- creases proportionately. 1 ?

Some of the very early workers noted that there seemed to be evidence that indicated an oxidation of CO to2 CO in the body. 1 3,

14, .15 Later investigators ^ felt that this view was mistaken and in all probability was due to the fact that the early investigators were m isled by the increased excretion of CO2 seen in the later stages of CO poisoning.

More recently, investigators have not only substantiated the early claims of the internal oxidation of CO to CO2 but have quantitated such reactions. Fenn and,Cobb 18 reported in 1932 an increased gas consumption of from one and one-half to three times the control values in frog muscle respiring in an atmosphere of 79 per cent CO and 21 per cent CO. This decrease in the R.Q. suggested an oxidation of CO to CO^. Such a reaction has an intrinsic R.Q. of 2.00, although the measured R.Q, of the re­ action was only 0.66. This reaction has been confirmed.

Recently Sjostrand ^ and Nicloux have presented data suggesting the endogenous formation of CO in man.

The absorption and excretion of CO by the lungs represents only

a portion of the whole picture of the mechanism of CO pickup, for

it is obvious that the reactions of CO with the blood and other tissues

or fluids of the body will determine the time course of both absorption

and excretion.

C. Oxygen, Carbon Monoxide and Hemoglobin

It is apparent from the rapidity with which CO is absorbed and

the slowness with which it is given off that, although the reaction is

reversible, the rate of the dissociation reaction differs from the rate

of association. It is upon this characteristic that the extreme toxi­

city and insidiousness of CO poisoning rests. On the one hand we see the rapid formation of the COHb with the resultant interference

in the hemoglobin's ability to transport O2 and, on the other, we

see the persistence of this disability over a prolonged period of

time after discontinuing CO breathing.

COHb has a bright cherry-red color which was first noted in A the middle of the sixteenth century by M arcellus Donato of Montua.

Troja ^ also noted the color and recorded that this color persisted

for some time after death. Similar observations were made in dogs

a 0 7 poisoned with CO by Piarrey in 1826, Maryl reported the bright

heaLthy appearing venous blood in cases of CO poisoning but it r e ­

mained for Bernard ^ and Hoppe-Seyler ^ to determine that this

red color was the result of the formation of a new pigment in the blood as a result of the combination of CO with hemogLobin. They

also noted that in spite of its heaLthy color, it was unable to sustain

life, it resisted oxidation and putrifacation to a greater degree than normal blood and that the 2 O carrying powers of the blood were

hampered by its' presence. They concluded that CO poisoning was

therefore a form of asphyxiation.

In regard to the resistance of COHb against by

9 c putrifacation, several reports can be cited. Weimann was able

to dem onstrate the presence of COHb in cadavers up to fifty days

post m ortem ; WiethoLd ^ after 122 days; and Heilmenn ^7 after

144 days. The presence of HbCO in exhumed bodies has been

demonstrated by chemical and spectrographic methods in ateLectatic

after two and one-half years and in blood kept in completely

2 ft full containers after twenty years or more, according to Piezarkowski.

It is apparent from the speed of the reaction between CO and hemo-

globin that there is a strong affinity of the one substance for the other.

From the work of Haldane *0 and others, it is now known that the

affinity of Hb for CO is in the order of 300 times greater than the

affinity of Hb for 0£. Although it was generally believed that this

affinity was due to the speed with which the reaction between CO and

Hb took place, H artridge ^ felt that both reactions occurred

practically instantaneously in both cases. Roughton, ^ however,

found that when the reaction rates of 2O and CO with Hb were com ­ pared, the Hb0 2 combination was found to proceed at ten times the 10 rate of the COHb combination. He therefore concluded that the affinity was due to a very low velocity constant of the COHb-^ CO + Hb reactions. He also found that although the reaction rates of CO and

O2 with Hb were not greatly affected by changes in hydrogen ion concentration, the shapes of both Hb02 and COHb dissociation curves were markedly affected.

31 32 3 3 It has been pointed out by , Haldane, and Nicloux that the reaction of Hb with O2 and CO, as when in contact with a given concentration of CO in air, follows the law of mass action.

It therefore follows that the speed and ultimate degree of saturation

Hb with CO depends largely upon the concentration of CO in the gaseous mixture exposed to the blood. It might also be noted that for every concentration of CO, there is an equilibrium level that cannot be exceeded regardless of the length of time during which the Hb is exposed to the mixture. See Figure 2.

The combination of CO with Hb is a completely reversible reaction although, as mentioned above, the dissociation velocity constant is very low. It has been shown 34, 35 that Hb binds equal quantities of both CO and 2O. It has been demon­ strated ^ that one atom of binds one molecule of CO in the manner shown on the following page. 11

H e

CH

It can be seen that a single moLecule of hemoglobin can carry from 0 to 4 molecules of CO or or combinations of the two substances.

However, the affinity of Hb for CO does not rest solely upon the low velocity constant of dissociation and the law of m ass action. Krogh, ^ and Later Hartridge, found a variation in affinities of

Hb for CO and which was species dependent. Douglas, HaLdane and HaLdane ^9 reported that not only was tlie affinity of Hb for and CO specifically variable but also the Hb from one individual varied in its degree of affinity from day to day. • It must be men­ tioned that the above data were compiled with hemolized blood Hb,

39 unhemolized blood Hb or Hb of questionable purity.

It is apparent then that the action of Hb as found in the living red blood differs from "freed" Hb and that the behaviour of the

40 former is of greater bioLogical significance.

A 1 As mentioned by HaLdane, *±I it would be difficult to understand some of the symptoms of CO poisoning if the action of CO were simply the production of an hypoxic state by interfering with the O^ transport by Hb. One of the actions of COHb is to shift the Hb02 dissociation curve to the Left, The greater the per cent saturation of Hb with CO, the further to the left the Hb02 curve is shifted. ^

See Figure 1. This in effect reduces the amount of O2 available to the tissues over a given difference in PO2 such as might be found between and blood. The shifting of the Hb02 dissociation curve to the left increases the slope of the Logarithmic phase of the curve. This means that fewer volumes per cent of 02 will be released over a given arteriolar-venule PO2 gradient. Thus, not only does CO dispLace 2 O in the Hb molecule at a ratio of about 13

3 00 to 1, but the very existence of COHb in the circulating blood decreases the ability of Hb0 2 to perform it's primary function, i.e ., that of supplying the tissues with O^.

In addition to the deleterious action of COHb on the function of

Hb0 2 > many investigators have reported a certain degree of hyper­ ventilation accompanies CO intoxication. ^

This hyperventilation results in a decreased a^-~ veolar air spaces and subsequently reflects itself in a decreased concentration of CO2 in the blood. This adversely affects the dissociation of COHb, as was demonstrated by Douglas, Haldane and Haldane. ^ In their experiments it was shown that increasing the CO2 content of the blood shifted the COHb dissociation curve to the right, resulting in a shallower slope which favors COHb dissociation.

The decreased concentration of CO2 mentioned above would naturally be accompanied by a decreased hydrogen ion concentration 44 as reflected by the Henderson-Hasselbach equation:

p H =p K + log HCQ3 (1) H2co3

However, respiratory alkalosis might be prevented by increased

45 lactic acid levels and the interference of CO with the HbO^-HHb buffering system which normally operates in the blood. The end result is an increase of hydrogen ion concentration which again 14 tends to shift the HbC>2 dissociation curve to the left.

When blood or a of Hb .is exposed to a m ixture of CO and O2 , an equilibrium is reached in accordance with Haldane's equati on:

HbCQ = M P CO (2) POT

The term s HbCO and HbC02 represent the number of moles of

CO and O2 combined with the Hb in one liter of blood or Hb solu­ tion. M is a constant which, as mentioned above, is species variable but which, in whole blood, is remarkably constant in any

46 individual of a given species. In humans it is normally about 2 50.

The 3?qq and PO^ refer to the respective partial exerted by the CO and O2 in accordance with Dalton's Law of Partial

P ressures:

P total = Pi + P2 + + ...... O)

For any given concentration of gas in a physiological gas sample under BTPS conditions, the following equation is used for determining the partial exerted by that gas:

P gas “ ^barometric” x F gas

Here the term P refers to the pressures in millimeters of mercury and F is the concentration of gas expressed as a decimal equivalent of 1.0. 15

Equation (2) has been repeatedly verified where the sum of P^q and P0 2 high enough to saturate the Hb so that the amount of reduced Hb present in the blood is negligible.

Equation (2) holds true in a strict sense only at equilibrium.

However, if the above-mentioned assumption is correct, i.e. , that

the combinedP q q and'PQ^ are high enough to insure complete satura­ tion of the hemoglobin with either O2 or CO, then at any given moment during the course of the CO intoxication an indication of the amount of COHb circulating in the blood can be obtained by utilizing the following empirical relationship:

COHb (1.00 - HbOz) (5)

D. Blood and Circulatory Changes in CO Poisoning

(a) Blood Changes

The blood changes noted in CO poisoning are in some cases striking, but when the overall picture is viewed these changes show an inconsistency. This is probably due to the varying concentrations of CO used to produce intoxication as well as varying time courses.

47 48 49 Many authors ’ ’ have reported increased red counts in cases of prolonged exposure to relatively low con­ centrations of CO.

The mechanism of the polycythemia remains obscure, although,

52 51 in acute cases of CO intoxication, reports on dogs and cats 16

indicate that a contracture of the splenic musculature is

responsible, although this would seem an unlikely mechanism for

maintaining the prolonged polycythemia sometimes seen following

moderate exposure to CO. von Oettingen ^ feels that this polycy­

themia represents an alarm signal caused by direct CO damage

upon vital structures rather than the reflection of a physiological

compensation.

(b) Blood Vessels and CO

The circulatory apparatus is unquestionably affected to various

degrees and in different fashions by exposure to CO but it may be

stated that the prim ary effect of CO is upon the

and most, if not all, of the other variations are the result of these circulatory disturbances.

'54 Ackerman in 1858 noted that CO and illuminating gas poisoning

always produced and hyperemia of the brain which was

more marked than if was produced by HCN or chloroform.

o c e c L This finding has been repeatedly substantiated. ’ ’ The vaso­

dilation found in CO intoxication is not uniformly distributed through-

57 out the body. Weimann notes that in spite of generalized cerebral

hyperemia some collapsed vessels could also be found. The direct

effect of CO on the blood vessels cannot be overlooked.

(c) Behaviour of the Heart

Investigators appear to be unanimous in the opinion that cardiac

irregularities are almost always observed in CO poisoning and that 17 they tend to be more frequent following prolonged exposure to Low concentration of CO. The irregularities seen following acute 58 exposures to CO may appear after a latent period of a few days,

Symanski ^ stated that existing compensated cardiac injury would be aggrevated by exposure to CO.

Chance and Jackson ^ found that in severe acute CO poisoning the heart is seriously affected and that once the heart has stopped beating it is extremely difficult to revive. This has also been demon- 61 strated in a series of 67 dogs by Schwerma et al who state that the condition of the heart is paramount for survival. In their total experimental series of 206 dogs those failing to survive the CO in­ toxication showed venous dilation, pulmonary and hepatic congestion and . These investigators attribute the deaths to cardiovascular failure. Zondek reported on four cases of acute dilation of the heart and referred to six other cases cited in the literature. This may indicate a weakening of the cardiac

6) 3 muscle, however Zondek felt as did Klebs that this was not a primary effect on the heart muscle but was secondary to vasomotor 64 failure. Haggard has shown that the vasomotor center retains its activity longer than the respiratory center. If dogs exposed to lethal concentrations of CO were supported in the latter stages of the intoxication by being given eight to ten per cent CO„ to far breathe, the respiratory center continued to function and there was 18 no decrement in the auriculo-ventricular conduction time. From this

Haggard concluded that the impairment of cardiac conduction is solely a function of anoxemia and that there is no direct effect of CO on the heart.

E 0 Effect of CO on Respiration 1 3 Haldane in his self experiments reported that an increase in ventilation was noted when the saturation of hemoglobin with CO

6 5 reached about 35 per cent. Chiodi, Dill, Consolazio and Horvath measured ventilatory responses in men and dogs at levels up to fifty per cent HbCO. They reported no significant changes, nor were variations of any magnitude noted in CO^ capacity or ^ con_ 66 trast, von Oettingen, Donahue, Valaer and Miller reported an increase in ventilation in anesthetized dogs which amounted to a

45 twofold increase over the control level. Swann and Brucer con­ firmed this finding in conscious dogs. Recent work in this labor a- 6 7 tory tends to confirm the finding of increased ventilation in CO poisoning.

The stimulus for this hyperventilation is obscure for the of the blood remains normal in CO poisoning and therefore it is reasonable to assume that the chemoreceptors of the carotid and aortic bodies would not be stimulated. This view agrees with the 68 findings of Comroe and Schmidt. F0 Recent Integrated Studies

The only report citing consistently increased during CO poisoning is that of Chiodi, Dill, Consolazio and Horvath,

Although definite trends in their data can be seen, it is questionable in some instances whether or not these changes reflect real or apparent conditions.

In their comprehensive investigation of rapid anoxic death, 4’5 Swann and Brucer devote one chapter to death by acute CO intoxi­ cation. Their results are too lengthy to be shown in tabular form, but a graphic time course for one dog is shown in Figure 3. This time course was selected by them apparently as representative data.

Their data show variations in ventilation which are definite but are said by the authors to be slight, in view of the tenfold increases possible. The arterial pH decreases steadily throughout the course of the experiment. No data concerning cardiac output is presented but the writers state that cardiac output is probably maintained until shortly before death. They further state that previous re­ ports which indicate a gradual decline in arterial are in error due to incomplete measurements and that although the diastolic pressure does decline slowly, the systolic pressure shows a great concurrent increase and as a result the mean blood pressure is defended. However, mean blood pressure calculated from their data shows that half of their animals did exhibit a gradual, progress­ ive decrease in mean blood pressure over the entire time course. This work was done on dogs with only LocaL anesthesia at surgical sites. The average initial mean blood pressure was 153 mm Hg.

Systolic pressures ranged up to 323 and diastolic up to 134 mm Hg.

This probably reflects cortical effects in that the dogs were fully conscious and no matter how well trained could be expected to be apprehensive. This fact would make interpretation of blood pressure and minute ventilation data extremely difficult. METHODS AND PROCEDURES

A. Anesthesia and Surgical Preparation

Healthy mongrel dogs of both sexes, females non-gravid, weigh­ ing from ten to fifteen kilograms each were utilized as experimental animals. General anesthesia was induced by the intravenous injec­ tion of thirty milligrams of pentobarbital per kilogram of body weight. The animals were prepared for the surgical pro­ cedure by shaving away the hair from the right side of the neck and from the medial surface of the right thigh.

An endotracheal tube equipped with an inflatable cuff was'intro­ duced into the under direct visual observation and advanced to the carina.

A cutdown was made on the right external jugular , which was then tied off distally. A number 8 cardiac was intro­ duced into the lumen and under direct fluoroscopic observation was advanced into the pulmonary . It was then tied securely into place in order to prevent it from dislodging. A three-way stopcock was attached to the end of the catheter to allow withdrawal of samples of mixed venous blood and of desired substances.

A second cutdown was made on the right femoral artery which was tied off distally. A blunted 13 gauge needle was then introduced into the lumen of the vessel and tied securely in place. This was used for the periodic withdrawal of and was also utilized 22. in monitoring the arterial blood pressure. The latter was accom­ plished by attaching a fluid-filled Statham pressure transducer having a range of from 0 to 75 centimeters of mercury. The output of the

transducer was fed into a Sanborn strain gauge amplifier and direct writing recorder. The Sanborn apparatus had been previously cali­ brated with a mercury column manometer.

Following the surgical procedure the animal received via the

cardiac catheter 4 milligrams of sodium per kilogram of body weight.

B. Respiration Measurements

The endotracheal tube was attached to a specially constructed

having a of less than 50 cubic centimeters. The

direction of the flow of the gases used was controlled by low r e ­

sistance check . The inspiratory side of the vaLve connected by means of a large bore corrugated tubing to three 130 liter capa­

city Douglas bags linked in series. See Figure 4. These bags were

filled with either room air or with a CO in air mixture. The differ­

ent gas mixtures were prepared as follows: For the 0. 3 per cent

CO in air mixture, 1170 cc. of commercially prepared 100 per cent

CO was measured out into a large Tissot gasometer. Room air was

then drawn into the gasometer and forced into the Douglas bag system

until a total of 390 liters, of gas occupied the system. The 0.4 per cent

CO in air was mixed in the same way except that 1560 cc. of CO was 23 initially placed in the Tissot gasometer. Prior to administration of the GO, the DougLas bags were manually agitated for several minutes to insure thorough mixing of the two gases. The concentration of the final mixture was checked by means of a Mine Safety Appliance CO

Detector. In our hands this detector had an accuracy of about 0.03 per cent in the range being utilized. The end concentration, i.e., after the dogs had died, of the CO in the bags was checked to insure that the CO had not diffused out of the bags in any significant amount during the procedure. There was no detectable difference in the CO

concentration between the two measurements.

The expiratory side of the valve on the endotracheal tube was attached by means of rubber hose to a large Tissot gasometer which was equipped with an ink-writer recording drum driven by a constant speed motor. In this manner the ventilation could be constantly monitored. The ventilatory spirogram and the continuous­ ly recorded blood pressure were observed until a steady state was

seen to exist, i.e., the spirogram slope remained constant and there was no persistent rise or fall in either systolic or diastolic pressure.

The expired air samples drawn at the three observation periods were transferred from the small Douglas bag to 50 cc. oiled equipped with three-way stopcocks and a small rubber-tipped capi­

llary tube. These syringes were then used to transfer the expired air samples into the reaction chamber of a Scholander Microgaa Analyzer for determination of the concentration C>of2 > and N. in the sample. Repeated analyses were done until checks on all

samples were obtained. The limit of error allowed in the analysis

of the expired gas samples was 0. 04 per cent.

Minute ventilation and the respiratory rate were obtained from

the recorded spirogram. From these data the inspired volume,

consumption, CC>2 output and R.Q. can be determined by the follow­

ing formulae:

( 6)

(7) V0 2= " (VE x FE q 2)

VC 0 2 = ^ E x F Ec0 2 )-(YI xFIc02) ( 8)

vco2 (9)

Vj = Volume inspired

VE = Volume expired

F e^ = Fraction of N2 expired Z 25

Ftp = Fraction of GO„ expired ^ C 0 2 2

Fj = Fraction of CC>2 inspired

u

V q 2 = Volume of 0 2 per minute

= Volume of C0 2 per minute

Throughout the ventilation measurements the Tissot gasometer

and the barometric pressures were noted and all ob~..

served gas volumes were corrected to BTPS or STPD as required.

C. Experimental Protocol

The dogs were divided into three groups. Group A, numbering

seven dogs, received 0.4 per cent CO in air. Group B, six dogs,

received 0.3 per cent CO in air. Group C, five dogs, served as the control group and received only room air.

During the initial steady state, the first sample series was taken.

This series consisted of a three-minute sample of the mixed expired

air collected in a 5-liter Douglas bag, which had been previously

evacuated by means of a vacuum pump, immediately followed by a

simultaneous withdrawal of 1 2 cc. samples of arterial and mixed

venous blood. During the withdrawal of the blood samples, the>2 C

uptake was measured directly by means of a Benedict-Roth basal

metabolism apparatus which was connected by three-way valves to

the inspiratory and exspiratory sides of the ventilation apparatus.

Immediately following the first series of observations, the dog 26- was connected to the bags containing either one of the CO mixtures or, in the case of the control series, room air. The time of talcing the second series of samples varied with the concentration of CO being used and to a certain extent from animal to animal. The criterion for the second sampling was in part subjective but in general depended upon the attainment of a new steady state after the onset of CO breath­ ing, as indicated by the spirogram.

The third and last sample series was taken when irregularities appeared in the respiratory pattern. These irregularities usually preceeded death by four or five minutes. See Figure 5. During the

O^ uptake measurement for the third sample, a brief ventilatory recovery period frequently was observed. This was due to the necessary interruption of the flow of the CO m ixture when the ani­ mal was placed on the 50 per cent O-, mixture utilized in the Bene- dict-Roth apparatus. This mixture was used in order to insure that all reduced hemoglobin which might possibly be present in the a r­ terial blood would be saturated with O^.

The recovery period mentioned above was always brief, for immediately upon completion of the one-minute period required for the O^ uptake determination the animal was turned back into the

Douglas bags containing the CO mixture and the progressive build­ up of COHb allowed to proceed to its lethal consequence. 27

D» BLood Analysis

The blood sampLes were withdrawn anaerobicalLy and the capped,

Heparinized syringes were placed in a container filled with iced water

until the blood gas determinations were done. The hematocrit was

determined by means of centrifugation in Heparinized tubes

50 mm. in length. The pH of the arterial blood was determined as

rapidly as the experimental procedure would admit and was accom­

plished in ail cases within 15 minutes of withdrawal. A Cambridge

pH meter was used for all determinations. Before and after each pH

determination a standard buffer was read to insure that the machine

was properLy adjusted.

The and CO^ contents of both arterial and venous blood were

measured by means of the Van Slyke manometric technique, as modi­

fied by Niell. ^ In addition, the initial capacity of the arterial

blood was measured on each dog by saturating an aliquot of blood utilizing room air.

The C>2 uptake data from the Benedict-Roth apparatus and the difference between the O^ contents of the arterial and venous bloods

permits calculation of the cardiac output by means of the Fick

70 equation: Uptake (cc/min) :______(io) C.Oo - A q 2 - V q 2 (V%) 28 .

Only the direct Fick method of determining cardiac output was

used in Group A, which received 0.4 per cent CO in air, and in

Group C, the control group. Group B, which received 0.3 per cent

CO in air had the cardiac outputs of the individual dogs simultaneously

determined by means of the dye dilution technique utilizing bromo-

sulfophthalein sodium, which will be described below, and also by

the direct Fick technique. It was decided that in view of the agree­

ment between the two methods and the relative ease with which the

Fick method can be employed that the latter method would only be

employed in Groups A and C.

The basis for the dye dilution technique7 0 used to measure

cardiac output in Group B is that the amount of dye injected into the

venous side of the system must pass through the heart and appear

in the arterial side of the circulation. This assumes a negligible loss of dye in transit through the . If I = the

amount of dye injected and O = the amount of dye appearing in the

arterial blood, then:

1=0 ( 11)

If the arterial blood is collected over a period of timed intervals,

then:

I = Ox + Oz + 0 3 + 0 4 + ...... On (12)

If the time intervals are one second apart, then the amount of

dye may be expressed as concentrations of dye in known volumes of 29 blood collected at the specified intervals and:

I = Ci x Q + C 7 x Q +...... C xQ (13) So d 60 n 60 where Q equals 1 / 6 0 th of the output of the heart for any given 60 sixty-second period. By factoring equation (13) we obtain:

I = Q (Cj + c2 + ...... Cn) (14) 60 and

I x 60 = Q (Cx + C 2 + ...... Cn) (15) or

Q - J a i l ------:— (1 6 )

x : • '

We see then that the cardiac output, Q, is equal to sixty times the amount of dye injected divided by the sum of the concentrations of the dye over one through n samples at t intervals. In this case, one sample per second was taken.

Using equation (1 6 ) it is possible to determine the cardiac out­ put by injecting a known amount of dye and then determining the con­ centration of the dye in successive samples of collected arterial blood. The concentration of the dye in the arterial blood was deter­ mined by centrifuging the blood and determining the optical density of the dye'in the supernatant plasma by means of the Beckman Model

DU Spectrophotometer. The concentrations so obtained were then plotted on diphasic semi-logarithm paper. Sufficient points were 30

plotted so that the exponential decline of the descending limb of the

concentration curve could be seen and the line extrapolated from

these points to zero concentration on the abcissa. The sum of the

concentrations was calculated and equation1 6() was then utilized for

determining the cardiac output.

Simultaneous Fick determinations were also made as described

above. The two methods agreed within less than 10%, showing ran­

dom variation.

E. Statistical Analysis

The standard deviations for the mean values of each of the

measurements and calculations were determined by use of the 73 following equation:

(17)

N - 1

where x is the mean value of the measurement or calculation and N

is the number of measurements or calculations comprising the mean.

The standard error of the difference between the sample means

71 was calculated by the equation on the foLLowing page: 31 .

Ni + N 2 S» E. = ( 18)

N1 N 2

Where x^ is the mean of the experimental group and ^x is the mean of the control group, Nj and N2'refer respectively to the number

of items represented in the experimental and control means.

The t ratio was obtained for any given pair of measurements by dividing the difference between the two means by the standard

error, S.E. Using this value and entering the t table for degrees

of freedom - 2 , the probability that the difference between

the means in question represents only a random difference can be determined. RESULTS

The results obtained from these series of dogs are tabulated in

Tables I through IV and are shown graphically in Figures6 through

27.

A. Ventilatory Responses

In Group A there was no change in the tidal volume between

Sample I and Sample It; however, a slight but statistically insignifi­

cant increase is seen in Sample III. The minute ventilation showed

a 54 per cent increase over pre-exposure values with a decline to

less than pre-exposure values as the animal approached death.

The respiratory rate showed an increase of 64 per cent; then a de­

cline to less than pre- expo sure rate.

Group B showed little if any ventilatory response except for a

decrease in minute ventilation during Sample III.

Group C showed no significant change. It should be stated that none of the variations mentioned above were statistically significant,

i.e., the probability levels were greater than 5 per cent.

® • Blood Changes

In Group A there was a decrease of 8 mm. Hg on the arterial blood F<30g followed by an increase of6 mm. Hg. as the animal

approached death. There was noticeable change in the arterial pH

in that there is an initial rise from7.27 to 7. 32 followed by a drop

to 7. 19 in Sample III. There was no change in hematocrit; however, 33- there was a very significant fall in the content of the arterial and venous bloods.

In Group B, the findings parallel those seen in Group A.

Group C showed no significant change.

C, Cardiovascular Changes

In Group A there was a slow but progressive decline in the , accompanied by an initial increase in volume , followed by a fall in in Sample III. The cardiac out­ put showed an initial increase of 54 per cent. This rise was not maintained, as the output fell to almost pre-exposure levels in

Sample HI. The above changes are not statistically significant, however there was a significant and progressive decrease in the mean blood pressure, indicating a decrease in peripheral resis- tance. The data on Group B parallel the changes seen in Group A with the exception that there was an initial slight decrease in cardiac output. Group G showed no significant changes from the initial values.

Although the time course differed rather sharply in Groups A and

B, the same general trends were seen whether 0. 3 per cent or 0.4 per cent CO was used. In the discussion of the ventilatory changes it is necessary to consider simultaneously the changes which have been observed in the chemical nature of the blood for, according to the multiple factor theory, these chemical changes provide the stimuli for altering the respiratory pattern or reflect any such change. The absence of changes in Group B in contrast to Group A after an average of 30 minutes of CO breathing can be explained on the basis that the lower concentration of HbCO had not yet provided sufficient degree of hypoxia to invoke an alteration in the respiratory pattern. However, it must be noted that the arterial blood reflected a decrease of 7 mm. Hg. in anc^3JX *ncrease 0. 03 pH units, changes suggesting hyperventilation. At the time of these findings the approximate concentration of COHb was 24 per cent. DISCUSSION

The enigma posed by exposure to lethal concentrations of CO

seems to be whether death is due to prim ary failure of the re sp ira ­ tory center or to circulatory failure which, in turn, causes the failure of the respiratory center.

At first glance it may appear that the answer is apparent.

When an animal dies as a result of CO poisoning, the breathing

ceases but a can still be felt; therefore, it can be said that

the respiratory center failed. The problem, However, is not so

simple, for there are numerous factors to be considered. The

concentration of CO to which the animal is exposed m ust be known, for this determines the survival time of the experimental animal.

The individual susceptibility of the animal also affects the time course of the intoxication and, finally, the use of anesthesia necessarily alters the experimental condition by depressing the

level of responsiveness of many reflexes.

In this series of anesthetized dogs no great change in tidal

volume or respiratory rate was seen when the animals were exposed

to either 0. 3 per cent or 0.4 per cent CO. In Group A there was an increase of about 54 per cent in minute volume, although the variance

of the individual minute volumes was so great as to render this in­

crease statistically insignificant. It may be considered as meaning­

ful in view of the fact that an increase occurred in six out of seven instances and that a real increase would be expected from the changes observed in the acid base balance of the blood. This increase was seen when the mean arterial O^ content was 6 , 2 volumes per cent.

This is in agreement with the findings of von Oettingen, Donahue,

6 6 3 5 Valaer and Miller and Miller but is in disagreement with Ghiodi 65 45 et al. Although Swann and Brucer state there was no significant or, at best, a slight increase in ventilation, their data show a doubling of the ventiLation at an O., saturation of between twenty and forty per cent.

The cause for this increase is puzzling, for the arterial blood theoretically remains normal throughout the course of GO poisoning and therefore should not stimulate the chemo- receptors of the carotid bodies or the aortic glomi. There is some 70 evidence of a decreased Pq2 as re;ftected ^y a lowered venous ^0 2 " This brought about by the increased arterial - tissue

Pc >2 grac^ erLt required to release a sufficient amount of O2 for tissue utilization. This increase in 3?Q2 grac^ en^ necessary because of the decreased oxyhemoglobin level coupled with the Haldane effect, i.e., a shift to the left of the HbC>2 dissociation curve found in CO poisoning. See Figure 1. Perhaps studies of various parts of the venous system, right heart and/or the pulmonary would shed some light on this problem. It is not beyond the realm of possibility that the respiratory center contains a 37 receptor' sensitive to decreased>2 C content in arterial blood, or a discrepancy between metabolism and oxygen supply may allow acid metobolites to accumulate in the center.

The apparent increase in ventilation is accompanied corres­by ponding blood changes which seem to indicate a hyper ventilatory

state. These changes tend to lend credence to the suggestion of an increase in ventilation for the data show an increase in pH0.04 of and a decrease of8 mm. Hg in the alveolar Pqq , the fact that 2 statistics fail to indicate significance for any one measurement is not necessarily conclusive. Non-significance, statistically speaking, may have meaning when applied to one set of measurements and may

reflect the true situation but when three dependent variables vary in known interrelationship then one questions the validity of single re­ gression analysis applied to individual terms of a multiple regression.

In contrast to the results obtained in Group A, Group B showed no increase in ventilation; on the contrary, a slight decrease was noted. Yet here there is also a slight increase in the mean pH of

the arterial blood and a slight decrease in the mean alveolar values. At the time that these changes were noted, the >2C content

of the arterial blood was 14.24 volumes per cent.

As was expected in both Groups A and B, there was a consistent

decrease in the content of both the arterial and venous blood.

This is due to the increasing amount of CO which is bound to the hemoglobin. As a consequence, the capability of the hemoglobin to transport O^ is progressively reduced. In both groups the >2C con­ sumption decreased and the output remained relatively constant, thus resulting in an increased .

The arterial and venous CO^ contentsaLso decreased as the CO intoxication progressed. The most probable explanation seems to be that with an increasing amount of carboxyhemoglobin in the blood there is a severe reduction in the amount of reduced hemoglobin available, not only for >2 C transport, but for CO2 transport as well.

As a result, the CO 2 content falls progressively but not nearly as rapidly as the O^ content. This then would imply not only an oxygen starvation at tissue levels but also a CO2 accumulation.

In regard to the hematocrit, numerous investigators have found

AO a polycythemia associated with both acute and chronic CO poisoning. °»

In this series of investigations there was no demonstrable variation in the hematocrit in Group A as measured by the centrifugation tech­ nique. Although hematocrit was not measured in Group B, the O2 capacity determinations did not appear to reflect anything but normal hematocrit ratios. In Group C no change was noted. However, it was noted upon autopsy of the experimental animals that the was pallid and contracted. This was not the case in the control group. In addition, it was noted in both experimentals and controls that occasional was seen in the third blood samples following centrifugation. The finding of normal hematocrit values associated with contracted and pallid becomes understandable

only if one postulates a concomitant fluid shift in the tissues of the

experimental animals. Unfortunately, measurements of this nature were not made, therefore, it is possible only to state that in this

series no change in hematocrit was noted in either experimentals or

controls.

Of particular interest are the data concerning cardiac output

and peripheral resistance. In both of the experimental groups there

is a gradual decline of mean blood pressure between sampling periods

II and III. This gradual decline is climaxed by a sudden drop to zero

pressure just prior to death. The striking feature of these data is

that no apparent effort is made by the animal's body to increase the cardiac output by a sufficient amount to defend the declining mean ai-terial pressure. None of the reflex mechanisms for increasing cardiac output appear to be initiated. For example, the body fails

to defend the mean blood pressure by the use of the cardioaccelera-

tory reflex. Ordinarily a declining mean pressure stimulates the

presso-receptors of the carotid and aortic bodies. These re­

ceptors initiate afferent impulses via the vagi or Hering's

to the cardioaccelerator centers of the medulla. These in turn re-

flexly increase the heart rate by increasing sympathetic nervous

activity in the heart as well as by decreasing vagal activity. 40

However, in this series of experimental animals there was no statis­ tically significant change in the heart rate, in fact, the observed change was a slight decrease.

This failure to defend the mean blood pressure by appropriate reflex action could be caused by any one of several factors:1 ) de­ pression of the sensory receptors in the carotid and aortic bodies;

2) failure of afferent or efferent fibers conveying impulses; 3) de­ pression of the vasomotor center in the medulla with resultant venous pooling; or 4) inability of the heart to respond to changes in sympa­ thetic or parasympathetic nervous activity. Of these four possi­ bilities the last two appear to offer the most reasonable explanations.

It should be noted that these factors have been repeatedly investi­ gated. Haldane ^ and Van Slyke and Neill ^ assumed the pressor

6 3 changes were caused by hypoxia. Klebs thought the decreased blood pressure was due to peripheral vasodilation while, contempo-

1 3 rarily, Pokrowsky felt that it was due to the weakening of the myocardium.

From this investigation it seems likely that the decrease in the comes about as a direct result of decreased peripheral resistance. Although venous pressure was not measured in this series of experiments, it seems reasonable to assume that

l, with decreased peripheral resistance the concomitant peripheral pooling would, result in a decreased venous pressure, therefore de­ creased venous return. 41-

In contrast to the consistent gradual decline of mean arterial pressure seen in Group A, it must be mentioned that in Group B, between Samples I and II, an increased peripheral resistance was noted. This may be accounted for by the fact that in Group B a 5 lower concentration of CO was administered, although von Oettingen states that generally speaking the blood pressure response to CO poisoning is a short rise followed by a subsequent decline.

From the experimental data cited here it seems that the course of CO poisoning using 0.4 per cent in anesthetized dogs is as follows: the initial response of the animal is increased ventilation possibly due to a decreased tissue ^0 3 ’ hyperventilation is accomplished by increasing the respiratory rate and is accompanied by an increased arterial pH and a decreased alveolar PcC>2‘ a^terial ^02 rernaans normal throughout the course of the intoxication. The O2 and CO^ contents decline steadily in both the arterial and venous blood as the

CO competes more successfully for the available hemoglobin. This results in a greater and greater amount of HbCO in the blood which progressively shifts the Hb0 2 dissociation curve to the left. This, in turn, means that in order for the tissues to obtain a sustaining amount of O2 the tissue P 0 2 raust be lowered so that a greater gradient exists. The vasodilation accompanying the CO intoxication brings about a progressive decrease in peripheral resistance. The falling blood pressure, accompanied by the lack 42

of available O2 in the arterial blood, produces a progressive hypoxia

of the vasomotor center which is reflected by a diminishing heart

rate and the failure to increase cardiac output in response to the

falling mean arterial pressure.

That the failure of the animal to increase the cardiac output is not due to any inherent weakness of the myocardium can be demon­

strated by allowing the failing animal to breathe room air or1 0 0 per

cent C>2 . Under these circumstances the blood pressure returns to­ wards the normal level and the animal improves. SUMMARY AND CONCLUSIONS

A course of investigation was undertaken for the purpose of cLearly elucidating the mechanisms involved in the course of lethal exposure to carbon monoxide fumes. Anesthetized dogs were used as the subjects and cardiovascular and ventilatory responses to the carbon monoxide intoxication were carefully measured. Of particular interest were the measurements of cardiac output and peripheral resistance.

Two concentrations of carbon monoxide were used in the course of the investigation; 0. 3 per cent and 0.4 per cent. Al­ though the time course of intoxication differed, the basic changes observed were of the same general nature, although the magni­ tudes differed somewhat.

The conclusion drawnfrom this series of experiments is that death as a result of carbon monoxide poisoning is due to the progressive failure of the vasomotor center. This produces a gradually increasing state of hypoxia in surrounding medullary centers so that, finally, the respiratory center ceases to respond to the impulses which it receives and respiration ceases. Sjjo/vi \^/ Jf/OO W 3 "raMft/nld s Q ot

= 3 Of

or

or

Figure 1

CO Concentration versus Time. w

00/ 0 6 09 09 oroi

o>

£

U o s

0 9 s o'x.

9HOO JOOA/OD (

Figure 2

The Haldane Shift Ul 46

PLASMA PfioTSWS

ftJ;/*0 6LO3tA/

eesr. e*T£r

PVLMOAtAfV V£A/r.

V*

v % oo coA/re<¥7

0 0* c o m t g m t

L4CTIC A-c-ro

- i d sA & r y \ Soo Z»0 A£Te£//)L P/t&ssujeg 10 0

40 \J £ W o o £

M t/JUTGS Figure 3

Representative Data From Swann andBrucer. DOG

5 m w m = A r e c o r d e d

Figure 4

Schematic Diagram of Experimental Set-Up aaia/. —)

Figure 5

Terminal Ventilogram

oo 20' A/I BA/v Val ues G rp ^

CO

3. z. cc. mm.

3.00

too

gas 7S 3 0

Figure 6

Data Plot. Mean values of dogs in Group A. 50

20 v%

So

CO, v%

cc. min.'

dyne cm. X /o' 3 80 SAT.

. p. m.

too

too

gas 30 GO 3 0

F ig u r e 7

Data Plot. Group A, Dog 2/4. 51

zo Gctp. A - Dog zfe /z.8Kg $ V%

So

C 0o V7o

cc. mm.

dyne cm. X /o' 3 80 CL SAT.

ZOO

too

30 60 gas t s 7S~ 9 0

F ig u r e 8

Data Plot. Group A, Dog 2/9. 20 QrR.P. A ~ Oog I2..G g 2

So

3.

cq. mm.

dyne cm. X /o'3 80 0 'SAT. +o

.p.m.

too

30 6 0 gas IS 9 0

F ig u re 9

Data Plot. Group A, Dog 2/11. 2 0 ' Gr*?' A-D o$ % //- v9 /

/o

So

C0 o v%

cc. mm.

dyne cm. X /o'3 80 0o SAT.

%oo

6 0 gas 50

Figure 10

Data Plot. Group A, Dog 2/12. 54

G ur.A -D og % /3. o <%■

So ecu v%

cc. mm.

dyne cm. X /O'3 90 CL SAT.

. p. m.

too cc. mm. gos 30 45- 6 0 7 S Time in minutes 9 0

Figure 11

Data Plot. Group A, Dog 2/13. 55

to G ap. A-Dog- t3.8K% a*

/ 2. So

COo V7o

cc. min.

dyne cm. X I O ' 3 80 SAT.

.p. m.

30 60 gas t S 3 0

F igure 12

Data Plot. Group A, Dog 2/16. 56

4 0 G gp- A~ %8 /o.oKg a* V% ‘ / 3 So

CO, v%

cc. mm.

dyne cm. X to '3 80 0o SAT.

/oo

60 gas t S 3 0

F igure 13

Data Plot. Group A, Dog 2/18. to A'Wa/'/ Vaiu^s G#P. Q

V%

dyne cm.0 X /O '3 80 Op ^AT. ^

too

gos IS 3 0 9 0

Figure 14

Data Plot. Mean values, Group B. 58

2 0 '20

ecu V%

cc. min.

dyne cm. X /O'3 80

0„ SAT.40

.p. m.

too

too

30 60 gas IS 7 S 3 0

Figure 15

Data Plot. Group B, Dog 8/20. 59

20 G-tfi 8-Oof //OKcf £ V% /6

cc. min.'

too

too

6 0 g a s i5 3 0

F igure 16

Data Plot. Group B, Dog 8/22. 60

2 0 Gr #P. B ~ D o cf 0 / . 6 K c j $

------17

3 .

cc. min.

dyne cm x /0 ' 3 80

0o SAT.40

200

too cc. min.

gos IS 3 0 45- 6o 7S~ Time in minutes 3 0

Figure 17

Data Plot. Group B, Dog 8/25. 2 0

So

3.

cc. mm.

dyne cm. X /O'3 80

CL SAT. 4 0

200

/OO cc. min. tS s~>

F ig u re 18

Data Plot. Group B, Dog 8/Z7. 62

40 &XF. 8 ' Do$ % 9 112 Ks $ v%

cc. min.

dyne cm. X /o' 5 80

CL SAT. 40

.p.m.

too

100

gas IS 30 GO 3 0

Figure 19

Data Plot. Group B, Dog 8/29. 63

2 0 Ctnp. 8 - Pog ^ /2 Kef v?

iTT------20

•TO

cc. min./ ,

dyne cm. X /o' 3 80 0„ SAT.

2 00

too cc. min. gas IS 30 Go 9 0

Figure 20

Data Plot. Group B, Dog 9/2. 64

to t-AA./ V^Loes V% 2-1 So

C 02 V7c

C . 0. cc. min/

P. R. dyne cm. X 10 -3 80 02 SAT. ^ °/'O

I. p. m. C .8 R.Q..4 0

V 0 2 i.-0 \sU cc. min.

gos is 1°.Time in +? minutes . ?r 9 0

F igure 21

Data Plot. Mean values, Group C, 65

G«p.C-Dog % /o.i Kg -£ o 2 V %

C0o V% o ■3-1 C . 0. t cc. mm. o a P. R. dyne cm. X /O'3 80 02 SAT.*, °//<9

I. p. m. 0 .8 R.Q..1- 0

200 O. too cc. min. gas I S 3 0 + S . 60 7 S Time in minutes 30

F igure 22

Data Plot. Group C, Dog 2/23. 20 Grup. c ~ Dog 5 ! ~- 0 C ? V% Z 3

cc. mm.

dyne cm. X /O'3 80 CL SAT. ■w

.p. m.

too

too cc. min. IS 30 + s 6O' gas Time in minutes 90

Figure 23

Data Plot. Group C. Dog 2/27. 67

2 0 C-Oo^ % 13.7 K$

C02 V% o 3. C . 0. 2.. cc. min.‘ O e P. R.< dyne cm., X 10' 80 0 2 SAT.^ °'o /

I. p. m. C .8 R.Q./f- o

V O. /oo cc. min. I S 30. A * . 6 0 ^ gas Time in minute s 90

F igure 24

Data Plot. Group C, Dog 3/2. zo

v%

So

CO

cc. mm.

dyne cm. X /O ' 3 80

0„ SAT.■w

. p. m.

gas t s 3 0 9 0

F igure 25

Data Plot. Group C, Dog 3/3. 69

ao G*P.C-Dog% /O-Otg $ 0. V% Zi> 0, So

COo V7c

C . 0. cc. mm. 0 0

P. R. 4 dyne cm.0 X /o' 3 80

° 2 SAT.^ °/'o

I.p.m. 0 ,8 R.Q./f-

0 too o . too cc. min. i s 30 4 S 60 7S' gas Time in minutes 90

Figure 26

Data Plot. Group C, Dog 3/6. 70 TABLE I

CARDIOVASCULAR RESPONSE TO CO

Group Cardiac Output Peripheral Res. Heart Rate Stroke Volume & Dog I II III I II III No. I II III L/M Dyne CM I II III

2/4 168 176 204 1185 2279 4654 2069 - 7 13 - 2/9 160 128 80 1411 995 14 5098 5542 285 9 8 0. 2 2/11 160 216 124 898 1288 832 5923 4219 5283 6 6 7 2/12 23 2 116 116 1576 2127 1281 4006 2893 312 7 18 11 2/13 132 110 40 1169 2712 1769 2666 1238 1988 9 25 4 2/16 134 128 140 1161 2659 3048 6677 2194 1835 9 21 22 2/18 146 128 134 1064 967 1331 4206 2066 900 7 8 10

Av. 162 143 119 1209 1861 1379 5262 2889 1767 8 13 9 S.D. + 34 39 51 223 762 149 1982 1492* 1870J 1. 2 7.6 6.6

B

8/20 156 164 120 1782 1654 1449 4664 3672 3585 11 10 12 8/22 176 108 120 1441 801 1503 5879 5587 2659 8 7 13 8/25 132 164 136 1479 1443 1762 7295 7477 3765 11 9 13 8/27 140 136 80 2494 2390 1985 3205 3143 2537 18 18 25 8/29 136 122 104 1327 1112 2393 4939 6468 1369 10 9 23 9/6 172 168 128 3230 2128 2964 3266 4957 2103 19 13 23

Av. 152 145 115 1957 1568 2063 4875 5217 2670 13 11 18 S.D. + 19 8 20 239 601 582 1568 1647 902^ 4.5 4.0 6.1

C

2/23 112 128 124 1050 1650 1348 5842 3875 3918 9 13 11 2/27 166 178 182 1656 1833 1862 7384 7064 6782 10 10 10 3/3 146 140 140 1811 1425 1500 6928 5721 5275 12 10 11 3/2 188 196 148 1395 1531 1776 1375 1357 1035 7 8 12 3/6 152 144 130 1550 1057 826 6950 11050 11920 10 7 6

Av. 153 157 145 1492 1499 1462 5696 5613 5786 10 10 10 S.D.+ 25 29 23 290 290 412 785 3278 4030 1.8 2.3 2.3

^Statistically significant TABLE II

BLOOD GASES

(Group A)

C ao 2 c v CA a co 2 Dog 2 Yc°2 No. I II III I II III I II III I II III

2/4 18.56 8.65 4.10 11.02 4.73 1.21 40.70 36,76 32. 65 45.86 39.30 34.46 2/9 17.76 6.70 3.35 12.71 3.11 0.73' 34.09 30.04 22. 68 38.77 31.15 23.41 2/11 19.13 4.80 3.89 8.84 1.93 0.75 33.77 22.93 24.40 42.49 26. 66 25.73 2/12 16.31 6.76 3.04 12.02 4.09 1.26 40.82 34.39 31.61 50.75 41.63 32.47 9/13 19.57 6.03 3.11 8.93 1.83 1.29 25.39 28.79 30.34 33.32 31.55 31.49 2/16 18.22 6. 14 4. 66 11.21 2.74 1.93 41.38 39.32 35. 00 45.78 41.93 37.06 2/18 15.45 4.30 4.19 5.57 1.87 1.47 40.51 37. 68 34.91 46.33 39.96 37.98

Av. 17.86 6. 20 3.76 10.04 2.90 1.23 36. 67 32.84 30. 23 43.33 36.03 31.’8" S.D.-f 1.49 1.42* 0.63* 6.86 5.54* 2.76* 5.96 5.86 4.89 7.15 8.26 6. 64

Statistically significant TABLE II (Continued) BLOOD GASES (Group B)

C C y cao 2 CA Dog 2 Aco 2 C°2 No. I II III I II in I II III I II III

8/20 15.20 14.70 7.03 10.82 9.90 5.37 44.47 38.82 39.38 47.39 41.97 41.97 8/22 17.29 15.09 8.44 12.29 7.63 3.93 40.97 35.85 33.26 41. 10 40.07 35.79 8/25 13.04 11.67 7.28 10.42 7.77 2.92 50.02 46.38 39.38 50. 00 44.24 41.38 8/27 15.92 13.58 4.44 12.05 11.11 2.29 40.61 39.04 40.01 43. 89 44.07 40.59 8/29 19.52 14.36 5.01 4.34 9.34 3.23 33.06 32.08 28.57 44. 70 36,42 31.56 9/2 20.50 16.01 4.42 17.04 12.58 2. 14 39.24 39.03 ? \,75 40. 72 42.16 37.33

Av. 16.92 14.24 6.10 11. 16 9.72 3.31 41.39 38. 53 36.06 44. 65 41.49 38.11 S.D.+ 1. 58 1.45* 1.45* 4.09 1.92* 1.18 5.62 4.72 4.52 10. 74 7.21 7.87

Statistically significant TABLE II (Continued) BLOOD GASES (Group C)

C-tr C a cao 2 vo C v Dog 2 Aco2 vco2 No. I II III I II III I II III I II hi

2/23 16.44 19.34 19.57 8.86 13.40 13.47 32.98 33.83 32.46 40.45 40.01 35.41 2/27 19.07 20. 24 20.24 13.58 14.20 14.38 32.59 27.18 28.16 40.22 36.33 36. 26 3/3 15.90 17.54 17.48 11.08 11.80 11.18 43.34 41.76 41.30 46.62 43.82 44.17 3/2 19.56 20.73 20.84 13.13 14.87 14.78 42. 24 37.98 37.58 45.27 42.35 42.07 3/6 20.11 22.29 22.33 15.88 15,66 13.83 40.74 39.23 39.25 42.41 38.93 43.38

Av. 18. 21 20. 03 20.09 12.51 13.99 13. 53 38.38 35.99 35.75 42.99 40.29 40.26 S.D.+ 1.92 1.76 1.79 2. 66 1.48 1.38 5.19 5.70 5.36 3.57 5.61 4.33 74

TABLE III

BLOOD CHANGES

Group Initial Cal. Arterial pH Hematocrit Art. COHb & Dog P C°2 °2 No. I II IH I II III I H III I II III Cap.

A

2/4 48 35 32 7. 25 7.35 7.33 -- - 0 55 79 19.11 2/9 43 31 32 7. 22 7.30 7.15 41 42 46 0 63 81 18. 12 2/11 39 38 49 7.26 7. 10 7. 01 47 61 59 0 76 80 19.85 2/12 42 21 38 7.30 7. 55 7.24 32 38 45 0 59 82 16.67 2/13 30 28 44 7. 22 7.32 7.14 42 46 43 0 70 85 20.16 2/16 53 47 46 7.25 7.28 7. 20 56 53 44 0 67 76 18.33 2/18 42 37 40 7.30 7.33 7.26 35 40 44 0 74 75 16.70

Av. 42 34 40 7.27 7.32 7.19 42 47 47 0 66 80.,. 18.42 S.D, + 7.2 8.3 6. 6 .44 .73 .44 8. 6 8.8 6.1 0 2.5* 1.4

B 8/20 51 37 42 7. 28 7.34 7.29 -- - 0 18 64 17.49 8/22 42 34 37 7.31 7.35 7.28 -- - 0 16 53 18.01 8/25 62 52 48 7. 22 7. 25 7. 22 --- 0 26 54 15.76 8/27 44 40 52 7. 28 7.31 7. 18 --- 0 17 76 16.45 8/29 37 33 33 7.29 7.32 7.26 -- - 0 33 76 21. 50 9/2 46 45 52 7. 26 7. 26 7. 15 -- - 26 80 21. 58 0

Av. 47 4C 44 7. 28 7.31 7. 23 — _- 0 24 64 18.57 S.D. + 10.7 7.2 7.9 . 20 .01 . 14 0 2. t 1.2

C

2/23 41 45 42 7. 21 7. 19 7. 20 41 44 45 0 0 0 19.85 2/27 42 34 37 7. 20 7. 22 7.19 43 44 41 0 0 0 20.84 3/3 50 49 48 7. 29 7. 26 7. 27 48 46 41 0 0 0 17.96 3/2 45 42 41 7.30 7.29 7.31 44 47 47 0 0 0 20.89 3/6 46 45 46 7.28 7. 28 7. 26 46 49 45 0 0 0 22.38

Av. 45 43 43 7. 26 7. 25 7.26 44 46 44 0 0 0 20.38 S0D. -f 3.6 5. 6 4.3 . 15 .01 .01 8.5 2.1 2.7 0 0 0

Statistically significant 75

TABLE IV

VENTILATORY RESPONSE

Re spiratory Group Tidal Volume Minute Ventilation & Dog Rate No. I II III I II in I II in

A 2/4 171 206 199 2222 5866 5466 13 29 28 2/9 159 160 365 3112 11033 6931 19 69 19 2/11 181 302 115 4886 5748 1150 27 19 10 2/12 305 156 179 2133 4528 2687 7 29 15 2/13 134 171 417 8960 7679 3334 67 45 8 2/16 137 157 134 5329 5755 4476 34 32 29 2/18 149 .91 108 3433 5722 3576 17 63 33

Av. 177 178 217 4296 6619 3950 26 41 20 S.D. + 165 161 196 2390 2154 1889 20 19 10

B 8/20 245 316 214 5137 2210 3868 21 7 18 8/22 500 139 166 1497 2652 2321 3 19 14 8/25 310 264 201 1205 1055 1611 4 4 8 8/27 295 244 268 2063 2443 1877 7 10 7

8/29 117 130 - 4036 4420 - 34 34 - 9/2 221 234 212 2652 2578 1688 12 11 8

Av. 281 221 212 2765 2726 2273 14 14 11 S.D. + 192 194 206 1535 1074 933 12 11 5

C

2/23 164 162 150 2288 2432 3001 14 15 20 2/27 256 271 261 2559 3 248 3390 10 12 13 3/3 149 121 126 2829 4244 3537 19 35 28 3/2 187 251 228 3183 4526 4102 17 18 18 3/6 281 294 232 1684 2060 1623 6 7 7

Av. 207 214 199 2509 3320 3131 13 17 17 S, D. + 192 193 188 566 1083 931 5 11 8 BIBLIOGRAPHY

Sayers, R. R. and Yant, W. P. Bureau of Mines Reports . of Investigations, Serial No. 2476, May 1923 =

Yant, W. P», Chornyak, J. , Schrenk, H. H. , Patty, F. A. and Sayers, R. R. Studies in Asphyxia. Pub. Bull., No. 211, 1934.

Sayers, R. R. and Davenport, S. J. Review of Carbon Monoxide Poisoning. Pub. Health Bull. , No. 195, March 1930.

Lewin, R. Die Kohlen oxydvergiftung. Berlin, 1920. . von Oettingen, W. F. CO; Its Hazards and the Mechanism of its Action. Pub. Health Bull., No. 290, 1944. '

Walton, D. C. and Witherspoon, M. G. Skin Absorption of Certain Gases. J. Pharmacol. , 26, 315, 1925.

Schutze, W. On the Danger for Man and Animal from High Concentrations of Some Toxic Gases on the Skin. Arch. Hyg., 98, 70, 1927.

Henderson, Y« and Haggard, H. W„ Noxious Gases. The Chemical Catalogue Co., Inc. New York, 1927.

Sayers, R, R0, Yant, W. P„, Levy, E. and Fulton, W. B. Effect of repeated daily exposure of several hours tp small amounts of automobile exhaust gas. Pub. Health Bull., No. 186, 1929.

Haldane, J. B. S. The Dissociation of Oxyhemoglobin in Blood During Partial CO Poisoning. J. Physiol. , 45, xxii, 1929.

Sayers, R. R. and Yant, W. P. The Elimination of CO From the Blood by Treatment with Air, O2 and a Mixture of CO 2 and 0 2. Pub. Health Bull. No. 38, 2053, 1923; Reprint No. 865.

Williams, I. R. and Smith, E. Blood Picture, Reproduction and General Condition During Daily Exposure to Illum­ inating Gas. Am. J. Physiol. , 110, 611, 193 5. 77

13. Pokrowsky, W. On Poisoning with. CO. Arch. Path. Anat., Virchow’s, 30, 525, 1864.

14. Kreis, E. On the Fate of CO During Detoxification after Exposure to CO. Arch. ges. Physiol., Pfluger's, 26, 245, 1881-1882.

15. ZaLeski, S. On a New Reaction for CO HemogLobin. Z. Physiol. Chem. , 9, 225, 1885.

16. Donders, F„C0 The Chemistry of Respiration, a Process of Dissociation. Arch. ges. Physiol., Pfluger's, 5, 20, 1872.

17. Haldane, J.S0 and Smith, J.L. The Absorption of O^ by the Lungs. J. Physiol., 22, 231, 1897-1898.

18. Fenn, W.O„ and Cobb, D.M. The Stimulation of Muscle Respiration by CO. Am. J. Physiol., 102, 379, 1932.

19. Schmitt, F.O. and Scott, M.G. The Effect of CO on Tissue Respiration. Am. J. Physiol., 107, 85, 1943.

20. Sjostrand, T. Nature, 164, 1949.

21. Nicloux, M. On the Excretion of CO after Severe Poisoning. Comp. Rend. Soc. Biol., 92, 174, 1925.

22. Bock, J. CO. Heffter's Handbuch exptl. Pharmacol., 1, 1, Julius Springer, Berlin, 1923.

23. Bernard, C. Lecons sur les effets des substances toxiques et medicamenteuses. J.B.Balliere and Sons, Paris, 1857.

24. Hoppe-Seyler, F. On the Effect of CO on Hemoglobin. Arch. path. Anat., Virchow's, 11, 288, 1857.

25. Weimann, W. On the Detection of CO in Exhumed Cadavers. Deut. Z. ges. gerichtl. Med., 17, 48, 1931.

26. Wiethoid, F. Demonstration of CO in the Exhumed Cadaver. Deut. Z. ges. gerichtl. Med., 14, 135, 1929.

27. Heilmann, P. Late detection of CO in the Exhumed Cadaver. Deut. Z. ges. gerichtl. Med., 23, 215, 1934.

28. Pieczarowski, M. Late Post Mortem Demonstration of CO. Abstr. J. Ind. Hyg. ToxicoL. , 19, 136, 1937. 78

29. Hartridge, H. Calibration of the Reversion Spectroscope for the Estimation of CO in Blood. J. Physiol., 57, 47, 1922-1923.

30. Roughton, F.J.W . Kinetics of Hemoglobin. Parts IV, V and VI. Proc. Soc., London, Series B, 115, 464, 1934.

31. Hufner, G. On the Distribution of the Blood Pigment Between CO and 0£; A Contribution to the Doctrine of Chemical Mass Action. J. prakt. Chem. , 30, 69, 1884.

32. Haldane, J.S. The Relation of the Reaction of CO to 2 O Tension. J. Physiol., 18, 201, 1895.

33. Nicloux, M. The Law of Absorption of CO by BLood in Vivo and in Vitro. J. Physiol. Path. Gen., 16, 145, 164,1914.

34. de Saint-Martin,' L.G. New Studies in the Absorption Power of Hemoglobin for O2 and CO. Comp. rend. acad. sci., 131, 506, 1900.

35. Muller, F. On the Ferricyanide Method for the Determination of O 2 in Blood Without Blood Gas Pump. Arch. ges. Physiol., Pfluger's, 103, 541, 1904.

36. Hoppe-Seyler, F. Contributions to the Knowledge of the Proper-ties of the Blood Pigment. Z. physiol. Chem., 13, 447, 1889.

37. Krogh, A. On the combination of Hemoglobin with mixtures of O2 and CO. Skand. Arch. Physiol., 23, 217, 1910.

38. Hartridge, H. The Action of Various Conditions on CO Hemoglobin. J. Physiol., 44, 22, 1912.

39. Douglas, C0G., HaLdane, J.S. and Haldane, J.B.S. The Laws of Combination of Hemoglobin with CO and 2O. J. Physiol., 44, 275, 1912.

40. Altschul, A.M., Sidell, A .E., Jr. and Hogness, T.R. Note of Preparation and Properties of Hemoglobin. J. Biol. Chem., 127, 123, 1939.

41. HaLdane, J.S. Respiration. YaLe University Press, 1922.

42. Haggard, H.W. and Henderson, Y. Hemato-respiratory functions. XII. Respiration and Blood Alkali During CO Asphyxia. J. Biol. Chem., 47, 421, 1921. I

79

43. Chartschenko, N.S. The Effect of CO on the Psychomotor Centers. Arbeits-physiol., 6, ^45, 1933.

44. Haldane, J.S, and Priestly, J.G. Re/spiration. Clarendon Press, Oxford. 1935.

45. Swann, H.G. and Brucer, M. The Cardiorespiratory and Biochemical Events During Rapid Anoxic Death. Texas Repts. Biol. Med., 7, 4, 1949.

46. Roughton, F.J.W . Respiratory Functions of Blood. Hand­ book of Respiratory Physiology. USAF Sch. Av. Med. , Randolph AFB, Texas.

47. 13dbbeling, C.H. Catatonic Condition in CO Poisoning. Abstr. Deut. Z. ges. gerichtl. Med., 14, 180, 1930.

48. Litzner, S. CO Poisoning and Polycythemia. Arch. Gewer- bepath, Gewerbehyg., 1, 749, 1930.

49. Barker, L .F . A Case of CO Poisoning from an Oil Stove. J. Ind. Hyg., 15, 238, 1933.

50. Beck, H.G. and Fort, W. Chronic CO Poisoning; its Effect On The Blood, With Report of Two Cases Simulating Pernicious Anemia. Ann. Clin. Med., 3, 437, 1924.

51. De , S. and Carroll, D..C. The Mechanism of the Splenic Reaction to General CO Poisoning. J. Physiol., 59, 312, 1924.

52. Barcroft, J. and Barcroft, H. Observations on the Taking Up of CO by the Hemoglobin in the Spleen. J. Physiol. , 58, 138, 1924.

53. Barcroft, J, Murray, C.D., Orahovats, D. , Sands, J. and Weiss, R. The Influence of the Spleen in CO Poisoning. J. Physiol., 60, 79, 1925.

54. Ackermann, T. Studies on the Effects of Suffocation on the in Brain and Lungs. Arch. path. Anat., Virchow's, 15, 401, 1858.

55. Pick, F. On the Effects of Vasomotor Agents on the Circu­ lating Blood Volume. Arch, exptl. Path. Pharmacol., 42, 399, 1899. 80

56. Claude, H. and Lhermitte, J. Experimental Studies on the Toxic Effects of CO on the . Gompt. Rend. soc. biol., 72, 164, 1912.

57. Weimann, W. Brain Findings in Death in the CO Atmosphere. Z. ges. Neurol. Psychiat. , 105, 213, 1926.

58. Biumberger, K. Occupational Disturbances of the Heart. Med. Klin., 35, 1245, 1939.

59. Symanski, H. On the Appraisal of Heart Muscle Damage Following CO Poisoning. Arztl. Sachverst. Ztg. , 44, 183, 1938.

60. Chance, O.G, and Jackson, D0E„ The Pharmacological Action of Some Poisonous Gases With Special Reference to CO. ,J. Pharmacol., 25, 145, 1925.

61. Schwerma, H., Ivy, A.C., Friedman, H. and Labrosse, E. Study of Resuscitation From Juxtalethal Effects of Exposure to Carbon Monoxide. Occup. Med., 5, 24, 1'948.

62. Zondek, H. Cardiac Findings in Illuminating Gas Poisoning. Deut. Med. Wochschr., 45, 678, 1919; ibid., 46, 235, 1920.

63. Klebs, N. On the Effect of CO on the Animal Organism. Arch. Path. Anat., Virchow's, 32, 450, 1865.

64. Haggard, H.W. Studies in CO Asphyxia. I. The Behaviour of the Heart. Am. J. Physiol., 56, 390, 1921.

65. Chiodi, H., Dill, D.B., Consolazio, F. and Horvath, S.M. Respiratory and Circulatory Responses to Acute CO Poisoning. Am. J. Physiol., 134, 683, 1941.

66. von Oettingen, W .F., Donahue, D.De, Valaer, P. J, and Miller, J.W. Studies on the Mechanism of CO ­ ing as observed in Dogs Anesthetized with Sodium Amytal. Pub. Health Bull., No. 274, 1941.

67. Unpublished work from this laboratory. Dissertation of Thiede, F.C,

68. Comroe, J0Hi,Jr. and Schmidt, C.F. The Part Played By the Reflexes from the in the Chemical Regu­ lation of Respiration in the Dog. Am. J. Physiol., 121, 75, 1938. 81

6 9. Van Slyke, D.D. and Neill, J.M. The Determination of Gases in the Blood and Other Solutions by Vacuum Extraction and Manometric Measurement. J. Biol. Chem., 61, 523, 1924.

70. Moore, J.W ,, Kinsman, J.M ,, Hamilton, W.F. and Spurting, R.G. Studies on the Circulation. II. Cardiac Output Determinations; Comparison of the Injection Method with the Direct Fick Procedure, Am. J. Physiol., 89, 322, 1929.

71. Snedecor, G0W, Statistical Methods. Ames, Iowa, Collegiate Press. 82

AUTOBIOGRAPHY

I, Kenneth Rae Coburn, was born in Windsor, Ontario,

Canada, I received my secondary school education at Redford

High School, Detroit, Michigan, My undergraduate training was taken at Western Michigan University, Notre Dame, University of Detroit and Hillsdale College. I received the degree Bachelor of Science in Biology from Hillbdale College in 1948. For the next three years I attended the University of New Mexico seeking the degree Doctor of Philosophy in , however in 1951 I was recalled to active service with the United States

Navy, In January of 1957 I was ordered to The Ohio State

University for post-graduate study in Physiology. During the ensuing three years I completed the requirements for the degree Doctor of Philosophy.