A Bipolar Theory of Living Processes

Variance of opinion as to the physiologic effect of alcohol suggested the use of this agent in this series of studies. In 1 rabbit 20 сс. of a 50 per cent solution of aleohol was introduced into the stomach and in 2, 8 сс. of a 25 per cent solution was given intravenously. In each instance the temperature of the brain fell with a rapidity corresponding to the fall in an animal in a moderate degree of shock. Periods of muscular activity occurred at almost equal intervals, each causing a momentary rise in the temperature of the brain and the liver amounting to about 0.07°C. The injection of adrenalin in these animals caused a rise in the temperature of the brain corresponding to that in normal animals with an abrupt fall after the maximum point had been reached to a point which in one instance was 1.7 degrees below the temperature when the adrenalin was given. In another case the animal died suddenly when the temperature of the brain had fallen to a point 0.35 degrees below that at which the adrenalin was given.

1. It is assumed that the effect of the injection of adrenalin upon the temperature of the brain in normal animals can be used as a unit of measurement whereby to estimate the effect of an agent upon the oxidative power of the brain. 2. From these experiments it would appear that the alteration in the temperature of the brain following the injection of adrenalin is not notably affected by the effect of the agent upon the blood supply of the brain.

a. When adrenalin is injected in the presence of morphin, the temperature response of the brain is diminished in direct relation to the depth of narcosis. | b. The injection of adrenalin in the presence of strychnin produces not only a characteristic rise in the temperature of the brain but also a marked decrease in the temperature of the liver. c. Alcohol of itself alone produces a fall in temperature corresponding to that observed in rabbits in shock, while the injection of adrenalin in these animals produces a characteristic rise in temperature; this rise is followed by a fall which exceeds that observed in normal rabbits.

Note: Since the presentation of this paper a further series of experiments has been initiated, in which thermocouples are placed within the jugular vein and the carotid artery. The first experiment is cited here only as a preliminary report of an investigation which may aid in the interpretation of the studies summarized above. In this experiment, after each injection of adrenalin the temperature of the blood within the vein rose above that of the blood within the artery.

In view of the findings by Cannon ^? and by other investigators that the secretion of epinephrin is increased by asphyxia, an observation which had also been made by one of us in a previous research; and since it had been demonstrated in the biophysical laboratories of the Cleveland Clinie that the injection of adrenalin in normal animals produces a typical increase in the temperature of the brain, it seemed that it would be of interest to measure the temperature of the brain during asphyxia, both in animals with intact adrenals and in animals from which the adrenals had been removed. A series of eleven experiments was performed (Table 1).

The effect о] asphyxia ироп the output of epinephrin as indicated by à variations im the temperature of the brain Double adrenalectomy under ether anesthesia 9 min. before asphyxia Double adrenalectomy under ether anesthesia 11 min. before asphyxia Double adrenalectomy under ether anesthesia 10 min. before asphyxia Abd. manip. and exposure approximately equiv. tothat of removing adrenals 13 min. before asphyxia Note. This was supposed to bea complete double adrenalectomy; the risein temperature accompanying asphyxia led to an autopsy which showed one-third of right adrenal in situ.

In two normal animals the temperature of the brain during the period of asphyxia rose 0.51°C., and 0.98°C. respectively. Subsequent periods of asphyxia in these animals were also accompanied by a rise in brain temperature to a lesser degree. One animal was heavily narcotized with morphin. Asphyxia while the animal was deeply narcotized caused an increase in the temperature of the brain of only 0.04°C. Three hours later, however, when the period of narcotization was nearly passed the temperature of the brain during the period of asphyxia rose 0.26°C. In marked contrast to the effect of asphyxia in animals narcotized by morphin was the effect in animals which had received a preliminary dose of strychnin. In each of two animals which had received lis of a grain of strychnin five minutes before each was subjected to asphyxia, the temperature of the brain rose 0.4? C. In three animals the adrenals were removed, the operation being completed ten minutes before the period of asphyxia. In each of these animals the temperature curve of the brain was unaltered by the asphyxia. Тһе injection of adrenalin in each of these caused an abrupt and large increase in the temperature of the brain of from 1 to 2.09°C. Since in each of these last three animals

the shock of the operation was necessarily great and the temperature of the brain was falling steadily, it was suggested that the lack of response to the asphyxia might be due to the degree of shock in each animal. In two animals, therefore, under ether anesthesia for the same period as that required for adrenalectomy, the abdomen was opened and the intestines were exposed and manipulated. At the conclusion of the manipulation the temperature of the brain of each of these animals was falling at approximately the same rate as in animals in which the adrenals had been removed; but in the two animals subjected to shock alone during asphyxia the temperature of the brain rose 0.64? C. and 0.36? C. respectively. In another animal in which an attempt was made to perform a complete double adrenalectomy the temperature of the brain rose 0.37? C. during asphyxia. Autopsy, however, showed that one-third of the right adrenal was in situ. ln none of these experiments was any alteration in the temperature of the liver observed, excepting in the animals which had received a preliminary dose of strychnin in which the temperature of the liver fell during the period of asphyxia.

Asphyxia produces an increased output of epinephrin which is manifested by an increase in the temperature of the brain. From The Journal of Laboratory and Clinical Medicine, St. Louis, Vol. IX, No. 5, February, 1924. In the biophysics laboratory of the Cleveland Clinic it has been shown that adrenalin exercises a specific effect on the oxidative power of the brain, which is manifested in normal animals by an increased temperature of the brain in direct relation to the amount of adrenalin injected." The establishment of this specific effect of adrenalin on the brain has provided us with a criterion whereby to test the capacity of certain agencies for increasing or diminishing the oxidative power of the brain—that is, one would expect that if the oxidative power of the brain is increased by the agent which is being tested, then the response of the brain to an injection of adrenalin given within the period of activity of the agent would be increased. Conversely, the response of the brain to the injection of adrenalin would be diminished in the presence of an agent which depresses the oxidative power of the brain.

In view of the variance of opinion regarding the value of gum acacia (Bayliss) solution as a means of conserving the failing energies of the organism after hemorrhage or after shock, and believing that failing powers presuppose among other things a diminished power of work, hence of oxidation within the brain, it seemed desirable to make a comparative study of the effects of gum acacia solution and of transfusion upon the power of the brain to respond to adrenalin. A six per cent gum acacia solution was used, prepared according to the formula of Bayliss,? the solution being used at a temperature of 38.75? C.

of from five to seven pounds. From 30 to 50 се. of blood was withdrawn from each, the quantity in each case being determined by the clinical condition of the animal. None was reduced to extreme prostration. The temperature variations in the brain and the liver were measured by especially constructed thermocouples. When the maximum effect of the hemorrhage had been produced, as registered by the cessation of the fall in temperature of the brain, four of the animals were given an intravenous infusion of the gum acacia solution, the remaining four receiving a transfusion of citrated blood taken from another rabbit. Table 1 shows the variations in temperature in each of these animals. The significant columns are those which show the maximum temperature attained after the injection of adrenalin as compared (a) with the temperature before the hemorrhage and (b) with the maximum temperature attained after the infusion or transfusion. In every animal which received the transfusion of blood the temperature after the injection of adrenalin rose to a level of from one to nearly two degrees above the temperature of the animal before it was subjected to the hemorrhage. On the other hand, in only one of the three which received the gum acacia solution did the temperature after the adrenalin injection rise even to its level before the hemorrhage. In no instance after the gum acacia infusion was the adrenalin able to raise the temperature of the brain to the maximum attained after the infusion, while in every animal which received the transfusion the maximum rise after the transfusion was surpassed after the injection of adrenalin.

Another significant feature is the length of time after the infusion or transfusion of blood was given, during which the temperature of the brain continued to rise. 'The longest period of rise after the gum acacia infusion was 14 minutes. On the other hand, the shortest period of rise after the transfusion of blood was 1 hour and 23 minutes, the longest 3 hours and 52 minutes. The clinieal symptoms of the animals throughout corresponded to the temperature observations. In all four of the animals subjected to the gum acacia solution the temperature of the brain fell to a very low level before the termination of the experiment, and the animals were practically in extremis. Of the four animals subjected to blood transfusion, all were in good condition at the end of the experiment, and the temperature of each was but little below that at the beginning of the experiment,

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A comparison of the response of the brain to the injection of gum acacia solution and to the transfusion of whole blood indicates that the oxidative power of the brain is markedly diminished by the gum acacia solution, while it is increased by the transfusion of whole blood. From the Department ој Biophysics, Cleveland Clinic Foundation Many extensive investigations have been made for the purpose of determining the electrical conductivity of cell suspensions such as blood, body tissues, bacterial suspensions, and the like; the biological importance of this constant being that it would give some idea of the permeability of the cells (or the cell walls) in question. The investigations of the conductivity of blood by Bugarszky and Tangl; Fraenckel? Oker-Blom,? Roth, Stewart, and others are well known. These investigators found that the

1 Bugarszky, S., and Tangl, F., Eine Methode zur Bestimmung des relativen Volums der Blutkörperchen und des Plasmas, Zentr. Physiol., 1897- 98, xi, 297. з Fraenckel, P., Ueber die Bestimmung des Blutkorperchenvolumens aus der elektrischen Leitfühigkeit, Z. klin. Med., 1904, lii, 470. з Oker-Blom, M., Thierische Säfte und Gewebe in physikalisch-chemischer Beziehung, Arch. ges. Physiol., 1900, Ixxix, 510. "Stewart, С. N., The behaviour of the hemoglobin and electrolytes of the coloured corpuscles when blood is laked. J. Physiol., 1899, xxiv, 211. The relative volume of weight of corpuscles and plasma in blood, ibid., 356. The mechanism of hemolysis with special reference to the relations of electrolytes to cells, J. Pharmacol. and Exp. Therap., 1909-10, i, 49,

red corpuscles act as perfect insulators,? so that the electrie current exclusively passes in the space between them. Oker-Blom, for instance, showed that the resistance of a blood cell suspension of а certain volume concentration was equal to the resistance of а suspension of quartz sand of equal volume concentration in the same suspending medium. The ratio of the resistance of the blood suspension to the resistance of the suspending medium in this case is, therefore, a function of the volume concentration of the corpuscles alone. Experience shows that this function is independent of the size and only slightly dependent on the shape of the suspended particles. In the case of suspensions of cells such as bacteria, part of the current generally goes through the intercellular liquid and part goes through the cells. The resulting conductivity of the suspension is, therefore, a function both of the specifie conductivity of the suspending medium and of the specific conductivity of the cellular substance as well as of the volume concentration.

Тһе purpose of the present paper is to derive a formula whereby the specific conductivity of the cellular substance may be calculated from observed values of the specific conductivity of suspensions of the cells in question and of the suspending medium. This formula will be here derived for the simple case of a dilute suspension of spherical homogeneous cells. Nevertheless, the formula can be applied also with fair accuracy to the more general case of. a suspension of homogeneous ellipsoids, for it is found that up to a quite large value for the eccentricity of the ellipsoids the same formula as that for homogeneous spheres is applicable. "This is in accord with the findings of Oker-Blom and Fraenckel mentioned above, that the conductivity of suspensions of red corpuscles and of quartz sand depends only on the volume concentration of the suspensions.

It may be well at this point to consider briefly the assumption that the cells are homogeneous. In general, a single cell comprises many regions of varying conductivities, as indicated, for instance, by the very large polarization capacity of all living cells. Thus, it is probable that all cells are surrounded by a thin membrane which is semipermeable and in consequence has a comparatively high resistance. However, the formula applicable to a cell consisting of several concentric layers of different conductivities will be identical with that which we shall develop for a homo-

*Tt is to be understood that this apparently high resistance to ап electric current of low frequency may be wholly due to polarization at the surfaces of the red corpuscles. geneous sphere, the specific conductivity of the homogeneous sphere being replaced by a certain average value of the conductivities of the different layers of the non-homogeneous cell.’ We shall, therefore, consider the case of a suspension of homogeneous spheres (radius a, specific conductivity of cell material kı) in а medium of specific conductivity kə. We shall assume that the suspension is so diluted that each sphere acts as if it were alone in the suspending medium—that is, that the spheres are so far apart that the current lines become parallel in the space between them. Аз will be shown below, a comparison with experimental data shows that within an accuracy of a few per cent this assumption is fulfilled with concentrations from zero up to 30 per cent.

Let us consider first the case of a single sphere suspended in the medium. We assume that the medium is placed in an electrolytic cell with a volume of 1 се. and that a constant electrical current, i, passes through the suspension. Тһе difference in potential of the electrodes we shall designate as V. Since the electrodes are 1 em. apart, V is equal to the electric force. The surface of the sphere is the seat of a certain distribution of surface charges. The surface charge, 5, at an arbitrary point can be developed into a series of surface harmonics.

S = do So + 04.8, + а. S, + Glo A neo On S, + “бе е ete == RW. The potential at any point outside the sphere is? r being the distance of the point from the center of the sphere. The component of the electric force perpendicular to the surface at a point situated on the external surface of the sphere is "Тһе treatment of the general case of an ellipsoid having a surface layer of a conductivity different from that of its interior will be taken up in a paper which will appear іп the Physical Review. This case includes that of a homogeneous polarizable ellipsoid.

* See Jeans, J. H., The mathematical theory of electricity and magnetism, Cambridge, 1920, p. 206. At a point situated on the internal surface of the sphere we have for the same component : These forces are the forces due to the electric charges on the spheres. 'The total forces are obtained by adding the component of the original electric force due to the surface charges on the electrodes of the electrolytie cell. 'This component is equal to — V sin q.

The electric force has a discontinuity at the surface of the sphere. At the moment when the current through the suspension is started, the electrie force is the same on the two opposite sides of the surface of the sphere. As soon as the current is started, since the conductivity of the substance of the sphere is different from the conductivity of the suspending medium, a different amount of electricity will be carried to the outside of an element of surface from that which is being carried away from the inside at the same instant.. Thus, an accumulation of charge on the surface begins and is continued until the difference between the electric forces on the two sides of the surfaces of the sphere becomes so large that the same amount of electricity is carried to the outside of the surface as is carried away from the inside in the same time interval. "That is, the equation of equilibrium is

(Fr ext. рп Ф) 2 (еи в Ф) kz n+ 1 А пазе ee БАР ee kı = or TS S vsing ) 1 n В A ор БЕ Е 3 ( RE TI Vsin p) в. (3) We have? 5, ==4 (8 sin q? — 1) In order that the right and the left side of equation (3) be identities it is necessary that o, be made zero for all values of n Substituting this value for 04, in equations (1) and (2) for the potentials Vext, and Vint, we now obtain The conductivity of the suspension can now be calculated by means of the following method. The space between the electrodes of the electrolytic cell is divided into an infinite number of volume elements, dS, dx; dS being a surface element parallel to the electrodes and dz a line element perpendicular to the electrodes. If Р, is the component of the electric force along x then by using Ohm’s law

k being the conductivity of the suspension. The first integration is taken over all volume elements outside the sphere, the second integration over all elements inside the sphere. In carrying through this integration on the left side we integrate all elements situated in the cylinder defined by a fixed dS. И this cylinder does not cross the sphere, its elements contribute solely to the first integral. This contribution is The contribution of the cylinders crossing the sphere to the left side of our equation is

erence ential | ЕА the ae RI P, е nx crosses the surface of the sphere. Then, what was found above — У Using the cane obtained for Contr., and Contr. equation (4) becomes - Calling the volume of the sphere р we obtain es 3 (kz — kı) ) i (1+ SBS | This formula with р being the total volume of spheres per cc. will hold for a suspension of spheres so diluted that each sphere deforms the current lines of the original current independently of every other.

Е а БҮТ k pex 8р The above formula shows that for diluted cell suspensions — 3 e A test of the theory here developed has been made by applying formula (6) to red corpuscles. As was stated above, red corpuscles are very nearly non-conductors; that is, b;—0. According to our theory ———— should therefore have the constant value of — 4. о given by Fraenckel for red corpuscles of man, dog, horse, and cow. It is seen that the experimental value of = has the theoretical

value of —0.50 up to concentrations of between 30 and 40 per cent. The formula here developed can therefore be expected to hold up to this limit of concentration. Experiments dealing with the applications of the theory here developed to colloids (especially graphite suspensions) and to suspensions of different living cells are at present being made in this laboratory and will be reported in a later paper. (Prom the Department of Biophysics, Cleveland Clinic Foundation)

In another paper, p. 341, I have presented a theory for the conductivity of a suspension of homogeneous spheroids * in which the following formula was derived. k, specific conductivity of suspension. с ы * suspending medium. "o ПИ + “ suspended medium. р, volume concentration “ ~ ы 1The theory for the simple case of a diluted suspension of spheres is given in the paper оп р. 341. ; b Le Muse eei kı kı 7 жалы (2) k * ky sn For a diluted solution 2 is approximately equal to 1; introducing this value of Ё. equation (2) reduces to та kı

This is the equation, the derivation of which was given in another paper.* It is of interest to note that according to equation (1), for a constant volume concentration of the suspended medium the conductivity of a suspension is independent of the size of the suspended particles and also nearly independent of the form of the particles when the difference between the conductivities of the suspended and the suspending media is not very large. This is especially true for suspensions of prolated spheroids which are less conductive than the suspending medium.