The use of artificial electric polarization of 
tissue during relatively short periods of time is there­
fore not necessarily comparable with what we can 
expect to encounter in many biologic situations. Nev­
ertheless , relatively brief application of direct current 
of high density may be helpful as a guide for under· 
standing structural and functional mechanism s of tis· 
sues. When a DC potential is applied between two plati­
num electrodes in a living tissue, several effects can be 
noticed. The quantity of charge transported between 
the electrodes can be determined as the product of 
Activation of VICC 173 current and time, expressed in coulombs (amp-sec). Thus, a current of one rnA over 24 hours gives 86 
coulombs and is equivalent with 10 rnA over 2.4 
hours. Current per unit· time can be increased by 
increasing the vohage between the electrodes. If the 
total quantity of current transported is held constant, 
differences of electrode potential may lead to differ· 
ences in quality of material in the tissue. Such effects 
can be explained by the fact that different materiab 
possess different specific energy levels of ionization. Moreover, different materials possess different molec­
ular and ionic adsorption characteristics, which each 
vary with electrode potential, actual concentrations 
and the properties of the tissue matrix. The covering 
of an electrode by adsorbed molecules is characteristic 
for all materials and solutes. In this way different 
electrode potentials can be expected to affect the com­
petitive adsorption s of different molecules and there­
fore the qualitative composition on the electrode sur­
faces and in the supporting media between the elec­
trodes. Increasing voltage will increase the speed of ioniza­
tion of tissue (see also Chapter XVII) and enhance the 
effects of polarization. Gas fonnation , for instance, 
will be more extensive and produce more pressure 
effects when electrode potentials arc large rather than 
small, under conditions of equivalent total charge 
transport. Experimental demonstration of these effects 
is described in Chapter IX in the studies of pressure 
changes and electrical induction of transport of water 
in cotton wool and lung tissue. Gas formed at the electrodes may interfere with 
conductivit y. Heat may also be liberated at relatively 
high voltages, leading to evaporation of water and 
coagulation of protein. Furtbermore, the catalyzing 
action of the material of an electrode will vary with 
varying electrolyte composition. A number of different factors in tissue influence the 
effects of different electrode voltages at equal total 
amounts of transported charge. Blood circulation , con­
vection, diffusion, regional changes in conductivity , 
matrix effects, buffering by fluids, etc., will each af­
fect ionic transports. These influences may be evident 
already from the fact that the circulation of blood and 
lymph will in varying degree supply and remove mate­
rial in the electric field. The total effect of these factors 
per unit time relates also to the current density in the 
electric field. ln this way a variable biologic influence 
of qualitative and quantitative changes can be estab­
lished between the electrodes. When anodic and cathodic reactions develop in a 
tissue, secondary effects can also be expected. For 
example, ionization product; in the electric field 
should interfere indirectly with zeta potentials of cells 
and modify the transmembranous ionic transports of 
individual cells. Experimental support for these as-
174 Activation of VICC sumptions will also be presented in terms of the behav­
iour of blood and fat cells (Chapter XVI). Our immediate interest now will turn to some gross 
morphologic expressions of the functions of the postu­
lated biologically closed electric circuit, as simulated 
by leading electric current between electrodes in tis­
sues. A. Materials and methods 
Initial experiments could not be performed easily in 
the lung because of the technical difficulties connected 
with direct visual inspection of this organ in vivo. Because the actual problems appear to be of a general 
biologic character, the easily accessible mesentety and 
omentum of dogs were used in a series of 20 experi­
ments. In this way it was hoped that some additional 
direct evidence might be obtained for the existence of 
the proposed VICC. ln these studies platinum elec­
trodes were used. Control experiments were also performed in kidneys 
and lungs of dogs. In these experiments as well as in 
the experiment$ on mesenteric tissue, Evans blue dye 
or a radiographic intravascular contrast medium (Urn­
gratin~, 50%) was injected intravenously or intraarter­
ially at the same time as direct current was applied 
between the electrodes. Dogs were anaesthetized with intravenous sodium 
pentobarbital. A radiopaque plastic catheter (internal diameter, 1.5 
mm) was introduced via a femoral artety or vein and 
placed in the aorta or inferior vena cava with its tip at 
the approximate level of the diaphragm. A platinum 
string was introduced through the catheter and used as 
one electrode at the tip. A midline incision was made in lhe abdomen and a 
piece of mesentery or omentum carefully pulled out 
through the incision and placed on a glass plate for 
support. Another platinum electrode was then placed 
against the mesentery . In other experiments both elec­
trodes were placed against the mesentery. A DC potential difference was then applied between 
the platinum electrodes. The potential was obtained 
by means of a ,-oltage generaror in series with a mil­
liampere meter of high accuracy. Voltage differences between 100 mV and 10 V were 
tested. When morphologically visible changes were 
produced , the experiments could be checked or modi­
lied rc~tedly in the same animal with a fresh piece of 
mesentery. The vessels and other tissues of the mesentery were 
inspected at different magnifications with an operating 
microscope. Photographs were taken of the mesenteric 
electrode and surrounding tissues. The mesentery un­
der and around the electrode was excised at the end of the experiments and fixed in formalin on a glass slide 
before fuation. Staining was usually performed with 
haematoxylin and eosin before microscopy. B. Charging and discharging 
of tissue 
When 100 mV DC voltage was applied between one 
electrode resting against the mesentery and one elec­
trode in the aorta or vena cava, one to several mi­
croamperes could be observed to pass between the 
electrodes . The current increased continuous ly, ftrst 
slowly and then more rapidly. After a wbile the Oow of 
current decreased or became interrupted, usually from 
gas produced at the electrodes . In this way the tissue became electrically charged, 
as shown in Chapter VI (Fig. VI: 13). When the exter­
nal power source was then disconnected and the two 
terminals short-circuite d over a microampere meter, 
such a charged "biologic battery" was found to dis­
charge. Similar discharge from a carcinoma and granu­
loma, spontaneous ly polarized, is shown in the pa­
tients in Figs. VI: II and 12. C. Diapedetic bleedings 
Diapedetic extravasation of red blood cells is one of 
the early visible changes around the electropositive 
electrode. This extravasation can be seen in Fig. XIV: I as many sites of bleeding along small arterial 
branches after the application of DC current in small 
quantity (1.0 pA, 30 min, 1.0 V). This voltage repre­
sents the potential difference actually measured be­
tween the electrodes. Focal arterial contractions a.nd 
associated intraarterial thromboses also interrupt the 
blood stream in many places. Sometimes these blood 
clots were seen washing away, resulting in temporary 
restoration of circulation through the artery. Diapede­
tic bleedings were also observed, but to a much lesser 
extent, around the electronega tive mesenteric elec­
trode. The normal transcapillary openings between blood 
and extravascular spaces, which contain water and 
electrolytes, may be considered as leaking junctions. When an electric potential is applied across capillaries 
some of them appear to open further, as shown by the 
transcapillary passage of red blood cells. After a while the diapedetic blood as well as the 
thrombosed blood in the vessels becomes brown. White foam (Fig. XIV: 2) produced by gas then devel­
ops around the electropositive electrode against the 
mesentery. The surrounding brown dots represent 
diapedetic blood. Fig. XIV: I. Effecrs of elec~ric curren< in small quan<iry (1.0 
pA, 30 min, 1.0 V), passed berween aorta (carhode) and me­
scnrery (anode) of an anaesrherized dog. (a) In vivo phoromi· 
crograph shows diapedetic bleedings as irregular dots around 
rhe plarinum anode. (b) Local diapedetic bleeding. Narrow 
empty arteriocapillaries , wide venocapillaries with granulo· 
cytes. Haematoxylin -eosin stain. Fig. XIV: 2. Foam of oxygen and chlorine gas develops at 
the electropositive electrode againsr dog mesentery. Diape­
detic blood appears as brown dots in the mesentery near the 
clccrrodc. Electrode size 4x 4 mm. Cathode in the aorta. Activation of VICC 175 D. Vascular pockets, 
ischaemic dystrophy and 
perifocal enhancement of 
radiographic contrast 
An in vivo photogra ph (Fig. XIV: 3) shows narrowing 
and discolouration of vessels in the lower part of the 
figure, close to the electropositiv e electrode. Blood at 
this moment was not flowing through the discoloured 
vessels but was flowing freely in the artery and vein 
seen in the upper part of the f~gure . The open branches , still red, of these vessels pro­
duce in this way vascular pockets around the electrode. Circulation is completely suspended around these 
pockets. Similar vascular changes of irregular narrow­
ing and thrombosis of vessels around spontaneo usly 
polarizing lesions have already been described (Chap­
ter III, G). The absence of pulsations sometimes ob­
served in the vessels around carcinomas, re.ported by 
Marchal and Marchal (26), and the frequ<nt presence 
of extensive thrombosis in vessels around carcinomas 
(II) may therefore possibly be detennined by bioelec­
tric eve;nts. Fig. XIV: J. Impairment of circulation induO<d after 25 
minutes at 5 V electrode potential around an anode (out of 
si&Jlt b<olow the: f.,ure) opinst the mesentery of an anaesthe­
tiud dec. Some vessels are narrowed. Brown discoloured 
•usels are blocked by thromboses . Thromboses contribute 
to the development of dystrophy of tissue adjacent to the 
electrode ("A" zone effect). Red blood continues to Oow in 
larger arteries and veins (upper pan of figure), which con­
nect with open sections of the blocked vascular branches. After injection, radiographic contrast medium Oowing in the 
large vessels was found to escape through the partly open 
vascular pockets. Some contrast mtdium was found 10 re· 
main for a prolonged time in the pocke1.s while some contrast 
leaked throu3h capillary membranes into the interstitial U.. 
sue, producilll! "radiogr2phiccontrasl enhanttment" . 176 Activation of VICC Fig. XIV:4. DemonStration of dys<rophy in !issue around 
site or OC electrode. (a) Platinum anode against dog mesen­
tery. Cathode in the aorta. Extensive 1hromboses, dis­
colouration and desiccation de•·eloped around the anode 
after a total of0.20 coulombs (5 V overpolential ). (b) After a 
clunge of polarity and passase of ano1her 0.06 coulombs , tis­
sue water eotered the newly cathodic rqpoo, whieb pre­
viously had become dry wben anodic. An "avascubr'' region 
is seen at the former site of the electrode. Vessels are blocked 
by thrombosis and probably also compressed by inflowing 
tissue water. Such vascular changes around a polarizing tis­
sue may subsequently lead to local dymophy and circular 
displacc:mcnt of tissue (creation of an ''A'' zone effect • ra· 
diographic halo formation, Fig. XI: 9). These mechanisms 
of incerfcrenc:e with circulation of blood create conditions 
whereby a 1umour may regress, either spontaneous ly or by 
anifidally induO<d polarization (see Chapt<r XVII on ru­
mour therapy). We will now focus our interest on a possible role of 
field-induced vascular effetu wirhin a VICC, leading to 
tissut dyscrophy. A positively charged body in tissue 
may be surrounded by extensive thrombotic changes 
which impair local circulation of blood. Such a mecha­
nism should then produce a radiolucent, dystrophic 
"A" zone around a rumour. Tlris mtclranism may also 
be om of uwraltxp/anations for the occasi<mal observa­
tiDtl of spontaneous regresri011 of malignant tumours. This 
aspect is parriculary important because it may represenl one 
of Narure's own ways w combat a mali~ma11t tumour, and Fig. XIV: S. Transport of fluid to region of mesenteric cath­
ode. Anode in aorta. (a) Vessels are seen before application 
of 4.2 V overpotential . (b) After 1.01 coulombs, fluid is seen 
to ~ttumulate agains-t the ~:lectrode. Vessels arc preserved. Circulation of blood continued both inside and ouuide the 
electrode loop. also be a process which can be simulated artificially in 
tissue by supply of electric energy owr impltJnud electrodes 
(Chapter XVII). Early effects of direct current on vessels around the 
anode are illustrated in Fig. XIV:4a. Five volts were 
applied between the electropositive mesenteric elec­
trode and the electronegative aortic electrode. Disco­
louration began in many small vessels after 0.05 cou· 
lombs. Rather extensive discolouration and thrombo· 
sis could be seen in the vessels around the anodic 
electrode after 0.20 coulombs. Tissue in this region 
became desiccated. Polarity was then reversed and 
0.06 coulombs were passed between the dectrodes . Tissue fluid reentered the region around the mescn ~ 
teric electrode. An "avascular" zone developed 
around the electrode (Fig. XIV: 4 b). After extensive 
inflow of water, one can imagine that circular displace· 
ment of tissue structures may occur. Such displace­
ment of tissue may later be enhanced by dystrophic 
changes due to nutritional disturbances. Fig. XlV: 5 a shows a platinum electrode against the mesemery. 4.2 V were then applied between this elec· 
trode and the electropositive platinum electrode in the 
aorta. Mter 0.01 coulombs, fluid is seen to accumulate 
against the cathode (Fig. XIV: 5 b), but the vessels are 
unchanged. No vessels narrowed, blood did not disco­
lour, no thrombi are seen. When the current is increased between the elec­
trodes, vessels narrow around the electronegative elec­
trode and local circulation slows markedly. Diapedetic 
blood is destroyed and becomes brown. This material 
will then disappear and the cathodic area looks empty 
(see also Fig. XIV: 9a). The arterial branches are then 
empty and collapsed . Some veins contain morphologi ­
cally changed, pale red blood ceUs, which do not stain 
with haematoxylin and eosin. In this way ischaemia is 
produced around the cathode as well as around the 
anode. When longstanding, such effects must inevita­
bly lead to ischaemic dystrophy of the affected tissues. The formation of vascular pockets, as seen in Fig. XIV: 3, may also explain the development of so-called 
perifocal contrast enhancement in angiography. Con· 
trast medium distributed in functioning vessels around 
a tumour will escape, e.g., by the force of gravity, into 
the blindly ending pockets formed by panly throm· 
bosed or contracted vessels. Because capillary permc· 
ability is increased, some molecules of the contrast 
medium will also probably leak through the capillary 
walls into the perirumoural interstitial tissue. E. Ionization and ionic 
recombination s 
When a few volts are applied between two electrodes 
placed against mesentery, gas is produced at the elec· 
trode surfaces. The gas can be seen as foam, more 
developed around the cathode than around the anode. Oxygen and chlorine are produced at the anode, hy· 
drogen at the cathode (20). Around the electronegative 
electrode the tissue becomes alkaline, around the elec· 
t.ropositive electrode it becomes acid. Acid-base effects were shown by placing wet strips 
of litmus paper on the mesentery after the passage of 
0.9 coulomb at one volt between the electrodes (Fig. XlV: 6). The red coloured part around the anode cor· 
responds to pH I, the blue-black around t.he cathode 
corresponds to pH 12. A thin uncoloured zone is aJso 
seen between the red and blue areas. The development 
of alkaline and acid regions in electrolysis of water 
within a matrix has been discussed earlier in connec­
tion with electric transpon of water (Chapter IX). The 
experiment in Fig. XIV: 6 shows how this process can 
present in a living tissue. It is apparent that a recom­
bination of hydroxyl and hydrogen ions must take 
Activation of VICC 177 10mm • 
Fig. XIV:6. Acid·b:asc: effects of clccuicity in mcstntcry be· 
tween elrctrodes. After I V and 0. 9 coulomb, strips of wet 
Utmus paper were placed on the tissue beside the electrodes , 
showing a pH of about 12 around t.be cathode and about I 
around the anode. place between the blue and red areas. The change of 
the yellow colour of the litmus paper of pH 7 into 
white indicates that other mechanisms are also in· 
volved. Thus, bleaching of the litmus might very well 
depend on local accumulation of chlorine, which is 
also liberal~ at the anode (20). For the distribution of material in an electric field 
the ionic mobility u, (cm2 v-•s-1) is of fundamental 
imponancc. Ionic mobility for simple ions at strong 
dilution and 2s•c (12) is shown in Table XIV: I. 
Migrating and diffusing ions produced at the elec· 
trodc surfaces tend to recombine when they meet. For 
the most rapid ions, t.bc hydroxyl ions and the pro­
tons, such recombination s should take place according 
to their respective mobilities , about V1 of the distance 
from the cathode ro the anode. In rhe same way other 
recombinations can be expected to rake place at differ· 
ent distances between the eJectrodes, depending on the 
relative speeds of transport of the reactants. The diffusion and migration of protons and by· 
droxyl ions produc~ at t.bc electrode-tissue interfaces 
is always greater than the diffusion of produe~ oxy· 
gen, chlorine and hydrogen gas, as shown in Chapter 
Table XIV: I. tonk mobiliriu (•,)for l<l"IM simple io"', tX· 
prow 1n rm' v-' ,-• ZS'C 
w u· Na' K-em a NO, 
II, • J(t' 36.2 4.0 ).2 7.6 20.7 7.9 7.~ 
I 78 Activation of VICC XIII. This fact means that a primary destructive effect 
on tissue around the electrodes is produced by cathod­
ic alkaJinity and anodic acidity. The chlorine gas, 
which diffuses rclatively slowly, will secondarily 
bleach some of the already acidic, devitaliz~ 
tissue around the anode. These effects are most easily 
demon strat~ when the electrodes are positioned at 
large distances from each other. For example, when 
the anode is placed in the lung parenchyma and the 
cathode only a few em distant in the pulmonary artery 
of a dog (see Figs. XJV: 20, 21), a recombination zone 
of protons and hydroxyl ions is not established because 
of tissue circulation, convection and buffering by tis­
sue fluids. Consequently, central bleaching of tissue is 
produced by chlorine diffusing slowly in the dark 
tissue already destroy~ by acidity. At a shon electrode distance, the propagation of 
protons and hydroxyl ions may stop at the creation of a 
recombination zone. In a tissue matrix, local accumu­
lations of acidic and basic material can also be seen 
against the recombination zone (Fig. XIV: 6). The 
rapidly diffusing and migrating protons and hydroxyl 
ions prevent eac.h ot.her from funher movement be­
cause they recombine as water. In this case the slowly 
diffusing chlorine gets its chance to reach the recom­
bination zone and bleach the material in this region. It 
is important to realize that the diffusion and migration 
of protons around the anode produce the devitalization 
of tissue both at "long" and "short" distances between 
the electrodes. These distinctions about elect.rochemi· 
cal destruction of tissue around the anode warrant 
emphasis because some incorrect interpretations have 
appear~ in recent literature (37), claiming that chlo­
rine produced around the anode causes the primary 
destruction o( tissue. The ionization and transpon of material after appJi. cation of an external source of energy means that 
energy is transfer~ to the tissue and tissue entropy is 
decrea~. Such changes can be observ~ directly in 
several ways. The possibility of discharging the tissue 
as ment ioncd in Section B is one, some mechanical 
effects are oth.er eJWDples . F. Transport and mechanical 
effects 
Particle size and structural properties of the tissue 
matrix are imponant mechanical factors in tissue elec· 
tropboresis. As always, temperature, pressure-volume 
and gravity factors will also influence the combined 
effects of electric field forces and chemical forces. The exchange of fluid in tissue by .-Jcctroosmosis, 
osmosis, convection and circulation of blood and lymph is also of fundamental importance for ionic 
transportS, possible recombinations of ions and distri­
bution of movable dielectric material in the tissue. The 
existence of narrow spaces lined with faxed charges is, 
for example, a prerequisite for the development of 
"true" (anomalous ) electroosmosis (see Chapter IX). An electrophoretic transport of water by recombina ­
tion of protons and hydroxyl ions will also need a 
"capillary" matrix to counteract hydrostatic recircula­
tion. Mesenteric water transport can easily be seen as 
accumulation of water around the cathode and a dehy­
d1'11tion of the tissue around the anode. In Fig. XIV: 7 
a dry zone in the mesentery shows a white spot at the 
former site of the anodic electrode. Such electroosmo­
tic dehydration gives an uneven distribution of water, 
seen as local oedema around the electronegative elec­
trode. ln the case of a distant cathode, a hydropic 1.one 
is also seen peripheral to the anodic hydropenic zone, 
due to the outflow of water from the anode. Accumu­
lation of water around both the dry anodic zone and 
the cathode produces mechartical effects by local in­
creases in turgor pressure. Similar uneven distribut.ion of material in the elec­
tric field is caused by ionic dissociation , but takes 
place also as a transport of dielectrics. By dipole induc­
tion, such particles will move to the anode as well as to 
the cathode, wheneve r they are located close to the 
electrodes and where the electric field gradient is 
steep. The gradual disappearance of brown mat<rial 
from intravascular blood cells and from diapedetic 
interstitial blood also appears to be an e.xpression of 
electrOphoret .ic transportS -
Gas, produced locally at the electrodes , increases 
pressure in the tissues. At the surface of the anode, 02 
and Cl2 are produced, while H2 is produced at the 
surface of the cathode. Around the cathode, the vol-
Fig. XIV: 1. A wrute, dry site of destroyed tissue and blood. Oxygen and chlorine gas d<n<>tes the fonner site of a mesen­
rericanodc . Fig. XIV: B. Streams of gas arising from the surface of plati­
num electrodes in human blood (direct microscopy). (a) Oxygen and chlorine at the anode after 0.138 coulomb, 
6 V. (b) Hydrogen at the cathode after 0.142 coulomb, 6 V. 
The oxygen--chlorine gas sho~"S thinner ''dendrites'' than the 
hydrogen gas. ume of H2 is twice as great as the volume of Oz 
around the anode (20). Chlorine gas adds its volu.me to 
the volume of oxygen around the anode. Living tissue, 
however, resorbs gas. An equilibriu m will therefore 
become created around the electrodes between the 
111tes of production and elimination of gas. When a 10 
V potential is applied to electrodes implanted in ''ivo 
in various tissues, e.g., liver, lung or fat, the gas 
produces within about an bour a r.~diographically ''isi­
ble cavity in the tissue around each electrode. The 
cavity around the cathode is always larger than that 
around the anode. The relative differences of gas pro­
duction arc in themselves a result of a complex series 
of events. Variations in anodic current density, e.g., 
by physical factors such as dehydration and electrode 
depositions, will to some extent lead to changes in the 
relative content of hydrogen , chlorine and oxygen in 
the tissue. A direct microscopic study of the gas produced at 
the electrodes is shown in Fig. XJV: 8. Two platinum 
strings each 0.2 mm thick were fixed on a glass slide. The space between the electrodes was filled with some 
drops of human blood. A 6 volt direct current poten-
Activation of VICC 179 tiaJ was applied between the electrodes. Dendritic 
bubbles of gas arise from the anode (oxygen<hlori ne, 
Fig. XIV: Sa) after 0.138 coulombs and from the cath­
ode (hydrogen, Fig. XIV: 8 b) after 0.142 coulombs. The oxygen-chlorine gas produces more delicately pro­
truding gas bubbles than does the hydrogen gas. In a 
suitable matrix it was shown (Chapter XIII) that gas 
bubbles are liberated and transponed away from the 
electrodes. This mechanism of electrogenic ergonar 
production and transport might be considered in fu­
ture research on vesicular development and function. G. Conductivity changes 
The mechanical effects of gas production, which is 
. most pronounced around the cathode, will influence 
conductivity withi.n the circuit. The spontaneous in­
crease in the amount of current in the early stages of 
tissue electrOlysis can be ascribed to the continuous 
increase in the amount of ionization of the tissue. As 
the process goes on, polarization at the electrode sur­
faces will counteract the process of ionization. Thus, 
gas produced at the electrodes may even completely 
interrupt the flow of current through the circuit. De­
hydration around the anode has also been found capa­
ble of interrupting the circuit in experiments with 
mesentery and lung tissue. The influence of gas produced at the electrodes in a 
tissue can be diminished by applying a relatively low 
current ove.r a correspondi ngly longer time. The rate 
of gas resorption is then relatively more prominent 
than the rate of gas production. The applkation of low rates of current flow through 
the circuit still cannot entirely abolish the effects of gas 
production and dehydration of the tissue. These ef­
fects depend on several factors. The adhesion of polar­
ized material against the electrode surfaces and gas 
adsorption cannot be avoided. Gas in a tissue will 
become trapped in the matrix. Secondary changes in 
morphology will also take place, e.g., within the ves­
sels. Thrombosis around the anode will interfere with 
the resorption of gas and other polarization products 
and with the supply of water to the dehydrated tissue. H. Effects on red blood cells 
and their distribution 
We will ftrst summarize brie.fly wme aspects of 
chemical transformation of haemoglobin . According to 
Lemberg and Legge (23), the prosthetic group (iron 
protoporphyrin ) called haem is rather unstable and 
180 Activation of VICC easily oxidizable. Haemoglobin is split into globin and 
a cationic haemin in acid s<;>lution and into globin and 
an anionic haemin in basic wlution. If haemin is dis­
solved in excess alkali and titrated with acid, electro­
neutral haematin will precipitate. From these data one may already anticipate that 
certain changes will take place in in vivo electrophore ­
sis. Acid haemin should develop around the anode 
(which easily reaches a pH of 2, while pH at the 
cathode reaches beyond 12). Base haemin should 
therefore be found around the cathode. On the migra­
tion of cells containing acid and base haemin in the 
electric field, these haemins should attract each other. Cells with haemin of equal polarity should repulse 
each other. As pH changes, charged haemin com­
pounds develop readily, while proteins are less 
changed. If a solution of haemoglobin is shifted as far 
as pH 12 (which may occur in applied in vivo electro­
phoresis), denatured globin haemochrome is formed 
(23). When this compound is allowed to autooxidize, 
or when dilute sodium hydroxide is added to haemo­
globin or to oxyhaemoglobin and the solution is partly 
neutralized , a denatured globin haemichrome is 
formed, which is called cathaemoglobin. Solutions of 
cathaemog lobin in weakly alkaline solution are brown­
ish and have a characteristic haemichrome spectrum , a 
weak band at 558 Jlm and a stronger one at 530 pm 
(23). In in vivo electrophoresis the material around the 
cathode may be characterized macroscopically by evo­
lution of hydrogen, alkaline reaction, oedema, and a 
brownish material, which should contain cathaemoglo ­
bin. When the pH of a solution of haemoglobin is made 
more acid than 3-4 (which is the case around the 
anode in tissue electrOphoresis ), the linkage of the 
prosthetic group with tbe protein is ruptured , while 
tbe protein is denatured. The purple colour of the 
haemoglobin then changes to reddish-brown and 
shows a rather indistinct, two-banded spectrum in the 
green of haem, protected by protein from flocculation 
(23). In the presence of oxygen the ferrous iron in the 
haem is oxidized to ferric iron. Acid haematin is then 
formed, which .has a hand in rhe red ar 660 prn and a 
feeble band in the green. When oxyhaemoglobin is 
acidified, oxiilation of the protein in addition to the 
iron atom takes place (23). As a preliminary survey of the morphologic distri­
bution and general appearance of field-induced 
changes in in vivo electrophoresis, acute experiments 
were performed first on dog mesentery and lung and 
also, after four weeks observation time, on dog 
lungs. Cathodic mesentery of a dog is shown in Fig. XIV: 9a (haematoxylin-eosin ). One coulomb of cur­
rent at 5 volts has passed between two mesenteric a • lb 
Fig. XIV: 9. Effects of in vivo electrophoresis on red blood 
ceUs in dog mesentery. Anode and cathode, 3 em apart, rest 
against the mesentery (5 V, I coulomb). Cathodic field, 
stained with haematoxylin and eosin after ftxation in formal­
dehyde. (a) The ti:i-:$uC appears empty or cc:lb. VCMC:b appear 
dark and depleted of normal blood ceUs. (b) The central dark 
narrow vessel is an artery containing decomposed blood. The 
electrodes , placed ab<)Ut 3 em from each other. The 
tissue looks "empty", as if no normal blood cell ele­
ments were present in the field. Higher magnification 
(Fig. XIV: 9b) shows an artery and a vein containing 
dark material but no normal c.ell elements. A vein, the 
larger vessel, contains numerous, clearly visible, round 
and oval bodies. Their nonstructured , transparent 
content is seen surrounded by dark material, which is 
also nonstructured . The light material does not stain 
with haematoxylin and eosin. Some interstitial cells in an adjacent part of the 
cathodic field appear as ring-structures, as seen in Fig. XIV: 9c. These cells have been interpreted as diapede­
tic red blood cells which have lost most of their pig­
ment, making their membran es visible. Dark material, 
like that seen in the vessel in Fig. XIV: 9 b, was found 
in the interstitial tissue and was also interpreted as 
modified blood pigment. Red blood cells in the anodic field revealed varying 
degrees of central accumulation of dark material, 
which presumably consists of transformed blood pig­
ment. These obscn,ations were therefore extended by 
studying human CPD-adcnin ® (Terumo, Japan) blood 
on glass slides. Direct current was applied between 
platinum electrodes at 6, 12 and 18 volts, with current 
densities of 1-5 rnA over 5 to 10 minutes. Material 
13t-8US86 Nmdrrmriim • 
0.1mm ._____, c 
• 
0.1mm 
larger vessel is a vein with dark material and round or oval 
bodies, which are likely to be red blood cells depleted of pig­
ment. (c) Some extravascular cells, presumably erythroc~· tes, 
stow cell membran es darker than the pale interiors. This 
change of the red blood cdl:s ha.s been int<:rprct<:d as an <:Cfcct 
of the cathodic field depleting the cells of pigment. treated in this way was sampled from different parts of 
the anodic and cathodic areas, smeared on glass slides, 
dried and stained (hacmatox ylin and eosin, poly­
chrome methylene blue, van Gieson or May· 
Grilnewald-Giemsa ). In these studies remarkable 
transformations could be seen in red blood cells. Cathodic erythrocytes contained multiple particles 
of birefringent material, located inside and often close 
to the cell membranes (Fig. XIV: lOa). In other areas 
particles were seen at the outer surface of cells or, even 
in in vivo experiment s, in the surround ing medium 
(Fig. XIV: 14a). Other cathodic red blood cells were 
pale, as if they contained a reduced amount of hae­
moglobin. They also showed crenations (Fig. XIV: lOb). Blood material from the area close to the anode also 
showed considerable morphologic changes. Many cells 
showed a concentration of centrally located, birefrin­
gent material connected with the interior cell walls 
with thin radiating structures, like spokes in a wheel 
(Fig. XIV: JOe). The cytoplasm of many anodic cells 
also contained round inclusions interpreted as vacuoles 
of varying sizes (Fig. XIV: lOd). These Structures 
seem to develop as small light areas in the cytoplasm, 
which appears granulated. Some of the vacuoles grow 
and take up more and more of the intracellu lar space. Activatio n of VICC 181 a 
> 
... 
c o· 
; . . . ; 
.... :o: ·--
-. \2; -;· 
J 
{~ -. ;; ~ 
(~/ ;> 
c 
... 
\_~ . ... Fig. XIV: 10. Erythrocytes after exposure m direct electric 
current (6 V, 0.5 coulomb). (a) Blood near cathode. Birefrin­
gent parucles, probably alkaline haemJltin, arc seen in cells. These particles can sometimes also appear outside cells: in the 
~urrounding medium. (b) Otbcr caLhoWc crythrocy1es are ci· 
I her d"rk or pale. Some show crcnations. {c) Anodic blood. The "spokes" seem tO develop at the contact surfaces 
of the vacuoles. At the same time the granulated mate­
rial seems to condense in the centre of the ceUs. Only a preliminary explanation of the field·induccd 
structural changes in the blood can be g:ivcn so far. 182 Activation of VICC d .. 
Hacmatin, probably acidic, accumulates centrally and is con· 
ncctcd with the interior cell waUs by th.in radiating struc· 
turcs, like spokes in a wheel. (d) Anodic blood. Large va· 
cuole·Like changes are seen in the cytoplasm, possibly pre· 
ceding the changes seen in c. 
Strong alkalinity is pre>ent in the vicinity of the cath· 
ode. As reported earlier, it is known that weakly acidic 
and alkaline solutions split haemoglobin into its globin 
and prosthetic parts. Alkaline (elecuonegative ) haem 
may then represent the particles of bire[ringent blood Fig. XIV: II. "Giant •erythrocytes" after exposure of blood 
to direct current. (a) Erythrocytes from the cathodic area 
seem to fuse into "giant cciJs" with characteristic arrange· 
me.nts of blood pigme:nt. In normal·sized erythrocy tes the 
haemoglobin is granulated or (b) arranged like Roman nu-~ ·""~ ,_ 
~-'\ ' ""t, 
~ 
~ T 6 ...;:.. 
~ 
@ ~~ -~ };1 
~ · ~· . ? 
"' " ,...., _J 
C) ' ~ 
tt:-i \..:! 'Y.r 0 
'Y) 
0 i 
""':! merals II and V. (c) The anodic area contains different "gi· 
ant cells" appearing as round structures which contain a fair· 
ly homogeneous , granulated material surrounded by a mcm· 
brane-like structure . Other blood cells appear smaller than 
normal. (d) Control. The same normal blood, untreated. Activation of VICC 183 ·~ · 
I' 
c 
• .. .. 
• 
• . ~ 
<D2mm. JF~m . Fig, XIII: 12. Transformotion of blood by direct current. (u) 
"Giant cdJstl ncur the cathode arc arranged in clusters. The 
"cellll" in cnch cluster are pr~:sumcd to adhere to 3 centrally 
located. undefined matcriol. (b) Dc:ail "icw of a cluster. The 
pigment in the "giant cells'· is arra1:gt. :d more or l.:ss in a cir-
184 Activation of VICC 50JJm 
de. (c) Similllrly structured conglomerates of blood material 
in the anodic region. (d) Demit \1iew of nn anodic COt'lglomer­
ate. A rncrnbrane- like, thin s1rucrure forms lln interface be­
tween this "intercellular space", which contains a fain d)' 
stainable m{tteriaJ, and the surrounding anodic cells. Fig. XIV: 13. Behaviour of 
pigment in red blood cells 
after electrophoresis. Ca· 
thodic alkalinity and anodic 
acidity produce electronega· 
tive and electropositive 
blood pigment, resptttively. Competitive concentra tion 
forces among cells and elec­
trostatic forces of haemin 
of equal polarity are inter­
preted to pro<JU(. "C mutual re­
[Hlling of pigment in adja­
cent cells, as seen in 2 
(which represents cathndic 
blood). Same arrangement 
of pigment was also ob-­
served in anodic blood. (b)Cathodic blood showing 
rows of erythrocytes with in· 
temal attraction of their 
blood pigment. This ftnding 
was also observed in anodic 
blood, (see Fig. XIV: !Oc). Such attraction of pipnent 
should take place when ad­
joining ceUs contain blood 
pigment of opposite polar­
ity. t/ 
II>< 
b -cf -I II' 
pigment, which move toward the cell periphery after 
the cells have moved electrophoret ically from the elec­
tronegative into the electropos itive electric Held. The 
particles even leave the cells and are seen attached 10 
the outer cell surface. Before these cells have left the 
electronegative fteld, however, the alkaline haem may 
concentrate inside the cells. In the anodic fteld acidic 
haem should be produced. This pigmen 1 should then 
also behave differ<ntly when the blood cells appear in 
the electroposit ive field and after an electrophoretic 
transport into the electronegative field. Associated 
mechanisms are complex and do not at present allow 
any definitive analysis. The varied behaviour of blood 
pigment in these studies clearly.show s, however, that 
chemical and structura l modificat ions do take place 
in~irle a~ well a~ Otlt!-:icfe cells in the elecrrlc field. Cathodic cell populations in the in vitro electrophor ­
esis of blood appeared partially fused, in some way, 
resulting in enormously large "cells" (Fig. XIV: I I a). These "cells" appeared in varying sizes ranging from 
the normal size of red blood cells to well beyond ten 
14t-824S86 NordiPisrriim times this diameter. These "giant cells" appear en­
closed in a structure which looks like a cell membrane 
and contains partly structured dark material. This 
material may often be seen as linear, parallel or angu­
lated linear structures (like Roman numerals II or 
V). The dark material appears not only in the "giant 
cells" but also in cathodic cells of lesser size (Fig. XIV: I I b) (see also the presumed early stages of linear 
formation of haemoglobin in Fig. XIV: 13a, b). The anodic area contained blood cells (Fig. XIV: II c) whicb were smaller than untreated blood 
cells (Fig. XIV: I ld). These small anodic cells showed 
granulated cytoplasm. Another kind of "giant cells" 
also appeared in the anodic area (Fig. XIV: I I c). These "cells" comained an evenly distributed granu­
lar substance surrounded by a dense membrane-like 
structure. Structural arrangeme nt of cathodic "giant cells" 
was also observed in smears from cathodic blood (Fig. XIV: 12a, b). In smears from anodic blood,local accu­
mulation of Structured material could also be seen 
Activatioa of VICC I 85 a 'V c. 
I 
) 0 , 
0 Ge 
0 /-
}) 
r'1 
b c 
around some lighter amorphous material, surrounded 
by a polycyclic, thin or membrane-like structure (Fig. XIV: 12c, If). Curious differences in local arrangements of hae­
moglobin were observed in the anodic and cathodic 
fields. Thus, haemoglobin became concentrated in 
cells near the cathode (Fig. XIV: 13a) or appeared as 
if mutually repelling haemoglobin in adjoining cells. To a lesser extent, haemoglobin also appeared as if it 
were attracting when cells were close together (Fig. XIV: 13 b). Haemoglobin in cells from ncar the anode (Fig. 186 Activation of VICC ... e oo 
' 
G C 
25pm 
Fig. XIV: /4. Effects of di­
rect current on dog blood. 1700coulombsat 10 vohs 
were applied in vivo be­
tween the aorta (cathode) 
and neck muscles (anode). A 
sample of venous blood 
from the animal was hepar· 
inized and centrifuged. Blood smears were made 
from (a) top layer, which 
showed birefringent pani· 
des inside and just outside 
the periphery of red blood 
cells (compare Fig. XIV: lOa), and (b) bottom 
layer' wruch showed mainly 
normal erythrocytes , and (c) 
mjnor changes similar to 
those produced in vitro 
(compare Fig. XIV: 11 b) 
were found in a few regions. XJV: JOe) showed similar differences in distribution 
("attracting and repelling haemoglobin "). Concentra­
tion of haemoglobin in separate cells or "repelling 
haemoglobin " in adjoining cells was common in both 
anodic and cathodic fields. Changes in erythrocytes could also be produced in 
in vivo experiments with direct current in dogs. Thus, 
blood is shown from a dog in Fig. XIV: 14 after it had 
received 1 700 coulombs at 10 vohs between the aorta 
(cathode) and the neck musculature (anode). Blood 
was taken from a vein and heparin added. After cen­
trifugation , blood from the top and bottom layers was smeared on slides. Erythrocytes from the top layer 
(Fig. XIV: 14 a) showed birefringent haemoglobin par­
ticles inside and just outside cells, very much like the 
cathodic cell material which was shown in Fig. XIV: lOa. Blood from the bottom layer (Fig. XIV: 14b) showed cells which have been interpreted 
as appearing normal, although they seemed possibly 
less stainable than control blood. Some changes were, 
however, found in this blood (Fig. XIV: 14c), which 
resembled cathodically transformed erythrocytes. The morphologic changes observed in blood from 
the anodic and cathodic fields varied considerably with 
the distance from the electrodes. Erythrocytes at the 
anodic electrode surface were completely destroyed. At a sufficient distance from the electrodes, normal 
cells were seen in both the anodic and cathodic fields. Thus far we may conclude that considerab le mor­
phologic changes can be observed in red blood cells in 
both the anodic and cathodic fields. As will be shown 
in Chapter XVI, fat and water in adipose tissue can be 
induced electrophoretically to move over cellular 
membranes. It is therefore likely that similar mecha­
nisms are also involved i.n the development of the 
morphologic changes just described for red blood 
cells. The experimental conditions in vivo and in vitro 
are certainly not comparable in many respects with 
spontaneous in vivo conditions. Nevertheless, an aug­
mented experimental activation of anticipated BCEC 
channels may give us a hint about the capability of this 
biokinetic mechanism to modify the structure of cells 
and tissues. With this possibility in mind we will now 
consider the in vivo behaviour of leukocytes in a 
VICC activated experimentally in mesentery. I. Accumulation of 
granulocytes 
One of the dramatic morphologic changes in the elec­
tric field is marked accumulation of granulocytes 
around the anode at "low" voltages. One volt, which must still be regarded biologically 
as a relatively high voltage difference, was app.lied 
between the electropositive platinum electrode placed 
against the mesentery of a dog and the electronegative 
platinum electrode in the aorta (Fig. XIV: 15). After 
0.002 coulombs (60 minutes, 0.6JIA), the current was 
interrupted and a piece of the mesenteric tissue excised 
around the site of the anode. The tissue was then 
placed o.n a glass slide, fixed in formalin and stained 
with hacmatoxylin and eosin. Dark brown material 
was found to have accumulated near the edge of the 
electropositive electrode (Fig. XIV: !Sa). The large 
dark vessel in the figure is a vein, which anastomosed 
15-814586 N(lfJnmriim Fig. XIV: IS. Accumulation or granulocytes around the an­
ode after application of direct current to dog mesentery . (a) 0.002 coulombs were delivered at I V overpotcntial be· 
tween an anodic mesenteric electrode and an aortic cathodic 
electrode. Mesentery then was flxed i.n formaldehyde and 
stained with haematoxyJin and eosin. Brown material accu· 
mula ted at the site or the anode (left). The blue \•esse I to the 
right is a vein. It is entirely fllled with granulocyte s. Brown 
material in lhe large vessel to the left consists mainly of de· 
compo<ed granulocytes. (b) Detail view shows white blood 
cells in vein. Adjacent artery (arrow) is collapsed and empty. with a blue vein in the right-band side of the figure. This blue material consists of granulocytes (Fig. XIV: I 5 b) completely ftlling the vein. No red cells 
could be seen in these vessels. Adjacent arteries are 
narrow and empty of blood cells. By following the blue 
vessel branches to the black ones it can be seen that the 
black material consists of granulocytes, increasingly 
decomposing from right to left. Fig. XIV: 16a shows a vein leaking granulocytes 
about one em from the cathode. Anode and cathode in 
this experiment were each placed on dog mesentery (2 
volts, 2.80 coulombs). Granulocytes arc attracted to the walls of veins be­
fore they pass into interstitial tissue. Tbis attraction is 
seen in Fig. XIV: 16 bas "margination of leukocytes" 
Activation of VICC 187 Fig. XIV: 16. Accumulation of granulocytes around elec­
trodes in dog mesentery . (a) G1'1ll1ulocytes leaking through 
veins into the interstitium (2 V, 2.8 coulombs) one em from 
the cathode. (b) White blood cells marsinatins to the walls of 
a vein (large vessel, left) near the anode. Right vessel is anal­
most empty artery. in a vein containing a moderate increase in quantity of 
these cells. Mesenteric electrode was anodic, aortic 
cathodic (2 volts, 1.2 coulombs ). The same collection 
of leukocytes in veins occurred when the polarity was 
reversed. While gl'llllulocytes were accumulating selectively in 
veins, adjacem aneries have been empty of blood cells. Leukocytes accumulated in veins also when the cath­
ode was on the mesentery and the anode placed in the 
inferior vena cava, or when the polarity of these elec­
trodes was reversed. This finding is illustrated in Fig. XIV: 17, which also shows that in bnth instances the Fig. XIV: 17. Accumulation of granulocytes and altered vas­
cular calibre induced by direct current in dog mesentery . (a) Mesentery cathodic and inferior vena cava anodic: a vein, 
containing granulocytes, is wide. An adjacent artery, empty 
of cells, is narrow. (b) Mesentery anodic and vena cava ca­
thodk a vein, packed with gramtlocyles, is narrow. The ad­
jacent artery, empty of cells, is wide. adjacent arteries were empty. (See also regional 
"arteriocapillary" contractions and "venocapillary" ac· 
cumulation of granulocytes in Chapter XU.) The distribution of individual cells may be taken as 
support for the theory that BCEC systems represent an 
additional circulatory system capable of selective 
transports . It may be possible that BCEC syste.ms can 
tl'llllsport selectively not ooly electrolytes but also 
charged particles as large as blood cells. The mechanism of electrophoretic transport of 
charged particles very likely depends on many factors. Thus, it is known that granulocytes make their way 
Table XIV: 2. Surface-charge characterisrics and nature of chemical groups contributing to surface charge per cell surface area for 
pliltekts, lymplw<ytes , and erythrocytes 
Negatively charged groups 
Anod!ic 
electro- Cor· Phosphate 
phoretic Apparent =ted Amino NANA susceptible to" Weak 
mobility electron electron groups a-carboxyl acidic -SH 
(pmlfi<d charges charges (+) (pK. 2.6)" RNase~· Alk. phos.' (pK-4) groups 
Cell type Vicm) (x Ill") (X !IJ") (X lo') (X lo') (X 10') {X 10'') (X II}') (xiO") 
PlateletS-" 0.85±0.04 1.8 2.04 2.42 8.9 5.0 6.5 0.28 
L)'liiphocyt<>• 1.09±0.08 9.34 10.29 9.5 29.0 8.7" 55.5 1.98 
Erythrocyu!s ~ 1.08±0.03 10.29 10.29 Not detected 62.0 40.9 Not detected 
• Surface areas o( cells: 2B.27 ~~m1; 113 pm1; 163 pm1. Electrokineti c data obtained in physiological saline (0. 145 M NaCJ; pH 7.2 at 250C). b May lx present but <5 x 10~ per cell. 
' ThrOiftb. DiJJth. lfaonorrh. Suppl. l6: 53 (1967); Z6: 370 (1971). • lnr. Arch. Allnr~42 : 69 (1972). • Arch. Bioch•m. Bioploys. /35: 356 (!969). 'Exp. C<ll Res. 50: 441 (1968). 188 Activation of VICC Fig. XIV: 18. Illustration of 
interaction between blood 
flow and cathodic field lead· 
ing to accumulation of gran. ulocytes. The flow of blood 
in veins is directed tOward 
the cathode. The electrone· 
gative electric field will se· 
lectiveLy counteract the flow 
of electronegatively charged 
granulocytes which leads to 
an increasing accumulation 
of granulocytes in these ves· 
sels. See also accumulation 
of an electronegative chemi­
cal compound (Fig. XIV: 19). through pores in the capillary membranes by means of 
pseudopods. Also various matrix factors in tissue may 
influence electrophoretic transport of different cells. The relation between their surface charges and the 
strength of the superimposed electric field might be 
important elements in the mechanism of selective 
transport of cells in tissue. In this connection, surface 
characteristics of blood platelets, lymphocytes and red 
blood cells arc of particular interest. A compilation of 
actual data from the literature (30) is presented in 
Table XIV: 2. A surplus of ftxed electronegative ~harg~ also ~bar· 
acterizes granulocytes. In a closed electric circuit any 
cell with a surplus of electronegative charges "\Viii tend 
to move toward the anode and be repelled by the 
cathode. This fact does, however, not necessarily 
mean that all electronegative cells wiU migrate to the 
same extent. Differences in excess of charge and steric 
position of charges in relation to the matrix should 
influence the transportS, as well as factors such as 
surface friction, size and pliability of the cell. The 
repellent force of a cathode or an electronegative phase 
of a degrarung tissue was also found to influence the 
distribution of blood cells. Thus, the area immediately 
around a cathode was always fr<'C of granulocytes and 
red blood cells when granulocytes were attracted to the 
anode. Distal to the zone free of granulocytes, collec­
tions of granulocytes could, however, be seen in ves­
sels and interstitial tissue. This finding might be ex­
plained as a selective repellent force on these cells flow­
ing in blood toward the electrode, resulting in a rela­
tive increase in their local concentrations. Fig. XIV: 18 
Illustrates the effect of such a flow and field interaction leading to the accumulation of granulocytes. Two 4x4 
mm large platinum electrodes were gently placed on 
the mesentery of a dog. After 100 mV, 10 t•A, 30 
minutes, veins directed toward the cathode fllled with 
granulocytes up to a certain distance from the cathodic 
surface. During the flow of blood in these veins, the 
electronegative field of the cathode must evidently 
selectively retard the electronegatively charged granu· 
locytes, which thereby will accumulate (see also Sec· 
tion K on selective Oow and field accumulation of an 
electronegative dye around the cathode). J. A revised view of 
so-called "chemotactic" 
accumulation of granulocytes 
in inflammation 
Van Lancker (22), in an excellent monograph on mo­
lecular and cellular mechanisms, states: "The nature 
of the mechanism that directS the movement of the 
leucocyte against a concentration grarueot of a chemo­
tactic substance is completely obscure. . . . There 
seems to be no obvious common denominator in all 
agents capable of eliciting chemotactism .... Little is 
known of the machinery which brings the cell to re­
spond to the cell of nhe chemotactic agent." A survey of the field of chemotaxis has been made 
by Harris (17, 18). His conclusion was almost a dam­
nation of the biological meaning of chemotaxis. In his 
Activation of VICC 189 view, chemotaxis is an in vitro phenomenon which 
probably has no relevance in vivo. In an authoritative survey in 1978 on leukocytic 
locomotion , Stossel ( 43) refers to several partial 
mechanisms but remarks that "whether any of the 
above "explanations" for the components of locomo­
tion and for the locomotion per se turns out to be 
correct, remains to be seen." Wilkinson and Allan also stated in 1978 (52, 53) that 
a number of researchers have found "the loose use of 
the word chemotaxis to describe migration of cells in 
the presence of an attractant is increasi.ngly unsatisfac ­
tory and confusing". They therefore propose a defini­
tion of chemotaxis as "a reaction by which the direc­
tion of locomotion of cells is determined by substances 
in their environment". Most remarkable about the concept of chemotaxis is 
the fact that this commonly used term is not related to 
any defmed bioltinetic mechanism. As long as the 
concept of chemotaxis varies from describing a specific 
effect to a whole bioltinetic process including antici­
pated but unknown chemical and humoural factors, 
confusion will persist. The reader may already have a presentiment of an 
alternate explanation for the attraction of leukocytes in 
inflammation: granulocytes are well known to carry a 
surplus of fixed electronegative charges on their sur­
faces (I, 2, 5, 50). Granulocytes should therefore be 
attracted to the anode within a closed electric circuit, 
as shown in the experiments earlier presented . Corre­
sponding in vivo to experimental circuitry powered by 
an external battery, is the VICC system, powered by 
physico(-electro-)chemical gradients between the in­
jured and surrounding normal tissues. We will discuss 
this alternate mechanism in more detail after reviewing 
some essential information from the literature on che­
motaxis and inflammation. Some of the relevant litera­
ture on chemotaxis is also presented in Chapter XVI, 
Section U, where a corresponding discussion is made 
on the background of spontaneous accumulation of 
lymphocytes in breast carcinomas. Leukocyte s are considered to respond to a variety of 
chemoattractants "most likely" by way of specific 
receptors on their surface (21, 46, 47). In this theory it 
is assumed that chemical substances released at the 
endothelial surfaceS< possess dJe ·capacity to ·'·'glue" 
leukocytes to the vessel walls, producing marginations 
(8). Alternatively, leukocytes in some way may be 
bound electrostatically to the endothelial cells, as de­
scribed for thrombocytes (38-42). The essential draw­
back in these prevailing theories on margination is that 
no "glue" has yet been identified. Moreover , electro­
static attraction of charged leukocytes to charged en­
dothelial sites should interfere with diapedetic migra­
tion through the vessel walls (27). It can be concluded that the inflammatory accumu-
190 Activation of VICC lation of leukocytes in blood vessels and, particularly, 
the adhesion of leukocytes to endothelia l walls have 
not yet been explained satisfactorily. The local accumulation of polymorphonuclear leu­
kocytes in inflammation appears first in the vessels. Some of these vessels are widened (24, 54). The leuko­
cytes either adhere to the vessel walls, a process called 
leukocytic marginati on (14), or fill the lumens of the 
small vessels. The leukocytes next migrate through the 
vessel walls via "diapedetic transport" (17) into the 
interstitial tissue. In the interstitium the leukocytes are 
thought to be attracted in some way to the cause of 
inflammation, e.g., bacteria. The mechanism of vaso­
dilatation in inflammation is only known to a certain 
extent (24). ·Focal liberation of serotonin, histamine 
and histamine-like substances (15, 35, 44, 45) are 
involved, e.g., as a result of stimulation of sympathetic 
nerves through axon reflexes (29). A direct effect by 
the injurious agent on the capillaries is often discussed. Various peptides and proteins, isolated in inflamma ­
tory tissue (4, 6, 10, 31), have been thought to produce 
an increase in capillary permeability. This group of 
substances includes leukotaxine, exudime, brady­
kinin, substance P, kallidine and plasmin (13, 32, 33, 
49). The prostaglandins (7) induce local signs of in­
flammation, e.g., erythema and oedema, and have also 
been suggested to function as mediators of inflamma­
tion (28). A large number of substances have been studied 
with regard to their chemotactic properties. Rivkin 
and Becker (36) have shown that cyclic AMP, hista­
mine, epinephrine, isoprotereno l and prostaglandin E 
inhibit chemotaxis of polymorphonuclear leukocytes 
in rabbits. Cyclic nucleotides, e.g., 3', 5' guanosine 
monophosphate, strongly attract polymorphonuclear 
cells (19, 36). N-formyl-methionyl-methionyl-methi­
onine, N-forrnyl-methionyl -leucylphenyl-alani ne and 
other peptides, as well as different suspensions of 
bacterial cultures, are often used as chemoattractant s 
in experimental chemotaxis (46). The diapedetic process has been studied by light 
and electron microscopy. These studies have revealed 
only that white blood cells either appear to be pushed 
somehow through the endothelial cells or that they 
actively pass between endothelial cells by means of 
pseudopods .. (25, 34). In this case the spaces between 
endothelial cells are enlarged or thought to be widened 
by some action of the leukocytes. The interstitial accumulation of leukocytes is gener­
ally believed to be caused by "chemical signals" from 
the injured tissue to the leukocytes. Bacteria are be­
lieved to attract leukocytes, as judged from in vitro 
experiments. Menkin (33) exrracted from inflamma­
tory tissue an unidentified polypeptide he called Jeuco­
tax:ine with uchemotaxic properties ". The attraction of leukocytes in inflammation and their •engulfment of bacteria and necrotic tissue materi­
al are still to a large extent not well understood. The present studies focus our interest on the fact 
that one essential part of inflammation, the accumula­
tion of granulocytes, can be produced experimentall y 
in abundance by in vivo electrophoresis without the 
use of bacteria, "chemotactic compound s" or by in­
ducing primary tissue injury {the platinum electrodes 
were placed gently on the mesentery ), inoculating mi­
croorganisms or using chemical agents. The present 
studies also include other component s of inflammatory 
responses: thrombosis , capillary leaking, diapedesis , 
transport of water, producing oedema, and, as we will 
also see in late phases of healing, the formation of scar 
tissue. In this view, the accumulation of granuiO<:ytes in in­
flammation may be regarded as a special case of an 
electrochemical field reaction within a biologically closed 
electric circuit (the vascular-interstitial closed circuil, 
VICC), combined with electrophoreti c transport of granu­
/O<:yte.s. To summarize, a damaged tissue presents a fluctuat­
ing electrochem ical potential in relation to surround­
ing normal tissue. This potential difference will pro­
duce a driving force in a vascular-interstitial closed 
circuit (VICC), attracting the electronegati ve granulo­
cytes. to the initially electropositive injured tissue. During this phase arteriocapillaries contract in certain 
regions while venocapillaries permit diapedetic passage 
of the electronegatively charged granulocytes . An elec­
trophoretic transport with accumulation of electrone ­
gatively charged cells, i.e., red blood cells, thrombo­
cytes and leukocytes should then take place. It is still 
not dear if the magnitude of the driving potential 
difference of the circuit, the specific surface charges of 
the cells or other mechanisms are responsible for the 
preferential attraction of a certain type of ceUs .. Mas­
sive attraction of granulocytes to the anode, for in­
stance, was observed preferably at "relatively low" 
voltages (in the tested region of I to 3 volts). In the 
region of 10 volts a minor increase in quantity of 
identifiable granulocytes was present around the an­
ode. This fmding has been interpreted as an effect of a 
too violent interference of the electric field with forces 
which in spontaneous inflammation probably interact 
very delicately . Several other factors may also influence the electro­
phoretically accumulated cells. When, for instance, 
the polarity of an injured tissue changes from anodic 
into cathodic, one must assume that the granulocytes 
attractro during the anodic phase will in the new 
cathodic surround ing become trapped and electro­
chemically changed. Electronegative ly charged blood 
cells flowing in vessels toward a cathodic tissue region 
shouad selectively become retarded in the blood stream 
and consequently increase in concentration . Certain extracts or chemicals added locally to a tissue should 
also be able to produce positive or negative "chemo­
taxis", depending on their polarity. It is therefore 
possible to include different partial factors connected 
with "chemotaxis" under one common mechanism: 
electrophor etic, selective lranspor:t of ewc(T(megative/y 
charged granulocytes in a biologically closed, activated 
electric circuit. Acceptance of such a mechanism for 
charged blood cells to enter a tissue suggests that 
therapeut ic accumulation of granulocyte s might be in­
duced by an external electric DC power source over 
electrodes . Another aspect may be to look for "leuko­
toxic agents" with regard to their properties to polarize 
a tissue. It should be evident that VlCC mechanisms 
may also be capable of influencing the distribution of 
other cell elements in blood, i.e., lymphocytes , plate­
lets and red blood cells. K. Local accumulation in 
tissue of a charged chemical 
compound 
The principle of the proposed mechanism for accumu­
lation of granulocytes in an injured tissue should also 
apply for charged chemical compounds. To confirm 
this hypothesi s, Evans blue dye, which is electronega ­
tive, was selected for investigation. An overpotential of 4.2 volts was applied between 
two platinum electrodes against a piece of mesentery of 
an anaesthetized dog. After one coulomb had passed 
through the tissue, the usual vascular changes were 
seen in the tissue in an area about 15 mm in diameter 
around each electrode. A few grains of Evans blue dye 
were dissolved in 20 ml of saline. This solution was 
then injected into the aorta via a catheter while current 
was applied to the mesenteric e!ectrodes. After a few 
minutes a ring of blue stain could be seen against the 
brown-yellow zone around the el.ectronegative electrode 
(Fig. XIV: 19). A very small amount of blue stain 
could also be seen around the electropositive electrode . Electrophoretic tests of the dye in vitro showed that it 
moves in water solution to the electropositive electrode. Nevertheless, upon repeated tests of injecting the dye 
into the dog aorta, the dye always accumulated in 
tissue around the electronegative electrode. These 
tests also included the simultaneous application of cur­
rent and injection. It is evidently possible to attract a charged com­
pound to a polarized part of a tissue. In the experiment 
described , the electronegative Evans blue dye possibly 
combined in the blood stream with some other com­
pound or compounds provided with a surplus of elec-
Activ·ation of VICC 191 Fig. XIV: 19. Electrophoretic anraction of a charged com­
pound, injected intravascularl y. After I coulomb at 4.2 V 
overpotential was passed between two mesenteric electrodes, 
dilute electronegative Evans blue dye was injected into the 
aorta. It accumulated as a blue ring around the cathode. This 
accumulation is explained as an interaction between blood 
flow toward the cathode and the repellant force of theca­
thodic field. tropositive charges compared to the magnitude of neg­
ative charges on the molecules of the dye. In this way 
the resulting combination might be amacted to the 
electronegative and not to the electropositive elec­
trode. A more likely explanation is the one illustrated 
in Fig. XIV: I8 for accumulation of granulocytes in 
veins directed toward a cathode. According to this 
mechanism the electronegative dye, flowing in arteries 
or veins in the direction of the cathode should accumu­
late in these vessels by the opposing electric field force 
of the cathode. This experiment demonstrates that artificial electric 
polarization can cause a chemical compound to accu­
mulate locally in a tissue. It also shows that the polari­
tie.s of the electric field and of the charge of the 
compound are not the only factors which determine 
where the compound will accumulate in an in vivo 
experimen t. 
L. Direct current studies 
in the dog's lung 
The directly observable tissue changes by direct cur­
rent in mesenteric tissue were also studied in the lun.gs 
of dogs. 192 Activation of VICC Platinum strings, 0.25 mm thick, were coated with 
Teflon. The coat was removed for a length of 0.5-2.5 
em at each end of each string. One end was then 
inserted for a distance of 0.5-1.0 em into the tip of an 
ordinary biopsy needle, I mm thick and 16 mm long. Bending the string at the tip of the needle formed a 
hook. The string-containing needle was then inserted 
percutaneously under fluoroscopic control into the 
lung of an anaesthetized dog. The needle was re­
moved, leaving the string electrode in place. The hook 
served the purpose of keeping the electrode in place.
631928626-Biologically-Closed-Electric-Ci-Bjorn-Nordenstrom-pdf.txt
Summarize the following text section accurately and concisely. Maintain specific details and numerical information, and use anti-repetition mechanisms:

        The use of artificial electric polarization of 
tissue during relatively short periods of time is there­
fore not necessarily comparable with what we can 
expect to encounter in many biologic situations. Nev­
ertheless, relatively brief application of direct current 
of high density may be helpful as a guide for under· 
standing structural and functional mechanism s of tis· 
sues. When a DC potential is applied between two plati­
num electrodes in a living tissue, several effects can be 
noticed. The quantity of charge transported between 
the electrodes can be determined as the product of 
Activation of VICC 173 current and time, expressed in coulombs (amp-sec). Thus, a current of one rnA over 24 hours gives 86 
coulombs and is equivalent with 10 rnA over 2. 4 
hours. Current per unit· time can be increased by 
increasing the vohage between the electrodes. If the 
total quantity of current transported is held constant, 
differences of electrode potential may lead to differ· 
ences in quality of material in the tissue. Such effects 
can be explained by the fact that different materiab 
possess different specific energy levels of ionization. Moreover, different materials possess different molec­
ular and ionic adsorption characteristics, which each 
vary with electrode potential, actual concentrations 
and the properties of the tissue matrix. The covering 
of an electrode by adsorbed molecules is characteristic 
for all materials and solutes. In this way different 
electrode potentials can be expected to affect the com­
petitive adsorption s of different molecules and there­
fore the qualitative composition on the electrode sur­
faces and in the supporting media between the elec­
trodes. Increasing voltage will increase the speed of ioniza­
tion of tissue (see also Chapter XVII) and enhance the 
effects of polarization. Gas fonnation, for instance, 
will be more extensive and produce more pressure 
effects when electrode potentials arc large rather than 
small, under conditions of equivalent total charge 
transport. Experimental demonstration of these effects 
is described in Chapter IX in the studies of pressure 
changes and electrical induction of transport of water 
in cotton wool and lung tissue. Gas formed at the electrodes may interfere with 
conductivit y. Heat may also be liberated at relatively 
high voltages, leading to evaporation of water and 
coagulation of protein. Furtbermore, the catalyzing 
action of the material of an electrode will vary with 
varying electrolyte composition. A number of different factors in tissue influence the 
effects of different electrode voltages at equal total 
amounts of transported charge. Blood circulation, con­
vection, diffusion, regional changes in conductivity, 
matrix effects, buffering by fluids, etc. , will each af­
fect ionic transports. These influences may be evident 
already from the fact that the circulation of blood and 
lymph will in varying degree supply and remove mate­
rial in the electric field. The total effect of these factors 
per unit time relates also to the current density in the 
electric field. ln this way a variable biologic influence 
of qualitative and quantitative changes can be estab­
lished between the electrodes. When anodic and cathodic reactions develop in a 
tissue, secondary effects can also be expected. For 
example, ionization product; in the electric field 
should interfere indirectly with zeta potentials of cells 
and modify the transmembranous ionic transports of 
individual cells. Experimental support for these as-
174 Activation of VICC sumptions will also be presented in terms of the behav­
iour of blood and fat cells (Chapter XVI). Our immediate interest now will turn to some gross 
morphologic expressions of the functions of the postu­
lated biologically closed electric circuit, as simulated 
by leading electric current between electrodes in tis­
sues. A. Materials and methods 
Initial experiments could not be performed easily in 
the lung because of the technical difficulties connected 
with direct visual inspection of this organ in vivo. Because the actual problems appear to be of a general 
biologic character, the easily accessible mesentety and 
omentum of dogs were used in a series of 20 experi­
ments. In this way it was hoped that some additional 
direct evidence might be obtained for the existence of 
the proposed VICC. ln these studies platinum elec­
trodes were used. Control experiments were also performed in kidneys 
and lungs of dogs. In these experiments as well as in 
the experiment$ on mesenteric tissue, Evans blue dye 
or a radiographic intravascular contrast medium (Urn­
gratin~, 50%) was injected intravenously or intraarter­
ially at the same time as direct current was applied 
between the electrodes. Dogs were anaesthetized with intravenous sodium 
pentobarbital. A radiopaque plastic catheter (internal diameter, 1. 5 
mm) was introduced via a femoral artety or vein and 
placed in the aorta or inferior vena cava with its tip at 
the approximate level of the diaphragm. A platinum 
string was introduced through the catheter and used as 
one electrode at the tip. A midline incision was made in lhe abdomen and a 
piece of mesentery or omentum carefully pulled out 
through the incision and placed on a glass plate for 
support. Another platinum electrode was then placed 
against the mesentery. In other experiments both elec­
trodes were placed against the mesentery. A DC potential difference was then applied between 
the platinum electrodes. The potential was obtained 
by means of a,-oltage generaror in series with a mil­
liampere meter of high accuracy. Voltage differences between 100 mV and 10 V were 
tested. When morphologically visible changes were 
produced, the experiments could be checked or modi­
lied rc~tedly in the same animal with a fresh piece of 
mesentery. The vessels and other tissues of the mesentery were 
inspected at different magnifications with an operating 
microscope. Photographs were taken of the mesenteric 
electrode and surrounding tissues. The mesentery un­
der and around the electrode was excised at the end of the experiments and fixed in formalin on a glass slide 
before fuation. Staining was usually performed with 
haematoxylin and eosin before microscopy. B. Charging and discharging 
of tissue 
When 100 mV DC voltage was applied between one 
electrode resting against the mesentery and one elec­
trode in the aorta or vena cava, one to several mi­
croamperes could be observed to pass between the 
electrodes. The current increased continuous ly, ftrst 
slowly and then more rapidly. After a wbile the Oow of 
current decreased or became interrupted, usually from 
gas produced at the electrodes. In this way the tissue became electrically charged, 
as shown in Chapter VI (Fig. VI: 13). When the exter­
nal power source was then disconnected and the two 
terminals short-circuite d over a microampere meter, 
such a charged "biologic battery" was found to dis­
charge. Similar discharge from a carcinoma and granu­
loma, spontaneous ly polarized, is shown in the pa­
tients in Figs. VI: II and 12. C. Diapedetic bleedings 
Diapedetic extravasation of red blood cells is one of 
the early visible changes around the electropositive 
electrode. This extravasation can be seen in Fig. XIV: I as many sites of bleeding along small arterial 
branches after the application of DC current in small 
quantity (1. 0 pA, 30 min, 1. 0 V). This voltage repre­
sents the potential difference actually measured be­
tween the electrodes. Focal arterial contractions a. nd 
associated intraarterial thromboses also interrupt the 
blood stream in many places. Sometimes these blood 
clots were seen washing away, resulting in temporary 
restoration of circulation through the artery. Diapede­
tic bleedings were also observed, but to a much lesser 
extent, around the electronega tive mesenteric elec­
trode. The normal transcapillary openings between blood 
and extravascular spaces, which contain water and 
electrolytes, may be considered as leaking junctions. When an electric potential is applied across capillaries 
some of them appear to open further, as shown by the 
transcapillary passage of red blood cells. After a while the diapedetic blood as well as the 
thrombosed blood in the vessels becomes brown. White foam (Fig. XIV: 2) produced by gas then devel­
ops around the electropositive electrode against the 
mesentery. The surrounding brown dots represent 
diapedetic blood. Fig. XIV: I. Effecrs of elec~ric curren< in small quan<iry (1. 0 
pA, 30 min, 1. 0 V), passed berween aorta (carhode) and me­
scnrery (anode) of an anaesrherized dog. (a) In vivo phoromi· 
crograph shows diapedetic bleedings as irregular dots around 
rhe plarinum anode. (b) Local diapedetic bleeding. Narrow 
empty arteriocapillaries, wide venocapillaries with granulo· 
cytes. Haematoxylin -eosin stain.
The application of direct current is not comparable to real-life biological situations, as it is a brief application, whereas in biological situations it would be a prolonged process.
What is the main difference between the application of direct current in the study and real-life biological situations?
The quantity of charge transported between the electrodes can be determined as the product of current and time, expressed in coulombs.
What happens when a direct current is applied between two platinum electrodes in a living tissue?
The electric potential difference can lead to differences in the quality of tissue material, as different materials possess different specific energy levels of ionization and ionic adsorption characteristics.
What is the effect of an electric potential difference on the quality of tissue material?
The mechanism of electrophoresis is the migration of ions and the recombination of protons and hydroxyl ions, which can lead to the formation of an "A" zone and ischaemia.
What is the mechanism of electrophoresis, and how does it affect the tissue?
The main focus of the studies is on the role of electric field-induced vascular effects in the development of "A" zones and the potential for tumour regression.
What is the main focus of the studies in this chapter?