The permeability of capillary vessels was investigated in order to determine if acceleration alone or following prolonged hypokinesia would induce changes in the vascular wall leading to the penetration by l-albumins and/or proteins with larger molecules. In rats undergoing action of +5 Gz accelerations, no increase in vascular permeability, as tested with the use of (Cr-5k)-globulin, was demostrated. In rats immobilized for 4 weeks before centrifugation, rather weak migration of (Cr-51)-globulin from the vessels was observed. Immobilization resulted also in lowering of conduction velocity in the sciatic nerve

Baranski, S.

Edelwejn, Z.

Wojtkowiak, M.

English

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NASA TD micAL ME mDRANDum 	 NASA 04-76195
DISPLACEMENT OF PLASMA PRIM IN AND C^ICTION VELOCITY IN RATS
UNDER ACTION OF ACCELERATION FORCES AND HYPOKINESIA
S. Baranski, Z. Bdelwejn and M. Wojtkowfak
(NASA-TH-76195) DISPLACEMENT OF PLASMA
	 h80-279n9
"PROTEIN AND CONDUCTION VELOCITY IN HATS
UNDER ACTION OF ACCELERAWON FORCES AND
HYPOKINESIA (Rational Aeronautics and Space
	 Unclas
Adainistratien) 10 p HC A02/MF A01 CsCL 06c u3/5 1 248028
t
Translation of "Badania nad przenieszczeniem sie bialek
MOM kiwi oraz szybkoscia przo4odnictwa nerAvego
u szczurow poddanych dzialaniu przyspeszen i hipokinezji,"
PostepY Astronautyki, Vol. 5, No. 2, 1972, pp 61-70.
NATIONAL ALIMAUTICS AND SPACE ADMINISTRATION
WASHINMON j D. C. 20546
	 JUNE 1980
tq , r9a	
. ar ' `
1t
1
STANDARD TITLE PAGE
P rr 
N16195 
	 2	 Govornm.nt Accession No,
D".
 '^'M-
3.	 Reciplent r s Carole# No.
4, Title and Subti t le 
DISPLAiLMMM OF PIASM^ PROTEIN
5,	 Report
June a1980
CONDUCTION VE	 CITY IN RATS UNDER AICrION OF
ACCELERATION
 
MRCES AND HYPOKIMIA
b,	 Performing Orounisotion Colo
7.	 Authors) s.	 Performing Organisation Report No.
S. Baranski, Z. Edelwejn and M. Wojtkowiak 10,	 work unit No,
11,
	
Contract or Grant No.
9, Performing Orlon se ► ion Pismo and Address
Leo Xannbr -Ars.socia tes
Redwood City, California 90463
13,	 Typo of Report and Period Covered
Translation
12. Sponsoring Agency Name end Address
National Aeronautics and space Administration
Washington, D. C. 20546
14,	 Sponsoring Agency Cade
15. Supplementary Notes
Translation of "Batiiani,a nad przemieszczenien sie bialek osocza krwi oraz
szybkoscia przewo&ictwa nerwvwego u szczurow poddanych dzialaniu przy-
speszen i hipokinezji," Postepy Astronautyki, Vol. 5, No. 2, 1972, pp
61-70. C A 7 ^ .^ 1 ^ ^ ^ ^^z
16.	 Abstract
In animals undergoing action of +5Gz aeceleratiffs, no increase in vas-
cular peremeability, as tested with the use of
	 Cr-globulin, was deron-
strated.	 In rats Fnobilized for 4 weeks before centrifugation rather
weak migration of	 Cr-globulin from the vessels was observed-.
	 Ilmabi-
lization resulted also in lowering of conduction velocity in the sciatic
nerve.
17. Key Words (Selected by Author(s)) Is.	 Distribution Statement
Unclassified-Unlimited
19.	 Security Closcii, (of this report) 20.	 Security Ciassif. (of this Pope) 21• No. of Pages 22,	 Price
Unclassified Unclassified 10
r
vA5A.HG
L
DISPLACEMW OF PIAM PH EIN ATw QONDUCHON VELOCITY IN RATS
UNDER ACTION OF ACCELEWION FORCES AND HYPOKINFSIA
S. Baranski., Z. Ddelwejn and M. Wojtkowiak
Military Academy of Aviation Medicine, Warsaw
Introduction	 61*
In previous studies V., it was established that an acceleration of +5Gz opera-
ting during a period of frcr^ 15 min to 3 hrs induces changes based on the displace-
ment of 131I-albumins in accordance with the direction of the accelerating action.
It was also shown that the observed changes were dependent on the migration of pro-
teins from the blood vessels. In later studies [2, 31, attention was directed to
hamodynamic [sic) disturbances occurring in animals affected by acceleration follow-
ing a prolonged period of hypokinessia,.
The survival rate of animals subjected to acceleration after prolonged hyp-)kine-
sia declined and such animals could not stand the stress of centrifugation for longer
than a perie3 of 37 min. Moreover, it was found that there was a siT.Aficantly more
marked migration of I-albumin through the blood vessels under the influence of longi-
tudinal acceleration. This change was accompanied by disturbances of a functional na-
ture within the peripheral neuron together with the motor cell of the anterior horn 62
of the spinal chord. Changes of this kind may be dangerous for the organism under
conditions of high altitude and space flights. A useful result of these points of
view has been resaarch conducted in the area of the permeability of the capillary
vessels. Authors carried out tests to find out, whether it was acceleration alone or
acceleration following prolonged hypokinesia which induced the occurrence of changes
leading to penetration of the vascular walls by I-albumins exclusively or also pro-
teins with larger molecules, i. e. globulin.
Method
The experiment was conducted on 70 white Wistar rats weighing about 200-250 g.
* Numbers in the margin indicate pagination in the foreign text.
1
The animals were divided into 2 groups: experiment and control. The experimental
group comprised 40 animals kept during 4 weeks under conditions of complete immobi-
lity by being placed in special immobility cages that prevented any movement (Fig. 1).
^ ^
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^}t	 'fI	 ..	 +•,n^'.'mq[/^';•
•'eR'h ^^. •	 '^	 •r	 r 	 7	 b>'s.r.^
Fig. 1. Rat in cage restricting mobility.
Feeding was done by means of specially adapted containers. Following 4 weeks /63
of immbilization the animals of this group were subjected to an acceleration of
+5Gz during periods of 15 min, 1 hr, 2 hrs and 3 hrs. Each of these subgroups num-
bered 10 animals. The control group was represented by 30 animals. This control
group was subjected to acceleration only for 15 min, 1 hr and 2 hrs. During the en-
tire experiment the animals in this group were maintained under normal vivarium condi-
tions.
A special centrifuge (Fig. 2) was used for carrying out the experiment. It was
adapted for experimental tests on animals, making it possible to centrifuge 4 rats si-
multaneously.
The animals were placed in special receptacles in such a way that the vector of
centrifuge force traversed the long body axis in a craniocaudal direction. Before
centrifuging was begun the animals of both groups, i. e, experimental and control,
received, intravenously 51Cr,-globulin in the amount of 100 units per gram body weight.
2
At the and of the designated time rotation was stopped, the animals subjected to
	 64
ether anesthesia, and dehematized., whereupon segements of the muscles of the fore-
limbs and hindlimbs were taken to measure radioactivity using the method presented
in previous works 11, 2, 31. Radioactivity of the selected samples was determined in
amounts of imp/Smin/mg of dry tissue. A percentage calculation was made of the differ-
ence in radioactivity between the muscles of the forelimbs and the hindlimbs, taking
the former as 100%. In several cases, both for the control and experimental groups,
a scintigram was taken.
For measuring conduction velocity we used the method proposed by Hodes at al.
(4). At two points along the course of the anatomic sciatic nerve we placed 2 sti-
mulating needle electrodes: adducent and educent. They were placed 1 cm. apart and
connected with a DISH stimulator (Multistim) with a transformer as an insulation u-
nit. The stimulating electrode used was a concentrating needle electrode of the
type DISH 13K 04, length 42 mm and cross section 0.65 mi. The muscular potential
produced was recorded from the gastrocnemius muscle using the educent concentrating
electrode DISH 13K 51. Supramaximal stimuli were used, t2iat exceeded maximal stimu-
li by 20 volts. Stimuli frequency was one cycle per second. Recording was done on
a three channel eleetromyogram DISH with an input impedance of 200 megaohms.
Conduction velocity measurement was done on a group of 40 animals subjected to
examination just before ,immobilization and at the end of the 4 week imrobilization
cycle. No assessment was made of the effect of centrifugation on conduction velocity.
Test Results and Evaluation
Table I shows the difference in radioactivity of the muscles of the forelimbs
and hindlimbs for both control and centrifuged animals following prolonged imrobili-
zation.
From the data presented above we see that in the case of the control animals, /65
,i. e. those kept in normal vivarium conditions, and then subjected to acceleration
for 15 min, l hr and 2 hrs, there was no occurrence of extravascular proteins la-
beled with chromium and consequently 51Cr-globulin. For this reason experiments
with this group were abandoned following 3 hrs of acceleration. on the other hand
in the experimental group the presence of Cr-globulins was not noted extravascularly
3
TABLE I. RADIOACTMTY CHANCES EN 51Cr-QABULIN IN MUSCILES OF CONTMI, RATS
AND GKW CObQ'LETELY U"WILIZED FOR 28 DAYS, FOL.LM -1r, HYPOKINESLA
Acceleration	 Con-	 Animals after
	 Diff-
time	 trol
	 immobilization	 erence
96,9
	 • U,9
111,4
11),4
TABLE II. EFFECT OF 8 WEEK IMMOBILIZATION OF RATS ST~JN
CONDUCTION VELOCITY IN THE SCIATIC NERVE
Mean standard	 Significance of
deviation, X + S
	 difference
Before immobilization	 „1Z Y	 v C'W1
After 8 weeks of in--
mobil i zation
ll?I: iIRICJtt l.^Ur\Lii,Yr.
.^	 `; •::'^	 ;^	 ..^	 ^'. `•;f:Jw..+'	 ass^,	 w 	 -pry	 ^	 ..	 ...^...	 v,• ..I: •... •^+•	 wr
 'tom ^ •^1 r\''''' ' i ^-^`^ R --•	 - __^.A' tF	 ^^:^ ^'w	 0 ^^^,,,.^.r•^r1
yak,\	 j ^•^•1 ..	 , _^a '3 . 3 i i^	 .	 !
Fig. 2. Centrifuge for studying the effect of
acceleration on small laboratory animals.
4
h-
following 15 min or 1 hr of acceleration at
+5 Gz. However, the data obtained after 2 and
3 hours of centrifugation support the hypothe-
sis, that 51Cr-globulins find their way out of
the vascular bed only to an insignificant de-
gree. The graph in Fig. 3 illustrates the dy-
namics of such changes.
This graph likewise shows the penetration
of th:: wall by 1312-labeled protein and there-
fore I-albumin [2]. In the course of these
curves there is a considerable difference in
17 5P
F	
' J1
x,	
r
k^Yi
	
rrr
1
r
is
40+J
Nr
N:m'yeW^'^.^	 y	 u.. : ,, .»«.a^:	 y^	 va^^ca-as ^•..m7a+s^
45mLq CT r.
	 420rnn	 %Orrm
------- albumins
--- 
globulins
permeability of the vascular wall for proteins
Fi.g. S^. Migration of, 131I-albumin
and Cr-globulin from the ves- of different molecular size. Previous studies
sels of animals centrifuged after, 	 [11 2, 31 made it possible to assert that these
4 weeks of immobilization.	
changes regress in a rather, significant degree
in line with the effect of experiment time. The tests above permit us to estimate
that 1 month of hypokinesia has only an insignificant effect on hemodynamic changes
in the organism during the time of acceleration.
Restricted mobility increases permeability of the vessels under the influence /66
of acceleration not only for microwleeular proteins but also for macrom>lecular
proteins, such as the globulins, which in a very insignificant degree may penetrate
beyond the vessels. The scintigrams produced no new data. They did not show any
place where the red coloring was evidence of much penetration of 51Cr-globulin be-
yond the vascular becl.
In testing the penetration of albumins by the use of 131I-albumin it was shown
that in ratio to the time increase in acceleration there is increased escape of such
proteins from the vascular bed amounting to 64% after 3 hrs. These results, in our
opinion, are evidence of significant heradynamic disturbances in peripheral circu-
lation. However, determination of the migration of 51Cr-globulin through the vas-
cular walls indicates only an insignificant amount of permeability for these pro-
teins. Even after 2-3 hrs of centrifugation the differences ranged from 5 to 10% /67
and were therefore significantly lower than for 	 I-albumin. The works of Czerski
et al. [5, 6], using the Keti tests, likewise showed increased permeability of the
5
vessels for 22Na even when acceleration was in the range of +3 G and continued for
several hours. These authors, using multiple acceleration over a long period of time,
even demonstrated the presence of histopathological changes in the walls of blood
vessels. These changes were characterized by distention of the cells that gave the
impression of transudate changes. Similar changes were observed by Senelar et al.
[71 in experiments on dogs subjected to the protracted effect of acceleration of +2.5
Gz. From the above data we may judge, that the time pea:iod of hypokinesia used in
our experiments did not produce irreversible changes in the vessels themselves as
well as in their muscular bed. On the other hand, the dissimilarity between the
present results and those of previous studies [1, 2, 31 as well as the studies of the
authors cited j5, 63 is in all likelihood based on t°.l;e size of the molecules pen(txa-
ting the vascular walls. In our experiments we tested this kind of penetrability
for large molecules and single acceleration. It is possible that multiple accelera-
tion, even under the conditions of our experiments, might produce the saris, changes
noted by other. authors. In the present cxperiment we did not use the method. of mul-
tiple acceleration because we wished_to have our ex-.rimental conditions approximate
reality as closely as possible. An astronaut who finds himself in conditions of limi-
ted mobility and reduced gravitation is subjected to the effect of acceleration once
and for a relatively short time. The same is true on plane flights where the pilot is
not exposed for a long time to acceleration.
An analysis of conduction velocity measurements in the sciatic nerve of rats sub-
jected to compelled immobility showed a statistically reliable reduction in velocity
in respect to the above values. Numerical data are presented in Table Ii.
It appears that the observed reduction in the velocity of impulse conduction was
occasioned by changed conditions in the blood supply to the nerve fibers and the also-
dated disturbances of a metabolic nature. Skorpil [8) called attention to the fact/68
that reduced motor activity may induce changes in conduction velocity. However, the
very strong views of this author in regard to this kind of effect are based upon high-
ly myelinized fibers and therefore on greater conduction velocity.
Final Conclusions
As a result of the experiments conducted it was shown:
6
1. Subjecting the animals to acceleration in the amount of +5 Gz for 15 min to
2 hrs did not induce penetration of the vascular wall by vac ►-cmolecular proteins such
as globulins.
2. Keeping animals for a long time under conditions of immobility does little
to favor penetrability of the vessels by macremolecular proteins. Permeability for
5'er-globulin is considerably less than for 1311-albtxiin.
3. Imrobilization of the animals nor 4 hours induces a statistically signifi-
cant reduction in the conduction velocity of the sciatic nerve.
7
r.
9
1. Baranski, S and M. Wojtkowiak, Postepy Astronautyki 4 (1970).
2. idem, Z. Edelwejn and M. Wojtkowiak, Space Life Sciences 2, 400 (1970).
3. idem and M. Wojtkowiak, Resume SynWzju po Kosmiczeskoj Biologii w Miedicinie
(Abstract of *A Symposium on Space Biology in Medicine), B. A. N. 9, 22 (1969)
4. Hodes, R., M. G. Larrabee and W. German, Arch. Neurol. Psychiatr. Chic. 60, 340
(1948) .	
._...
5. Czerski, P. M. Wojtkowiak and H. Zaremba, Patologia Polska L9/3, 395 (1968).
6. idem, idem, Postepy Astronautyki 4, 113 (1969).
7. Senelar, R., R. Loubiere and F. V olette, BioassV Techniques for Human Centri-
uges and Phsyiological Effects of Acceleration  Bergext, Oxford	 r
B. Skorpil, V., Rozpr. Cesk. Ak. Ved. 75, 1 (1965).
8
G


Displacement of plasma protein and conduction velocity in rats under action of acceleration forces and hypokinesia

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