JPRS ID: 10227 EAST EUROPE REPORT ECONOMIC AND INDUSTRIAL AFFAIRS

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1~OR OHNI('IAI. I1SE ONI,Y
- ~ JPRS L/ 10227
31 December 1981
U~SSR Re~ ort
p
llFf SCIENCES
EFFECTS OF NONIONIZING ELECTROMAGNETIC RADIATION
CFOUO 2/81~
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JPRS L/10227
31 December 1981
USSR RFPORT
LIFE $CIENCES ~
EFFECTS OF NONIONIZING ELECTROMAGNETIC RADIATIC~N
(FOUO 2/81)
CONTENTS
Study of SHF Microwave Effect on Sex and Somatic Cells of Mammals..~..'.... 1
Immunomorphological Changes in the Testes Unde.r I~?fluence of
Superhigh-Frequency Electromagnetic Field 9
Effect of Industrial Current Electromagnetic Fie1d Upon Nature of
Growth and Mttotic Activity of Human Fibrob~.ast Cell Cultures 17
Acute Ex~erimental Emotional Stress in Rabbite (Phyaiologic-
Cytochemical Aspects) 23
Ultrastructur~ of Chick Embryo Skeletal Musc1~ Tisaue With Microwave
Damage 34
Effect of 50 Hz Frequency ElActromagnetic Fie1d on Ce11 Passage af
Certain Mir_otic Cycle Periods 40
Investigat:ions of Fluctuations ir~ Operator Sensory Sensitivity 45
Problems of Space B~ology, Vol k0: Biolo~ical Effects of Electromagnetic
Radiation in Microwave Range 50
- a- [III - USSR - 21m S&T FOUO]
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k'
UDC ~75.24:849:599.323.4
, STUDY OF SHF MICROWAVE EFFECT ON SEX AND SOMATIC CEL"LS OF MAMMALS
Kiev TSITOLOGIYA I GENETIKA in Russ~an Vol 14, No 6, Nov-Dec 80
(manuscript received 7 Feb 79) pp 3-8
- /Article by L. K. Ramayya, M. D. Pomerantseva, G. A. Vilkina and V. S. Tikhonchuk,
Institute of General Genetics ~f the USSR Academy of Sciences, Moscow/
/Text/ Introduction. Many studies showing the harmful ei~e~t of SHF el.ectroma.g-
netic waves on the r~productive system and its function in m~mmals have appeared
recently /1-3/. As yet, however, the problem of the genetic effect of SHF range
microwaves remains little studied. The data on th.~ mutagenicity of a SHF electro-
magnetic field available in the literature are contradictory. There are data on a
weak muta;;enic effect of SHF microwaves diiring a fracLionated irradiation of bone
marrow cells /4, 5/ and sex cells of male mice /6, 7/. At the same time, other
studies did not reveal changes in the properties of nucleic acids of th.e sperma-
togenic epithelium l8/ or a mutagenic effect under the effect of SHF micro�~avps on
yeast cells /9/. 1"ne results of studies of the combined effect of SHF mi~~rowaves,
chemical mutagens and radiation are difficult to co~npare and often contrudictory
/io, ii/.
Material and methods. The genetic effecti of a single and fractioned action of SHF
range microwaves with energy flux density of 60 and 800 mW/cm2 and a wa~~e len.gth
of 12.6 cm (oscillation frequency 2,400 MHz) was studied. This range encompasses
the upper and lower limits of the values of energy of a SHF electromagnetic field
_ most frequently used in practice. Hybrid sexua~ly mature male mice F1 (CBAXC57BL)
were subjected to the action. Nonline white females were use~ in the variant with
a single action and both white nonline and hybrid females aged 2.5 to 4 months, in
the variant with a ~ractionated action. Every case had its own biological i~~lenti-
cal control, which ruled out the effect of the female genotype on embryonal mor-
tality.
To detect the. mutagenic eff ect of SHF microwaves, three tests were used: frequen-
cy of dominant lethal mutations in sex cells of male mice; frequency of anomalous
spermium heads and frequenryo uf chromosome aberrations in bone marrow ce11s.
Males were irradiated on the Parus unit in an anechoic chamber under a single ac-
tion for 12 min with energy flux density of 60 mW/cm2 and for 21 s with energy
flux density of 800 mW/cm~. 'i7ie time of irradiation was chosen so that the level
" of lethal effect of mi.crowaves did not exceed 10%. In this investigation the mor-
- tality of animals depending on the series of experiments ranged f~om 0.1 to 10%.
1
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Uncler a fractionated action the conditions of irradiation were as follows: energy
flux density $00 mW/cm2 and time of irradiation Z1 s once a day for 10 days at an
ambient tempe~ature of 21-22 �C.
To detect dominant lethal mutations, intact females were exposed to irradiated
males for a week. Under a single effec~ the succession of six implantations
mad~ guaranteed participation in fertilization of gametes irradiated at the stage
of mature spermia (first week), late spermatids (second week), early spermatids
(th~ird week), late spermatocytes (fourth week), early sperma.~_~cytes (fifth raeek)
and spermatogon3a (sixth week). In case of a fractionated action irradiation
~asted 10 c?ays. lfierefore, spermia irradiated at the sta.ges af spermia and sper-
- matid~ (first week), spermatids and spermatocytes (second and third wee~-s), sper-
matocytes (fourth week) and spermatogonia (iifth week) took part in fertilization.
The number of implantations in this va-riant was five. Eighteen days after the be-
- ginning of crossing the pregnant females were anesthetized and cut open. The per-
cent of embryonal death was determin~d on the basis of the ratio of the number of
yellow bodies in the ovary, places of implantation and live embryos in the uterus.
The frequency of induced dominant lethal mutations was determined on the basis of
- a comparison of tha following indicators: survival rate of embryos (ratio of the
number of live embryos to tl-,e number of yellow '~~dies); death before implantation
(ratio of the difference between the number of yellow bodies and places of implan-
_ tation to the number of yel~.ow bodies); mortality of embryos after implantation
(ratio of the number of dead embryos to the z~umbe-r of places of implantation) /12~.
Furthermore, ~he percent of effective crossings and t.he weight o~ tes;:es 45 days
after the action were determinEd. These indicators m~de it possible to judge the
effect of SHI~' micro~vaves on the re.productive capacity of the males.
To count anomalous spermium heads, smears from the content of the epididymis were
prepared on the 35~h day after the action. The frequency of anomalous heads of
mature spermia developed_from irradiated ~permatogonia attests to the mutagenicity
of the action factor /13/. No less than 300 spermatozoids from each ~f the six
males were investigated.
Chromosome disturbances in bone marrow cells were analyzed in metapha~es on per-
manent preparations /14/ on compl.etion of a 10-day fractionated irradiation.
- It should be noted that standard methods of statistical analysis of the data on
dominant lethal mutations have not been developed conclusively to.this day /15/.
The msthod XZ for an evaluation af the differences in postimplantation mortality
between the experiment and control was used in th-?s work. Another method of sta-
tistical processing was used in some cases /16/. According to t.his method the
evaluation of errors was calculated on the basis of ehe following formula:
~y! n I E~y1- y)~ (x~ - x)s _ 2 E(x~ - x) E(yi - y) 1
Exi d/ n- t ~E'_`yt~~ -F- (Exi)' ~EX~) IEJ~) j
where n is the number of investigated females; x,~ is the number of yellow bodies;
yti is the number of embryos in some females; x, y are the corresponding average
values calculated for all females.
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The evaluation of the significance of differences was made according to Student`s
~-criterion.
Results of investigatior~s and their discussion. Table 1 presents data on embryonal
death caused by genetic disturbances arising in sex cells of males as a result of a
single action of SHF microwaves. As can be seen fram table l, the average number
of yellow bodies, places of implantation and live embryos per female did not dif-
fer significantly between the experiment and control. Nor did we f ind statisti-
cally signif icant differences between the Qxperiment and control in postimplanta-
~ tion mortality, which, as is wzll known, is the basic indicator of the frequency .
of dominant lethal mutations. We would like to note that nongenetic. factors, such
as the level of fertility of sperm~.a, the number of ovulated ova and so forth, can
also have an effect on the amounts of pre{mplantation losses and, accordingly, the
survival rate of embryos /12/. A certa~.n. incrPase in postim~lantation mortality
- observed under the effect on late sperm~tids (fourth week) proved to be statistic-
cally in~ign:Lficant (X'=2.4; P>0.05). Thus, induced dominant lethal mutations were
not detected at any stage of spermatogenesis.
_ The percent of effective crossings in the experimental groups of mice was even
- slightly higher than the control level. Forty-f ive days af cer the action the
weight of testes did not differ from the control level. In cantrol and experimen-
tal groups it avEraged 207, 210 and 213 mg respectively.
~ Table 2 presents the results of analysis of embryonal mor~ality in the offspring
of males subjected to a fractionated irradiation. As f ollows from the data, in
this experimental variant, as well as under a single action, there were no differ-
_ ences between the experiment and control in the average number of yellow bodies,
. places of implanta.tion and live embryos per female. Under the effect on gametes
realized during the first 2 weeks after processing a small increase in postimplan-
tation mortality in the experiment wa~ observed. In one case the differences bet-
ween the experiment and control proved to be staristically si~nif icant (X2=5.2; P
0.5). If the data on the combined eff ect of SHF microwaves on all the
stages of spermatogenasis are presented, as fc~llows from table 2, the lack of a
mutagenic effect can be seen clearly.
_ Under a fractionater~ action of SHF microwaves di~turbances in the form of spermium
heads were studied. According to the data in the literature, the period of 35 days
is optimal for the use of this indicator during a study of the effect of ionizing
radiation on spermatogenesis, beca+ise the largest number of anomalous heads is ob-
~ ser-;ed dtiring this period. It is assumed that anomalous spermium heads are formed
as a resu].t of genetic disturbances of the type of point mutations in sex cells
subjected to some mutagenic action /13/. As an analysis of the preparaticros .
showedy the frequency of anomalous spermium heads in the experiment and control
was on the same le~el and averaged 1.4 and 1.6So resp~ctively.
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Table l. Embryonal Mortality in the Offspring of Males Subjected to a Single Ac-
tion of SHF Microwaves
.e..=--..~ .
tl I K ~ ~ Y o I , r ~ ~ ~L ~ ~ o~~` ~ ~o~ , CMCpTHOC7b .
~Y v ~ o
/ ~ z
\l~ 2~ n r' n a ~ FCl r. ~(11> ,
n- o $ K ~y ~ ~ ~ I ~7
K F 4 a p c
~ ~ m Y a s~ ~f m GC ~
S e~ 7 F~ a ~
r O K u = ~ * p ~ _ ~
n 1
U= CA o x ci C= 7c0* 7 ~ i O C q~ C la- ,
(22~ 1-~ I CBy-1'(1 ) 36 I&3,3 ~ 9,5 8,3 I 7,8 81,8 13,0 6,0
(;13~B1l-I1'� 30 97,2 8,9 8,2 7,3 82,0 8,0 l0,8
(1 ~Koarponb 36 83,3 ~ 9,7 8,8 7,8 80,5 9,2 11,3
- (15) 2�~ ~~CB4-I 36 91,7 9,8 8,9 8,2 83,7 9,5 7,5
CB~I-I1~13 36 97.2 I 9,8 9,2 8,5 86,8 5,9 ',8
~(~~KOHTpO]Ib 36 80,5 9,6 8,9 8,3 86,0 7,9 6,6
(16) 3-a CB4-I 35 94,3 10,1 8,6 7,8 77,8 14,4 9,1
(~CBy-II 36 86,1 9,7 8,7 7,8 80,1 10,9 10,0
( 4)Koxrponb 36 77,8 10,5 9,6 8,6 82,0 8,5 ]0,4 '
(17) 4-s CBy-I 36 86,1 10,6 IO,Q 9,l 85,4 6,4 8,7
( 3~By-I] 36 69,4 10,8 10,1 8,6 80,3 5,9 14,6
(1 ~ KOHTPOJIb 36 80,6 ]0,1 9,2 8,3 82,2 ~ 8,9 9,8
(18~ 5-fl ~ 3~CB4-I 36 69,4 ]0,5 9,8 9,3 88,9 6,1 5,3
CBU-II 36 83,3 10,5 9,7 8,7 83,4 7,6 9,7
(14) KoxTponb 36 50,0 9,9 9,6 8,6 86,6 3,4 16,4
(19) 6-~ ~~B~3-1 36 72,2 9,9 9,2 8,5 85,6 6,6 �8,3
B~!-II 3.3 66,7 9,9 9,3 8,4 84,9 6.4 9,3
~(1 ~COHTpOIIb 29 58.6 9,8 ~J,1 8,4 85,6 7.2 7,7
~20) B c e r o CB~1-I 215 82,8 10,1 9,1 8,4 83,6 9,5 7,6
~~B~I-lI 213 83,6 9,8 9,1 I 8,2 83,d 7,5 10,3
~ (1 ~(oNrponb 209 72,2 10,0 9,2 8,3 83,4 7,9 9,5
- *Super high frequenr_y-I--energy flux density--60 mW/cm2, irradiation 12 min. **9u-
per high frequency-Il--energy flux density--800 mW/cm2, irradiation 21 s.
Key:
1. Mating periods, weeks 11. After implantation, percent
2. Effect variant 12. First
3. Number of implanted 13. Super high frequency
- 4. Percent of effective crossings 14. Control
5. Yellow bodies per 15. Second
6. Places of implantation per 16. Third
7. Live embryos per 17. Fourth
8. Survival rate, percent 18. Fif th
9. Mortality 19. Sixth
10. Before implantation, percent 20. Total
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Table 2. Embryonal Mortality in tha Off.spring af M~.les S~.bjected to a Fra:Ltionated
- Action of SH~' Microwaves
m
_ 1 ~ (2) ~ )T I ~e(7)I ~8~~ . c.we
~ac~(10)
( ) . 4 0 of 8' K ~ ~ I " ; ~ ,~11 ~ ~ (12)
e 5 e x Y w xs �e
z ~
u~ Q ~ 'x a9, ~X ~ ~Q~ ~O* ~j.
K s ~ * g u
~ a~ x ~ ~ ~ s ~ ~ _ :F = ~ a npoa. 1 g ~
(14)�1-s 15)C84 ~1~B/n 215 46,5 ~ 10,2 8,9 T,7 76,3 12,0 13,3
(1 ) Koxrponb 159 34,6 ~10,3 9,0 8,1 T8,9 12,1 10,3
_ ~1 ~ CB4 CH6pN~tw l32 77,3 10,7 9,6 R,9 83,2 10,6 6,9
~l ~ KOH1'~10d6 ~18J 117 80,3 10,~ 9,4 9,0 85,7 9,8 5,0
~I~~ 2-A 1~)CB~ (()S/a 215 56,7 l0,6 9,4 8,3 78,1 13,8 9~4
li~oxrpom~~ 159 69,8 9,9 8,9 8,1 81,9 10,8 8~2
S~CB~i IH6 x A~ 135 8J, 5 10,6 9,3 8.8 82, 5 12,2 6, 0
_ ~1 ) Koxrpo~ ~18 ) t17 82,0 L0,5 9,1 8,8 83,5 13,4 3~7
- (2p) 3.A (15) CB~I B/n lt4 ?1,9 10,8 9,5 8,8 8i,3 11,9 7,7
(1 ) Koxrponb ~16) ~9 73,7 10,5 9,8 8.9 84,7 7,0 9,0
~ ~21~ 4-s 15)CB~I B/n ll7 65,8 11,5 10,4 9,5 82,5 (0,0 8,4
(1 ~ KOH'1'pOdb 99 68,7 12,3 10,7 9,0 ~7~,9 13,4 15,8
- (22) 5-s 15~CB~{ B/n !06 79,2 11,9 10,1 9,3 ?8,2 14,8 8,1
(17 KoHrpona ~16~ 89 73,0 12,0 l0,1 8,9 74;5 i6,2 I1,2
- ~23~ Bcero CB4 (1 ) 1034 65,5 10,8 9,5 8,7 I 80,2 12,3 8,5
(].7 Koxrpom~ 839 67,6 10,8 9,5 8,7 8U,? 11,8 8,5
Key:
1. Mating period, weeks 13. Percent
2. Fxperimental variant 14. First
- 3. Genotype 15. Super high frequency
4. Number of implanted 16. Sterile
S. Percent of effecti~e crossings 17. Control
6. Yellow bodi.es per 18. Hybrids
- 7. Places of imglantation per 19. Second
= 8. Live embryos per 20. Third
9. Percent of survival rate 21. Fourth
10. Mortality 22. Fif th
11. Before implantation 23. Total
~ 12. After implan.tation
- The resul ts of study of the effect of SHF microwaves on bone marrow cells are pre-
sented in table 3. As follows from the data, the percent of aberrant metaphases in
_ the exper iment and control comprised the same values.
Thus, not a single test used by us disclosed a mutagenic effect of SHF microwaves
under the g;ven experimental conditions. A comparison of our results with the
data on this problem in the literature often is diff icult, because the conditions
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~ of irradiation ar.: not cc?mparable. Thue, with a local irradiation of mouse testes
a weak increase in tre frequency of induced dominant lethal mutations was obtained
/6, 7/. Howevsr, no pattern in a rise in the level of mutations depending on the
stage of spermatogenesis subjected to the effect was observed. If another method
of statistical processing is used, that is, the number of yellow bodies, places of
implantation and live embryos is presented in the form of an average nuu~ber per
female, the d~.fferences between the experiment and control in these studies wi].1
prove to be insignif icant. Another author /S/ obtained a mutagenic effect of SHF
- ~nicrowaves under a fractionated action on bone ma.rrow cells. However, the condi-
tions of irradiation in our experiment differ from those used in the above-mentirned
~tudy, which makes a comparison of results difficult. An analysis of the data from
- source5 in the literature available to us makes it possible to assume that, appar-
ently, th~ degree of a mutagenic effect or its absence depends prima.r~ly on the can-
ditions of the action. In the opinion of some authors, to this d~ there are no
- direct proofs of a specific nonthermal effect of SHF micrawaves /17/, because there
are no data on the existence of a molecular interaction of the biosystem.with the
absorbed energy of a SHF electromagnetic f ield. Proceeding from the above-stated,
it is possible to explain the lack of a mutagenic effect in our experiments.
Table 3. Frequency of Cl-~romoso:ne Aberrations in Bone Marroo~ Cslls of Mice as a
Result of a Fract~onated Irradiation With SHF Range Microwaves ~
Action~ Number of Examined Metaphases Percent of Aberrant Metaphases
- Total Aberrant
SHF 740 12 1.62
Control 372 5 1.37
When ~his axticle had already been prepared for the press, a study canfirming the
data obtained by us on the lack of a mutagenic ~ffect of SHF microwaves in sex
cells of mammals appeared. The authors, varying the ti~ne nf a chronic irradiation of
male rats with SHF microwaves with energy flux density of S and 10 mW/cmz, did nat
detect induced dominant lethal mutations or a decrea.se in the fertility of ani-
mals /18/.
Conclusions. As a result of the study of a single action of SHF microwaves with
energy flux density of 60 and 800 mW/cm2 and of a fractionated aetion of SHF mic-
rowaves with energy flux density of ~00 mW/cm2 on ssx and somatic cells of male
mice, a mutagenic effect of this factor was not revealed by any of the three used
- tests (frequency of induced dominant lethal mutations; frequency of anomalous sper-
mium heads and frequency of chromosome aberrations in bone marrow cells).
The authors are grateful to V. S. Lysenkova and N. V. Chernysheva for their help
in this study.
BIBLIOGRAPHY
1. "Vliyaniye SVCh-Izlucheniy r.a Organizm Cheloveka i Zhivotnykh" /Effect of SHF
Radiation or.i the Human and Animal Organism7, edited by I. R. Petrov, Leningrad,
Meditsina, 1970, 214 pages.
6
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2. Gorode~skaya, S. F., Kerova, N. I, and Rappoport, M. B., "Effect of SHF Radis-
tion on the Function, Morphology and Nucleic Metaboli;m of Sex Organs of Ani-
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nykh Voln Radiochastot" /Labor Hygiene and Biological Effect of Electromagnetic
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Moscow, June 1968, Moscow, 1968, p 41.
3. Bereznitskaya, A. N. , Kazbekov, I. M. and Rysir~a, T. Z. ,"Effe~.t of M3crowaves
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spring," in the book "Gigiyena Truda i Biologicheskoye Deystviye Elektromagnit-
nykh Voln Radiochastot," Abstracts of Reports of the Fourth A~.1 Union Symposium,
Moscow, June 1972, Moscow, 1972, p 35.
4. Sokolov, V. V. anc~ Chulina, N. A., "F'roliferation and Chro~osome Disturbances
in ~3one Marrow Cells of Individuals Working Under Conditions of Action of SHF
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gicheskoye Deystviye Elpktromagnitnykh Voln Radiochastot," Abstracts of Reports
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5. Ka.pustin, A. A., Rudnev, M. I., Leonskaya, G. I. and Konobeyeva, G. I., "(?n the
_ Cytagenetic Effect of the Action of a SIiF Range Alternating Electromagnetic
Field," TSITOLOGIYA I GENETIKA, 1976, 10, No 5, pp 400-402.
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ical Effec ts of Electromagnetic Waves," selected papers of the USNC/URSI Annu-
al Meeting, Maryland, Public Health Service, 1975, 1, pp 38b-396.
7. Varma, M. M., Dage, E. L. and Joshi, S. R., "Mutagenicity Induced by Nonioniz-
ing Radiation on Swiss Male Mice," in "Biologi.cal Effects of Electromagnetic
Waves," selected papers of the USNC/URSI Anniial Meeting, Maryland, Public
Health Service, 1975, pp 397-406.
Loshak, A. Ya. and Vedernikova, N. N., "Effect of Chronic SI~' Irradiation of
Nonthermal Intensity on the Genetir. Activity of Cells," in the book "Gigiyena
Truda i Biologicheskoye Deystviye Elektromagnitnykh Voln Ra.diochastot," Ab-
- stracts of Reports of the Fourth A11-Union Symposium, Moscow, June 1972, p 35.
9. Madhavana.th, U., Subrahmanyam, P., Sankaranarayanan, N, and Singh, D. R., "Mu-
tagenic Ac tion of Nonionizing Radiations: Its Implication in Radiation Pro-
tection," RECUEIL ~ES COMMU[dICATIONS, 1977, 2, pp 333-336.
10. Danilenko, I. I. and Mirutenko, V. I., "Effect of an Alternating Magnetic and
SHF Electromagnetic Fields on Mutagenesis," in the book "Molekulyarnaya Gene-
tika i Biofizika" /Molecular Genetics and Biophysics/, Kiev, Vishcha Shkola,
1976, Issue 1, pp 117-123. �
11. Bikkutov, R. I, and Orobey, V. V., "Effect of a Combined Fractionated Effect
of SHF an d of a Long-Wave Roentgen Irradiation on Cytogenetic Ch2.nges 3n Bone
Marrow Cells of Mice," RADIOBIOLOGIYA (Information Bulletin), 1975, No 18, pp
28-29.
7
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1'L. 1'ouierantsev~, M. D. and 12amayya, L. K. ,"Mutagenic Effect of Irradiatiorxs of
Various Types on Sex Cells of Male Mice. Re;~ort I. ~omparative Genetic Ra-
diosen~itivity of Spermatogonia and Other Stages of Snermatogenesis," GENETIKA,
1969, 5, No S, pp 103-112.
13. Bruce, W. R., F~rrer, R. and Wyrobec, A. J., "Ahnorm3lities in the Shape of
P~urine Sperm After Acute Testicular X-irradiation," *NTAT. RES., 1974, 23,
N 4 , pp 381-386 .
14. Ford, F. and Woolan, D. H. ,"A Study of ~h~ Mit�otic Chromosomes of Mice of the
SGrong A Line," EXP. C~.LL RES., 1963, 32, pp 320-324.
15. Ehling, U. H., Machemer, L~, Buselmaier, W. et al., "Standard Protocol for
the Dominant Lethal Test on N1zle rlice Set up by the Work Group 'Dominant Le-
thal Mutations of the ad hoc Com~aittee Chemogenetics,"' ARCH. TOXIKOL., 1978,
39, pp 173-185.
16. Shapiro, N. I., Plotnikova, Yt. D., Strashnenko, S. I. and Suslikov, V. I.,
~ "Relative Gezietic Ra.dio Sensitivity of Varlous Types of Mammals," in the book
"Radiatsionnaya Genetika "/Radiation Genetics/, Moscow, Izd-vo AN SSSR, 1962,
pp 63-78.
17. Cleary, S. F., "Uncertainties in the Evaluation of the Biological Effects of ,
Microwave and Radio Frequency Radiation," HEALTH PHYS., 1973, 25~ N 4, pp 378-
404 .
18. Berman, E. and Carter, H., "Mutagenic and Repr~ductive Tests in Male Iiats Ex-
posed to 425 or 2450 MHz (CW) Microwaves," Abst. of Scient. Papers. Open Sym-
posium of the Biological Effects of Electromagnetic Waves. XIX General Assem-
bly of the Internationa~ Union of Radio Science with the Cooperation of th~
International Iladiation Protection Association, 1978, p 97.
COPYRIGHT.: Izdatel'stvo "Naukova dumka", "Tsitologiya i genetika", 198a
?1,439
= CSO: 1840/329
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IMMUNOMORPHOLOGICAL CHI~NGES IN THE TESTES UNDER r~UENCE OF SUPERRIGH--FREQUENCY
ELECTROMAGNETIC FIELD
Leningrad ARKHIV ANATOMII, %ISTOLOGIt I II~BRIOLOGII in Russian Vol 80, No 2, Feb 81
pp 69-75
[article by V. V. Grigor'yev, R. P. Ogurtsov and Yu. N. Zubzhitskiy, Department
of Histology and Embryology (headed by Prof A. A. Klisho~ ~ Military Medical Academy
J imeni S. M. Kirov; Department of Microbiology and Immunology (headed by Froi B. N.
Sofronov), Scientific Research Institute of Experimental Medicine, USSR Academy of
Medical Sciences; Scientific Resea-rch Laboratory of Electron Microscopy and Histo-
chemistry (headed by K. K. Zaytseva, doctor of inedical sciences), i~ilitary Medical
Academy imeni 5. M. Kirov, submitted 14 Feb 80]
[Text) The reactions of organs isolated from the effects of immunological factors
by histohematic barriers are of special interest to experimental morphology.. It is
� possible to investigate the role of antibodies and sensitized lymphoid cells in
development of autoi~umune processes on models of damage to the brain. peripheral
nervous system, testes, thyroid and eye (V. V. Serov, 1968; A. D. Ado, 1972; A. I.
Strukov, 1~72).
Heretofore, the s~.udy of the testes by iimnunomorphological methods had been conducted
- in experiments where traumatic factors (S. S. Raytsina, 1970), immunization with
antigens of homologous testes (J. Freund et al., 1955: B. H. Waksman, 1959; P.
Brown et al., 1963) and cross-reacting antigens of microorganisms (R. P. Ogurtsov
and Yu. Nv Zubzhitskiy, 1978) were the triggering mechanisms of the process.
A superhigh-~requency electrom~gnetic field (SHF IIKF) of nonthermal intensity elicits
aspermatogenesis and development of lymphoid infiltrates in testicular interstitial
Cissue, even with brief exposure (V. V. Grigor'yev, 1978). Since this effect cannot
be attr~~-�:r_ed to the thermal effect of the field on cells, there is reason to
believe that tlie damage is possibly related to lesion to thp hematotesticular
barrier (HTB) and is associated with development of an immunopathological process.
Our objective here waa to make a study of the possibility of development of an ~
immune reaction to r~sticular antigens and its r~lation to impairmer.t of structure
and germeability of the HTB under the influence of SHF EMF of nonthermal intensity,
- using immunomorphological, histological methods and electron microsccpy.
- Material and Methods
Experimer~ts were conducted on 77 puberal male rabbits weighing 2.5-3 kg. The
testes were submitted to local irradiation for 6 min delivered from a Luch-58
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radiotherapy machine (wavelength 12.6 cm, intensity 8.8 mW/cm2). The contro 1 group
consisted of 15 nonirradiated ra~bits. The animals were weighed 1, 3, 5, 7, 14, 21,
30, 60, 90 and 210 days after exposure, sacrificed by decapitation, after which the ~
testes, blaod and spleen were taken for examination. We determined the ratio of
red to white pulp mass of the spl.een by the me*_hod of Gammar, as well as weight of
the testes and the animal. Various parts of the same testis wE~re fixed according
to Carnoy and Colfield, the spleen according to Carnoy and imbedded in paraff in,
and the testes were also imbedded in epor_ Q12. Part of the testis was fro~.en in"a
mixture of ice and petroleum ether to obtain cryostat sections. The paraff in sec-
' tions were stained with hematoxylin-eosin, while the ultrafine sections were con-
trasted with uranyl acetate and lead nitrate. The preparatians were viewed under
a light microscope and type JEM 100 CX electron microscope. ~
For d~monstration of antibody gZobulirl fixed in the testes, the cryostat sec tions
were eluated for 10 min in phosphate buffer (pH 7.2) and, after drying, incubated
with FITTs [fluorescein isothiocyanate)-labeled antirabbit serum (produced by the ~
Scientific ResearcAl Institute cf Epidemiology and Microbiology imeni N. F. Gamaleya,
- USSR Academy of Medical Sciences). Antibodies tio antigens of rabbit testes were
- assayed in blood serum with the complement fixation test under refrigeration (V. I.
Ioffe and K. ri. Rozental', 1943) with homologous testicular extract in a dosage
of 1 mg/mQ protein and by the indirect immunofluorescence method (T. H. Weller and
A. H. Coons, 1954), using cryostat sections of homologous testes fixed in 96 �
ethanol as antigen.
Hypersensitivity of the delayed type was tested after 9, 14, 30 and 90 days, for
which purpose a group of 17 experimental and 6 control animals was given intra-
. cutaneous injections of homologous testicul~r extract (in doses of 1-0.01 mg/mR
per rabbit). The reaction was evaluated 24 h after the injection.
In order ro examine permeability of the HTB 1 and 14 days after irradiation, we
gavz intravenous injections of 3 mQ ox serum albumin in a concentration of ~
100 mg/mQ/kg an.imal weight, conjugated with rhodamire chlorideor Evans b~ue dye.
Af ter 2 h, the testes were removed, cryostat sections prepared and examined under
an ML-4 fluorescence microscope with UFS-3 and SZS-14 light filters, and ZhS-3
ocular filter.
Results and Discussion
The experimental animals were divided into three groups, according to tl~e individual
reactions to the deleterious effects of SHF EMF.
The first graup (25 rabbits) developed mild reactive changes in the testes, mani-
- fested by death of only a few cells of the spermatogenic epithelium in both cen-
trally situated tubules and near the testicular membrane after 1-3 days. Af ter
S-6 days, spermatogenesis was virtually normal.
- Already 1 day after expasure, the second group (.22 rabbi;ts? presented microscopiG
hemorrhages, which were the most extensive in the region of the testicular r ete.
Lymphoid infiltrates were formed in areas with hemorrhages after 2-5 days. The
damage to the testis was notably nonuniform: complete atrophy of spermatogenic epi-
thelium in the region of hemorrhages and adjacent zone, confined to the lobules,
whereas in tubules of adjacent lobules removed from the lesion site the process of
- spermatogenesis c~ntinued.
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