INDUCTION OF COLICINE SYNTHESIS BY MEANS OF MILIMETER RADIATION

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Article from "Elektronnaya Tekhnika," I, Electronika SVCh,
(Electronic Engineering, Series I, Microwave Electronics)
[Volume no., issue no., and year are not given on this article.]
pages 52-54.
INDUCTION OF COLICINE SYNTHESIS BY MEANS OF MILLIMETER RADIATION
by
R.
L.
Vilenskaya,
A.
Z.
Smolyanskaya,
V.
G.
Adamenko,
Z.
P.
Bulvasheva,
E.
A.
Gel'vich,
M.
B.
Golant,
d
an
D. Ya.
Gol'vgaver
UDC 576.881.48.097.29.095.14
For an institute of experimental and clinical oncology, Academy of Medical Sci-
ences of DUSSOR and Scientific Research Institute of the Ministry
of the Electronic Industry. (Submitted for publication March 17, 1971. Pre-
sented by a of the Academy of Medical Sciences of DUSSOR, M.M.
Vlokhain.)
Abstract
The irradiation of the colicinogenic strain E. coli C. 600 (El) with
electromagnetic waves in the millimeter range having a nonthermal intensity is
capable of causing the induction of colicine synthesis. The dependence of the
induction effect on wavelength, irradiation time and temperature of the test
object is noted. (Bulletin of Experimental Biology, No. 4, p. 52, 1972).
A series of authors [1,3,5] have demonstrated that irradiation with
millimeter waves having a nonthermal intensity may lead to perishing of a bac-
terial cell.
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The present work studies the action of waves in the millimeter range
on the phenotypical expression of the genetic elements of bacteria which cause
lethal synthesis.
Experimental Procedure
A colicogenic factor--The extra chromasomic genetic element of bacteria
which causes lethal synthesis of the protein substrate colicine which exerts an
antibacterial action relevant to bacteria of the same or related forms was
chosen as the object of the investigation.
The investigations were carried out with the colicogenic strain E. coli
C600 (El) and the E. coli K12S strain which is sensitive to the produced colicine
and was obtained' from the genetics and collection laboratory of the I.I. Mechnikov
Institute.
The activity of colicine synthesis was determined by the method of
Lacunae [15] in which a count is made of the individual bacteria which synthesize
colicine. A two-hour culture of bacteria grown in Martin broth. was irradiated in
a Teflon vessel that was "transparent" to millimeter.
The oscillator producing the electromagnetic wave radiation consisted of
a -wave tube of the 0V612 type with an electron wave-tuning range extend-
ing from 5.7 to 8 nm and a maximum power in the continuous-wave mode that was
close to 100 mW [4]. The irradiation was carried out from a horn antenna at wave-
lengths of 5.8, 6.15, 6.5, 6.57, and 7.1 nm. The power plus density amounted to
1 mW/cm2.
The optimal dosages of culture and nutrient medium were selected which
allowed reliable measurement of the transmitted power under the conditions requir-
ing preservation of biological activity of the culture. The thickness of the bac-
terial suspension in the Teflon vessel amounted to 0.6 mm. The effect was estimated
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according to the magnitude of the induction coefficient:
Ki =
Ilk. irr./nce. irr.
nk. nonirr./nck. irr.
Where NL.irr. is the average number of lacunae in the irradiated culture;
nk.nonirr. is the average number of lacunae in the control for.the same
dilutions; nce.irr. is the average number of irradiated cells; nce.nonirr.
is the average number of cells in the control for the same dilutions.
We have noted the dependence of the induction of lacunae formation
by means of microwave irradiation on the irradiation time (see figure). The
irradiation during the first thirty min did not affect the synthesis of colicine.
Irradiation for a period of one hour caused an abrupt growth in the number of
cells synthesizing colicine at the corresponding wavelengths. An increase in
irradiation time to two hours caused a further growth of the induction coefficient.
However, for a subsequent increase in irradiation time no clearly defined results
were obtained.
Figure and Table RP53
Figure Caption: RP53, Dependence of the Induction Coefficient
of Colicine Synthesis on Irradiation Time.
The Irradiation Time is Plotted along the
Axis of Abscissas (in hours); the Induction
Coefficient is Plotted Along the Axis of
Ordinates. 1)1 = 6.5 mm; 2)X = 5.8 mm; 3)A =
7.1 mm.
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Title of Table 1: The Induction Coefficient for the Synthesis of
of Colicine Under Microwave Irradiation.
Legend for Table 1: 1 - Indicator, 2 - Wavelengths (in mm), 3 -
Number of Experiments, 4 - Kiav + 6K1 5 -
Index of Statistical Reliability, 6 - Error
Probability (in %)
In the main the induction activity of the microwave radiation depended
on the wavelength (see Table). Waves having a length of 5.8, 6.5 and 7.1 mm
caused induction of colicine synthesis; wavelengths of 6.15 and 6.57 mm no
substantial effect was observed. Statistical handling of the results obtained
demonstrated their reliability which was likewise checked by a series of control
experiments without irradiation. In this case the values of the induction co-
efficient remained constant and close to unity.
It is well-known that microwave irradiation may cause an increase in
temperature in the irradiated object. Kohijaman Nomura [13] demonstrated the
possibility of inducing colicine synthesis with an increase in temperature of
the thermally sensitive mutant of the colicinogenic strain containing colicine
E2. Although we used an fortiori low irradiation intensitites which did not ex-
ceed 1 mW/cm2 at which an increase in temperature was improbable and the strain
E. cola C600 (El) was not thermally sensitive, the indicated circumstances com-
pelled us to monitor the temperature in the irradiated and nonirradiated systems.
The temperature of the investigative system (the medium with the suspension of
bacteria) was measured by means of a thermocouple. For a reading accuracy of
0.1 to 0.2? a temperature difference could not be successfully revealed.
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This allowed the conclusion to be drawn that the effect is of a nonthermal charac-
ter.
In order to intensify the effect obtained irradiation was carried out
in a thermostat at a temperature of 37?. In this case induction was also observed
when irradiation lasted for 30 min. However, the effect could not be increased
substantially.
Thus, it was demonstrated that irradiation with electromagnetic waves
.in the millimeter range is capable at specified wavelengths of inducing the syn-
thesis of colicine in colicinogenic bacteria.
Heretofore the ability of various substances to induce colicine synthesis
that is lethal for a colicinogenic cell has been associated mainly with their dis-
integration properties with respect to DNA or with their capability of blocking
the synthesis of DNA. Ultraviolet radiation has such properties [17,10,12], and
so do certain chemical preparations which disrupt the synthesis of DNA in a cell
and are capable of causing damage to the DNA molecule [2,6,8,9,11,14]. The
mechanism of induction by means of millimeter waves must be qualitatively differ-
ent from the induction caused by these agents which are tropic with respect to
DNA, since the energy of a millimeter-wave quantum is too small (104 times
smaller than the energy than an ultraviolet-radiation quantum) to produce the
rupture of any chemical bonds or to damage a DNA molecule. From this point of
view millimeter radiation may be treated as a fundamentally new agent which
without causing direct damage in a DNA molecule leads to disruption of the
mechanism by which the function of the episomic genes are regulated in a cell.
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[1] V. G. Adamenko, R. L. Vilenskaya, M. V. Golant, et al. Electronic Engineering,
Series 1, No. 12, p. 132, 1966.
[2] S. F. Vorunova and B. M. Girdo. Antibiotics, No. 6, p. 513, 1966.
[3] M. V. Golant, V. G. Adamenko, and R. P. Vilenskaya. Abstracts of Papers
Delivered at the 4th Interuniversity Conference on Microwave Electronics,
Minsk, 1969, (p. 316).
[4] M. V. Golant, R. L. Vilenskaya, E. A. Zyulina, et al. In the books "Instru-
ments and Experimental Techniques," in Russian, Moscow, p. 136, 1965.
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Continuation of Electronics Engineering, Series No. 1
[5]. V. P. Komov, V. F. Kondrat'eva, V. N. Shvedova, et al. Transactions of
the Leningrad Chemical-Pharmaceutical Institute, Issue No. 20, Part 1,
91, 1967.
[6]. D. Kudlai, V. G. Likhoded, V. M. Girdo, T. B. Padloko, et al. Abstracts
of Papers Delivered at the 9th International Microbiology Congress. In
Russian. In Moscow, 1966, p. 15.
[7]. V. G. Likhoded, Microbiology, No. 7, p. 116, 1963.
[8]. V. G. Likhoded, T. V. Padloko. Transactions of the Moscow Naturalist
Society, Volume 22, p. 114, 1966.
[9]. A. Z. Smolyanaskaya. Microbiology, Issue No. 4, p. 707, 1968.
References 10 through 15 pick up from the Russian
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