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Effects of Weak ELF on E. Coli Cells and Human Lymphocytes: Role of Genetic, Physiological, and Physical Parameters

Authors:
Igor Belyaev at Biomedical Research Center Slovak Academy of Sciences
Igor Belyaev
  • Biomedical Research Center Slovak Academy of Sciences
Yevgeny D. Alipov
Yevgeny D. Alipov
Yevgeny D. Alipov
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Mats Harms-Ringdahl at Stockholm University
Mats Harms-Ringdahl
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Abstract

An increasing number of investigations has shown that weak ELF alternating (AC) magnetic fields (MF) affect biological systems (Goodman et al., 1995). The effects of AC fields have been observed within relatively narrow frequency bands at so-called resonance frequencies (Smith et al., 1987, Belyaev et al., 1994; Blackman et al., 1994; Prato et al., 1995). Relatively narrow windows were also observed in the amplitude dependencies of the AC field effects (Liboff et al., 1987; Lednev, 1991; Blackman et al., 1994; Prato et al., 1995). It has been found by Blackman et al.(1985) that the ambient static magnetic fields (DC) can significantly influence the effects of alternating magnetic fields. The importance of static MF for the ELF effects was confirmed in several papers (Lednev, 1991; Belyaev et al., 1994; Blackman et al., 1994; Fitzsimmons et al., 1994; Prato et al., 1995). Therefore, the effects of weak ELF are observed under specific combinations of DC/AC exposure. Several physical mechanisms were suggested to explain these observations (Liboff et al., 1987; Chiabrera et al., 1991; Lednev, 1991; Belyaev et al., 1994; Binhi, 1997). The dependence of ELF effects on some physiological factors such as concentration of ions during exposure of cells has been shown (Smith et al., 1987; Karabakhtsian et al., 1994).
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EFFECTS
OF
WEAK
ELF
ON
E.
COLI
CELLS
AND HUMAN LYMPHOCYTES:
ROLE
OF
GENETIC,
PHYSIOLOGICAL,
AND PHYSICAL PARAMETERS
Igor
Y.
Belyaev
'.2,
Yevgeny
D.
Alipov
2,
Mats Harms-Ringdahl
'.3
, Department
of
Radiobiology
Stockholm University, S- 10691
Stockholm, Sweden
2 Department
of
Radiation Physics, Biophysics and Ecology
Moscow Engineering -Physics Institute
115409, Moscow, Russia
3 Biomedical Unit
Swedish Radiation Protection Institute
117
16,
Stockholm, Sweden
INTRODUCTION
An
increasing number
of
investigations has shown that weak ELF alternating (AC) magnetic fields (MF) affect
biological systems (Goodman et
aI.,
1995). The effects
of
AC
fields have been observed within relatively narrow
frequency bands at so-called resonance frequencies (Smith et
aI.,
1987, Belyaev et
aI.,
1994; Blackman et aI., 1994;
Prato et aI., 1995). Relatively narrow windows were also observed
in
the amplitude dependencies
of
the AC field effects
(Liboff et
aI.,
1987; Lednev, 1991; Blackman et aI., 1994; Prato et aI., 1995).
It
has been found by Blackman et
al.( 1985) that the ambient static magnetic fields (DC) can significantly influence the effects
of
alternating magnetic
fields. The importance
of
static MF for the ELF effects was confirmed
in
several papers (Lednev, 1991; Belyaev et aI.,
1994; Blackman et
aI.,
1994; Fitzsimmons et
aI.,
1994; Prato et aI., 1995). Therefore, the effects
of
weak ELF are
observed under specific combinations
of
DC/AC exposure. Several physical mechanisms were suggested to explain
these observations (Liboff et aI., 1987; Chiabrera et
aI.,
1991; Lednev, 1991; Belyaev et aI., 1994; Binhi, 1997). The
dependence
of
ELF effects on some physiological factors such
as
concentration
of
ions during exposure
of
cells has
been shown (Smith et aI., 1987; Karabakhtsian et
aI.,
1994).
The effects
of
weak ELF and microwaves
of
millimeter range (MW) on the genome conformational state (GCS)
of
E.
coli KJ2 cells were described recently (Alipov et
aI.,
1994; Belyaev et aI., 1993). Both ELF and MW effects
depended on frequency, DC magnetic field, post-exposure time before analysis, growth stage and cell density during
exposure.
In
particular, the MW resonance frequencies were shown to be different for different strains (Belyaev et aI.,
1993).
In
recent
s,tudy,
the different frequency response to ELF was observed for two
E.
coli K12 strains within a range
of
6-37 Hz (Alipov et aI., 1996).
In
the present investigation,
we
tested whether the resonance ELF frequencies are
different for strains AB1157 and EMG2 within 6-69 Hz. According to some models, the probable targets for ELF
resonance effects are ions, radicals or charged molecular complexes such
as
DNA-protein complexes.
If
the same targets
respond to ELF
in
cells
of
different types, the similar spectra
of
resonances would
be
observed
in
these cells.
To
test this
hypothesis, the effects
of
ELF on chromatin conformation
in
human lymphocytes from healthy donors were studied
under the same conditions
of
exposure
as
for
E.
coli cells.
It
has been shown, that cell response to ELF correlated positively with cell density during exposure (Belyaev et aI.,
1995). The possible role
of
chemical messengers such
as
radicals or ions was suggested
to
explain the cooperative
response.
In
this study,
we
performed experiments exposing the cells to ELF with a specific scavenger
of
calcium,
EGTA, and the radical scavenger glycerol.
Electricity
and
Magnetism in Biology
and
Medicine
Edited
by
Bersani,
Kluwer
AcademiclPlenum
Publishers,
1999
481
MATERIALS
AND METHODS
The strains
of
E.
coli
KI2
were: wild-type EMG2 and
ABl157
F thrI aral4 leuB6 pro A2 lacGI tsx33 supE44
galK2 hisG4 rfbDI
mgl51
rpsL31
xyl5
mtii argE3 thiI
').:
rac-.
The cells were grown
as
previously described (Alipov et
aI., 1994) and then exposed at a concentration
of
4X\07 cells/ml
in
the M9 buffer. The human lymphocytes were
obtained from peripheral blood
of
healthy donors. The cells were diluted to a concentration
of
2x
I 06 cells/ml
in
RPMI
1640 medium. Cells were exposed to ELF at a pre-set frequency for 15-20 min, and then incubated
in
the same media
before lysis. Sinusoidal magnetic field,
21
liT
r.
m.
s.,
was
applied using the Helmholtz coils
by
means
of
an AC signal
generator. The intensities
of
DC and
AC
magnetic fields were control\ed
by
means
of
a magnetometer and a
microteslometer. The calculated
AC
magnetic field did not differ from the measured field. The col\inear and
perpendicular components
of
the static magnetic field were equal to 43±1 liT and
19±1
liT, respectively. The
E.
coli
cells were lysed as previously described with some modifications (Belyaev et
aI.,
1993). Briefly, solutions
of
1.5
mglml
lysozyme (Sigma, 0.3 ml),
2%
sarcosyl (Serva, I mi) and 3 mg/ml papain (Merck,
in
\0% glycerol, 0.7 ml) were added
sequentially to I
ml
of
the cell suspension. All solutions were prepared
in
a lysing buffer: 0.25 M Na2EDTA, 0.01 M
Tris, pH 7.1. The human lymphocytes were lysed
by
addition
of
3 mllysis solution (0.25 M Na2EDTA,
2%
sarcosyl,
\0
mM Tris-base, pH 7.4) to 0.4
ml
of
a cell suspension. The A VTD method was used
as
described previously (Belyaev et
ai, 1993). The control cells were concurrently subjected
to
the same manipulations except for exposure. Sham-exposed
cells were run under exactly the same conditions
as
exposed cells except for disconnection
of
wires between generator
and the Helmholtz coils. Control and sham-exposure were run
in
each experiment. Comparison
of
samples were
performed using the Student's t-test. Maximum relative viscosity was used to determine the ELF effect.
RESULTS
Four frequency windows were observed
in
19
independent experiments with
AB
1157 cells (Fig. I).
In
all
of
these
windows the effect showed a pronounced resonance structure and fitted well to a Gaussian distribution. The resonance
frequencies were 8.9±O.l Hz, 15.5±0.5 Hz 29.4±0.5 Hz and 62±1 Hz. Each resonance was reproduced
in
4-5
independent experiments. Within the same frequency range,
we
found only 3 resonant frequencies for EMG2 cells:
8.3±0.1 Hz, 27.0±0.5 Hz and 56.5±0.5
Hz.
With these cells,
we
did not observed any effect
of
ELF near
15
Hz. The 8.3
Hz, 27 Hz and 56.5 Hz resonance frequencies for EMG2 cells were shifted significantly (p<O.OI)
in
comparison with
correspondent resonances
of
AB
1157 strain: 8.9 Hz, 29.4 Hz and 62
Hz.
The half-widths and peak values
of
resonances
did not change significantly from strain
to
strain. The influence
of
5 mM EGTA and
3%
glycerol on ELF effect was
studied.
In
these experiments, the ABI157 cells were diluted to a concentration
of
5-6x\08 cells/ml
in
M9 with EGTA
or glycerol and then exposed
IS
min
at 8.9 Hz. The cells were lysed each 20
min
during 160 min after exposure. The
same ELF effects were observed
as
previously described (Belyaev et
aI.,
1995). The radical scavenger glycerol totally
abolished the ELF effect and calcium scavenger EGTA reduced the ELF effect
by
half. These data confirm the previous
findings about the role
of
calcium
in
ELF effects (Smith et
aI.,
1987; Karabakhtsian et
aI.,
1994) and provide evidence
for possible role
of
radicals
in
the cooperative response
of
cells to ELF (Belyaev et
aI.,
1995).
Human lymphocyte from 4 healthy donors were exposed to ELF
in
the range
of
5-12 Hz under the same
combinations
of
ACIDC fields
as
for
E.
coli cells.
In
human lymphocytes, the condensation
of
chromatin increased
in
response to ELF
in
contrary to the chromatin decondensation which
was
observed after ionizing irradiation or treatment
with DNA specific compounds such
as
ethidium bromide and etoposide VP-16. Statistically significant (p<O.OI)
decrease
in
the A VTD peaks was observed for two donors while cells from other donors did not respond to ELF. The
sensitive donor A and the insensitive donor B, were analyzed in respect
of
the reproducibility
of
ELF effect during one
year. The analysis
of
pooled data produced significant effect (p<0.OO5-0.05) with cells from Donor A at
6,
7, 8 and 9 Hz
with maximum around 8 Hz. The cells
of
Donor A were affected significantly by ELF around 58 Hz
in
6 independent
experiments within 52-65 Hz. The kinetics
of
the ELF effects at 58 Hz and 8 Hz were similar, with maximum around 60
min after exposure. Within the same frequency range,
we
did not observed significant ELF effect
in
lymphocytes
of
Donor B.
In
all cases, the ELF effects were transient and disappeared 2-3 h after exposure.
DISCUSSION
The data
of
present paper provide evidence that ELF resonances frequencies,
as
well
as
MW resonance frequencies
(Belyaev et
aI.,
1993), differ from strain to strain and depend on the genotype. Bacterial strain ABI157 has several
mutations.
In
particular, these cells have a mutation
in
the lac operon, which
is
responsible for the catabolism
of
the
disaccharide lactose
in
E.
coli, as well
as
mutations affecting metabolism
of
arabinose, arginine, threonine, leucine,
proline and galactose (Taylor and Trotter, 1974). The data obtained lead to the suggestion that at least one
of
these
mutations resulted
in
the appearance
of
a 15.5 Hz resonance in
AB
1157 cells which did not appear
in
the wild-type
482
1,40
>-
1,35
I-
en
1,30
0
0
CJ)
1,25
:>
w 1,20
>
~
« 1,15
....I
w
II:
1,10
::2:
:::>
1,05
::2:
X 1,00
«
::2:
0,95
0,90
0 10
20
30
40 50 60 70
FREQUENCY,
Hz
Figure
1. Frequency dependent changes in relative viscosity for two different
E.
coli strains exposed to
ELF
(15 min,
21
J.lT
AC magnetic flux
density, collinear
DC
field 43
J.lT).
Cells were lysed for measurements
70
min after exposure.
strain. This mutation is possibly involved in response at one
of
the intermediate steps between interaction
of
ELF with
intracellular target and changes in GCS. The data suggest also, that these mutations resulted in changes
of
target for
resonance response to ELF since shifts in resonances in comparison to wild-type strain were observed. Such shifts
of
resonances could be explained by the mechanism
of
phase modulation
of
high-frequency oscillations in chromosomes
(Belyaev
et
aI., 1994). According to this mechanism, the effective ELF frequencies are determined by the frequencies
of
natural oscillations in different parts
of
chromosomes due to DNA-protein interactions.
If
the strains are different in
some oscillations, a rearrangement in frequency spectra may be expected. A detailed analysis
of
data within the
framework
of
the phase modulation mechanism will be given elsewhere. By analogy with
E.
coli cells, the difference
between donors can be explained by individual genetic traits affecting the primary interaction
or
subsequent steps. As
the differences in resonance frequencies between
E.
coli and human lymphocytes is not higher than difference between
different
E.
coli strains, we concluded that these frequencies are determined by very similar targets. It was shown in
previous studies, that effects
of
AC magnetic fields on the GCS depended on collinear but not perpendicular component
of
the DC magnetic field (Belyaev
et
aI.,
1994).
We
compared our resonance conditions with predictions
of
several
models for collinear
ACIDC
fields. In two models, the resonance-like effects were discussed regarding to the
"cyclotron" frequencies for some ions
of
biological relevance such as calcium, magnesium and potassium (Liboff et aI.,
1987; Lednev, 199\). The first mechanism (Liboff
et
aI., 1987, 1991) predicted effects
at
cyclotron frequencies and their
harmonics, while the second one (Lednev, 1991) dealt with cyclotron frequencies and their subharmonics. According to
the ion cyclotron model, a good correspondence is observed for our resonance
of
29.4 Hz and the predicted value
of
29.3
Hz
for the cyclotron frequency
of
45
Ca
+2
at the DC field
of
43
/LT.
On the other hand, there is no coincidence
between other experimental and predicted resonances. Moreover, this model cannot explain our resonance frequencies
around 8-9 Hz because the ions
of
4OCa+
2
, magnesium and potassium
don't
have such harmonics at the DC field
of
43
/LT.
As we did not see the effects at the cyclotron frequencies
of
the biologically important ions, our data
don't
fit to the
model
of
Lednev as well. Nevertheless, there was partial correspondence
of
resonance frequencies and first
subharmonics for
K+
(8.4 Hz),
40Ca+2
(16.4 Hz), and
Mg2+
(27.1 Hz). A better agreement was observed between the
experimental data for AB 1157 strain and theoretical predictions
of
the ion interference mechanism for
4OCa+
2 (Binhi,
1997).
In conclusion, the ELF effects on the conformation
of
chromatin were observed under magnetic flux densities
which are close to fields
of
occupational and residential exposure. The ELF effects on chromatin conformation depend
on several physical parameters such as frequency, magnetic flux densities and orientation
of
AC/DC fields, genotype
and on several physiological parameters as well. This dependence may explain poor reproducibility
of
ELF effects in
some studies. Under specific combination
of
AC/DC exposure, the ELF resonance frequencies for cells
of
different
types were closely situated. This suggests similar target/targets for interaction
of
ELF
with different types
of
cells.
483
ACKNOWLEDGMENTS
These
studies
were
supported in
part
by
the Swedish Council for
Work
Life Research, Swedish Electrical Utilities
Research
and
Development
Company, Swedish Radiation Protection Institute
and
Grant
from the Russian
Foundation
for
Basic
Research.
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484
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Full-text available
  • Feb 2019
  • BMC CANCER
Background It has been demonstrated that relatively small variations of the parameters of exposure to extremely low frequency magnetic fields (ELF-MF) can change significantly the outcome of experiments. Hence, either in trying to elucidate if these fields are carcinogenic, or in exploring their possible therapeutic use, it is desirable to screen through as many different exposures as possible. The purpose of this work is to provide a proof of concept of how a recently reported system of coils allows testing different field exposures, in a single experiment. Methods Using a novel exposure system, we subjected a glioblastoma cancer cell line (U251) to three different time modulations of an ELF-MF at 60 different combinations of the alternated current (AC) and direct current (DC) components of the field. One of those three time modulations was also tested on another cell line, MDA-MB-231 (breast cancer). After exposure, proliferation was assessed by colorimetric assays. Results For the U251 cells, a total of 180 different exposures were tested in three different experiments. Depending on exposure modulation and AC field intensity (but, remarkably, not on DC intensity), we found the three possible outcomes: increase (14.3% above control, p < 0.01), decrease (16.6% below control, p < 0.001), and also no-effect on proliferation with respect to control. Only the time modulation that inhibited proliferation of U251 was also tested on MDA-MB-231 cells which, in contrast, showed no alteration of their proliferation on any of the 60 AC/DC field combinations tested. Conclusions We demonstrated, for the first time, the use of a novel system of coils for magnetobiology research, which allowed us to find that differences of only a few μT resulted in statistically different results. Not only does our study demonstrate the relevance of the time modulation and the importance of finely sweeping through the AC and DC amplitudes, but also, and most importantly, provides a proof of concept of a system that sensibly reduces the time and costs of screening. Electronic supplementary material The online version of this article (10.1186/s12885-019-5376-z) contains supplementary material, which is available to authorized users.
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... However, the mechanism of interaction and its consequences in living systems is ambiguous and need more work to be clarified. Effects of exposure to ELF-EMF were spotted to be dependent on signal physical characteristics, frequency and field intensity, exposure time and growth stage [9][10][11][12][13][14]. Several studies were performed to evaluate the bio-effects on bacterium as a result of exposure to such fields and different consequences were approached. ...
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... This letter is in clear line with these previous publications, aiming not only at succinctly retrieving already well-known recommendations, but also at doing so from an updated perspective and, perhaps most importantly, at contributing with some points that, to the best of our knowledge, have not yet been given sufficient attention. Of note, our recommendations deal with fields and temperature generation, monitoring, shielding, and reporting, while many other important variables such as genotype, physiological state, cell density, and concentrations of ions and radicals [Belyaev et al., 1998;Belyaev et al., 1999;Belyaev and Alipov, 2001;Sarimov et al., 2011] are out of scope of this letter. Table 1 summarizes the recommendations that we present in full detail in the following sections of this letter. ...
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... These include induction of oxidative stress, DNA damage, epigenetic changes, altered gene expression and induction including inhibition of DNA repair and changes in intracelluar calcium metabolism. Both low-intensity ELF-EMF and non-thermal RF-EMF effects depend on a number of physical parameters and biological variables and physical parameters, which account for the variation in health outcomes (Belyaev, 2015;Belyaev et al., 1999). Importantly, the most severe health effects are observed with prolonged chronic exposures even when intensities are very low (Belyaev, 2017). ...
Thermal and non-thermal health effects of low intensity non-ionizing radiation: An international perspective
Article
  • Jul 2018
Exposure to low frequency and radiofrequency electromagnetic fields at low intensities poses a significant health hazard that has not been adequately addressed by national and international organizations such as the World Health Organization. There is strong evidence that excessive exposure to mobile phone-frequencies over long periods of time increases the risk of brain cancer both in humans and animals. The mechanism(s) responsible include induction of reactive oxygen species, gene expression alteration and DNA damage through both epigenetic and genetic processes. In vivo and in vitro studies demonstrate adverse effects on male and female reproduction, almost certainly due to generation of reactive oxygen species. There is increasing evidence the exposures can result in neurobehavioral decrements and that some individuals develop a syndrome of "electro-hypersensitivity" or "microwave illness", which is one of several syndromes commonly categorized as "idiopathic environmental intolerance". While the symptoms are non-specific, new biochemical indicators and imaging techniques allow diagnosis that excludes the symptoms as being only psychosomatic. Unfortunately standards set by most national and international bodies are not protective of human health. This is a particular concern in children, given the rapid expansion of use of wireless technologies, the greater susceptibility of the developing nervous system, the hyperconductivity of their brain tissue, the greater penetration of radiofrequency radiation relative to head size and their potential for a longer lifetime exposure.
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... The evident reason for this eventual inconsistency is strong dependence of the EMF effects on a number of physical and biological parameters, which significantly varied between studies. These dependencies were established for both ELF (70)(71)(72) and RF effects (24,27). ...
EUROPAEM EMF Guideline 2016 for the prevention, diagnosis and treatment of EMF-related health problems and illnesses
Article
Full-text available
  • Jul 2016
  • Rev Environ Health
Chronic diseases and illnesses associated with non-specific symptoms are on the rise. In addition to chronic stress in social and work environments, physical and chemical exposures at home, at work, and during leisure activities are causal or contributing environmental stressors that deserve attention by the general practitioner as well as by all other members of the health care community. It seems necessary now to take "new exposures" like electromagnetic fields (EMF) into account. Physicians are increasingly confronted with health problems from unidentified causes. Studies, empirical observations, and patient reports clearly indicate interactions between EMF exposure and health problems. Individual susceptibility and environmental factors are frequently neglected. New wireless technologies and applications have been introduced without any certainty about their health effects, raising new challenges for medicine and society. For instance, the issue of so-called non-thermal effects and potential long-term effects of low-dose exposure were scarcely investigated prior to the introduction of these technologies. Common electromagnetic field or EMF sources: Radio-frequency radiation (RF) (3 MHz to 300 GHz) is emitted from radio and TV broadcast antennas, Wi-Fi access points, routers, and clients (e.g. smartphones, tablets), cordless and mobile phones including their base stations, and Bluetooth devices. Extremely low frequency electric (ELF EF) and magnetic fields (ELF MF) (3 Hz to 3 kHz) are emitted from electrical wiring, lamps, and appliances. Very low frequency electric (VLF EF) and magnetic fields (VLF MF) (3 kHz to 3 MHz) are emitted, due to harmonic voltage and current distortions, from electrical wiring, lamps (e.g. compact fluorescent lamps), and electronic devices. On the one hand, there is strong evidence that long-term exposure to certain EMFs is a risk factor for diseases such as certain cancers, Alzheimer's disease, and male infertility. On the other hand, the emerging electromagnetic hypersensitivity (EHS) is more and more recognized by health authorities, disability administrators and case workers, politicians, as well as courts of law. We recommend treating EHS clinically as part of the group of chronic multisystem illnesses (CMI), but still recognizing that the underlying cause remains the environment. In the beginning, EHS symptoms occur only occasionally, but over time they may increase in frequency and severity. Common EHS symptoms include headaches, concentration difficulties, sleep problems, depression, a lack of energy, fatigue, and flu-like symptoms. A comprehensive medical history, which should include all symptoms and their occurrences in spatial and temporal terms and in the context of EMF exposures, is the key to making the diagnosis. The EMF exposure is usually assessed by EMF measurements at home and at work. Certain types of EMF exposure can be assessed by asking about common EMF sources. It is very important to take the individual susceptibility into account. The primary method of treatment should mainly focus on the prevention or reduction of EMF exposure, that is, reducing or eliminating all sources of high EMF exposure at home and at the workplace. The reduction of EMF exposure should also be extended to public spaces such as schools, hospitals, public transport, and libraries to enable persons with EHS an unhindered use (accessibility measure). If a detrimental EMF exposure is reduced sufficiently, the body has a chance to recover and EHS symptoms will be reduced or even disappear. Many examples have shown that such measures can prove effective. To increase the effectiveness of the treatment, the broad range of other environmental factors that contribute to the total body burden should also be addressed. Anything that supports homeostasis will increase a person's resilience against disease and thus against the adverse effects of EMF exposure. There is increasing evidence that EMF exposure has a major impact on the oxidative and nitrosative regulation capacity in affected individuals. This concept also may explain why the level of susceptibility to EMF can change and why the range of symptoms reported in the context of EMF exposures is so large. Based on our current understanding, a treatment approach that minimizes the adverse effects of peroxynitrite - as has been increasingly used in the treatment of multisystem illnesses - works best. This EMF Guideline gives an overview of the current knowledge regarding EMF-related health risks and provides recommendations for the diagnosis, treatment and accessibility measures of EHS to improve and restore individual health outcomes as well as for the development of strategies for prevention.
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... Overall these studies demonstrated that static magnetic fields, ELF-EMF and pulsed electromagnetic fields cause alterations in growth rate of bacteria (Cellini et al. 2008, Obermeier et al. 2009, Gu et al. 2012, Segatore et al. 2012, Ahmed et al. 2013, Ibraheim et al. 2013, morphological changes (Volpe et al. 2002, Inhan Garip et al. 2011 and that bacteria sense these fields as stress factors and thus express heat shock proteins (Del Re et al. 2006. It was also shown that ELF affected conformation of chromatin in Escherichia coli cells in dependence on a number of biological and physical variables including genotype, ELF frequency and static magnetic field at the location of ELF exposure (Alipov and Belyaev 1996, Belyaev et al. 1998, 1999, Belyaev and Alipov 2001. ...
Effect of extremely low frequency electromagnetic fields on bacterial membrane
Article
Full-text available
Purpose: The effect of extremely low frequency electromagnetic fields (ELF-EMF) on bacteria has attracted attention due to its potential for beneficial uses. This research aimed to determine the effect of ELF-EMF on bacterial membrane namely the membrane potential, surface potential, hydrophobicity, respiratory activity and growth. Materials and methods: Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli were subjected to ELF-EMF, 50 Hz, 1 mT for 2 h. Membrane potential was determined by fluorescence spectroscopy with or without EDTA (Ethylenediaminetetraacetic acid) with DisC3(5) (3,3-dipropylthiacarbocyanine iodide), zeta potential measurements were performed by electrophoretic mobility, hydrophobicity of the membrane was measured with MATH (Microbial Adhesion to Hydrocarbons) test, respiratory activity was determined with CTC (5-Cyano-2,3-ditolyl tetrazolium chloride), colony forming unit (CFU) and DAPI (4',6-diamidino-2-phenylindole, dihydrochloride) was used for growth determinations. Results: ELF-EMF caused changes in physicochemical properties of both Gram-positive and Gram-negative bacteria. Hyperpolarization was seen in S. aureus and EDTA-treated E. coli. Surface potential showed a positive shift in S. aureus contrariwise to the negative shift seen in EDTA-untreated E. coli. Respiratory activity increased in both bacteria. A slight decrease in growth was observed. Conclusion: These results show that ELF-EMF affects the crucial physicochemical processes in both Gram-positive and Gram-negative bacteria which need further research.
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... It is possible that this same mechanism is at work in bacterial cells, as bacterial cells also possess an equivalent T-type channel that displays similar properties and functions (Matasushita et al. 1989). This is also supported by researched conducted using a Ca 2+ chelator, where when it was added to growing cultures of E. coli the effects of the EMF were significantly decreased (Belyaev et al. 1995(Belyaev et al. , 1999. ...
Bacterial growth rates are influenced by cellular characteristics of individual species when immersed in electromagnetic fields
Article
Full-text available
  • Jan 2015
Previous studies have shown that exposure to extremely low-frequency electromagnetic fields (ELF-EMFs) have negative effects on the rate of growth of bacteria. In the present study, two Gram-positive and two Gram-negative species were exposed to six magnetic field conditions in broth cultures. Three variations of the 'Thomas' pulsed frequency-modulated pattern; a strong-static "puck" magnet upwards of 5000G in intensity; a pair of these magnets rotating opposite one another at ∼30rpm; and finally a strong dynamic magnetic field generator termed the 'Resonator' with an average intensity of 250μT were used. Growth rate was discerned by optical density (OD) measurements every hour at 600nm. ELF-EMF conditions significantly affected the rates of growth of the bacterial cultures, while the two static magnetic field conditions were not statistically significant. Most interestingly, the 'Resonator' dynamic magnetic field increased the rates of growth of three species (Staphylococcus epidermidis, Staphylococcus aureus, and Escherichia coli), while slowing the growth of one (Serratia marcescens). We suggest that these effects are due to individual biophysical characteristics of the bacterial species. Copyright © 2015 Elsevier GmbH. All rights reserved.
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Possible mechanisms by which extremely low frequency magnetic fields affect opioid function
Article
Full-text available
  • Jun 1995
Although extremely low frequency (ELF, <300 Hz) magnetic fields exert a variety of biological effects, the magnetic field sensing/transduction mechanism (or mechanisms) remain to be identified. Using the well-defined inhibitory effects that magnetic fields have on opioid peptide mediated antinociception or "analgesial' in the land snail Cepaea nemoralis, we show that these actions only occur for certain frequency and amplitude combinations of time-varying sinusoidal magnetic fields in a manner consistent with a direct influence of these fields. We exposed snails with augmented opioid activity to ELF magnetic fields, which were varied in both amplitude and frequency, along with a parallel static magnetic field, When the peak amplitude (0-547 mu T) of a magnetic field of 60 Hz was varied systematically, we observed a nonlinear response, i.e., a nonlinear reduction in analgesia as measured by the latency of a defined response by the snails to a thermal stimulus. When frequency (10-240 Hz) was varied, keeping the amplitude constant (141 mu T), we saw significant inhibitory effects between 30 and 35 Hz, 60 and 90 Hz and at 120 and 240 Hz. Finally, when the static field was varied but the amplitude and frequency of the time-varying field were held constant, we observed significant inhibition at almost all amplitudes. This amplitude/frequency "resonance-like" dependence of the magnetic field effects suggests that the mechanism (or mechanisms) of response to weak ELF fields likely involves a direct magnetic field detection mechanism rather than an induced current phenomenon. We examined the implications of our findings for several models proposed for the direct sensing of ELF magnetic fields.
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Evidence for Dependence of Resonant Frequency of Millimeter Wave Interaction with Escherichia coli K12 Cells on Haploid Genome Length
Article
Full-text available
  • Jan 1993
The method of anomalous viscosity time dependence (AVTD) was used to study the influence of millimeter electromagnetic radiation (EMR) on the genome conformational state (GCS) of Escherichiu coli K12 cells. Strain N99 of wild-type, lysogenic strains N99(A) and N99(λ,λimm434bio10) were used. In the 41.28-41.37 GHz and 51.73-51.79 GHz ranges the resonance effect of low-intensity EMR (10-10 W/cm2) on the GCS of N99 cells was shown. The resonance frequencies were 41.324 ± 0.001 GHz and 51.765 ± 0.002 GHz, respectively. The insertion in bacterial chromosome of prophage λ [strain N99(λ)] and prophage λimm434bio10 [strain N99(λ, λimm434bio10)] reduced both resonance frequencies considerably. The decrease of both resonance frequencies was proportionate to the increase of the haploid genome length. Shifts of the resonance frequency were not accompanied by change in the sign of effective circular EMR polarization. The results indicate that the frequencies of resonance interaction of E. coli cells with low-intensity millimeter waves are determined by the genome structure. A physical model was suggested to describe the observed shift of the resonance frequency.
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A Linkage Map and Gene Catalog for Escherichia coli
Chapter
  • Jan 1974
This contribution contains a diagram which illustrates the position of about 460 genes on the circular linkage map of this bacterium. In Figure 1 the inner circle, which bears the time scale from 0 through 90 minutes is based on the results of interrupted conjugation experiments. The map is graduated in 1-minute intervals beginning arbitrarily with zero at the thr locus. Certain parts of the map (e. g., the 9- to 10-minute segment) are displayed on arcs of the outer circle to provide an expanded time scale for crowded regions. The genetic symbols in this figure are defined in Table 1. Markers in parentheses are only approximately mapped at the positions shown. A gene identified by an asterisk has been mapped more precisely than the markers in parentheses, but its orientation relative to adjacent markers is not yet known. The arrows which are placed next to the phoA and argI genes and next to certain operons show the direction of messenger RNA transcription for these loci. The table provides an alphabetical listing of these genes and also shows the various specific enzyme activities and other phenotypic traits ascribed to the loci. In Table 1 the numbers refer to the time scale shown in Figure 1. Parentheses indicate approximate map locations. Citations to the scientific publications containing detailed descriptions of each locus under study are given in Taylor (1970) and Taylor and Trotter (1972). Figure 1 and Table 1 are reproduced with the permission of the American Society for Microbiology.
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The model of phase modulation of high frequency oscillations of a nucleoid for the effect of weak static and alternating magnetic fields on E.coli cells
Article
  • Jan 1996
The method of anomalous viscosity time dependences was used to investigate the influence of extremely low frequency alternating and static magnetic field on the genome conformational state of E.coli K12 AB1157 cells. The physical model was developed on the base of obtained results to describe effects of weak static and alternating magnetic fields on the living cells. In this model the high frequency oscillations of a charged nucleoid are considered with a classical approach. The wave-like dependence of maximum effect on magnetic induction within the range from 0 to 110 μT was established.
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Interference of Ion Quantum States Within a Protein Explains Weak Magnetic Field's Effect on Biosystems
Article
  • Jul 2009
A consistent physical model is proposed that explains weak ELF magnetic field effects on biological systems. An interference is described of quantum states of bound ions in protein cavities for the first time. The ion quantum dynamics is investigated at idealized case in the colinear static and alternating magnetic fields. There is taken into account a nuclear spin of ion and a nonlinear protein response on redistribution of the ion probability density. A formula is developed for the magnetic-dependent part of an ion-protein dissociation probability. Typical frequency and amplitude spectra are calculated for the interference in various magnetic conditions including the “magnetic vacuum” and compared with the known experimental data.
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Cooperativity in E. coli cell response to resonance effect of weak extremely low frequency electromagnetic field
Article
  • Apr 1995
  • Bioelectrochem Bioenerg
The response of Escherichia coli cells to extremely low frequency electromagnetic fields (ELF EMFs) was investigated at a magnetic induction amplitude of 30 μT. The influence of the EMF with a frequency of 9 Hz on the previously revealed resonance of the genome conformational state (GCS) has been evaluated. The resonance effect, which was measured by the method of anomalous viscosity time dependence, depended on the concentration of exposed cells. The effect was normalized to its own sham control at each concentration. The GCS changes increased with the concentration of exposed cells and reached a plateau at a value of about 6 × 103 cell ml−1. This means that cells interact during the resonance reaction to EMF, thus enhancing the effect. Saturation of the effect at concentrations close to those in the tissues of higher eukaryotes may reflect the importance of this parameter for the regulation of living systems. The experimentally established dependence of the resonance EMF effect on the concentration of exposed cells was described well by the theoretical model. This model was obtained by assuming that the cooperative effect was produced by the electromagnetic interaction of cells in the millimeter-submillimeter range of EMF.As in the case of the resonance reaction of E. coli cells to low intensity millimeter waves, the GCS changes were shown to correlate with the changes in the spectrum of the DNA-bound proteins.
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Systemic reaction of Escherichia coli cells to weak electromagnetic fields of extremely low frequency
Article
  • Jun 1994
  • Bioelectrochem Bioenerg
The method of anomalous viscosity time dependence was used to study changes in the genome conformational state (GCS) of Escherichia coli K12 AB1157 cells after their exposure to electromagnetic fields (EMFs) with an amplitude of magnetic induction equal to 30 μ T in the frequency range 7–12 Hz. A resonance effect which had good reproducibility and disappeared 100–170 min after the exposure of the cells was demonstrated. The direction and magnitude of the effect depended on the growth stage of the bacterial culture at which the cells were exposed to the EMF. The resonance frequencies and half-widths of the resonance dependences of the effect in the early stationary and middle logarithmic phases did not differ significantly. It was shown that the EMF resonance effect on the GCS is accompanied by considerable changes in the dynamics of cell division, DNA synthesis and protein synthesis. A comparative analysis of the systematic reactions of E. coli cells to the resonance effect of extremely low frequency (ELF) EMF and electromagnetic millimeter waves was carried out.
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Possible mechanism of the effect of weak magnetic fields on biosystems
Article
  • Jan 1991
A physical mechanism is suggested for a resonant interaction of weak magnetic fields with biological systems. An ion inside a Ca(2+)-binding protein is approximated by a charged oscillator. A shift in the probability of ion transition between different vibrational energy levels occurs when a combination of static and alternating magnetic fields is applied. This in turn affects the interaction of the ion with the surrounding ligands. The effect reaches its maximum when the frequency of the alternating field is equal to the cyclotron frequency of this ion or to some of its harmonics or sub-harmonics. A resonant response of the biosystem to the magnetic field results. The proposed theory permits a quantitative explanation for the main characteristics of experimentally observed effects.
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Calcium Cyclotron Resonance and Diatom Mobility
Article
  • Jan 1987
The hypothesis that movement of biological ions may be predicted by cyclotron resonance theory applied to cell membranes is tested in these experiments. Diatoms (Amphora coffeaeformis) were chosen as the biosystem since they move or don't move, depending on how much calcium is transported across the membrane. The experiments demonstrate that a particular ion (calcium) is apparently moved across the cell membrane in response to the DC and AC values of magnetic flux densities (B) and the frequency derived from the cyclotron resonance theory. A clear resonance is shown and a rather sharp frequency response curve is demonstrated. The experiments also show a dose response as the AC value of the flux density is varied, and that odd harmonics of the basic cyclotron frequency are also effective.
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