Stage 1: High-Frequency EMR Impact and Aires Resonator Influence on Rats and Honey Bees
2
Saint Petersburg, 2016
REPORT
ON AGREEMENT FOR SCIENTIFIC COOPERATION BETWEEN FEDERAL STATE BUDGETARY SCIENTIFIC ESTABLISHMENT PAVLOV INSTITUTE OF PHYSIOLOGY OF THE RUSSIAN ACADEMY OF SCIENCE AND AIRES HUMAN GENOME RESEARCH FOUNDATION “research of high frequency electromagnetic radiation effect and Aires resonators influence on behavior, genetic and epigenetic processes in cells of central and peripheral organs (model organisms: rat (Rattus norvegicus) and honey bee (Apis mellifera L.)”
THE FIRST STAGE (April – September 2016): research of router’s electromagnetic radiation effect and Aires resonators’ influence on stability of genetic apparatus in marrow cells of tistar male rats
In the context of technological advancement, growth of power generation, development of
communications systems there is an increasing negative influence of non-ionizing
electromagnetic radiation (EMR) on human organism. Disturbance of environmental
electromagnetic background is recorded everywhere, causing steady increase of different
diseases, including microwave sickness, electromagnetic hypersensitivity (EHS). The literature
on EMR influence on human and animal organism is quite extensive:
http://www.bioinitiative.org/report/wp-content/uploads/pdfs/RFR-11_28-research-summary.pdf,
http://www.bioinitiative.org/report/wp-content/uploads/pdfs/RFR-free-radical-abstracts.pdf . It
has been established that EMR enhancement influences genetic apparatus of cells in different
human and animal organs. For the first time mutagenic effect of certain electromagnetic
frequencies was demonstrated in 1959 in the paper by Heller, Teixeria-Pinto, published in the
journal Nature (Heller, Teixeira-Pinto, 1959). At present, evidence has been collected that
electromagnetic fields of different ranges including mobile phones and ti-Fi can induce a broad
spectrum of genetic damage, modify gene expression, affect structural and functional
characteristics of cell nuclei (Kryukov, 2000; Deinekina, 2002; Boyko et al., 2010, et al.). For
example, it was proven that the effect of microwave radiation (frequency of 7.7 GHz, power of
0,5,10,30 mt/cm2) with an exposure interval from 10 to 60 minutes increases chromosome
aberrations in human lymphocytes (Garaj-srhovac et al., 1992). tith long periods of exposure,
signals with a specific absorption rate (SAR) of 5 t/kg also damage chromosomes in blood cells
(Tice et al., 2002). However, the mechanisms of those processes are still insufficiently
understood.
There is a pressing need to create a system for protection from harmful effect of EMR and
study the mechanisms of its influence on genetic processes in the cells of central and peripheral
organs of model organisms. At present, Aires has created devices on the basis of the fractal-
3 matrix EMR conversion nanotechnology that efficiently redistribute EMR and have curative
properties
(http://www.aires.spb.ru/nano.html, http://airestech.ru ). However, the mechanisms of protective
effect of those devices on the cell chromosome apparatus have not been studied. Purpose and objectives of the first stage:
1) research of standard ti-Fi router’s EMR effect in different modes of operation on
destabilization of the genetic apparatus in dividing marrow cells in order to identify the
conditions of induction of mitotic disturbances by high-frequency EMR;
2) evaluation of the protective effect of Aires Defender fractal-matrix resonators on the stability
of the genetic apparatus in dividing marrow cells of tistar male rats under router's damaging
EMR effect.
MATERIALS AND METHODS.
The research was performed on tistar male rats weighing 250-300 g received from the
animal quarters of the Federal State-Funded Establishment of Science Pavlov Institute of
Physiology of the Russian Academy of Sciences. Rats of an outbread tistar population are a
generally recognized model used in medical and biological research. Upon arrival, the animals
were maintained in a laboratory animal quarters for at least two weeks for adjustment. The males
were kept in groups of 6 in standard cages on a standard food ration.
The research employed a standard ti-Fi router (LinkSys E1200-EE/RU wireless router)
with the following characteristics: wireless frequency: 2.4 GHz, number and type of antennas: 2
internal antennas, standard antenna gain factor, dBi: 4 dBi.
To study router's EMR effect, the «home» cage with animals was placed in a Faraday cage
(Fig. 1), a router was placed under the top lid of the cage in the center of a removable shelf. The
experimental groups were exposed to the router's effect in the following modes:
1) once for 2 hours (8:00-10:00);
2) 4 days, 6 hours a day (8:00-14:00);
3) 3 weeks, 6 hours a day (8:00-14:00).
The reference was groups of rats placed in a Faraday cage at the same time, but with no router,
as well as intact animals.
The experiments also employed Aires Defender fractal-matrix resonators (special circular
diffraction grating), which are a universal space-wave Fourier filter (whabrev et al., 2005). Due
to interaction of the electromagnetic field with the Aires Defender, its structural transformation
occurs. To evaluate resonators' influence on the router's damaging EMR effect, 6 resonators were
4 used. They were placed in the center of each side of the Faraday cage (Fig. 1). One of the
experimental groups was exposed to router's effect in the Faraday cage as per the schedule, 6
hours a day for 4 days.
Making human marrow cell preparations. 24 hours after exposure, human marrow cells were
fixated in freshly made fixative (1 part of glacial acetic acid : 3 parts of ethanol) for at least an
hour. The material was kept at a temperature of +4ᴼC until specimens were prepared following
the routine method (Makarov, Safronov, 1978; Dayev et al., 2009).
Analysis of marrow preparations. Squash marrow preparations were analyzed using a
Micromed-3 microscope with x640–1600 . Chromosome aberrations at the anaphase–telophase
stage were factored in (the standard ana-telophase method) with consideration of additional
recommendations (Dayev et al., 2009). At least 200 cells from each animal were analyzed. The
study considered the number of normal and aberrant anaphase-telophases with the following
types of disturbances: single reorganizations (a fragment, a bridge, a lagging chromosome),
multiple reorganizations (two or more disturbances of any type per cell) (Fig. 2).
Statistical processing. The data was checked for homogeneity using the nonparametric method
of multi-field χ2. Based on the check, individual data was united within groups. To present the
findings in tables, frequencies of detected disturbances in mitosis were calculated and expressed
as per cent with and error percentage. Among cells with chromosome reorganization, shares of
cells with different types of disturbances were determined, which characterized the spectrum of
chromosome aberrations (Dayev et al., 2009). Significance of differences between the versions
was determined and the spectra of mitotic disturbances were compared by means of the χ2
method (Glotov et al., 1982), as well as A ranking test and ANOsA using Statgraphics Centurion
us11 Statistica 6.0 software.
Fig. 1. The diagram of the Faraday cage used in the research showing the location of resonators.
Removable lid
2-level shelf for the ti-Fi router
Faraday cage for research
The placement zones are the centers of all sides
Defender
5
A b c
d e f
g Fig. 2. Dividing marrow cells in rats at the ana-telophase stage. There are examples of analyzed reorganizations (labeled with arrows): a- normal ana-telophase, b- lagging chromosome, c- bridge, d- fragment, d,e-multiple reorganizations with the bridge and lagging chromosomes, f- double bridge.
RESULTS
It is demonstrated that high frequency electromagnetic radiation of a router in the exposure
mode of 4 days, 6 hours a day cause d the highest destabilization of genetic apparatus of the
dividing marrow cells in tistar male rats: the overall frequency of mitotic disturbances rises by
4.5 times as compared to Reference2 (Faraday cage, 4 days x 6 hours) and by 3.9 times as
compared to Reference1 (Table 1), which has a high degree of significance in all statistical
criteria applied. One-time 2-hour long exposure to router's effect raises the number of
chromosome aberrations by 1.9 times as compared to the intact Reference 1 (Table 1),
significance of differences is confirmed by the multiple range test (Multiple Range Test ,
6 Diff.=6,03, +/-Limits=3,33) and ANOsA (F= 19,65 , p< 0,004), but not χ2. At the same time, as
compared to the group of animals exposed to router's effect for 4 days, 6 hours a day, the
frequency of disturbances reduced by 2.1 times (Table 1). After the animals were exposed for 3
weeks, 6 hours a day in the conditions of router's electromagnetic radiation, the frequency of
mitotic disturbances rises by 1.8 times as compared to Reference 3 (Faraday cage, 3 weeks, 6
hours a day) (Multiple Range Test , Diff.=5.02, +/-Limits=3.63; ANOsA , F= 3,13 , p< 0,01)
and by 1.5 times as compared to the intact Reference 1 (Multiple Range Test, Diff.=4,1, +/-
Limits= 3,9; ANOsA , (F= 2,49 , p< 0,04), but it decreases by 2.6 times as compared to the
group of animals exposed to the router's effect for 4 days, 6 hours a day (Table 1).
The effect of Aires Defender fractal-matrix resonators was evaluated using the pattern of 4
days, 6 hours a day, which caused the most damage to the marrow cell chromosome apparatus.
Using resonators in combination with router's effect reduces fourfold the frequency of damage to
mitosis in dividing marrow cells as compared to router's effect without protection, and that level
is comparable to Reference 2 (Faraday cage, 4 days, 6 hours a day) and intact Reference 1
(Table 1).
Comparison of the spectra of spontaneous mitotic disturbances and those induced by
router's EMRs revealed that router's effect of 4 days, 6 hours a day changes the spectrum of
chromosome disturbances due to increase in the share of lagging chromosomes and decrease in
the share of bridges among all types of disturbances, as compared to Reference 2 and Reference
1 (Table 2). Here it is important to note that the Reference 1 and Reference 2 groups differ by the
share of lagging chromosomes and multiple reorganizations among all types of disturbances
(Table 2). Using resonators changes the spectrum of disturbances owing to all types of
aberrations in question as compared to router's effect alone (Table 2).
7 Table 1. The frequency of mitotic disturbances in marrow cells of tistar male rats after exposure to router's electromagnetic radiation in different modes. sersions of exposure
Number of analyzed cells (those with disturbances among them)
Overall frequency of disturbances in mitosis
Significance of differences χ2 (ν= 1, r< 0,01)
Reference1 (intact)
1378 (92) 6.7 ± 0.7
16.8
Reference2 (Faraday cage, 4 d x 6 hrs)
1986 (114) 5.7 ± 0.5
Router (4 d x 6 hrs)
1360 (354) 26.0 ± 1.2
Router+resonator (4 d x 6 hrs)
1961 (127) 6.5 ± 0.6
Reference3 (Faraday cage, 3 weeks x 6 hrs;
1714 (96) 5.6 ± 0.5
Router (3 weeks x 6 hrs)
1789 (180) 10.1 ± 0.7*#
Router (2 hours)
1175 (149) 12.7 ± 0.9*
Legend: vertical lines mean significantly different values (χ2 criterion), # – differences from Reference3 are significant (Multiple Range Test, ANOsA (r<0,01), *- differences from Reference1 are significant (Multiple Range Test, ANOsA (r<0,01) .
8 Table 2 The spectrum of mitotic disturbances detected in the marrow cells of tistar male rats after exposure to router's electromagnetic radiation in different modes (%).
sersions of exposure
Number of cells with mitotic disturbances
Fragments Bridges Lagging
chromosomes
Multiple reconstructions
Significance of differences χ2 (ν= 3, r< 0,01) Reference1 (intact)
92 5 37 46 12 10.3 9.6 16.5 22.1
Reference2 (Faraday cage, 4 d x 6 hrs)
114 8 32.5 32.5 27
Router (4 d x 6 hrs)
352 9 18 59 14
Router+ resonators (4 d x 6 hrs)
127 5 33 30 32
Reference3 (Faraday cage, 3 weeks x 6 hrs;
96 9 26 40 25
Router (3 weeks x 6 hrs)
180 8 28 44 20
Router (2 hours)
149 7 29 33 31
Legend: vertical lines mean significantly different values.
DISCUSSION
The findings of the evaluation of chromosome aberrations in marrow cells of tistar male
rats revealed that router's electromagnetic radiation using different exposure modes (2 hours, 4
days x 6 hours a day, 3 weeks x 6 hours a day) has cytogenetic activity, can induce mitotic
disturbances. They can affect the functioning of immune system links related to the marrow's
operation.
The most significant changes in the chromosome apparatus were detected after exposure
of the animals in a Faraday cage with a plugged in router for 4 days, 6 hours a day – from 8 am
till 2 pm. That is the very mode to use for pronounced induction of mitotic disturbances in
9 tistar rats by means of high-frequency EMR for the purpose of subsequent research of the
efficiency of protective devices, as well as mechanisms underlying their protective properties.
Differences between the spectra of spontaneous EMR caused by exposure of animals in the
Faraday cage, as well induced EMR of a router in combination with the effect of matrix
resonators of chromosome disturbances can indicate a specific response of the animals'
organisms to the applied effects.
Change in stability of the genetic apparatus of dividing marrow cells under the influence
of a router's EMR can be viewed as the result of cell oxidation stress, whose mutagenic activity
is based on the genotoxic effect of internal causes of humoral nature and free-radical products of
peroxidation (Achudume et al., 2010). Those mechanisms can cause immunosuppression,
inhibition of immunopoesis and haematopoesis, which will affect the state of the organism.
However, information that long-term, 3-week-long exposure of animals to EMR conditions
decreases the level of mitotic disturbances relative to a 4-day-long session indicates that there
may be adaptive mechanisms causing elimination of cells with disturbances and/or activation of
reparative processes.
It should be noted that we were the first to demonstrate that Aires Defender fractal-matrix
resonators have a protecting effect on the genetic apparatus of dividing marrow cells in tistar
male rats under router's EMR effect. Its interaction with an Aires Defender causes structural
transformation of electromagnetic field (whabrev et al., 2005). In a counting number of
directions, electromagnetic field strengths concentrate, and in the other directions the
electromagnetic field reduces significantly, which in general weakens the sum-total
electromagnetic effect exceeding the organism's sensibility threshold to electromagnetic
radiation. Thus, resonators' protective properties are evidently based on restructuring
(conversion) of incident EMR, which reduces its damaging effect on dividing cells.
Significantly, among rats from the group using Aires Defender resonators there were
several subjects whose chromosome aberration level reduced even in comparison with the intact
reference. It is a very interesting fact that calls for further research and special experiments to
evaluate the effect of resonators proper on stability of the genetic apparatus in marrow cells of
rats.
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