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MARIUS A. SILAGHI*, ULRICH L. ROHDE* º, OVIDIU C. FRATILA**
,
HELGA SILAGHI*, TIBERIA IOANA ILIAS**
*Faculty of Electrical Engineering, University of Oradea
ROMANIA
masilaghi@uoradea.ro, hsilaghi@uoradea.ro º Brandenburgische Technische Universität Cottbus
GERMANY
rohdeu@tu_cottbus.de
** Medicine and Pharmacy Faculty , University of Oradea
ROMANIA
ovidiuf@rdslink.ro , tiberia_ilias@yahoo.com
Abstract: - Applications of microwave have been increased in the last years due to radars and police communication
systems, high power satellite and TV transmitters, mobile phones, microwave ovens and medical devices. Exposure to microwave emissions has a negative effect upon the general biological welfare of humans. The measurement of the
dielectric properties of biological tissues would play a significant role in any well-founded effort involving tissue
interaction with electromagnetic energy. Therefore we want to see whether current safety standard recommendations
are inappropriate as far as human blood function is concerned. Thirty age and gender matched blood samples will
serve as control. For increased accuracy detection of any possible alteration and to help us orientate towards the
details that are to be studied ultra structurally, we use a special stain. An electronic microscope analysis will be
performed on all the blood samples after specific preparation to see whether such microwave exposure might damage in some way the ultra structure of the blood cells. After modeling and sectioning the blocks we contrast them using
acetate-uranyl and lead-citrate and study them with an electronic microscope. The obtained images are processed
using Viewer Imaging Analysis software for further analysis. In the end we shall use scanning microscopy to detect
any surface dismorphisms that might appear due to the microwave exposure.
Key-Words: Exposure to microwave emissions, microscope analysis, ultrastructure of the human blood,
biological effects, modeling , simulation
1. Introduction Exposure to microwave emissions has a negative effect
upon the general biological welfare of humans. The
effects of microwaves might be divided in three
categories: cancer-causing effects, destruction of
nutritive value and biological effects of direct
exposure[2,8,10].
Applications of microwave have been increased in the
last years due to radars and police communication
systems, high power satellite and TV transmitters,
mobile phones, microwave ovens and medical devices.
The biological effects of this type of exposure on living
organisms have been studied by different
investigations[1,4,11]. Few studies were performed to
determine whether blood was affected by exposure to
microwave energies that equal or even exceed current
safety standard recommendations and most studies were
made on animals[9,12]. Thus, there is a need to study the
interaction of microwave with living organisms,
especially, its effect on biological materials . Therefore we want to see whether current safety
standard recommendations are inappropriate as far as
human blood function is concerned.
Approach on the High Frequency Electromagnetic Field
Effects on Human Blood
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ISSN: 1790-5117 50 ISBN: 978-960-474-193-9
2. Methods Our study involved blood samples from 30 otherwise
healthy subjects who were asked to attend the Clinical
County Hospital from Oradea, Romania. All participants
had had their history taking a physical examination to
detect symptoms and signs of any disease. All the data was recorded and further analysis was assessed.
Fig. 1. Microwave laboratory desk
After informed consent, a blood sample was taken from
all patients using syringes and it was later stored in
EDTA-treated sealed test tubes at 4°C in order to
prevent coagulation[3,5]. Afterwards we exposed
the blood from our patients to a microwave field using
the laboratory desk(Figure 1).
Fig. 2. Slightly abnormal erythrocytes
The experimental model for microwave processes is
composed from one microwave generator at 2450MHz,
and 1KW variable power as presented , which is
connected with interface to PC and printer ; the
magnetron type is TOSHIBA 2M248.
Fig. 3. Basophiles cells with normal ultrastructural
aspect
The electronic balance for the measure of weight is
for maximum 3100 g, with 0.1g precision and is
connected to PC. It is predicted also a dummy load
connected to water network from laboratory and
electrical generator for the microwave/hot-air (750W).
Fig. 4. Neutrophil cells with normal ultrastructural
aspect
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ISSN: 1790-5117 51 ISBN: 978-960-474-193-9
The experiments are to be performed in the
microwave system, the test glasses subsequently
subjected to different holding times. The control
treatment is performed in the same recipient using the
same temperatures and holding times as in the case of
the microwave treatment. In most cases we use the
microwave treatment at holding times above 100 s, and
temperatures as low as 50oC; for the capture of
temperature we use MIKRON infrared thermometer.
We follow the next assumptions: working frequency 2450 MHz dimensions to assure enough space for glass
recipient’s movement due to a non-stressful condition,
possibility to measure reflected and transmitted power
and homogenous electromagnetic field distribution to
assure an accurate exposure. To accomplish these
requirements we decided to design two types of
exposure chambers and compare their properties in order to find the best solution for our future work. As the most
appropriate chambers we choose parallel plate and
waveguide structures. The temperature responses during
each experiment are recorded. Samples are collected at
the various sampling sites, corresponding to different
retention times. Thirty age and gender matched blood
samples will serve as control[6,7]. So, in the end we
formed 2 groups:
Group 1, control blood, not treated with microwaves,
called CHB (control human blood)
Group 2, microwave treated blood called MTHB
(microwave treated human blood)
Fig. 5. Neutrophil cells with normal ultrastructural
aspect
An electron microscope analysis was performed on
all the blood samples after specific preparation to see
whether such microwave exposure might damage in
some way the ultrastructure of the blood cells.
After separating the serum from the blood cells, the
latter was prefixed with 2.7% glutaraldehide solution in
0.1M, pH 7.2 phosphate buffer resulting tiny tissue
blocks[9,12]. The postfixing process was done with 1%
osmium tetraoxide (OsO4). Then we added the second
fixing agent 1% OsO4 in 0.1M phosphate buffer at a 7.4
pH for 15 minutes. Using the aforementioned fixing
procedure we intended to stop the metabolic processes of
the tissue, to preserve the fine structure of the cells for
future preparing and optimal visualizing with the transmission electron microscope. For the inclusion
process we used an epoxidic pitch –Epon 812. For
dehydration we used acetone in water solution in rising
concentrations.
Fig. 6. PMN cells with degranulation process
The water extraction was done gradually, in short
times to ensure that the process is not followed by
structural alterations. The specimen encapsulation was
made using special plastic capsules followed by a
polymerizing process (temperature 50-60 C degrees for
72 hours) from which we obtain hard transparent blocks, containing inside the black pieces. We mention that at
every liquid changing during the processing, the blood
samples were spinned at 1000 rotations/minute in order
to remove the supernatant. From the aforementioned
blocks we first obtained semifine sections measuring
500 nm for light microscopy and later from the selected
zones of the same blocks we cut ultrathin sections measuring 40-60 nm for ultrastructural assessment. The
sections were made using a Leica UC6 ultramicrotom
which has DDK diamond knives.
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ISSN: 1790-5117 52 ISBN: 978-960-474-193-9
After modeling and sectioning the blocks we
contrasted them using acetate-uranyl and lead-citrate and
studied them with a JEM-1010 electron microscope. The
pictures were captured using a Megaview III camera.
The obtained images were processed using Viewer
Imaging Analysis software for further analysis.
3. Results
Electron microscopy of plasma membranes,
cytoplasm matrix, nuclei, cell organelles and non-
cellular components of peripheral blood was carried out
in both groups.
The overview images obtained from the semifine sections showed us the general situation of red blood
cells in CHB group, compared to the red blood cells
images in the MTHB group. We observed that in MTHB
group the red blood cells tend to became spherical,
phenomena probably due to the alteration of the
permeability membrane capacity.
Fig. 7. PMN cells altered structure
The ultrastructural images obtained at successive
magnification showed us the normal conformation of the
red blood cell population in CHB group, with
electrondense cytoplasm due to the presence of hemoglobin.
In CHB group the red blood cells are almost normal
in shape, the majority having the shape of a biconcave
disk from side view. There are also slightly abnormal
erythrocytes probably due to a long period from blood
collection till fixing processes (Fig2).
Among leucocytes we observe the apparition of
some basophiles (Fig.3) and neutrophil (Fig.4 and Fig.5)
cells which have a normal ultrastructural aspect.
In the MTHB group all the erythrocytes are oval or
round, many of them having a diluted content and some
of them being destroyed (hemolysis) so that fragments
from them are dispersed in the plasma serum. We also
observed that the PMN cells have altered structure
(Fig.6), abnormal plasmatic membrane so that they
suffer a degranulation process (Fig.7 and Fig.8).
Fig. 8. PMN cells altered structure
4. Conclusion
The results of the present study indicate that
microwaves per se might be harmful to human blood and
that poor penetrance of microwaves, together with
insufficient blood mixing during warming, are the
critical factors leading to hemolysis.
It is obvious that the applied microwaves have
induced alterations of the ultrastructure of the human
blood cells and organelles, processes that appear
especially due to the alteration of membrane
permeability capacity.
The ultrastructural assessment done in our research comprised especially the modifications which appear in
the human blood and red blood cells distinctively before
and after microwave treatment.
Using various and complex methods we can obtain
significant results which can therefore lead to
investigation methods for a better understanding of the
action mechanisms and evolution of our blood when it is submitted to microwave exposure so we think that future
studies are needed for a better clarification of the above
mentioned matters.
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Acknowledgment The authors would like to direct their warmest thanks to
Prof.Dr.H.C.Dr.Eng. Ulrich L.Rohde for the donation
which made the work possible.
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ISSN: 1790-5117 54 ISBN: 978-960-474-193-9