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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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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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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ISSN: 1790-5117 53 ISBN: 978-960-474-193-9

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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