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Research Article
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The Effects of Electromagnetic Field on the Microstructure
of Seminal Vesicles in Rat: A Light and Transmission Electron Microscope
Study |
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Amir Afshin Khaki,
Sina Zarrintan,
Arash Khaki
and
Afshin Zahedi
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ABSTRACT
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In the industrial world, almost everyone is unavoidably
exposed to ambient electromagnetic field (EMF) generated from various
technical and household appliances. Controversy exists about the effects
of EMF on various tissues of the living bodies. Seminal vesicles as one
of these accessory glands play an important role in natural seminal fluid
formation and the effects of EMF on its tissue is worthy of investigation.
In order to examine this 30 rat were selected and kept for one weeks in
quarantine and 15 (experimental group) were exposed to 50 Hz (non-ionizing
radiation) during postnatal life for 2 months. The materials were processed
and observed under a light and transmission electron microscope. In the
experimental rats epithelial and basal cells showed significant destructions
presented by heterochromatin and dense nuclei. Cell debris and abnormal
areas was recognizable in the stromal connective tissue. Obvious vacuolization
was present within the epithelial cell cytoplasm and also between the
cellular organelles. The nuclei of the endothelial cells of the blood
vessels were more rigid and endothelial cell cytoplasm contained a lot
of vacuoles and pinoctotic vesicles. The results suggested that EMF exposure
may cause profound changes in the vesicle seminal tissues. Therefore exposure
to EMF may result in pathological changes that lead to sub fertility and
infertility.
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INTRODUCTION In a modern world countless number
of people is exposed to an elevated electromagnetic field (EMF) with a
wide frequency range because of the ever increasing rise of utilization
of electric power for running domestic appliances and industrial gadgets.
The transport and use of electricity generates both electrical and magnetic
fields with a wide spectrum of frequencies, intensities and waveforms.
There are two types of EMF, ionizing and non ionizing. We usually associate
EMF being generated in relation to electrical substations, transformers,
overhead transmission and distribution lines, but significant other sources
of EMF exposure include endless list of house-hold gadgets and appliance
of everyday use such as laser printers, vacuum cleaners, electric shavers,
hair dryers, microwave ovens, television transmissions, cellular phones,
Video Display Terminals (VDT) etc. Accordingly, most mammalian reproductive
research has focused on these frequencies because of their ubiquitous
presence in the environment (Bracken et al., 1995; Chiang et
al., 1984; Quaglino, 2000).
One critical issue is whether EMF may potentially affect
the reproductive system. Many studies have reviewed the numerous outcomes
of the potential effects of EMF on infertility, miscarriage, premature
births, intrauterine growth retardation, low birth weight, congenital
malformations, genetic diseases and prenatal deaths. The possibility of
an association of early pregnancy loss with residual exposure has been
investigated by case-control studies (Juutilainen et al., 1993).
It has been shown that exposure to EMF adversely affects
spermatogenic cells, Sertoli cells, Leydig cells and boundary tissue of
the seminiferous tubules of the male reproductive system (Dym and Fawcett,
1970; Forgacs et al., 2004; Khaki et al., 2004, 2006; Lee
et al., 2004; Shafik, 2005). Although all the mentioned structures
play important roles in spermatogenesis, mechanical support and sperm
discharge (Chung et al., 2005; Lacy and Rotblat, 1960; Leeson and
Leeson, 1964; Yamamoto et al., 1987).
The seminal vesicles are male accessory sexual gland found
in many species of more than 4000 mammalian species alive on the earth
today. They lie inferior and lateral to the ampullae`s of the ducts deferens
against the fundus of the bladder. After puberty, the gland secretes a
fluid called Seminal Vesicle Secretion (SVS), which accumulates in its
lumen. SVS contains both protein and no protein components. When ejaculated,
SVS squirts into the urethra, contributing the major part of the liquid
portion of seminal plasma, which is the complex biological fluid formed
from mixing of various fluid in the male reproductive tract. It has been
found that extirpation of the seminal vesicle from mice and rats greatly
reduces fertility (Pang et al., 1979; Peitz and Olds-Clarke, 1986),
demonstrating the importance of SVS to sperm modification under natural
circumstances. It has been also demonstrated that protein and enzyme production
is dependent to testosterone, which is formed mainly by testes in males
(Mansson et al., 1981; Koenig et al., 1976). Moreover, epithelial
cell proliferation in the seminal vesicles seems to be testosterone dependent
in male mice (Tsuji et al., 1991; Justulin et al., 2006).
It is assumed that EMF exposure may have destructive effects on cytoarchitecture
and microstructure of seminal vesicles and accordingly, cause abnormal
or inadequate seminal fluid production and subsequent male sub fertility
and infertility. In addition, as mentioned earlier, EMF exposure could
have destructive effects on the testes (Dym and Fawcett, 1970; Forgacs
et al., 2004; Khaki et al., 2004, 2006; Lee et al.,
2004; Shafik, 2005) and thus, a deficiency in blood testosterone can alter
epithelial proliferation and protein synthesis in seminal vesicles. There
are little known about the effects of non-ionizing EMF on microstructure
of seminal vesicles and that is why the authors wanted to investigate
the possible effects of exposure to 50 Hz EMF (non-ionizing radiation)
on the cytoarchitecture and microstructure of the seminal vesicles during
postnatal periods no light and transmission electron microscopy. The harmful
effects of EMF ionizing radiations (e.g., X-rays and gamma rays) have
previously been demonstrated on gonadal tissues (Lee et al., 2004;
Cecconi et al., 2000; De Vita et al., 1995; Lokmatova, 1993;
Parsons, 1962).
Elbetieha et al. (2002) demonstrated that exposure
to EMF (50 Hz, 25 mT for 90 days) had no significant effect on the weight
of the testes or the number of implantation sites and viable fetuses.
The result of this study is of potential use, as exposure
to electromagnetic waves is ubiquitous; a large portion of the world`s
population is constantly exposed to a variety of this radiation as a result
of professional, residential, medical, industrial or other uses.
MATERIALS
AND METHODS
Animals and maintenance: A total
of 30 male Wistar rats (of approximately 5 weeks old) were used for the
study. Rats were housed in cages and kept in quarantine for one week to
rule out any disease. Rats were fed on compact food in the form of granules
and water. This food consisted of all the essential ingredients, including
vitamins and minerals. The environmental conditions (temperature and humidity)
in all the animal holding areas were continuously monitored. Temperature
was maintained in the range of 20-30°C and relative humidity was monitored
at 35-60%. Fluorescent light was provided on a 12 h light/dark cycle and
kept turned on from 8 am till 8 pm. Lights (electric fluorsent) were located
at a distance of three meters from the cages so that these did not interfere
with EMF of the experimental design.
EMF-producing system: The equipment was based on the Helmholtz
coil, which works following Fleming`s right hand rule. This produced an
alternate current of 50 Hz, creating an EMF of 80 G. The intensity of
the EMF could be controlled by a transformer. The equipment had two main
parts. In the first there were two copper coils placed one above the other
and separated by a distance of 50 cm. Between the coils (the exposure
area) there was a cylindrical wooden vessel, the interior of which had
a chamber for holding the cages of the experimental animals. The second
part was the transformer, which checked the input and output voltage with
a voltmeter and the current with an ampere meter. To prevent increases
in temperature inside the chamber a fan was utilized as necessary. Five
cages at a time were placed within the chamber with seven or eight rats
per cage (Fig. 1).
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Fig. 1: |
Electromagnetic
field producer system |
EMF exposure: Of the total of 30 rats of the experiment, 15 were
selected as experimental group and 15 as control group. In experiments
group, rats were exposed to EMF 8 h per day for two months. The control
group had the same biological and nutritional condition as the experimental
group during these two months and the difference was only EMF exposure.
The entire rats, both experimental and control groups, were anaesthetized
and sacrifices at the end of the two-month EMF exposure and were sent
to tissue fixation laboratory.
Tissue fixation: At the termination of the stipulated exposure
period as laid down in the experimental design the rats were anaesthetized
with chloroform and 10% formalin was then injected through the inferior
vena cava. The seminal vesicles were removed and fixed in formalin for
light microscopy. Haematoxylin and eosin were used to stain the 6 mm thick
histological sections.
Transmission electron microscopy: For Transmission Electron Microscopy
(TEM) the tissue samples were cut into pieces (2x2 mm) and fixed in 2.5%
glutaraldehyde and 1% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4)
for 6-8 h at 4°C. They were washed and postfixed in 2% OsO4
for 1 h at 4°C. The tissue was dehydrated through ascending grades of
ethanol and embedded in araldite CY212. Semithin sections (1 µm) were
cut and stained with toluidine blue. Ultrathin sections (60-70 nm) were
cut, mounted onto copper grids and stained with uranyl acetate and alkaline
lead citrate. Sections were observed under a Philips CM10 transmission
electron microscope.
Data analysis: All data were expressed as means±SD. All statistical
analyses were performed using SPSS software, version 13.0, on the basis
of the student`s t-test. A p-value less than 0.01 were considered significant.
Leven`s test for the equality of the means was also done prior to the
t-test.
RESULTS
Light microscopy Control group: In morphological investigation
of the light micrographs of the control group, seminal vesicles were seen
like highly twisted tubules (Fig. 2A). Each glad had
a musculoelastic capsule (Fig. 2E), the smooth muscle
of their wall had organized and recognizable fibers their nuclei were
euchromatic and visible (Fig. 2B). The musculoelastic
capsule was lined by two layers of cells. Secretary epithelial cells and
basal cells. The basal cells seemed to act as proliferate cells and the
pseudo stratified epithelial cells had secretary, protein synthesizing
appearance (Fig. 2D). The nuclei of the basal and secretary
cells were detected euochromatic (Fig. 2D). Inside of
the glands were seen twisted and honeycomb by low magnification, because
of penetration of capsular connective tissue. The transverse sections
of blood vessels were seen in the spaces between the secretary tubules,
the endothelial cells of the vascular walls and their nuclei were also
visible and the Red Blood Cells (RBCs) were recognizable (Fig.
2C).
Experimental group: Light micrographs of the experimental group
illustrated cell and tissue damage in musculoelastic capsule. Muscular
fibers were destructed and pathological, abnormal areas were numerous
in the capsule (Fig. 3). Smooth muscle fibers did not
demonstrate their organized arrangement. They were spread out in various
directions. The nuclei of the smooth muscle and epithelial cells were
heterochromatic and dense (Fig. 3B, C
and D, Table 1). The epithelial cell
layers were unorganized abnormal spaces were seen between the cells mentioned
above clearly. Spaces between the smooth muscle cells seemed abnormal
(Fig. 3A). The epithelial and basal cells nuclei were
significantly destruct ore presented by heterochromatin and dense appearance
(Fig. 3C and D, Table 1).
It seemed that the number of basal cells had been reduced (Fig.
3C). The cytoplasm of the epithelial cells showed an obvious decline
(Fig. 3D) Cell debris was recognizable in the stromal
connective tissue (Fig. 3E). Abnormal areas and frothy
spaces were seen together with the cell debris. Blood vessels showed a
thinner wall than normal and the nuclei of endothelial cells were dense.
Hyper perfusion of blood and an increase in RBCs were seen within the
vessels (Fig. 3F).
Transmission
electron microscopy findings
Control group: Epithelium of the seminal vesicle is in fact consisted
of high columnar cells. Apical parts of epithelial cells were stretched
to the inside of lumen and the free border had numerous microvilli in
different sizes. There were specific intercellular canals between the
apexes of epithelial cells. Lateral and basal parts of these cells were
interdigitated into each other (Fig. 4A). The nuclei
of the epithelial cells were located in range of base to the middle. Each
nucleus had a nucleolus together with its condensed chromatin (Fig.
4B). Rough endoplasmic reticulum was seen widely in the basal part
of the secretary epithelium. Their cistern were also wide, long and parallel
to each other. Ribosomes were spread in the cytoplasm. Golgi complexes
were seen to occupy a vast area at the top of
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Fig. 2: |
Light micrographs
of the seminal vesicle tissue of the control group, epithelial
cells, basal cells and smooth muscle cells are seen (A), smooth
muscle cell and its nucleus is shown by an arrow (B), blood
vessel and nucleus of the endothelial cell is shown by an arrow
(C), red blood cells (RBCs) are seen within the blood vessels
(D) and the smooth muscle cell in the seminal vesicle wall is
shown by a strike (E). Bar : 100 µm |
 |
Fig. 3: |
Light micrographs
of the seminal vesicle tissue of the experimental group, abnormal
spaces between the smooth muscle cells of the seminal vesicle
wall is shown (A), heterochromatinism is seen in the nuclei
of the smooth muscle cells are heterochromatin (B), the nuclei
of the epithelial cells are dense and exhibit heterochromatinism
(C), dense and heterochromatin nuclei of the epithelial cells
(D), cell debris in the connective tissue of the |
Table 1: |
Effects of
60 Hz EMF on vesicle seminal tissue |
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EG: Experimental Group; CG:
Control Group; ECs: Epithelial Cells; BCs: Basal Cells. The
number of heterochromatinic epithelial cells, ratio of dilated
to dense mitochondria and numbers of cytoplasmic vacuoles within
the epithelial and basal cells have been shown; a,b,c,d:
Comparison between groups show significant statistical differences
with p-value <0.01 |
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Fig. 4: |
Electron micrographs of the seminal
vesicle tissue of the control group, epithelial and basal cells
are shown by a strike and microvilli are shown (A), nuclei of
the epithelial cells and their nucleoli, mitochondria are also
seen (B), mitochondria of the epithelial cells (C), smooth muscle
cells and their nuclei (D) and blood vessels and nuclei of the
endothelial cells (E). Bar : 1 µm |
the epithelial cells. A number of vacuoles were also present and those
that were located at the apical border of the cells contained a dark and
dens material (Fig. 4A). Mitochondria were recognized
dense nearby the endoplasmic reticulum. They had clear walls together
with numerous cristae. The spaces between the mitochondrial cristae were
also normal and clear (Fig. 4C). Basal cells were visible
nearby the basal part of the secretary epithelial cells. These cells are
small, satellite shaped and their nuclei are located centrally. Basal
cell nuclear chromatin was condensed and there were not any clear nucleoli
within the nuclei. Golgi complexes were seen to be located near the cell
nucleus and there were some ribosome`s within the cytoplasm. The rough
endoplasmic reticulum, meanwhile, was wide and dilated. Mitochondria were
found in a large amount within the cytoplasm and they contained a number
of fatty particles. Basal cells act as supplementary and proliferate cells
to differentiate into epithelial cells when necessary (Fig.
4A). The smooth muscle of the capsule had spindle-shaped cells, which
contained oval, central nuclei. Meanwhile, myofilamants were located longitudinally
within the muscular tissue. Smooth muscle cells` cytoplasm contained a
small Golgi complex and the mitochondria were seen clearly. Ribosome`s
were seen as specific masses within the cytoplasm (Fig. 4D).
Blood vessels had clear walls and endothelial cells and their nucleoli
were visible. Basement membrane and the connective tissue were also seen
in the vascular walls. The nucleoli of the endothelial cells were oval
shaped and wide and they were curved into the lumen of the vessels. The
endothelial cells contained mitochondria and a Golgi complex. There were
also a
 |
Fig. 5: |
Electron micrographs
of the seminal vesicle tissue of the experimental group, epithelial
and basal cells and their nuclei, note the microvilli, vacuoles
and lysosomes (A), nuclei of the epithelial cells, note the
vacuolization (B), basal cell (is shown by the letter B in part
C) increasing of dilated mitochondria in basal cells is seen
and blood vessels, vacuolization is also seen in the cell cytoplasm
(C), reduction of microvilli (mic) and increasing of vacuoles
(vac) at the apical border of the epithelial cells, note that
the Golgi complex (G) is dilated (D) and nuclei of the smooth
muscle cells (N) and their nucleoli, note the abnormal spaces
between the cells (E). Bar : 1 µm |
number of ribosomal masses in the cytoplasm. A great amount
of pinoctotic vesicles were seen in the cytoplasm. These vesicles might
probably have played a role in the transport of various materials (Fig.
4E).
Experimental group: Secretary epithelial cells were seen to save
their columnar shape but the microvilli located in their apical border
were reduced significantly (Fig. 5A). Intercellular connections
were seen normal and as described for control group i.e., interdigitated.
The nuclei of the epithelial cells were located the same as the control
group. The nucleoli were absent or unclear in the experimental group.
The chromatins were dense and located marginally near the nuclear membrane
(Fig. 5B). Rough endoplasmic reticule were dilated and
located in the basal part of the epithelial cells Golgi complex was located
in the apical part of the cells and it was dilated too (Fig.
5D). Obvious vacuolization demonstrated within the cytoplasm which
was more significant in the apical part of the epithelial cells. A number
of the vacuoles contained the large liposome`s. Moreover, the cytoplasm
itself had a number of liposomes. Mitochondria were electron opaque which
were considered dilated or none energized (Fig. 5B, Table
1). Basal cells were present among the epithelial cells in the experimental
group. These cells were tiny and satellite-shaped. Their nuclei were lied
centrally; they were heterochromatic and picnotic. A number of basal cells
showed fragmented nuclei. An abnormal area was illustrated around the
nuclei of the basal cells, which contained a great number of mitochondria.
The mitochondria were dilated. Endoplasmic reticulum and Golgi apparatus
seemed dilated too. The cytoplasm contained a lot of liposome`s (Fig.
5C). Vacuolization was seen as a clear appearance within the cellular
cytoplasm both in epithelial and basal cells. A lot of vacuoles were seen
in the cytoplasm and intercellular spaces (Fig. 5A, B,
C and D, Table 1). The smooth muscle
cells were spindle-shaped and their nuclei lied in the central localization.
Heterochromatic and dense nuclei were seen in the marginal zone of the
nucleus i.e., near the nuclear membrane. There were numerous spaces between
the muscular cells. Cytoplasm of the muscular cells had a lot of small
vacuoles and in turn, it indicates the presence of vacuolization in the
smooth muscle cells. Myofilamants were disoriented from the longitudinal
axis. Muscular cell mitochondria showed dilation and were decreased in
amount (Fig. 5E). Within the blood vessels, oval-shaped
nuclei of the endothelial cells presented dense chromatins in the marginal
zone of the nucleus. The cytoplasm of the endothelial cells contained
a lot of vacuoles and pinoctotic vesicles were present. Basement membrane
was ruptured in some areas. Vascular connective tissue showed vacuoles
and abnormal spaces (Fig. 5C).
DISCUSSION
A number of studies have assessed the harmful effects of
X-irradiation on vesicle seminal tissue and their secretary activities
in rat (Gupta and Bawa, 1970; Kotscher and Voelkel, 1957; Melampy et
al., 1956; Roeske et al., 1995) evaluated the effects of radiation
therapy on the size and location of the prostate, seminal vesicles, bladder
and rectum in patients with localized prostate carcinoma and they found
that changes in the location of the prostate, seminal vesicles and normal
tissue volumes during the course of radiation therapy occur and have dosimeter
consequences that may impact tumor control and normal tissue complication
probabilities. Chan and Kressel (1991) have also evaluated the effects
of pelvic irradiation on prostate and seminal vesicle tissues. They revealed
that in the irradiated patient, the prostate and seminal vesicle can develop
several patterns of signal intensity abnormalities; in particular, diffuse
low signal intensity in the prostate and seminal vesicle should establish
radiation fibrosis as an important differential diagnosis to consider.
McGivern et al. (1990) revealed that low-frequency intermittent
EMF exposure during the critical prenatal period for neurobehavioral sex
differentiation can demasculinise male scent-seeking behavior and increase
the weight of accessory sex organs in adulthood. Lundesberg et al.
(1995) found no association between occupationally related categories
of EMF exposure and male sub fertility as evaluated by sperm morphology,
motility and concentration. Chung et al. (2005) showed that exposure
to EMF (from 60 Hz up to 500 mT), both prenatal and postnatal, did not
alter offspring spermatogenesis in the rat. Khaki et al. (2004,
2006) assessed the effects of 50 Hz EMF (non-ionizing radiation) during
in utero development and postnatal life on rat testicular tissue and revealed
that exposure to EMF have a destructive effect on Sertoli cells and the
boundary tissue of the seminiferous tubules. In this study we demonstrated
the effects of EMF (non-ionizing) on seminal vesicle tissue of rat investigated
by light and transmission electron microscopy. Light Micrographs (LM)
studies in experimental group revealed that epithelial cells had been
unorganized, damaged and destructed containing heterochromatin zed and
dense nuclei. The number of basal cells was reduced in amount (Hayward
et al., 1996). Transmission Electron Micrographs (TEM) of the experimental
group showed in turn heterochromatizim and dense nuclei in the epithelial
and basal cells. Obvious vacuolization was present inside these cells
and within the intercellular spaces. Mitochondria were dilated significantly,
which were considered none energized i.e., nonfunctioning. Rough endoplasmic
reticule had dilated cistern. Hence, epithelial cells changed to present
abnormal mitochondria, rough endoplasmic reticule and nuclei. Seminal
vesicles secret an exocrine viscous yellowish fluid composed of fructose,
citrate, prostaglandins and a number of proteins. These secretions are
essential for a normal seminal fluid formation. Thus, the destructed epithelial
cells would be deficient in their contribution in the normal seminal fluid
formation and cause subsequent sub fertility and infertility (Curry and
Atherton, 1990; Brewster, 1985). Our ultrastructrual findings confirmed
that the following reduction of organelles i.e., SER, mitochondria, Golgi
apparatus, synthesis of testosterone probably suppressed. Similar effects
demonstrated by other investigators previously. It was concluded that
the reduction of gondotropins resulted in reduction of testosterone that
impair the spermatogenesis. Seminal vesicle products` decline could also
occur in response to the lack of testosrone resulting from damaged testicular
tissue esp. Leydig cells (in addition to the seminal vesicles, EMF exposure
adversely affects the testes (Dym and Fawcett, 1970; Forgacs et al.,
2004; Khaki et al., 2004, 2006; Lee et al., 2004; Shafik,
2005), because secretary activity of the seminal vesicle epithelial cells
is testosterone dependent (Tsuji et al., 1991; Justulin et al.,
2006). The damaged basal cells would also fail to perform their proliferate
and supplementary activities. Smooth muscle cell nuclei were also heterochromatin
zed and dense. Mitochondria were dilated and vacuolization were present,
in their cytoplasm. The muscular cells showed pathological spaces between
them. The myofilaments were disoriented from their longitudinal axis.
Pathologic changes may lead to the muscular dysfunction of the seminal
vesicles. Thus it caused deficient release of its products and abnormal
seminal fluid formation. Myoid cells, which are modified smooth muscle
cells and probably maintain a certain pressure in order to facilitate
sperm discharge (Lacy and Rotblat, 1960), undergo destruction under the
EMF exposure. Similar smooth muscle cell destruction was seen in the seminal
vesicle tissue of the experimental animals of the present study. In the
study done by Khaki et al. (2006) it was found that the fourth
layer of the boundary tissue of the seminiferous tubules of the EMF exposed
rats was thin and composed of lymphatic and endothelial cells, which formed
an extensive system of per tubular lymphatic sinusoids. The irregular
gaps and formation of blisters with a break in the endothelium of the
lymphatic could be responsible for lack of lymph drainage and the resultant
edema, which was evident from the frothy spaces among the seminiferous
epithelial cells under LM. In this study, equivalent endothelial cell
destruction was present as well evidenced by dense chromatin nuclei, vacuolization
and pinoctotic vesicles. In addition, hyper perfusion, RBCs accumulation,
basement membrane rupturing and vascular connective tissue vacuolization
were present. EMF has determinately effect in the suppression of immune
suppressive affect which my lead to increase the harmful effects of EMF
by increase the free radicals, whatever had destructive effects on the
cells and tissues. The present study revealed the harmful effects of EMF
on the seminal vesicle tissue in rats. At the molecular level EMF produces
biological stress and free radicals, which can make the susceptible animal
population prone to congenital malformations, tissue and cell damage or
death (Lai and Singh, 2004; Soeradi and Tadjudin, 1986; Wolf et al.,
2005). Free radicals released can cause oxidative stress at the cellular
level, interfering with protein synthesis. These elements also play an
important role in acute inflammation, endothelial destruction, increased
vascular permeability and exudation of plasma, resulting in tissue edema.
It has been postulated that short-term exposure to EMF produces high levels
of oxidative stress as a result of its effect on the immune response (Zhitkevich
et al., 2001) and long-term exposure to EMF may be linked to even
higher levels of oxidative stress (Fernie et al., 2000).
CONCLUSION
Although we can not surely connect non-ionizing
radiation (EMF) to various disease or damage of biological system, this
study suggested that to achieve a real measure for evaluation the effects
of EMF on glands of male genital system, more and extended investigations
in laboratories are necessary. Generally these findings indicated that
exposure to EMF had a deleterious effect on seminal vasicule gland in
rat, thus result in irreversible effects which may lead to sub fertility
and infertility in male.
It seemed that if results of animal and epidemiologic
studies can be combined, we will have a vivid conclusion about the connection
of EMF that is used in electricity transfer, transportation and etc, with
public health.
ACKNOWLEDGMENTS
The electron microscope work was carried out at
the SAIF, AIIMS and New Delhi, India. The assistance of Dr. T.C. Nag is
acknowledged.
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