The semen quality of the mobile phone users

Abstract

Background:
The increased use of mobile phones, the media's attention for general health, and the increase of idiopathic male infertility suggest to investigate the possible consequences of an excessive use of mobile phones on semen quality.

Aim:
To evaluate the conventional and some of the main biofunctional sperm parameters in healthy men according to the different use of the mobile phone.

Subjects and methods:
All the enrolled subjects in this study were divided into four groups according to their active cell phone use: group A= no use (no.=10 subjects); group B= <2 h/day (no.=16); group C= 2-4 h/day (no.=17); and group D= >4 h/day (no.=20). Among the subjects of the group D (>4 h/day), a further evaluation was made between the "trousers users"(no.=12) and "shirt users"(no.=8), and they underwent semen collection to evaluate conventional and biofunctional sperm parameters (density, total count, morphology, progressive motility, apoptosis, mithocondrial membrane potential, chromatin compaction, DNA fragmentation).

Results:
None of the conventional sperm parameters examined were significantly altered. However, the group D and the trousers users showed a higher percentage of sperm DNA fragmentation compared to other groups.

Conclusion:
These results suggest that the sperm DNA fragmentation could represent the only parameter significantly altered in the subjects who use the mobile phone for more than 4 h/day and in particular for those who use the device in the pocket of the trousers.

Full text

970
J. Endocrinol. Invest. 36: 970-974, 2013
DOI: 10.3275/8996
ABSTRACT. Background: The increased use of mobile phones,
the media’s attention for general health, and the increase of
idiopathic male infertility suggest to investigate the possible
consequences of an excessive use of mobile phones on se-
men quality. Aim: To evaluate the conventional and some of
the main biofunctional sperm parameters in healthy men ac-
cording to the different use of the mobile phone. Subjects
and methods: All the enrolled subjects in this study were di-
vided into four groups according to their active cell phone
use: group A= no use (no.=10 subjects); group B= <2 h/day
(no.=16); group C= 2-4 h/day (no.=17); and group D= >4
h/day (no.=20). Among the subjects of the group D (>4
h/day), a further evaluation was made between the “trousers
users”(no.=12) and “shirt users”(no.=8), and they underwent
semen collection to evaluate conventional and biofunctional
sperm parameters (density, total count, morphology, pro-
gressive motility, apoptosis, mithocondrial membrane po-
tential, chromatin compaction, DNA fragmentation). Results:
None of the conventional sperm parameters examined were
significantly altered. However, the group D and the trousers
users showed a higher percentage of sperm DNA fragmen-
tation compared to other groups. Conclusion: These results
suggest that the sperm DNA fragmentation could represent
the only parameter significantly altered in the subjects who
use the mobile phone for more than 4 h/day and in particular
for those who use the device in the pocket of the trousers.
(J. Endocrinol. Invest. 36: 970-974, 2013)
©2013, Editrice Kurtis
INTRODUCTION
There is a growth of interest about the problem of the
mobile phone’s use and its potential impact on the male
fertility (1, 2). The increased use of mobile phones (3),
the increased attention of the media for the general
health (2), and in particular, in the case of the androlog-
ical diseases, the increase in the rate of idiopathic male
infertility (without apparent causes) (4) represent in our
opinion a justified reason to investigate this problem from
a clinical point of view.
The data search on Medline using the following key-
words, “mobile phone and male infertility” or “mobile
phone and sperm” or “mobile phone and semen pa-
rameters”, found a limited number of articles although
well distributed among several studies concerning ex-
perimental models (5-12) and a series of clinical evi-
dences (13-19). Recently, our group published a review
on this topic (20), confirming the need to improve the
knowledge through the clinical trials.
On the basis of these premises, the aim of this study was
to evaluate the quality of the conventional and some of
the main biofunctional sperm parameters of a selected
series of healthy men comparing the results between the
mobile phone “regular users” and “non-users”.
SUBJECTS AND METHODS
Design of the study
Observational study.
Subjects recruitment
Clinical evaluation of all patients consecutively referred to our
center of Andrology in the year 2012.
Subjects selection
Sixty-three healthy and fertile men (all studied subjects induced
pregnancy in the last year), with normal weight [body mass index
(BMI) range 19.0-24.5 kg/m2], aged between 18 and 35 yr, and
non-smoking were carefully selected for enrollment in this study.
The examined patients were selected from an original popula-
tion (consisting of 250 men, aged between 18 and 35 yr and
with BMI range 19.0-32.0 kg/m2) observed with the opportuni-
ty of the andrological screening (regular prevention of male in-
fertility) made in the past 2 yr in the institutes involved in the
study. To exclude subjects with the concomitant presence of an
andrological disease known as able to alter sperm convention-
al and biofunctional sperm parameters, a complete medical his-
tory was collected from each of them. All men with a negative
anamnesis underwent a careful physical examination and labo-
ratory (routine blood testing, sperm analysis, sperm culture, and
urethral swabs) and ultrasound instrumental (scrotal and tran-
srectal scans) evaluation. Men with systemic (21) and endocrine
diseases (22), male accessory gland infection (23), past or present
cryptorchidism (24) or varicocele (25), microrchidism (26),
cigarette smoking (27), alcohol (28) and/or drug abuse, and re-
cent hormonal treatment were excluded.
In particular, the threshold values used to exclude cases of hy-
pogonadism and hypothyroidism were the following: total
testosterone <3 ng ml–1 or 10.4 nmol l–1 (29); thyrotropin (TSH)
>4.5 µU ml–1 (30).
Overweight or obese subjects and smokers were excluded from
Key-words: Mobile phone, semen quality, users.
Correspondence: S. La Vignera, Section of Endocrinology, Andrology and Internal
Medicine, Department of Medical and Pediatric Sciences, University of Catania, Poli-
clinico “G. Rodolico”, Via S. Sofia 78, Building 4, Room 2C18, 95123 Catania, Italy.
E-mail: sandrolavignera@unict.it
Accepted May 9, 2013.
First published online May 30, 2013.
The semen quality of the mobile phone users
R. Rago1, P. Salacone1, L. Caponecchia1, A. Sebastianelli1, I. Marcucci1, A.E. Calogero2, R. Condorelli2,
E. Vicari2, G. Morgia3, V. Favilla3, S. Cimino3, A.F. Arcoria2, and S. La Vignera2
1Unit of Andrology and Pathophysiology of Reproduction, S.M. Goretti Hospital, Latina; 2Section of Endocrinology,
Andrology and Internal Medicine, Department of Medical and Pediatric Sciences, University of Catania; 3Department of Urology,
University of Catania, Catania, Italy
©2013, Editrice Kurtis
FOR PERSONAL USE ONLY

Cellular devices and male infertility
971
the study for the possible consequences that these conditions
may have on conventional and unconventional sperm parame-
ters, as demonstrated in other studies of our and other groups
(31-33).
Table 1 shows the clinical, hormonal and ultrasound character-
istics of the sample and the four subgroups.
All subjects enrolled in this study underwent semen collection to
evaluate conventional and biofunctional sperm parameters.
The patients were divided into four groups according to their ac-
tive cell phone use (assessed through a specific questionnaire):
group A= no use (no.=10 subjects); group B= <2 h/day (no.=16
subjects); group C= 2-4 h/day (no.=17 subjects); and group D= >4
h/day (no.=20 subjects). Among the subjects of the group D (>4
h/day) a further evaluation was made between the “trousers
users”(no.=12 subjects) and “shirt users”(no.=8 subjects)
(“trousers users” were considered the men who usually carry the
phone in the pocket of the pants and “shirt users” were consid-
ered the men who usually carry the phone in the pocket of the
shirt). All the examined subjects have used the same mobile
phone device [UMTS/HSDPA/HSUPA (850, 900, 1900, 2100 MHz)]
during the last year and reported that the device was turned on
during the 24 h of the day (data obtained by questionnaire).
The protocol was approved by the internal Institutional Review
Board and an informed written consent was obtained from each
men.
Sperm analysis
Two semen samples (7-10 days apart) were collected by mas-
turbation after 3-5 days of sexual abstinence. After liquefaction,
they were analyzed according to the World Health Organization
criteria (34). The remaining spermatozoa were used for flow cy-
tometry analysis.
Measurement of serum hormone concentrations
The hormone assays were performed by electrochemilumines-
cence with a Hitachi-Roche device (Cobas 6000, Roche Diag-
nostics, Indianapolis, IN, USA). The reference intervals were as
follows: TSH= 0.3-4.2 mUI ml–1, luteinizing hormone (LH)= 1.6-
9.0 mUI ml–1, follicle stimulating hormone (FSH)= 2.0-12.0 mUI
ml–1, 17β-estradiol= 8.0-43.0 pg/ml, total testosterone= 2.8-
8.0 ng ml–1, prolactin= 4.0-15.0 ng ml–1.
Ultrasound evaluation
The testicular and epididymal regions were carefully assessed
by scrotal ultrasound using a 7.5 MHz linear transducer. The
prostate-vesicular region was assessed by transrectal ultrasound
using a 7.5 MHz biplan biconvex transducer (Esaote GPX Megas,
Genoa, Italy). All patients underwent ultrasound evaluation be-
fore and after ejaculation, after sexual abstinence of 4 days.
The ultrasound parameters evaluated were: 1) Testicular volume
evaluated by using the formula of the ellipsoid (35); 2) Cranio-
caudal diameter of the epididymal head; 3) Cranio-caudal di-
ameter of the epididymal tail; 4) Prostate volume measured by
using the planimetric method (36-38). The three maximum di-
ameters (lateral-lateral, anterior-posterior and longitudinal) of
the prostate were calculated and prostate volume was expressed
using the mathematical formula of the ellipsoid (diameter 1 ×
diameter 2 × diameter 3 × 4/3 × π); 5) Anterior-posterior diam-
eter of the body of seminal vesicles.
Sperm flow cytometry evaluation
Flow cytometry was performed using the flow cytometer EPICS
XL (Coulter Electronics, IL, Italy), as previously reported (39) to
evaluate sperm mitochondrial function (after 5,5’,6,6’-tetra-
chloro-1,1’,3,3’-tetraethyl-benzimidazolylcarbocyanine chloride,
JC-1, staining), phosphatidylserine (PS) externalization (follow-
ing annexin V/propidium iodide (PI) double staining), chromatin
compactness (following PI staining), and DNA fragmentation (us-
ing the TUNEL assay).
JC-1 staining: Mitochondrial membrane potential (MMP) was
evaluated by staining with JC-1 (Space Import-Export, Milan,
Italy), as previously reported (39). Briefly, the sperm suspension
was adjusted to a density of 0.5-1 × 106cells/ml and incubated
with JC-1 for 10-15 min at 37 C in the dark.
Annexin V/PI assay: Staining with annexin V/PI was performed
using a commercially available kit (Annexin V-FITC Apoptosis
detection kit, Beckman Coulter, IL), as previously reported (39).
Briefly, an aliquot containing 0.5 × 106spermatozoa/ml was re-
suspended in 0.5 ml of binding buffer, labelled with 1 µl of an-
nexin V-FITC plus 5 µl of PI, incubated for 10 min in the dark,
and immediately analyzed. Signals were detected through FL-1
(FITC) and FL-3 (PI) detectors. The different labeling patterns in
the bivariate PI/annexin V analysis identified different cell pop-
Parameter Group A Group B Group C Group D
no use (no.=10) <2 h/day (no.=16) 2-4 h/day (no.=17) >4 h/day (no.=20)
Age (yr) 29.0±6.0 27.5±5.5 30.0±5.0 28.5±4.0
BMI (kg/m2) 21.0±3.0 23.5±2.0 22.0±3.0 22.5±2.5
FSH (mUI ml–1) 4.0±2.0 3.6±2.2 5.2±2.2 3.6±2.6
LH (mUI ml–1) 5.5±2.0 4.5±2.2 5.2±1.3 4.7±2.0
Total testosterone (ng ml–1) 6.6±1.6 6.3±2.0 7.0±1.4 6.7±1.2
17β-estradiol (pg/ml) 30.2±6.0 26.2±5.5 25.0±12.0 27.5±8.5
Prolactin (ng ml–1) 12.2±6.0 14.5±5.5 11.0±6.0 14.5±4.0
TSH (mUI ml–1) 2.2±1.2 2.4±1.6 1.9±1.3 1.7±2.0
Testicular volume (ml) 22.5±3.4 21.8±4.2 24.2±2.6 22.3±4.2
Cranio-caudal diameter of the epididymal head (mm) 11.2±0.6 10.5±1.2 11.0±0.5 10.8±1.0
Cranio-caudal diameter of the epididymal tail (mm) 4.6±0.6 5.2±0.3 5.3±0.6 4.9±1.0
Prostate volume (ml) 26.6±4.5 24.8±5.6 25.0±7.0 24.2±8.0
Anterior-posterior diameter of the body of the seminal vesicles (mm)
9.2±3.0 10.6±2.3 8.8±2.2 9.5±2.7
BMI: body mass index; FSH: follicle stimulating hormone; LH: luteinizing hormone;TSH: thyrotropin.
Table 1 - Clinical, hormonal and ultrasound characteristics of the sample and the four subgroups.
©2013, Editrice Kurtis
FOR PERSONAL USE ONLY

R. Rago, P. Salacone, L. Caponecchia, et al.
972
ulations: annexin negative and PI negative were designated as
alive cells; annexin positive and PI negative as PS externalized
spermatozoa (early apoptotic cells).
PI staining: Sperm PI staining was performed as previously re-
ported (39). Briefly, semen samples were centrifuged at 500 g for
10 min at room temperature, the supernatant removed and sper-
matozoa collected. An aliquot of about 1 × 106spermatozoa
was incubated in LPR DNA-Prep Reagent containing 0.1% potas-
sium cyanide, 0.1% NaN3, non ionic detergents, salts and sta-
bilizing (Beckman Coulter, IL, Milan, Italy) in the dark, at room
temperature for 10 min and then were incubated in Stein DNA-
Prep Reagent containing 50 µg/ml of PI (<0.5%), RNAsi type A
(4 Kunits/ml), <0.1% NaN3, salts and stabilizing (Beckman Coul-
ter, IL) in the dark, at room temperature for 30 min.
TUNEL assay: TUNEL assay was carried out using the Apoptosis
Mebstain kit (Beckman Coulter, IL, Milan, Italy), as previously re-
ported (39). The negative control was obtained by not adding
TdT at the reaction mixture; the positive control was obtained by
pre-treating spermatozoa with 1 µg/ml of RNAse-free deoxyri-
bonuclease I (Sigma Chemical) at 37 C for 60 min before label-
ing. The debris was eliminated following the same procedure
described above.
Statistical analysis
Results are reported as mean±SEM thoughout the study. Con-
ventional sperm parameters were submitted to statistical analy-
sis as the mean of the two determinations obtained from each
men enrolled. The data were analyzed by 1-way analysis of vari-
ance (ANOVA) followed by the Duncan Multiple Range test. Cor-
relation analysis was conducted by Pearson correlation test. The
software SPSS 9.0 for Windows was used for statistical evaluation
(SPSS Inc., Chicago IL, USA). A statistically significant difference
was accepted when the p-value was lower than 0.05.
RESULTS
Among the examined groups were not statistically sig-
nificant differences in age and BMI as well as in the group
D between trousers users and shirt users (Table 2). Sem-
inal fluid volume, sperm density, sperm total count,
sperm morphology and sperm progressive motility did
not show any significant variation among the examined
groups, as well as any of the other conventional sperm
parameters (Table 1). None of the selected ultrasound
parameters (testicular and prostate volume, cranio-caudal
diameter of the cephalic and caudal portion of the epi-
didymis, anteroposterior diameter of the seminal vesi-
cles) were significantly different between the examined
groups (Table 1). Relatively to the biofunctional sperm
parameters: the percentage of spermatozoa with low
MMP, the percentage of spermatozoa with PS external-
ization, and the spermatozoa with decondensed chro-
matin did not show any significant variation among the
examined groups (Table 2).
The percentage of spermatozoa with fragmented DNA
was significantly higher only in group D compared to the
other groups, and in group D the percentage of sper-
matozoa with fragmented DNA was significantly higher in
trousers users compared to the shirt users (Table 2).
The correlation analysis showed that the percentage of
spermatozoa with fragmented DNA correlated positively
with the duration of use of the mobile phone (Table 3).
Finally, none of the examined ultrasound parameters
showed a significant correlation with the duration of use
of the mobile phone (Table 3).
Group A Group B Group C Group D Trousers Shirt
no use <2 h/day 2-4 h/day >4 h/day users users
(no.=10) (no.=16) (no.=17) (no.=20) (no.=12) (no.=8)
Conventional sperm parameters
Volume (ml) 3.0±1.2 2.8±1.4 3.0±1.1 2.9±1.5 2.8±1.3 3.0±0.8
Sperm density (106/ml) 71.0±7.3 68.5±6.0 72.0±6.0 67.0±11.0 65.0±5.0 68.5±8.0
Total sperm count (106/ejaculate) 213.0±8.8 191.8±8.4 216.0±6.6 194.3±16.5 182.8±6.5 205.5±6.4
Progressive motility (%) 46.0±3.0 42.5±6.0 41.0±6.0 44.0±3.0 42.0±2.0 44.0±6.0
Normal forms (%) 11.0±6.0 10.0±5.0 12.0±4.0 10.0±7.0 9.0±7.0 11.0±5.0
White blood cells (106/ml) 0.64±0.09 0.77±0.06 0.60±0.03 0.55±0.05 0.33±0.03 0.60±0.06
Biofunctional sperm parameters
PS externalization (%) 3.5±3.0 4.1±2.0 3.9±1.8 4.3±2.5 4.8±1.6 3.9±1.8
Abnomal chromatin compactness (%) 17.0±3.0 16.7±2.6 18.0±6.0 18.8±3.0 18.8±3.0 17.2±6.0
DNA fragmentation (%) 3.0±1.2 3.2±1.6 3.1±2.2 6.6±2.2* 6.7±1.8^ 5.1±1.3
Altered (low) membrane mitochondrial potential (%) 5.0±1.5 4.4±2.0 4.6±4.0 4.2±3.0 4.6±3.0 4.0±2.0
*p<0.05 vs other groups; ^p<0.05 vs shirt users.
Table 2 - Conventional and biofunctional sperm parameters in the four examined groups.
Parameter r p
Sperm density (mil/ml) –0.20 ns
Total sperm count (mil/ejaculate) –0.19 ns
Progressive motility (%) –0.22 ns
Normal forms (%) –0.30 ns
Low mitochondrial membrane potential (%) 0.42 ns
Alive spermatozoa (%) –0.33 ns
Phosphatidylserine externalized spermatozoa (%) 0.23 ns
Spermatozoa with abnormal chromatin (%) 0.22 ns
DNA fragmentation (%) 0.82 <0.001
ns: not significant.
Table 3 - Correlation analysis between duration of use of the
mobile phone and conventional and biofunctional sperm pa-
rameters.
©2013, Editrice Kurtis
FOR PERSONAL USE ONLY

Cellular devices and male infertility
973
DISCUSSION
Our clinical study evaluated the conventional sperm pa-
rameters and some of the main biofunctional sperm pa-
rameters of a series of healthy men, suggesting that the
fragmentation of sperm DNA could represent the only pa-
rameter significantly altered in the subjects who use the
mobile phone for more than 4 h a day and in particular for
those who use the device in the pocket of the trousers. The
study was carried out on healthy fertile men, without other
risk factors for male infertility, with an age range between
18 and 35 yr, non-smokers, without difference between
groups for age and BMI. Moreover, all the examined sub-
jects used the same device and were classified according to
the duration and mode of use of the mobile phone. Also
from the ultrasound point of view, the examined patients
showed no significant differences in the testicular volume
(expression of testicular function) and the other ultrasound
parameters of the prostate, seminal vesicles and epi-
didymis, that are frequently altered in the chronic inflam-
matory process (38). In our opinion, this aspect is very im-
portant for the correct selection of the patients.
Several studies have addressed the link between mobile
phones and the potential consequences on male fertility.
In particular, previous experimental experiences have
highlighted the following evidences: a reduction of
testosterone, an increase in caspase-3, the overproduc-
tion of reactive oxygen species (5), a significant decrease
in protein kinase C (6) and a decrease of the antioxidant
enzymes glutathione peroxidase and superoxide dismu-
tase (7). Another study showed a significant drop in fruc-
tose levels (9). In addition, RF-EMR exposure is associat-
ed with an increase in lipid peroxidation in the testis and
epididymis (10). Moreover, the histological examination
showed also a significant decrease in the diameter of
seminiferous tubules in the phone group vs the controls
(11). Finally, a significant genotoxic effect on epididymal
spermatozoa was demonstrated (12).
From the clinical point of view, a significant difference was
observed in sperm morphology between mobile phone
users and non users. Moreover, the same study showed
higher T levels and lower LH levels in the mobile phone
users (13). In another clinical study, samples exposed to RF-
EMW showed a significant decrease in sperm motility and
viability, increase in ROS level, and decrease in ROS-TAC
score (15). In step with increasing SAR, motility and vitality
are significantly reduced after RF-EMR exposure, while the
mitochondrial generation of reactive oxygen species and
DNA fragmentation are significantly elevated. Furthermore,
there is an highly significant relationship between SAR, the
oxidative DNA damage bio-marker, 8-OH-dG, and DNA
fragmentation after RF-EMR exposure (17).
The sperm DNA fragmentation is the presence of breaks
or damage in the genetic material of the spermatozoa.
Possible causes are considered the defects of maturation
(40, 41), the apoptosis of the spermatozoa (42) and the
oxidative stress (43). The sperm DNA fragmentation can
be aggravated by altered mechanism of cellular repair or
environmental insults such as hyperthermia (44).
Recently, other studies have analyzed the effects of the
use of the mobile phone on sperm DNA quality, with dif-
ferent study designs and conflicting results. In the exper-
imental study of Aitken et al., mice were exposed to 900
MHz radio-frequency electromagnetic radiation (RF-EMR)
and after exposure, DNA damage to caudal epididymal
spermatozoa was assessed and a detailed analysis of DNA
integrity using QPCR (quantitative PCR) revealed statisti-
cally significant damage to both the mitochondrial
genome and the nuclear beta-globin locus (12). In a sub-
sequent study, Agarwal et al. examined the sperm DNA
damage of neat semen samples [one experimental aliquot
was exposed to cellular phone radiation (in talk mode),
and the unexposed second aliquot served as the control]
showing no significant differences from the unexposed
group (15). In another study, De Iuliis et al. evaluated pu-
rified human spermatozoa exposed to RF-EMR, showing
that in step with increasing SAR (specific absorption rates),
the mitochondrial generation of reactive oxygen species
and DNA fragmentation were significantly elevated. More-
over, it was also observed a highly significant relationship
between SAR, the oxidative DNA damage bio-marker, 8-
OH-dG, and DNA fragmentation (17). More recently, Fal-
zone et al. showed that the mobile phone radiation had
no statistically significant effect on any of the sperm pa-
rameters examined. In particular in this study, ejaculated,
density-purified, highly motile human spermatozoa were
exposed to mobile phone radiation at SAR of 2.0 and 5.7
W/kg and, at various times after exposure, the following
parameters were examined: caspase 3 activity, external-
ization of PS, induction of DNA strand breaks, and gen-
eration of reactive oxygen species (14).
Finally, another similar study of Agarwal et al. (16) shows
that, among the users >4 h/day, an alteration of the con-
ventional sperm parameters is observed; in particular sperm
motility, viability, and normal morphology appear signifi-
cantly different in cell phone user groups within two sperm
count groups. However, even this study does not appear
comparable for the following reasons: a) infertile patients
were examined; b) the biofunctional sperm parameters
were not examined; c) it did not compare the different
modalities of use of the mobile phone (trousers or shirt).
The results of the different studies are difficult to com-
pare owing to the different methods used. However, none
of the other studies selected the patients from the clinical
point of view as in the present study, focusing on the
healthy and fertile subjects users of the same device. In
our opinion, this aspect is a strong point of the present
study compared to other studies. Moreover, the results
of this study, in agreement with the recent study of Fal-
zone, suggest that apoptosis is not associated with sperm
DNA damage. However, the present study has limitations:
the small number of examined subjects and the lack of
explanation of the mechanism responsible for the appar-
ent damage of sperm DNA, that deserves further study.
REFERENCES
1. Depinder F, Makkler K, Agarwal A. Cell phones and male infertility:
dissecting the relationship. Reprod Biomed Online 2007, 3: 266-70.
2. Conclusions on mobile phones and radio frequency fields. European
Commission Scientific Committeeon Emerging and Newly Identified
Health Risks (SCENIHR). http://ec.europa.eu/health/opinions2/
en/electromagnetic-fields/l-3/5-conclusions-mobile-phones.htm
3. Worldwide mobile phone sales fell in 2012: Gartner. February 13,
2013. http://www.reuters.com/article/2013/02/13/us-mobile-
phones-gartner-idUSBRE91C0G120130213
©2013, Editrice Kurtis
FOR PERSONAL USE ONLY

R. Rago, P. Salacone, L. Caponecchia, et al.
974
4. McLachlan RI. Approach to the patient with oligozoospermia. J
Clin Endocrinol Metab 2013, 98: 873-80.
5. Kesari KK, Behari J. Evidence for mobile phone radiation exposure
effects on reproductive pattern of male rats: role of ROS.
Electromagn Biol Med 2012, 31: 213-22.
6. Kesari KK, Kumar S, Behari J. Mobile phone usage and male in-
fertility in Wistar rats. Indian J Exp Biol 2010, 48: 987-92.
7. Kesari KK, Kumar S, Behari J. Effects of radiofrequency electro-
magnetic wave exposure from cellular phones on the reproductive
pattern in male Wistar rats. Appl Biochem Biotechnol 2011; 164:
546-59.
8. Imai N, Kawabe M, Hikage T, Nojima T, Takahashi S, Shirai T.
Effects on rat testis of 1.95-GHz W-CDMA for IMT-2000 cellular
phones. Syst Biol Reprod Med 2011, 57: 204-9.
9. Salama N, Kishimoto T, Kanayama HO, Kagawa S. The mobile
phone decreases fructose but not citrate in rabbit semen: a longi-
tudinal study. Syst Biol Reprod Med 2009, 55: 181-7.
10. Mailankot M, Kunnath AP, Jayalekshmi H, Koduru B, Valsalan R.
Radio frequency electromagnetic radiation (RF-EMR) from GSM
(0.9/1.8GHz) mobile phones induces oxidative stress and reduces
sperm motility in rats. Clinics (Sao Paulo) 2009, 64: 561-5.
11. Salama N, Kishimoto T, Kanayama HO. Effects of exposure to a
mobile phone on testicular function and structure in adult rabbit. Int
J Androl 2010, 33: 88-94.
12. Aitken RJ, Bennetts LE, Sawyer D, Wiklendt AM, King BV. Impact
of radio frequency electromagnetic radiation on DNA integrity in
the male germline. Int J Androl 2005, 28: 171-9.
13. Gutschi T, Mohamad Al-Ali B, Shamloul R, Pummer K, Trummer H.
Impact of cell phone use on men's semen parameters. Andrologia
2011, 43: 312-6.
14. Falzone N, Huyser C, Franken DR, Leszczynski D. Mobile phone
radiation does not induce pro-apoptosis effects in human sper-
matozoa. Radiat Res 2010, 174: 169-76.
15. Agarwal A, Desai NR, Makker K, et al. Effects of radiofrequency
electromagnetic waves (RF-EMW) from cellular phones on human
ejaculated semen: an in vitro pilot study. Fertil Steril 2009, 92:
1318-25.
16. Agarwal A, Deepinder F, Sharma RK, Ranga G, Li J. Effect of cell
phone usage on semen analysis in men attending infertility clinic:
an observational study. Fertil Steril 2008, 89: 124-8.
17. De Iuliis GN, Newey RJ, King BV, Aitken RJ. Mobile phone radia-
tion induces reactive oxygen species production and DNA dam-
age in human spermatozoa in vitro. PLoS One 2009, 4: e6446.
18. Erogul O, Oztas E, Yildirim I, et al. Effects of electromagnetic ra-
diation from a cellular phone on human sperm motility: an in vitro
study. Arch Med Res 2006, 37: 840-3.
19. Fejes I, Závaczki Z, Szöllosi J, et al. Is there a relationship between
cell phone use and semen quality? Arch Androl 2005, 51: 385-93.
20. La Vignera S, Condorelli RA, Vicari E, D'Agata R, Calogero AE.
Effects of the exposure to mobile phones on male reproduction: a
review of the literature. J Androl 2012, 33: 350-6.
21. Mahmoud A, Comhaire F. Systemic causes of male infertility. In:
Andrology for the Clinician. Springer, 2006, 57-63.
22. Jarow JP. Endocrine causes of male infertility. Urol Clin North Am
2003, 30: 83-90.
23. La Vignera S, Vicari E, Condorelli RA, D’Agata R, Calogero AE.
Male accessory gland infection and sperm parameters (review). Int
J Androl 2011, 34: 330-47.
24. La Vignera S, Calogero AE, Condorelli R, et al. Criptorchidism and
its long term complications. Eur Rev Med Pharmacol Sci 2009, 13:
351-6.
25. La Vignera S, Condorelli R, Vicari E, D’Agata R, Calogero AE.
Effects of varicocelectomy on sperm DNA fragmentation, mito-
chondrial function, chromatin condensation, and apoptosis. J
Androl 2012, 33: 389-96.
26. Sakamoto H, Ogawa Y, Yoshida Y. Relationship between testicular
volume and testicular function: comparison of the Prader orchido-
metric and ultrasonographic measurements in patients with infer-
tility. Asian J Androl 2008, 10: 319-24.
27. Calogero AE, Polosa R, Perdichizzi A, et al. Cigarette smoke ex-
tract immobilizes human spermatozoa and induces sperm apop-
tosis. Reprod Biomed Online 2009, 19: 564-71.
28. La Vignera S, Condorelli RA, Balercia G, Vicari E, Calogero AE.
Does alcohol have any effect on male reproductive function ? A
review of literature. Asian J Androl 2013, 15: 221-5.
29. Dandona P, Rosenberg MT. A pratical guide to male hypogonadism
in the primary care setting. Int J Clin Pract 2010, 64: 682-96.
30. Cooper DS. Subclinical hypothyroidism. N Engl J Med 2001, 345:
260-5.
31. La Vignera S, Condorelli RA, Vicari E, Calogero AE. Negative effect
of increased body weight on sperm conventional and nonconven-
tional flow cytometric sperm parameters. J Androl 2012, 33: 53-8.
32. Lotti F, Corona G, Colpi GM, et al. Elevated body mass index cor-
relates with higher seminal plasma interleukin 8 levels and ultra-
sonographic abnormalities of the prostate in men attending an an-
drology clinic for infertility. J Endocrinol Invest 2011, 34: 336-42.
33. Calogero A, Polosa R, Perdichizzi A, et al. Cigarette smoke extract
immobilizes human spermatozoa and induces sperm apoptosis.
Reprod Biomed Online 2009, 19: 564-71.
34. WHO laboratory manual for the examination and processing of hu-
man semen, Fifth ed. Geneva: WHO, 2010.
35. Pilatz A, Rusz A, Wagenlehner L, Weidner W, Altinkilic B. Reference
values for testicular volume, epididymal head size and peak sys-
tolic velocity of the testicular artery in adult males measured by ul-
trasonography. Ultraschall in der Medizin 2012, 34: 349-54.
36. Vicari E. Seminal leukocyte concentration and related specific re-
active oxygen species production in patients with male accessory
gland infections. Hum Reprod 1999, 14: 2025-30.
37. Behre HM, Kliesch S, Schädel F, Nieschlag E. Clinical relevance of
scrotal and transrectal ultrasonography in andrological patients.
Int J Androl 1995, 18: 27-31.
38. Lotti F, Corona G, Mancini M, et al. Ultrasonographic and clinical
correlates of seminal plasma interleukin-8 levels in patients at-
tending an andrology clinic for infertility. Int J Androl 2011, 34:
600-13.
39. Perdichizzi A, Nicoletti F, La Vignera S, et al. Effects of tumour
necrosis factor-alpha on human sperm motility and apoptosis. J
Clin Immunol 2007, 27: 152-62.
40. Manicardi GC, Bianchi PG, Pantano S, et al. Presence of endoge-
nous nicks in DNA of ejaculated human spermatozoa and its rela-
tionship to chromomycin A3 accessibility. Biol Reprod 1995, 52:
864-7.
41. Sailer BL, Jost LK, Evenson DP. Mammalian sperm DNA suscepti-
bility to in situ denaturation associated with the presence of DNA
strand breaks as measured by the terminal deoxynucleotidyl trans-
ferase assay. J Androl 1995, 16: 80-7.
42. Sakkas D, Moffatt O, Manicardi GC, Mariethoz E, Tarozzi N,
Bizzaro D. Nature of DNA damage in ejaculated human sperma-
tozoa and the possible involvement of apoptosis. Biol Reprod
2002, 66: 1061-7.
43. Moustafa MH, Sharma RK, Thornton J, et al. Relationship between
ROS production, apoptosis and DNA denaturation in spermato-
zoa from patients examined for infertility. Hum Reprod 2004, 19:
129-38.
44. Ahmad G, Moinard N, Esquerré-Lamare C, Mieusset R, Bujan L.
Mild induced testicular and epididymal hyperthermia alters sperm
chromatin integrity in men. Fertil Steril 2012, 97: 546-53.
©2013, Editrice Kurtis
FOR PERSONAL USE ONLY