“THE INFLUENCE OF EXTREMELY LOW FREQUENCY-
ELECTROMAGNETIC FIELDS (ELF-EMFS) AND LIGHT AT
NIGHT (LAN) ON ESTRADIOL (E2) LEVELS, OXIDATIVE
STRESS AND DNA CONFIGURATION IN FEMALE NIGHT
SHIFT WORKERS” “
Kavitha Varak Dr.Ravindra Tiwari Research Scholar Supervisor
UGC Research Unit UGC Research Unit Bhavan’s New Science College Bhavan’s New Science College Hyderabad Hyderabad Telangana State Telangana State
An Electro Magnetic Field (EMF) is an array of waves
that arise with the gathering of electric and magnetic
fields, which oscillate at a specific frequency and at
definite intervals with each other.
INTRODUCTION
INTRODUCTION
Phones and wireless radiofrequency
devices are the most common EMF
sources that are used in close
proximity to the human body.
Several epidemiological studies have
found that long-term night working
women exposed to EMFs have a
higher risk of breast cancer than
those who do not work at night.
Lack of Concentration
Estradiol is a female sex hormone
produced by the ovaries, adrenal
gland and also the placenta during
pregnancy.
Estradiol is the most important
hormone during a female’s
reproductive years, and is required
for reproductive and sexual
function as well as having an
impact on various organs and
tissues.
A woman's risk for breast
cancer is supposed to
associated with increased
levels of estrogen.
ESTRDIOL & BREAST CANCER
BACKGROUND
INTERNATIONAL and NATIONAL
Data inconclusive
Controversial
Lacks scientific evidence
Literature Review
Prompts our study to rule out the
conflicting reports of ELFs
WORLD HEALTH ORGANISATION (WHO)
International Commission on Non-ionizing Radiation
Protection (ICNIRP)
Epidemiological studies give no consistent evidence of causal
relation between RF exposure and any health effect.
Bio-Initiative reports are – Inconclusive.
The RF Research Agenda indicates “high priority"
research for future health risk assessments.
The International Agency for Research on Cancer
(IARC) classified RF field as group 2B carcinogen .
OUR STRENGTH
Completed two UGC Major Research Projects in the same field.
1. Study of genomic stability and hormonal
configuration in human subjects exposed to extremely
low frequency electromagnetic waves from different
sources like mobile phones, computers and high
tension wires (2005-2008).
2. Risk assessment of human subjects occupationally
exposed to ELF-EMFs and light at night (LAN) (2011-
2014).
• Corresponding Author - Published papers in the field of EMR
in journals of good impact factors. Written chapters in books on
effects and applications of EMR.
•Referee for various research articles in journals of good impact
factors and project – ICMR.
•Invited speaker recently for an international conference in
Beijing-China, Chicago.
•One of the Papers is on the top of 20 Articles related to the
same field.
•Ministry of Science and Technology on behalf of society for
environment and development collected data of the outcome
from our major project (UGC - 2008).
OUR STRENGTH
India Population
1.25 billion
Mobile
subscribers
980+ Million
Mobile Phones and Female Night Shift -Statistics in India
Female
Night Shift
Workers
App 11 %
PICO
Population: Individuals exposed to Electromagnetic Fields
and Light at Night (LAN).
Intervention: To bring awareness among public, if the
findings indicate a positive effect.
Comparator: a a) Women subjects occupationally exposed to
EMFs and LAN:400
b) Controls (not occupationally exposed to EMFs and LAN) : 200
Outcome: To rule out the existing debate and controversy based on
our results to evaluate the mechanism of changes if any,
particularly related to Estrogen levels and DNA integrity
OBJECTIVES
To study DNA damage in leukocytes.
Chromosomal aberrations.
Malondialdehyde [MDA] in Plasma.
Estimation of Serum Nitric Oxide.
Estimation of Estradiol levels.
STUDY DESIGN
• Survey – women working in night shift in It software companies and
hospitals.
•Social determinants of health problems if any with the help of
questionnaire.
Qualitative (Observational):
Quantitative (Experimental):
•Estimations of the parameters as per the objectives
•Data Interpretation.
•Statistical Analysis.
METHODOLOGY
E S T R A D I
O L
L E V E L
N I
T R I
C
O X I
D E
M D A
A S S A Y
• Subjects –
a) Women subjects occupationally
exposed to EMFs and LAN : 400
b) Controls (not occupationally exposed
to EMFs and LAN) : 200
• Sampling -
Peripheral blood in a sterile heparinized
Vacutainer tubes.
• Comet assay -Single Cell Gel
Electrophoresis.
• Chromosomal Aberrations - MNT-
Micronucleus Test.
• Malondialdehyde [MDA] Assay.
• Estimation of Serum Nitric Oxide.
• Estradiol Levels- CLIA .
C O M E T
A S S A Y
MN T
A S S A Y
METHODOLOGY
E S T R A D I
O L L
E V E L
MN T
C O M E T
A S S A Y
Comet Assay
Sensitive test to study the degree of
DNA damage devised by Singh et al
(1988) with some modification after
McGlyann et al (1999) Regular Agarose
Lysing
Alkali Treatment Electrophoresis
Buffer
Sample + LMA Low Melting Agarose
MD A
A S S A Y
N I
T R I
C
O X I
D E
METHODOLOGY
GE
NE
EX
PR
ES
SIO
N Micronucleus Test (MNT)
To estimate chromosomal aberrations by slide observation method.
M N T
A S S A Y
E S T R A D I
O L L
E V E L
MD A
A S S A Y
N I
T R I
C
O X I
D E
C O M E T
A S S A Y
WORK DONE SO FAR
MD A
A S S A Y
MALONDIALDEHYDE [MDA] ASSAY:
The oxidative stress marker,
Malondialdehyde [MDA] is the stable
end product of lipid peroxidation and an
efficient parameter for lipid oxidative
damage.
Formation of MDA was assessed by
quantification of Thiobarbituric Acid
Reactive Species (TBARS).
E S T R A D I
O L
L E V E L
N I
T R I
C
O X I
D E
C O M E T
A S S A Y
MN T
WORK DONE SO FAR
ESTIMATION OF NITRIC OXIDE:
As Nitric Oxide (NO) is an unstable
compound, it is rapidly converted to
nitrates and nitrites in the body, hence their
concentration is parallel to NO levels.
Total production of NO was determined by
assaying for nitrite and nitrate. Nitrate and
nitrite concentrations were then estimated
by the Griess method.
E S T R A D I
O L
L E V E L
[
M D
A]
A S S A Y
C O M E T
A S S A Y
MN T
N
I
T
R
I
C
O
X
I
D
E
WORK DONE SO FAR
ESTRADIOL LEVEL:
Estrogen is a steroid hormone that is
necessary for the normal development
and growth of the breasts, menstrual
cycle and reproductive organs essential
for perpetuation of species.
Exposure to EMFs increase the level of
Estrogen.
Estradiol levels were estimated by
CLIA (CHEMILUMINESCENCE
IMMUNO ASSAY) –[Ratcliffe et al.,
1988]
N I
T R I
C
O X I
D E
[M D
A]
A S S A Y
C O M E T
A S S A Y
MN T
A S S A Y
E
S
T
R
A
D
I
O
L
L
E
V
E
L
WORK DONE SO FAR
N I
T R I
C
O X I
D E
M D A
A S S A Y
C O M E T
A S S A Y
MN T
AS SAY
E
S
T
R
A
D
I
O
L
L
E
V
E
L
Results of DNA Damage, MDA, Serum Nitric Oxide, Estradiol levels and MNT in the exposed compared with control group
DNA Damage Length (arbitrary units) Subjects N Mean + SD Range Variance
EMF-Exposed 400 8.76 + 1.31 1.72 - 9.23 1.723 Control 200 4.30 + 1.13 1.47-7.49 1.287
MDA Levels (nmoles/ml) Subjects N Mean + SD Range T value P value
EMF-Exposed 400 2.67 + 0.82 0.90 - 9.41 2.045 0.042
Control 200 1. 87 + 1.47 0.94 - 5.23 NO Levels (µmoles/ml)
Subjects Mean + SD Range T value p value EMF-Exposed 400 2.25 ± 0.09 0.42 - 4.58
2.148 0.033 Control 200 2.00 ± 0.07 0.66 - 4.30
Estradiol levels (pg/ml) Subjects Mean + SD Range T value p value
EMF-Exposed 400 284.4 + 89.08 130.56-736.27 22.99 < .00001
Control 200 72.10 + 34.08 33.7 - 169 % of Micronuclei
Subjects N Mean + SE EMF-Exposed 400 1.39 + 0.63
Control 200 1.24 + 0.67
0
5
10
15
20
25
30
35
40
0 50 100 150 200 250 300 350400450500 550600650700 750800
Ag
e
Estradiol concentration (pg/ml) Group
exposed
control
The t-value for
estradiol estimation
was 22.99. The p-
value was <00001.
The result was
significant at p < .05.
The alkaline comet assay
indicated DNA damage
ranging between 1.72µm to
9.23µm. The maximum tail
length (9.923 + 0.67µm) was
observed in night shift female
workers.
CONCLUSION
CONCLUSION
