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PSGCAS Search: A Journal of Science and TechnologyVolume: 4 No. : 2 ISSN: 2349 – 5456 1
Correlation between Dietary Pattern, Work Environment and Incidence
of PCOD in Working Women
Bhuvaneswari 1 , Kanjana K.
2, Madhan Shankar S R.
3
1,2 Department of Clinical Nutrition and Dietetics, PSG College of Arts & Science, Coimbatore, Tamil
Nadu, India 3Department of Biotechnology, PSG College of Arts & Science, Coimbatore, Tamil Nadu, India
*Corresponding author: vasukanjana@gmail.com
ABSTRACT
Polycystic ovarian disease (PCOD) is the commonest endocrine disease in women of
reproductive age. It affects 5 to 10% of women of reproductive age. PCOD now proves to be a
significant factor in female infertility. Women with PCOD are at increased risk of diabetes,
hypertension, cardiovascular disease, hyper estrogen related cancers. The investigation was pursued to
assess the dietary pattern and study work environment of the working women, and test the incidence
of PCOD in selected women by PCR based methods. Out of the 83 working women (Information
Technology, administration, labour, medical coding and Teaching) from Coimbatore city between the
age group of 20- 35 years, 10 subjects who did not express having irregular menstrual cycle and 10
subjects who complainant of having irregular menstrual cycle, treated as Non-PCOD and PCOD
samples respectively were subjected to blood analysis – Polymerase chain reaction (PCR) techniques.
Consumption of non-vegetarian food against the regular meal pattern, Occupational stress,
Consumption of high fat foods and skipping of regular meals and Age related dietary / life style
changes were found to render the working women to fall into a high risk group with respect to the
incidence of PCOD.
Key words: PCOD, PCR techniques, menstrual cycle, obesity, fat intake
INTRODUCTION
Polycystic Ovarian were described as
early as 19th
century. In1935 Stein &
Leventhal described syndrome of
amenorrhea associated with polycystic
ovaries [1]
. Polycystic ovary syndrome is a
condition in which a woman has an
imbalance of female sex hormones. This
may lead to menstrual cycle changes, cysts
in the ovaries, trouble getting pregnant and
other health changes. PCOD is linked to
changes in the level of certain female
hormones, estrogen and progesterone, help
a woman's ovaries release egg. Androgen, a
male hormone is found in small amounts in
women. The pathophysiology of PCOD
appears to be multifactorial with multiple
genes and environmental factors
contributing to the disease
susceptibility. Multiple genetic pathways
have been implicated in the pathogenesis of
PCOD, including steroid hormone
metabolism, gonadotropin action, obesity
and energy regulation, and insulin action [2]
.
Normally, one or more eggs are released
during a woman's period, called ovulation.
In PCOD, mature eggs are not released
from the ovaries. Instead, they can form
Correlation between Dietary Pattern, Work Environment and Incidence of PCOD in Working Women
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 2
very small cysts in the ovary. These
changes can contribute to infertility.
Symptoms of PCOD include Secondary
amenorrhea, Virilization, male like
characteristics like body hair growing on
the chest, belly, face, and around the
nipples, decreased breast size, enlargement
of the clitoris, thinning of the hair on the
head called male-pattern baldness and
deepen voice.
The women those who had PCOD, may
have the following common health
conditions like Diabetes mellitus, High
cholesterol level, High blood pressure and
Weight gain and obesity. The woman who
has PCOD would more likely to develop
the complications like Infertility, Breast
cancer (slightly increased risk) and
endometrial cancer [3]
.
Approximately 60% of women with
PCOD have weight management issues
which can lead to obesity with only normal
caloric intake. Energy in the form of
glucose (food) is stored right away as fat,
instead of being made available for other
functions within the body. This can lead to
chronic fatigue and undernourishment,
despite the fact that there is adequate food
intake and even an appearance of over-
nourishment.
However, it is important to note that
40% of women with PCOD are of normal
weight, or even fall under a normal weight
range [4]
.
Weight loss and dietary changes appear
to affect all parameters of hormonal
fluctuation. Follistatin is a single-chain
glycoprotein that is expressed in many
tissues including the ovary, adrenal cortex,
pituitary and pancreas. This polypeptide
specifically binds activin, neutralizing its
biological activity. Activin causes ovarian
follicular development, inhibits theca cell
androgen production, increases pituitary
FSH secretion and pancreatic insulin
secretion. Since follistatin inhibits the
activity of activin, its altered activity due to
over-expression would therefore be
expected to reduce serum follicle
stimulating hormone (FSH), impair ovarian
follicular development, increase ovarian
androgen production and insulin release
which turn out to be characteristic features
of PCOD. Based on its functional
relevance and evidence of strong linkage
with PCOD, the follistatin gene emerged as
the strongest candidate of PCOD. There
have been a limited number of studies that
attempted to identify sequence variants
within this gene and their association with
PCOD. The results of these studies have
been inconsistent [5, 6]
.
Activin promotes ovarian follicular
development, inhibits androgen production
and increases FSH and insulin secretion.
Follistatin, an activin-binding protein,
neutralizes activin bioactivity. Therefore, a
decrease in the ratio of activin/follistatin
Bhuvaneswari , Kanjana K. and Madhan Shankar S R.
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 3
might encourage characteristic features of
polycystic ovary syndrome (PCOS) [7]
.
The known association between insulin
resistance and polycystic ovarian disease
(PCOD) has been studied by determination
of the prevalence of a positive family
history of diabetes in a consecutive series of
oligomenorrheic women with polycystic
ovaries and eumenorrheic women with
normal ovaries who served as Non-PCODs.
Paternal and maternal family members
affected were in similar proportions, there
being no evidence of preferential
transmission through the female line in this
study 8.
PCOD is a metabolic disorder affecting
multiple organs. An interdisciplinary
approach of gynaecologists together with
endocrinologists specialized in metabolic
and nutritional disorders is needed to test
whether proposed interventions as weight
loss, treatment of hyper insulinemia,
regulation of menstrual cycle and others can
avoid long-term squeal [9]
. Hence the
present investigation is taken up with the
following objectives:
• To assess the dietary pattern of working
women between age group 20-30years
before joining and after joining for work
• To study the work environment of
subjects
• To test for PCOD in these women by
PCR based methods
• To ascertain a correlation between the
dietary pattern and PCOD levels as
obtained by molecular method
MATERIALS AND METHODS
Eight three working women involved in
Information Technology, administration,
labour, medical coding and Teaching) from
Coimbatore city between the age group of
20- 35 years volunteered to participate in
the study.
Out of 83 subjects, 10 subjects who did
not express having irregular menstrual
cycle and 10 subjects, complainant of
having irregular menstrual cycle, treated as
Non-PCOD (Non-PCOD) and PCOD
(PCOD) subjects respectively were
subjected to blood analysis- PCR
techniques using Case-Non-PCOD study
design.
PCR Technique
The polymerase chain reaction is a
repetitive bidirectional synthesis of DNA
via primer extension of nucleic acid. PCR
technique is used to amplify a region of
DNA between two oligosaccharide primers.
PCR technique was used to find out
whether PCOD gene is present or not for
those who indicated complainant of having
and not having irregular periods from the
past years.
Protocol for DNA amplification:
For the reaction, the mentioned reagents
(Table -1) are added to a PCR tube.
Correlation between Dietary Pattern, Work Environment and Incidence of PCOD in Working Women
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 4
Table 1
Reagents used for DNA amplification
Reagents Quantity
10X Assay Taq Pol Assay
buffer 15 mM MgCl2
5 µl
dNTP Mix 2 µl
Template DNA ( 50 ng/µl) 2 µl
Forward primer (50 ng/µl) 2 µl
Reverse primer (50 ng/µl) 2 µl
Taq DNA Polymerase (3 u/µl) 1 µl
Sterile Water 36 µl
Total Reaction Volume 50 µl
The contents were mixed gently.
The reaction mixture was layered with 50
µl of mineral oil and the amplification was
carried out using the reaction condition
for 30 cycles and the details of which are
given in Table 2. After the reaction got
over, the reaction mix was taken out and
10 ml of the aqueous layer was run in 1%
agarose gel for 1 to 2 hours at 100 volts.
The sample was run along with marker and
the amplified product was located by
comparing with the 0.8 kb fragment of the
marker. Once the amplification was
completed, the sample was loaded on the
agarose gel and subjected to
electrophoresis.
Table 2 Reaction conditions for
DNA amplification
Reaction Tempr
Time
Initial denaturation 94oc 1min
Denaturation 94 o
c 30 secs
Annealing 48 o
c 30 secs
Extension 72 o
c 1 min.
Final extension 72 o
c 2 mins.
Preparation of 1.5 % Agarose Gel and
Set Up of Electrophoresis
• Prepared 1X TAE by diluting
appropriate amount of 50X TAE buffer
with distilled water.
• Took 50 ml of 1X TAE in a 250 ml
conical flask and added 0.75 g of
agarose. Boiled to dissolve agarose (till a
clear solution results).
• Placed the combs of the electrophoresis
set such that the comb was about 2 cm
away from the cathode (black electrode).
• When the agarose gel temperature was
around 60oC, added 2ml of 10mg/ml
ethidium bromide and swirled to mix.
• Poured the warm agarose solution slowly
into the central platform of the gel tank.
• Made sure that the gel was 0.5 to 0.7 cm
thick, without air bubbles. Kept the set
undisturbed till the agarose solidified.
• Once the gel had solidified, poured 1X
TAE buffer slowly into the gel tank till the
buffer level stood 0.5-0.8 cm above the gel
surface.
• Gently lifted the combs. Ensured that the
wells were intact. Connected the power
cords, the red cord to the red electrode and
the black cord to the black electrode.
• Loaded the samples into the well. Set the
voltage to 50 V. Run till the second dye
(purple dye) from the well had reached
3/4th of the gel.
Bhuvaneswari , Kanjana K. and Madhan Shankar S R.
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 5
• Viewed under UV trans illuminator and
recorded the observation
Assessment of Nutritional profile
Socio demographic information of the
working women participated in the study
were collected through questionnaire
covering the details regarding working
hours, leisure time, working environment,
dietary pattern before and after joining to
work, and the kind of health problems they
are diagnosed.
The nutritional profile of the selected
subjects were assessed through
anthropometry (weight, height, waist and
hip circumferences), Percent body fat
using Omron digital body fat analyser (bio
elective impedance method), arterial blood
pressure and pulse rate using Digital
Blood pressure monitor and nutrients
intake through 24 hours Recall method).
Body mass index (BMI), a simple health
indicator of body fatness was calculated
from the height and weight of the selected
subjects using the formula9:
BMI= Weight (in kg) / Height (in meters
square)
Waist-Hip Ratio (WHR) 10
of the
subjects was assessed using waist
circumference measured at the umbilicus
and hip circumference measured at the most
extended part of the buttocks. WHR =
Waist circumference/Hip circumference
Dietary intake of the selected subjects
was assessed through 24 hour recall method
and the nutrients intake was calculated
using the food guide of ICMR (2010) 11
.
RESULTS AND DISCUSSION
Demographic profile of selected subjects
Majority of the subjects were from the
age group of 20-25 years (72.28%),
whereas more than 14% of the subjects
were from 26-30 years (16.86%) and 31- 35
years (10.84%) age groups. Highest
percentage (48.19%) of the subjects
responded was from information
technology. There were 66.26% of married
and 33.73% unmarried working women.
Dietary pattern
Most of the subjects were Non-
vegetarians (85.54%) and negligible
numbers of subjects were vegetarians
(7.22%), ova- vegetarians (6.02%) and
lacto-ova vegetarians (0.01%). 55.42% of
the subjects were having their meals 3 times
per day; 40.96 % of the subjects followed
two meals a day; and 3.61% of the subjects
were having only single meal per day.
Higher percentage of the subjects was
found to be skipping their meals (53.01%).
Most of the working women preferred
meals which are prepared at home (48.19%)
and secondly, they preferred fried foods
(21.68%). However 8.43% preferred to
have foods outside their home.
Work environment and Meal
Majority of the selected women were
involved in working for 8-10 hours
(57.83%) and 28.91% of them were
Correlation between Dietary Pattern, Work Environment and Incidence of PCOD in Working Women
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 6
working below 8 hours (28.91%). 55.42%
of the selected women were having leisure
time and 44.57% of them were not having
leisure time. Out of the subjects who had
leisure time, maximum (36.95%) number
had duration of 15mins whereas 23.91% of
the subjects had only 20 minutes of leisure
duration.
Before joining to work, majority of the
selected subjects took breakfast (96.38%),
lunch (96.38%) and dinner (96.38%) at
home. Even after joining to work, most of
the selected subjects had their breakfast
(56.62%), lunch (40.96%) and dinner
(57.83%) at home but exhibiting 30-40%
reduction; however 37.34% of the selected
subjects had their breakfast from hostel
mess, more than 30% had their lunch at
office canteen (31.32%) and dinner
(32.53%) in the hostel.
Past Medical History
Majority of the selected women were
found to have stress (62.65%), followed by
back pain (60.24%) and spinal pain
(34.93%). The next 39.75% and 30.12% of
the selected subjects were obese and
hypertensive respectively. 24.09% of them
were found to be suffering from irregular
periods and surprisingly none of the
selected women had Diabetes mellitus.
Nutritional Profile of selected subjects
based on age category
The selected subjects from different age
groups were found to be significantly
differing on Weight (F = 3.637**), waist
circumference (F = 4.974**) and diastolic
pressure (F=5.259**). There is a significant
difference between the intake of fried
snacks (F=7.867**
) and fried salty snacks
(F= 6.074**
) when analysed based on age
category. There is no significant difference
between the nutrient intakes based on age
category.
PCR Technique - Banding Pattern based
on PCR amplification
The PCOD subjects who volunteered
(N=10) to undergo DNA profile PCOD
(N=10) were studied for the demographic
profile, occupation category, Dietary
pattern, BMI category, Skipping meals, Fat
category, Irregular menstrual cycle and Past
medical History along with DNA profile
(Interleukin 1β polymorphisms) and
compared with that of Non-PCOD group
(Table 4). Banding pattern obtained in Non-
PCOD and PCOD groups based on PCR
amplification is presented in Figure 1 and 2.
Table 4 Degree of polymorphism for Non-PCOD and PCOD samples
Selected samples Degree of polymorphism band
Non-PCOD 64%
PCOD 66.65%
Bhuvaneswari
PSGCAS Search: A Journal of Science and
Figure1
Figure 2
62%
64%
66%
68%
Non
Pe
rce
nta
ge
(%)
Figure3
Bhuvaneswari , Kanjana K. and Madhan Shankar S R.
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456
Result of PCR for Non-PCOD samples
Figure 2 Result of PCR for PCOD samples
Non-PCOD PCOD
64%
66.65%
Selected samples
Degree of polymorphism
band
Non-PCOD
PCOD
5456 7
PCOD
Correlation between Dietary Pattern, Work Environment and Incidence of PCOD in Working Women
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 8
Interleukin-1 beta polymorphism is a
way to locate many disease incidences.
The higher the degree of polymorphism
indicated the greater the chance of
progression through the disease. In the
present study, Non-PCOD sample (N=10)
showed a 64% degree of polymorphic
bands when compared to PCOD (N=10)
which showed a slightly higher degree of
polymorphism (i.e.) 66.5%. This pattern
can be correlated to the dietary pattern
amongst the individuals.
Degree of polymorphism as per age group
The majority of the selected Non-
PCOD (70%) and PCOD (80%) samples
belonged to the age group of 20-25 years
which revealed that the individuals in this
age group inhibited greater degree of
polymorphisms which indicate that this
age group is more vulnerable for more
erratic food habits which could culminate
in incidence of being obese.
Degree of polymorphism as per marital
status
Majority of the selected Non-PCOD
(90%) and PCOD (60%) individuals were
unmarried which showed that they had a
higher degree of polymorphism. 40% of
the PCOD group and 10% of the Non-
PCOD group individuals were married.
Though they were married, they are also
had an equal degree of the same which can
lead to ovarian disorders. Despite the food
habit of married / unmarried, which
showed almost equal probability of
indicating a higher degree of
polymorphism bands which refers the
impact of eating habits does have
substantial influence on the chance of
processing symptoms related to PCOD or
similar ovarian disorders even before
marriage.
Degree of polymorphism as per
Occupation
Most of the Non-PCOD (40%) and
PCOD (70%) group of IT professionals
were showed a higher degree of
polymorphisms; whereas equal percentage
of both Non-PCOD and PCOD samples of
administrative workers showed higher
degree of the same; 20% of the selected
subjects who were working as lecturers
also had the higher degree of
polymorphism. This outcome showed that
IT professionals had a higher chance of
incidence of PCOD. Added to the changes
in food patterns among individuals;
occupational stress could also be a
cumulative factor which strictly dictate the
incidence of PCOD. The results shown are
chemical representations of these
phenomena.
Degree of polymorphism as per dietary
pattern
Most of the Non-PCOD (90%) and
PCOD (70%) subjects were non
vegetarians indicating the possibility of
polymorphism irrespective of subjects
Bhuvaneswari , Kanjana K. and Madhan Shankar S R.
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 9
complaining of having or not having
irregular menstrual disorders.
Degree of polymorphism as per BMI
20% of selected Non-PCOD and 60%
of PCOD samples belonging to overweight
group showed that they have higher degree
of polymorphism. Hence, it can be
understood that these selected samples
were at the risk of incidence of PCOD.
Degree of polymorphism as per skipping
meals
50% and 70% of the Non-PCOD and
PCOD samples were found to be skipping
their meals respectively whereas 50% and
30% of the Non-PCOD and PCOD groups
did not skip their meals. The risk of
incidence of PCOD was exhibited among
the subjects who skipped meals frequently.
Degree of polymorphism based on fat
intake
Majority of the Non-PCOD (10%)
and PCOD (30%) subjects were found to
have consumed 45-55g of fat per day
which showed that consumption of high
fat foods could eventually lead to
incidence of PCOD. In the present study, it
is deciphered that an average of 45-55g of
fat consumption per day would increase
the chance of incidence of being PCOD
positive.
Degree of polymorphism based on
irregular menstrual cycle
All the selected PCOD individuals had
irregular periods which correlate with the
pattern of polymorphism obtained
herewith.
Degree of polymorphism as per Past
medical history
Obese subjects (60%) expressed
irregular menstrual disorders and were
likely to show higher degree of
polymorphism; whereas equal percentage
of both Non-PCOD and PCOD subjects
having the past medical history of
hypertension and spinal pain also showed
equal degree of the same; and anxiety
present in 100% of the Non-PCOD
subjects represented higher degree of
polymorphism.
Factors contributing Polymorphism
Figure 4 reveals that age (70%),
anxiety (100%), increased fat intake (10%)
and dietary pattern (Non vegetarian) (90%)
are the factors that contribute towards a
greater degree of polymorphism among the
individuals in Non-PCOD groups.
Figure 5 indicates that age (80%),
increased stress (30%), irregular menstrual
cycle (100%), dietary pattern (Non
vegetarian) (70%) and Obesity (60%) are
the factors contributing towards a higher
degree of polymorphism among the
individuals in PCOD group.
Correlation between Nutritional Profile
and Degree of Polymorphism
In general positive relationship was
observed between “% body fat (r = 0.506*)
and hip circumference(r = 0.458*)” and
Correlation between Dietary Pattern, Work Environment and
PSGCAS Search: A Journal of Science and Technology
Fig. 4 Factors contributing to higher degree of polymorphism among
Fig. 5 Factors contributing to higher degree of polymorphism among
“degree of polymorphism”; whereas
negative relationship was found between
“WHR (r = -0.012) and degree of
polymorphism”.
Dietary Pattern
(Non veg)
90%
Dietary Pattern
(Non veg)
70%
Obesity
60%
Correlation between Dietary Pattern, Work Environment and Incidence of PCOD in Working Women
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456
Factors contributing to higher degree of polymorphism among Non-PCOD
Factors contributing to higher degree of polymorphism among PCOD
“degree of polymorphism”; whereas
negative relationship was found between
0.012) and degree of
To summarize, Consumption of
vegetarian food against the regular meal
pattern, Occupational stress, Consumption
of high fat foods and skipping of regular
meals and Age related diet
Degree of polymorphism
Age (20-25 yrs)
70%
Anxiety
100%
Increased fat intake
(10%)
Degree of polymorphism
Age (20-25 yrs)
80%
Increased stress
30%
Irregular menstrual
cycle
100%
Dietary Pattern
(Non veg)
70%
Incidence of PCOD in Working Women
5456 10
PCOD group
PCOD group
To summarize, Consumption of non-
vegetarian food against the regular meal
pattern, Occupational stress, Consumption
of high fat foods and skipping of regular
meals and Age related dietary/life style
Anxiety
Bhuvaneswari , Kanjana K. and Madhan Shankar S R.
PSGCAS Search: A Journal of Science and Technology Volume: 4 No. : 2 ISSN: 2349 – 5456 11
changes could contribute to the incidence
of PCOD in working women and it can be
concluded that all the above mentioned
factors would render working women to
fall into a high risk group with respect to
the incidence of PCOD; which can have a
post-marital impact on reproductive health
of the women.
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