EFFECT OF OVARIAN CYCLE ON BODY

COMPOSITION EVALUATED BY MFBIA

A THESIS
Submitted for the award of Degree
of

Doctor of Philosophy
in

Biomedical Engineering
by

Poonam Khalsa
SU/Ph.D./P.T/Biomedical/2015/01
Under the Supervision
of
Prof. (Dr). Jayanand Manjhi
Supervisor
Shobhit University, Meerut (U.P.)

School of Engineering & Technology (Department of BME)

Shobhit Institute of Engineering & Technology
(A NAAC Accredited-Deemed to be University)
MEERUT, U.P. (INDIA)
2021
 CANDIDATE’S DECLARATION

I, hereby, declare that the work presented in this thesis entitled “Effect of
Ovarian Cycles on Body Composition Evaluated by MFBIA”for the award of the
degree of Doctor of Philosophy, submitted to Faculty of Biomedical Engineering,
Shobhit University, Meerut, a Deemed-to-be-University, established by GOI u/s 3 of
UGC Act 1956, is an authentic record of my research work carried out under the
supervision of Dr. Jayanand Manjhi.

I also declare that the work embodied in the present thesis

(i) is my original work and has not been copied from any
Journal/Thesis/Book, and

(ii) has not been submitted by me for any other degree or diploma.

(Name and Signature of the Candidate)

ii
 CERTIF ICATE

This is to certify that the thesis entitled “Effect of Ovarian Cycle on Body
Composition Evaluated by MFBIA which is being submitted by Poonam Khalsa for
the degree of Doctor of Philosophy in Biomedical Engineering to the Faculty of
Biomedical Engineering, Shobhit University, Meerut, a Deemed-to-be-University,
established by GOI u/s 3 of UGC Act 1956, is a record of bonafide investigations and
extensions of the problems carried out by him under my supervision and guidance.

To the best of my knowledge, the thesis embodies the work of the candidate
himself and has not been submitted to any other University or Institution for the
award of any degree or diploma.

It is further certified that he/she worked with me for the required period in the
Faculty/School of Biomedical Engineering, Shobhit University, Meerut, a Deemed to
be University.

Date: NAME:DR.JAYANAND MANJHI

(Supervisor)

iii
 ACKNOWLEDGEMENT

I express all my gratitude to Almighty God who directly or indirectly inspired me and
helped me in choosing and accomplishing this work. All praises to God for blessing
me with opportunities abound and showering upon me his sympathy and guidance all
through my life.

I pay gratitude and thanks to former Prof. (Dr.) Amar P.Garg & Vice-Chancellor,
Shobhit University, Meerut, for his help during the dissertation. My heartfelt thanks to
Prof. (Dr.) Jayananad Manjhi Coordinator, Faculty of Biological Engineering, Shobhit
University, for providing all the facilities in the Department and for guiding at every
stage in the course of preparation of this thesis. His involvement has triggered my
intellectual maturity which will benefit us for a long time to come.

The work presented in this thesis could not have been done without the help of all of
whom I want to thank. First, It gives me immense pleasure to express my deepest
gratitude and indebtedness to Dr. Jayanand Manjhi., my thesis supervisor,
Assistant Professor, Centre for Biomedical Engineering, Shobhit University, Meerut,
for his patient guidance and support under whose inspiration& guidance I have
completed this thesis work. I am truly very fortunate to have the opportunity to work
with him. I found his guidance to be extremely valuable.

Besides my supervisor, I would like to thank the rest of my thesis committee:

Dr.Shobhit Kumar ji, Hon’ble Chairman and Shri Kunwar Shekhar Vijendra ji,
Hon’ble Chancellor, Shobhit Institute of Engineering & Technology Meerut and
Shobhit University, Gangoh, U.P who thoroughly devoted to empower people through
education and providing facilities for research and high quality teaching in their
institutions.

My Sincere thanks to Prof. (Dr.) Ajay Rana Vice Chancellor, Shobhit Institute of
Engineering & Technology for giving me the opportunity to work diligently.

Special thanks to Dr.Ganesh Bharadwaj, Registrar, Shobhit Institute of Engineering &

Technology for their valuable suggestions time to time related to my work.

iv
I also greatly appreciate the support receive from Dr.Rajiv Datta,Dean school of
Engineering &Technology and Dr.M.N.Bahugua,Associate Dean, School of
Engineering & Technology for his immensely to complete my work.

My deep thanks to Dr. Niraj Singhal, Director, IQAC, Shobhit Institute of Engineering
& Technology for giving me opportunity to work sincerely.

Advices given by Dr.Anuj Goel coordinator, Research degree program have also
helped me to complete my work. I am very thankful.

I extend thanks to Mr.V.D Sharma Office Superintendent, for providing me the

requisite institutional facilities and academic rules throughout my research work.

I submit my gratitude to my parent Mr.Karanjit Singh and M rs.Harmit Kaur and

my Brother Simranjit Singh without their support, nothing could be done. Also I
would like to thank Mr.Jas jot Singh my husband, who has been my side throughout
this Ph.D.I, would also like to say a special thanks to my father- in- law, mother- in law
and sister- in law for their unconditional love and support at all times.

Also, I would like to thank all those who have been part of my journey, my friends,
colleagues, teachers and other staff members for the guidance and support.

Finally, yet importantly, I would like to thank the almighty, GOD.

Poonam Khalsa…….

v
 LIST OF FIGURES

Figure 1.1 Schematic Representation of Bio-Impedance measurement system ............ 2

Figure 1.2: Principles of BIA from physical characteristics to body composition (Kyle
et al., 2001). ................................................................................................................... 4

Figure 1.3 Resistor and capacitor in the human body (Kyle et al., 2004). .................... 4

Figure 1.4: Types of BIA (Barbosa et al., 2005). .......................................................... 5

Figure 1.5 Placement of electrode for single and multiple frequencies BIA (Cornish et
al., 1993, Segal et al., 1991). ......................................................................................... 5

Figure 1.6 Deviation of phase angle and Relation between (R, Z, Xc) (Deurenberg et
al., 1993). ....................................................................................................................... 6

Figure 1.7: Experimental setup for Segmented- BIA (Bracco et al., 1996) .................. 7

Figure 1.8 Endometrial Cycle and Phases (Lukaski, 2000). .......................................... 7

Figure1.9: Ovarian Cycle (Yanovski et al., 1996). ........................................................ 8

Figure 1.10 Ovarian Cycle (Kutac and Kopecky, 2015). ............................................. 9

Figure 1.11 Phases of Uterine Cycle (Garlie et al., 2006). .......................................... 11

Figure 1.12 Hormone variation throughout the ovarian cycle (Deurenberg et al.,
1988). ........................................................................................................................... 13

Figure 1.13 Different models for body composition (Wells and Fewtrell, 2006). ...... 18

Figure 1.14 Body Composition Levels (Wang et al., 1992). ....................................... 19

Figure 1.15 Schematic diagram of Fat-Free Mass (FFM) (Kyle et al., 2003). ........... 24

Figure 1.16 Body Fluid Compartments (Clegg, 1984). ............................................... 25

Figure 1.17DXA scan (Bachrach, 2000)...................................................................... 33

Figure 1.18 Measurement of hydrostatic weight (courtesy of INCAP, Guatemala)

(Weight, 1996). ............................................................................................................ 34

Figure 1.19Skin Fold Method (Womersley and Durnin, 1977). .................................. 36

vi
Figure 1.20 Multi- frequency Bioelectrical impedance measuring tool (Aleman-Mateo
et al., 2010). ................................................................................................................. 37

Figure 4.1Body stat Quad Scan 4000 .......................................................................... 64

Figure 4.2Applications of Electrodes for Impedance Measurements .......................... 68

Figure 4.3Weighing Machine ...................................................................................... 69

Figure 4.4Height Measurement by Healthcare Practitioner ........................................ 70

Figure 4.5 Methodology used ...................................................................................... 72

Figure 4.6 Electrode placement ................................................................................... 73

Figure 5.1 Participants’ Distribution............................................................................ 77

Figure 5.2 SD and mean range values of different body composition in females. ...... 82

Figure 5.3Variation in impedance with age in females ............................................... 83

Figure 5.4Variation in phase angle with females age group........................................ 85

Figure 5.5 Correlation of ovarian cycle phases with female age group & FM% ........ 88

Figure 5.6Correlation of ovarian cycle phases with female age group & FFM (Kg) .. 89

Figure 5.7Correlation of ovarian cycle phases with female age group & impedance . 90

Figure 5.8 BMI variation with Ovarian Cycle ............................................................. 92

vii
 LIST OF TABLES

Table 1.1ACE Body Fat % Norms for Body Composition (Wells and Fewtrell, 2006).
...................................................................................................................................... 17

Table 1.2 Information deduced at Different Body Composition Levels (Wang et al.,
1992). ........................................................................................................................... 20

Table 4.1 Ideal Height to Weight Variations ............................................................... 70

Table 5.1 Participants’ Distribution ............................................................................. 77

Table 5.2Body composition (MFBIA) in Ovarian Phase ............................................ 81

Table 5.3 Measurements driven from (MFBIA) impedance and phase angle (Females)
at different frequencies................................................................................................. 83

Table 5.4 Measurements driven from MFBIA phase angle at different frequencies ... 85

Table 5.5 Phase Distribution for Analysis ................................................................... 86

Table 5.6 % FM variation with Ovarian Cycle ............................................................ 87

Table 5.7 FFM variation with Ovarian Cycle .............................................................. 89

Table 5.8 FM (kg) variation with Ovarian Cycle ........................................................ 90

Table 5.9 BMI variation with Ovarian Cycle .............................................................. 91

viii
 LIST OF ABBREVIATIONS

Abbreviation Description

BIA Bioelectrical Impedance Analysis

TIA Trans-Impedance Amplifier

D/A Digital-to-Analog

A/D Analog-to-Digital

BIS Bioelectrical Spectroscopy

FSH Follicle Stimulating Hormone

LH Luteining Hormone

GnRH Gonadotrophin –Releasing Hormone

ACE American Council on Exercise

FM Fat Mass

FFM Fat-Free Mass

ECS Extracellular Solids

ECF Extracellular Fluid

MRS Magnetic Resonance Spectroscopy

DEXA Dual-Energy X-Ray Absorptiometry

CT Computer Tomography

MRI Magnetic Resonance Imaging

BMI Body Mass Index

BFP Body Fat Percentage

LBM Lean Body Mass

ix
FFM Fat-Free Mass

TBW Total Body Water

ICW Intra Cellular Water

ECW Extra Cellular Water

BCM Body Cell Mass

VPs Visceral Proteins

DEXA Dual Energy X-Ray Absorptiometry

BMD Bone Mineral Density

BMC Bone Mineral Content

ADP Air Displacement Plethysmography

BVI Body Volume Indicator

MFBIA Multi-Frequency Bioelectrical Impedance Analysis

MAC Mid Arm Circumference

MAMC Mid Arm Muscle Circumference

CHI Creatinine Height Ratio

OC Oral Contraceptives

PCOS Polycystic Ovary Syndrome

OCPs Oral Contraceptive Pills

DXA Dual-X-Ray Absorptiometry

VAT Visceral Adipose Tissue

PAL Physical Activity Level

BIVA Bioelectrical Impedance Vector Analysis

x
 TABLE OF CONTENTS

CANDIDATE’S DECLARATION ............................................................................ II

CERTIFICATE ......................................................................................................... III

ACKNOWLEDGEMENT.........................................................................................IV

LIST OF FIGURES ...................................................................................................VI

LIST OF TABLES ..................................................................................................VIII

LIST OF ABBREVIATIONS ...................................................................................IX

TABLE OF CONTENTS ..........................................................................................XI

CHAPTER 1: INTRODUCTION............................................................................... 1

1.1 BIOELECTRICAL IMPEDANCE ANALYSIS (BIA)........................................ 2

1.2 PRINCIPLE BEHIND BIA .................................................................................. 3
1.3 METHODS OF BIA ............................................................................................. 4
1.3.1 Single- frequency BIA .................................................................................... 5
1.3.2 Multifrequency BIA....................................................................................... 5
1.4 BIOELECTRICAL SPECTROSCOPY (BIS) ...................................................... 6
1.5 SEGMENTAL-BIA.............................................................................................. 6
1.6 MENSTRUAL CYCLE........................................................................................ 7
1.6.1 Ovarian cycle ................................................................................................. 7
1.6.2 Uterine Cycle ................................................................................................. 8
1.6.2.1 Endometrium........................................................................................... 9
1.6.2.2 Menstrual Cycle ...................................................................................... 9
1.6.2.3 Decidua ................................................................................................... 9
1.7 PHASES OF OVARIAN CYCLE ........................................................................ 9
1.7.1 Follicular Phase ............................................................................................. 9
1.7.2 Ovulatory Phase ........................................................................................... 10
1.7.3 Luteal phase ................................................................................................. 10
1.8 PHASES OF UTERINE CYCLE ....................................................................... 11
1.8.1 Menstrual Phase ........................................................................................... 11
1.8.2 Proliferative Phase ....................................................................................... 11
1.8.3 Secretory Phase............................................................................................ 12

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1.9 HORMONAL CHANGES DURING OVARIAN CYCLE ................................ 12
1.9.1 Progesterone ................................................................................................ 13
1.9.1.1 Functions of progesterone (Damavandi, 2008)..................................... 13
1.9.1.2 Complications due to low progesterone production ............................. 13
1.9.1.3 Signs and symptoms of low progesterone levels .................................. 14
1.9.2 Estrogen ....................................................................................................... 14
1.9.2.1 Main types of estrogens produced by the body (Lobo, 1987): ............. 14
1.9.2.2 Functions of estrogens (Schutze et al., 1992) ....................................... 15
1.9.2.3 Complications due to low estrogens production in the body ................ 15
1.9.2.4 Complications due to high estrogens production in the body ............... 15
1.9.3 Follicle Stimulating Hormone ..................................................................... 15
1.9.3.1 Complications due to a high level of follicle-stimulating hormone ..... 16
1.9.3.2 Complications due to low level of follicle-stimulating hormone ......... 16
1.9.4 Luteinizing Hormone ................................................................................... 16
1.9.4.1 Feature and functions of luteinizing hormone: ..................................... 16
1.9.4.2 Complications associated due to high LH production .......................... 16
1.9.4.3 Complications due to low levels of luteinizing hormone ..................... 17
1.10 BODY COMPOSITION................................................................................... 17
1.10.1 Models of body composition ..................................................................... 17
1.10.1.1 Two-Compartment Model .................................................................. 18
1.10.1.2 Three-Compartment Model ................................................................ 18
1.10.1.3 Four-Compartment Model .................................................................. 18
1.10.2 Multi-compartment Models ....................................................................... 19
1.10.2.1 Atomic Level Multi-compartment Model........................................... 20
1.10.2.2 Molecular Level Multi-compartment Model ...................................... 21
1.10.2.3 Cellular Level Multi-compartment Model .......................................... 21
1.10.2.4 Tissue Level Multi-compartment Model ............................................ 21
1.10.2.5 Whole-body......................................................................................... 21
1.10.3 Significance of measuring body composition............................................ 22
1.11 BODY MASS MEASUREMENTS.................................................................. 22
1.11.1 Body Mass Index (BMI) ............................................................................ 22
1.11.2 Body Fat Percentage (BFP) ....................................................................... 22
1.11.3 Lean Body Mass (LBM) ............................................................................ 23
1.11.4 Fat-Free Mass (FFM)................................................................................. 23
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 1.11.4.1 Visceral Proteins (VPs) ....................................................................... 24
1.11.4.2 Intra Cellular Water (ICW) ................................................................. 24
1.11.4.3 Extra Cellular Fluid (ECW) ................................................................ 25
1.11.4.4 Bone Mineral....................................................................................... 27
1.11.4.5 Total Body Water (TBW) ................................................................... 27
1.12 FACTORS INFLUENCING BODY COMPOSITION .................................... 27
1.12.1 Genetic Composition ................................................................................. 27
1.12.2 Age............................................................................................................. 28
1.12.3 Hormones................................................................................................... 28
1.12.4 Menopause ................................................................................................. 28
1.12.5 Gender........................................................................................................ 28
1.12.6 Type of Physical Activity .......................................................................... 28
1.12.7 Nutrition..................................................................................................... 28
1.13 RELATIONSHIP BETWEEN OVARIAN CYCLE AND BODY
COMPOSITION....................................................................................................... 29
1.14 CHANGES IN BODY COMPOSITION DURING OVARIAN CYCLE ........ 31
1.15 TECHNIQUES TO MEASURE BODY COMPOSITION............................... 32
1.15.1 Dual Energy X-ray Absorptiometry (DEXA)............................................ 32
1.15.2 Field Methods ............................................................................................ 33
1.15.2.1 Hydro-densitometry / Under-Water Weighing ................................... 33
1.15.2.2 Air Displacement Plethysmography (ADP)........................................ 35
1.15.2.3 Body Volume Indicator (BVI) ............................................................ 35
1.15.2.4 Skin Fold Method ............................................................................... 35
1.15.3 MFBIA (Multi-Frequency Bioelectrical Impedance Analysis) ................. 36
1.15.3.1 Applications of MFBIA ...................................................................... 37
1.15.3.2 Challenges faced by MFBIA .............................................................. 38
1.15.4 Circumferences and other measurements .................................................. 39

CHAPTER 2: LITERATURE REVIEW................................................................. 40

2.1 VARIATION OF BODY COMPOSITION WITH OVARIAN CYCLE ........... 40

2.2 EVALUATING TECHNIQUE FOR MEASURING BODY COMPOSITION . 48

CHAPTER 3: AIMS AND OBJECTIVES .............................................................. 58

3.1 AIM OF THE STUDY........................................................................................ 59

xiii
 3.2 OBJECTIVES..................................................................................................... 59

CHAPTER 4: MATERIALS AND METHODS ..................................................... 61

4.1 MATERIALS ..................................................................................................... 61

4.2 SUBJECTS ......................................................................................................... 62
4.2.1 Exclusion Criterion ...................................................................................... 62
4.2.2 Inclusion Criterion: ...................................................................................... 62
4.3 INSTRUMENT USED (BODY STAT QUAD SCAN 4000) ............................. 64
4.3.1 Bodystat Quadscan 4000 measurements ..................................................... 65
4.3.2 Body stat Quadscan 4000 Window software:.............................................. 66
4.3.3 Protocol used with Bodystat Quadscan 4000 .............................................. 67
4.3.4 Features of Bodystat Quad scan 4000: ........................................................ 67
4.3.5 The principle involved in the study with Bodystat Quad scan 4000: .......... 67
4.4 IMPEDANCE DISPOSABLE ELECTRODE.................................................... 68
4.5 BLUETOOTH .................................................................................................... 69
4.6 WEIGHING MACHINE .................................................................................... 69
4.7 HEIGHT MEASUREMENT SCALE ................................................................ 70
4.8 MFBIA CALIBRATOR ..................................................................................... 71
4.9 METHODOLOGY ............................................................................................. 71
4.9.1 Estimation of Body Hydration using MFBIA ............................................. 74
4.9.2 Principles of used parameters ...................................................................... 75

CHAPTER 5: RESULTS .......................................................................................... 77

5.1 SUBJECT DISTRIBUTION .............................................................................. 77

5.2 PARAMETERS USED FOR ANALYSIS OF BODY COMPOSITION ........... 78
5.2.1 Body Mass Index (BMI) .............................................................................. 78
5.2.2 Body Fat (BF) .............................................................................................. 78
5.2.3 Fat Mass (FM) ............................................................................................. 78
5.2.4 Fat-Free Mass (FFM)................................................................................... 79
5.2.5 Body Water .................................................................................................. 79
5.2.5.1 Extra Cellular Water(ECW).................................................................. 79
5.2.5.2 Intra Cellular Water (ICW) ................................................................... 79
5.2.6 Body Cell Mass (BCM) ............................................................................... 79
5.2.7 Extra Cellular Mass (ECM) ......................................................................... 80

xiv
 5.3 BODY COMPOSITION ANALYSIS USING MFBIA IN OVARIAN PHASE 80
5.4 IMPEDANCE ANALYSIS AT DIFFERENT FREQUENCIES ........................................... 82
5.5 PHASE ANGLE ANALYSIS AT DIFFERENT FREQUENCIES .................... 84
5.6 BODY MEASUREMENTS VARIATION WITH PHASES OF OVARIAN
CYCLE ..................................................................................................................... 86
5.6.1 %FM variation with Ovarian Cycle............................................................. 87
5.6.2 FFM variation with Ovarian Cycle .............................................................. 88
5.6.3 FM (kg) variation with Ovarian Cycle ........................................................ 90
5.6.4 BMI variation with Ovarian Cycle .............................................................. 91
5.7 STATISTICAL ANALYSIS .............................................................................. 93

CHAPTER 6: DISCUSSION .................................................................................... 95

CHAPTER 7: SUMMARY AND CONCLUSION................................................ 106

7.1 CONCLUSION ................................................................................................ 107

7.1.1 Parametric Analysis of Body Composition for different Age Groups ...... 108
7.1.2 Impedance and Phase Angle Analysis at Different Frequencies ............... 108
7.1.3 Body Composition variation with Phases of Ovarian Cycle ..................... 108

REFERENCES......................................................................................................... 110

PLAGIARISM REPORT ........................................................................................ 127

PUBLICATIONS ..................................................................................................... 128

xv
CHAP TER 1: INTRODUCTION

Using correct data to deal with your body has grown more popular in recent decades.
If "beauty standards" were established and everyone hoped to match them, now
people are starting to think more personally, with ideas like fat percentage, drying,
whey protein, and of course bio-impedance becoming more common.

Bio- impedance is a diagnostic approach that uses electrical resistance and

anthropometric data to determine the body's capabilities and illness risks (Hannan et
al., 1995).

People who wish to reduce weight effectively without dehydration or muscle loss, or
develop muscle mass with little fat are the target audience.

The main capabilities of the BIA include measurements (Hannan et al., 1995):

 Fat mass;
 Body mass index;
 Percentage of body fat;
 The amount of muscle tissue;
 Percent of active cell mass;
 The amount and distribution of fluid in the body;
 Basal metabolic rate;
 Waist-to-hip ratio;
 Biological age.

We are made up of resistances and capacitance. The BIA approach assumes that fat is
a conditional insulator (does not conduct / weakly conducts current owing to lipid
composition) and that the conductor is a fat-free mass. Muscles are an excellent
conductor because they contain a lot of water (up to 75%). (fat-free mass will have
less resistance to electric current). The quicker the signal, the more muscles. The
electrical signal goes fast through hydrated muscle tissue but encounters resistance
when it “hits” adipose tissue. It is termed impedance (Helleputte et al., 2014)

1
The conductivity is influenced by the ratio of ions, the state of the bone tissue, and
other important processes in the body. The bio- impedance procedure itself consists in
placing electrodes on certain parts of the body (lower leg and forearm) and passing a
small (50 kHz) alternating current through them. Sensitive sensors record the
necessary indicators, and the computer produces a finished result.

1.1 BIOELECTRICAL IMPEDANCE ANALYSIS (BIA)

BIA is a non-invasive, low-tech method of evaluating body composition. Because of

these features, BIA is widely employed in clinical and fitness settings. BIA works by
sending an electrical signal into the body and detecting impedance or current
resistance (Kyle et al., 2004)

The fat- free bulk is higher in water and electrolytes, making it more conductive.
Water and electrolytes in adipose tissue reduce conductivity (Janssen et al. 2000).
Contact electrode BIA analyzers vary from traditional BIA systems tha t need gel
electrodes to be placed at precise anatomical sites. Utilizing preprogrammed
prediction models, contact electrode BIA analyzers automatically calculate body
composition parameters including BMI and FFM (Kushner et al. 1996). These
proprietary formulae employ impedance and other data like height, weight, sexual
category, age, and body type to estimate body composition (Barbosa et al., 2005).

A schematic of the electrical model of Bio-impedance is illustrated in Figure 1.1.

Figure 1.1 :Schematic Representation of Bio-Impedance measurement system

2
 https://www.analog.com/media/en/technical-documentation/tech-
articles/Bioelectrical-Impedance-Analysis- in-Monitoring-of-the-Clinical-Status-and-
Diagnosis-of-Diseases.pdf

In the diagram above, RE represents fluid resistance, RI represents intercellular fluid

resistance, and C m represents membrane capacitance. An electrode connects the
equipment to the human body. The gadget supplies electricity to the electrode and
measures the current it produces. A D/A converter converts the produced signal into
an alternating signal that is linked to the driver. A microprocessor controls the signal's
amplitude and frequency. It is configured to send a regulated signal to the D/A
converter. A Trans- impedance Amplifier (TIA) linked to the A/D converter measures
current. The data is transmitted to a microcontroller for additional processing to
extract useful information.

The human body is separated into five parts for bio- impedance measurements: upper
limbs (2), lower limb (2), and torso. Several characteristics must be considered during
BIA analysis. Height, weight, skin thickness, and others. Other characteristics include
age, gender, and health.

1.2 PRINCIPLE BEHIND BIA

We know that resistance is related to the length of the conductor (L) and inversely
proportional to the cross-section area (A), as shown in Figure 1.2. Because the human
body is not a single cylinder and its permeability is unstable, an empirical relationship
between impedance and electrolyte volume may be established (Kyle et al., 2001).

3
 Cylinder

Cross
Current sectional
Area (A)

Length (L)

Figure 1.2: Principles of BIA from physical characteristics to body composition

(Kyle et al., 2001).

Practically, it is easier to determine the height, usually greater than the transmitter
length from the wrist to the neck. Another difficulty that is observed in BIA is that the
body comprises two different kinds of resistances named capacitive resistance and
resistive resistance (Barbosa et al., 2005). The capacitive resistance has been arising
from the cell membrane whereas simple resistance exists due to intra and intercellular
fluid. The composition of resistance and capacitive/ reactive resistance is shown in
figure 1.3(Kyle et al., 2004).

Figure 1.3:Resistor and capacitor in the human body (Kyle et al., 2004).

1.3 METHODS OF BIA

BIA analyzer can be categorized into single or multiple frequencies depending upon
the electrical current which is used to determine the impedance of the body tissues
(Barbosa et al., 2005).

4
 BIA

Single Frequency Multiple Frequencies

Figure 1.4: Types of BIA (Barbosa et al., 2005).

1.3.1 Single-frequency BIA

Single- frequency analyzers use a 50 kHz electric signal to identify the body's tissue
composition. Figure 1.4 shows the electrodes on the hand and foot. The average
weighted total of intracellular and extracellular water. Single- frequency analyzers at
50 kHz estimate total body water but do not discriminate between intracellular and
extracellular water. To solve this, numerous frequency analyzers are needed (Kushner
et al., 1992).

1.3.2 Multifrequency BIA

On the contrary, multiple frequency analyzers send frequencies more than 1MHz to
achieve body composition measurements. It works with multiple frequencies such as
0 kHz, 1 kHz, 5 kHz, 50 kHz, 100 kHz, 200 kHz up to 500 kHz. The reproducibility
above and below 5 kHz and 200 kHz is small (Cornish et al., 1993, Segal et al.,
1991).

Figure 1.5:Placement of electrode for single and multiple frequencies BIA

(Cornish et al., 1993, Segal et al., 1991).

5
1.4 BIOELECTRICAL SPECTROSCOPY (BIS)

Unlike MF-BIA, BIS incorporates mathematics and mathematical models (such as the
Cole-Cole diagram in Figure 1.5). The Hanai formula is used to establish the link
between resistance and bodily fluid area. A hybrid model is created using prediction
equations and experience. This hybrid model's equations are regarded as accurate with
little bias in non-physiological individuals. But the modeling system has to be
improved.

Impedance Z
Reactance (Xc)Ohm

(Ohm)
Frequency
Phas Increase
e
Angle

Resistance R (Ohm)

Figure 1.6: Deviation of phase angle and Relation between (R,Z, Xc) (Deurenberg
et al., 1993).

Body cell mass (BCM) comprises muscles and electrical pathways. The cells are not
spherical, but they may be made cylindrical by following the current course. Several
academics studied how the resistivity constant affects the population size. Using the
mixing equation, some studies reported better accuracy than regression, while others
found no gain or worse accuracy. To improve BIS efficiency, data must be understood
using a trustworthy data fitting technique and a proper fluid distribution model
(Deurenberg et al., 1993).

1.5 SEGMENTAL-BIA

Segmented –BIA is carried out by positioning two electrodes on the opposite side of
both wrist and foot. The electrodes can also be placed on the upper iliac spine,

6
shoulder, and ankle. During experiments, a few researchers have placed electrodes on
the forearm, shoulder, lower leg, and upper leg. The experimental setup for segmented
BIA is presented in Figure 1.7.

Figure 1.7: Experimental setup for Segmented- BIA(Braccoet al., 1996)

1.6 MENSTRUAL CYCLE

A menstrual cycle is much more than a period. In actuality, the term period is just the
beginning part of the main cycle. It is made up of two interconnected cycles that work
together. The ovarian cycle occurs in the ovaries, whereas the uterine cycle occurs in
the uterus. The human brain, ovaries, and uterus all work together to control these
cycles. Figure 1.8 depicts the two cycles' principal stages.

Figure 1.8:Endometrial Cycle and Phases(Lukaski, 2000).

1.6.1 Ovarian cycle

The ovarian cycle is a normal cycle of changes in the uterus and ovary. This paper
examines the most modern and widely used methods for facilitating sexual
reproduction (Heyward and Wagner, 2004). Endogenous (internal) biological cycles

7
regulate its timing. The menstrual cycle is required for egg production and uterine
preparation for pregnancy.

The cycle only occurs in reproductive female humans and primates. The menstrual
cycle occurs in females from the age of menarche to menopause. The menstrual cycle
lasts from 21 to 35 days in humans, with 28 days being the average (Lukaski, 2000).
Each cycle includes three stages depending on ovarian or uterine events (Yanovski et
al.,1996).

Figure1.9: Ovarian Cycle (Yanovski et al., 1996).

Changes in hormones during pregnancy disrupt the menstrual cycle, which typically
sheds the functional layer of the uterine lining. A widespread misconception is that
follicle development happens in a single cycle (Nunez et al. 1997). In reality, an
ovulating follicle requires three menstrual cycles to form. The menstrual cycle is the
set of changes that a female body undergoes to prepare for conception. The menstrual
cycle repeats from menarche to menopause (Lohman et al., 2005). The menstrual
cycle lasts from 21 to 35 days in humans, with 28 days being the average.

The ovary divides each cycle into three phases (ovarian cycle or uterus cycle). The
ovarian cycle has three phases: follicular, ovulatory, and luteal, whereas the uterine
cycle has three: menstruation, proliferative, and secretary. The endocrine system
regulates both cycles, and hormonal contraception may interfere with natural
hormonal changes (Slinde and Rossander, 2001)

1.6.2 Uterine Cycle

The uterine cycle describes changes in the uterine lining (endometrium) during
menstruation. The endometrium thickens, supplying the expanding tissue lining with
abundant blood supply. If the environment is favorable, the blastocyst is implanted
and causes various modifications. The keywords are as follows:

8
1.6.2.1 Endometrium
It is the innermost layer of the uterus that is composed of a layer of glandular cells.
This is characterized as the mucous cells that guard and cover the mammalian uterus.
This is the part where the fertilized egg is initially implanted within the uterus.

1.6.2.2 Menstrual Cycle

It is a regularly reoccurring cycle in females that is associated with some typical
physiological changes. This has a close association with the reproductive ability of
mammalian females.

1.6.2.3 Decidua
It is the mucous membrane that is regularly shed out during menstruation and is
replenished over time. It gets modified to support pregnancy.

1.7 PHASES OF OVARIAN CYCLE

Figure 1.10 :Ovarian Cycle(Kutac and Kopecky, 2015).

1.7.1 Follicular Phase

The days of a menstrual cycle are numbered beginning with the first day of monthly
bleeding. During the follicular phase, estrogen steadily increases, causing menses to
cease and the uterine lining to thicken (Kutac and Kopecky, 2015). The ovary's
follicles begin forming under the influence of hormones, and after a few days one or
two become prominent (non-dominant follicles atrophy and die).

Salient features exhibited by the follicular phase:

9
  Very low levels of estrogen and progesterone hormones
 Endometrial lining starts to shed away
 FSH start to stimulate the development of new follicles in ovaries
 By the end of the stage, only one follicle develops

1.7.2 Ovulatory Phase

Luteinizing hormone rises throughout the ovulatory period. LT hormones cause the
dominant follicle to swell and then burst, releasing the egg. Ovulation lasts 16-32
hours. It terminates after the egg is released, 10-12 hours following the LT hormone
spike (Heyward and Wagner, 2004). After 12 hours, the egg may fertilize. The LT
hormone surge may be detected in urine (Buchholz et al., 2004). This measurement
helps indicate a woman's fertility. In the reproductive tube, before the egg is released,
sperms are more likely to fertilize (Cole, 1940). Most pregnancies occur 3 days before
ovulation.

Salient features exhibited by ovulatory phase:

 Increases flow of LH and FSH

 Estrogen levels start decreasing
 Progesterone level starts increasing

1.7.3 Luteal phase

On ovulation day, the egg is released from the ovary's egg follicle. It lasts 7–22 days
during a regular menstrual cycle. After ovulation, the egg follicle converts into the
corpus luteum, which produces progesterone and estrogen (Miller et al., 2016). Mid-
phase progesterone levels are linked to menstrual symptoms. During ovulation, some
women have pelvic or abdominal pain. These are typical ovulation signs (Garlie et al.,
2006).

Salient features exhibited by luteal phase:

 It stretches for 14 days

 The formation of the corpus luteum occurs
 The increased amount of progesterone hormone is secreted

10
  Estrogen level also remains high

1.8 PHASES OF UTERINE CYCLE

Figure 1.11: Phases of Uterine Cycle (Garlie et al., 2006).

1.8.1 Menstrual Phase

This stage is characterized by the following:

 Discharge of fluid- filled with blood, serous fluid and shed out endometrial
tissue.
 The Corpus luteum gets transformed to corpus Albicans.

1.8.2 Proliferative Phase

This phase begins in the anterior pituitary gland following the release of Follicle
Stimulating Hormone (FSH). The number of sequential events that take place in this
stage is as follows:

 After the release of FSH from the pituitary gland, it enters the ovaries and
stimulates the Graafian follicles to mature.
 Now follicles also begin to mature and are called theca cells. This releases
another hormone called estrogen.
 The estrogen hormone causes the thickening of the uterine lining and prepares
the endometrial for the next stages.
 With the accumulation of enough estrogen, another hormone called
Luteinizing Hormone (LH) is also released.

11
 All this leads to the beginning of the next phase of the uterine cycle.

1.8.3 Secretory Phase

After the completion of the ovulation process number of changes start to take place

 The ruptured follicles begin to collapse

 The granulosa cells begin to enlarge in size
 Internal thecal cells also enlarge
 Corpus luteum is formed

The freshly developed corpus luteum secretes progesterone and estrogen. In the
absence of fertilization, the corpus luteum degenerates after 10 days, producing a
small scar known as the corpus Albicans. However, if pregnancy occurs, the corpus
luteum begins generating hormones for 3 months before the placenta begins fulfilling
its designated job.

1.9 HORMONAL CHANGES DURING OVARIAN CYCLE

Menarche occurs at the ages of 12-13. Menopause is the end of a woman's

reproductive period and occurs between 45 and 55. Enumenorrhea is a 3 to 5day
menstrual cycle (Kyle et al., 2004). In animals, a GnRH spike precedes the LH surge,
indicating estrogen's major influence is on the hypothalamus, which regulates GnRH
production (Esco et al., 2015). Estrogen receptor alpha is responsible for the negative
feedback estradiol-LH loop, whereas estrogen receptor beta is responsible for the
positive estradiol-LH interaction (Deurenberg et al., 1988).

12
Figure 1.12: Hormone variation throughout the ovarian cycle (Deurenberg et al.,
1988).

1.9.1 Progesterone

Progesterone is an endogenous steroid hormone involved in human menstruation,

pregnancy, and embryogenesis (Dixon et al., 2008). From adolescence till menopause,
the corpus luteum produces large levels of progesterone. Progesterone is a naturally
occurring steroid hormone (Damavandi, 2008).

1.9.1.1 Functions of progesterone(Damavandi, 2008)

 It prepares the endometrial lining of the uterus for potential pregnancy

 It stimulates the thickening of endometrial
 It prevents muscular contractions of the uterus that may reject the egg
 Its high levels prevent ovulation
 If the body conceives, it triggers the formation of blood vessels in the
endometrium
 Its levels remain elevated throughout the pregnancy
 It also assists in the production of breast milk

1.9.1.2 Complications due to low progesterone production

 It results in the existence of an abnormal menstrual cycle

 It makes it hard to conceive
 It further increases the risk of miscarriage or a pre-term delivery

13
1.9.1.3 Signs and symptoms of low progesterone levels

 The presence of missed or irregular periods

 Occasional existence of abnormal uterine bleeding
 Pregnancy characterized by abdominal pain or spotting
 Increased frequency of miscarriages
 The presence of a very high level of estrogens results in increased weight gain

1.9.2 Estrogen

During premenopause, oestrogen levels increase and decline unevenly. Premenopause

symptoms may appear in women aged 25 to 70. Estrogens like estriol (E4) are only
created during pregnancy (Gomez et al., 1993).

The enzyme aromatase synthesizes oestrogen from androgens, especially testosterone

androstenedione. Premarin, a popular estrogenic medication, comprises the steroidal
estrogens equilin and equilenin (Mitchell et al., 1993). Ovulation causes a discomfort
termed mittelschmerz in certain women (in Germen meaning middle pain). The abrupt
shift in hormones during ovulation induces mid-cycle blood flow.

Progesterone is generated by the ovaries and placenta.

Estrogens are a class of chemicals that have a role in both menstruation and estrous
cycles. Female sex hormones. All vertebrates and certain insects produce estrogens.
Oral contraceptives employ estrogens. During pregnancy, estriol takes up this job, and
in postmenopausal women, estrone takes over (Teilmann et al., 2006).

1.9.2.1 Main types of estrogens produced by the body(Lobo, 1987):

 Estrone: It is also abbreviated as the E1 form of estrogen that is released after

the cessation of periods.
 Estradiol: It is also referred to as E2 and is usually found in women who are in
the age group of childbearing.
 Estriol: It is an E3 type of estrogen that forms the major proportion of the
estrogen that is found in the body during pregnancy.

14
1.9.2.2 Functions of estrogens(Schutze et al., 1992)

 Maintains bone health in both men and women

 Controls cholesterol levels in the body
 It is a very important hormone for childbearing
 Governs the growth of breast tissue
 Triggers menstrual cycle in females
 Rules the growth of underarm and pubic hairs

1.9.2.3 Complications due to low estrogens production in the body

 Existence of less frequent periods

 Causes night sweats or hot flashes in women
 Thinning or dryness of the vagina
 Causes in low sex desire
 Frequent mood swings
 Dryness of skin

1.9.2.4 Complications due to high estrogens production in the body

 Results in heavy or light menstrual bleeding

 The premenstrual syndrome gets worsen
 Occurrence of Non-cancerous lumps called fibrocystic
 Formation of fibroids in the uterus as non-cancerous tumors
 Results emotion arousal and feeling of anxiousness and depressed
 However, it results in poor erections and infertility in men(Dakin et al., 2015)

1.9.3 Follicle Stimulating Hormone

A flow towards the follicle-stimulating hormones is seen during the ovulation phase
that considerably decreases in the luteal phase. It is one of the most essential
hormones that is secreted by the pituitary gland and circulated via the bloodstream. In
women, it triggers the growth of follicles and in men, it stimulates sperm
production(Kumar et al., 1997).

15
1.9.3.1 Complications due to a high level of follicle-stimulating hormone

 It signals the malfunctioning in testis or ovaries

 Increases hormone level in the bloodstream rarely causes ovarian
hyperstimulation syndrome in women

1.9.3.2 Complications due to low level of follicle-stimulating hormone

 Incomplete development during puberty

 Causes ovarian failure or characterized by poor ovarian function
 Challenged growth of follicles resulting in infertility
 Improper sperm production in men
 Partial hormone deficiency results in delayed puberty and loss of
fertility(Oktay and Bedoschi, 2014).

1.9.4 Luteinizing Hormone

The anterior pituitary gland produces LH, a gonadotrophic hormone that regulates the
activity of the gonads, ovaries, and testes. This is a critical hormone for both men's
and women's reproductive health.

1.9.4.1 Feature and functions of luteinizing hormone:

 It triggers the production of oestradiol from ovaries

 In the case of fertilization, LH stimulates the production of progesterone from
the corpus luteum to maintain pregnancy.
 In men, LH triggers the production of testosterone in testes from the Leydig
cells to stimulate sperm production characterized by deep voice and facial hair
growth.

A high level of LH may be due to the existence of some genetic conditions such as
Klinefelter syndrome or Turner syndrome.

1.9.4.2 Complications associated due to high LH production

 A high level of LH may result in infertility

 In women, it results in polycystic ovary syndrome

16
1.9.4.3 Complications due to low levels of luteinizing hormone

 Low levels of LH also causes infertility

 It further stops ovulation
 In men, it also results in deficient gonadotrophin-releasing hormone
secretion(Dufau, 1988)

1.10 BODY COMPOSITION

Body fat percentage is measured by body composition, whereas leanness is measured

by weight. Water, protein, fats, and lipids are examples of chemical or molecular
constituents. Water, fat, connective tissue, bone, and muscle make up the body. The
human body has hundreds of different cell types, however, the majority aren't human,
resulting in GI bacteria. The body composition test evaluates the percentage of fat in
the body. Each of these components is found in equal amounts in the human body.
The table below illustrates the ACE healthy body composition requirements for
different demographic groups.

Table 1.1:ACE Body Fat % Norms for Body Composition (Wells and Fewtrell,
2006).

Description Women Men

Essential Fat 10% to 13% 2% to 5%

Athletes 14% to 20% 6% to 13%

Fitness 21% to 24% 14% to 17%

Acceptable 25% to 31% 18% to 24%

Obese Over 32% Over 25%

Athletes with minimal body fat tend to perform better in sports. However, low body
fat causes several health issues and injuries (Wells and Fewtrell, 2006).

1.10.1 Models of body composition

The human body is majorly made up of water in addition to bones, tissues, and
musculature.

17
 Figure 1.13: Different models for body composition (Wells and Fewtrell, 2006).

In body composition, the proportion of these constitutes is measured using different

models. Various types of models are illustrated in following Figure 1.13.

1.10.1.1 Two-Compartment Model

The most common and basic level used for the measurement is two-compartment
models that access the following components (Wells and Fewtrell, 2006):

 Fat Mass (FM)

It measures all the fat stores present within the human body.

 Fat-Free Mass (FFM)

It measures all the body components, except fats. This includes skeletal muscles,
bones, internal organs, and water present within the body.

1.10.1.2 Three-Compartment Model

This model further compartments the FFM into body water, minerals, and proteins
portions.

1.10.1.3 Four-Compartment Model

In this type of model, the minerals and proteins are measured as separate
compartments for better and more accurate measurement of body composition. It is

18
observed that as the number of compartments increases, there is a considerable
decrease in the error observed in the estimation of body composition (Wells and
Fewtrell, 2006).

1.10.2 Multi-compartment Models

Body mass is defined as the sum of all components at the atomic, molecular, cellular,
tissue-organ (functional), and whole-body levels. It contains potassium, sulfur,
sodium, chlorine, and magnesium, all of which are essential for life. Based on
scientific data, trace elements with over a dozen are necessary for life. When bodily
mass and energy balance are steady, the key components stay stable and have
predictable connections with each other. Extracellular materials, extracellular fluids,
and cells make up the cellular level. Components do not overlap (Wells et al., 1999).

Figure 1.14:Body Composition Levels (Wang et al., 1992).

Abbreviations: Extracellular solids (ECS) and extracellular fluid (ECF), 1 Nitrogen

and other elements2 Minerals, carbohydrates and other molecules 3 Other
tissues(Wang et al., 1992).

19
 Table 1.2:Information deduced at Different Body Composition Levels(Wang et
al., 1992).

Sr.
Levels Information Methods Used
No.

Elementals such as

o Oxygen
o Carbon Whole-body counting of total body
Atomic
1 o Calcium potassium Neutron Activation
Level
o Phosphorus analysis Isotope dilution
o Sulphur
o Chlorine
o Sodium, etc

o Isotope Dilution
o Water o Magnetic Resonance
Molecular o Protein Spectroscopy (MRS)
2
Level o Lipid o Multicomponent models
o Minerals o Dual-Energy X-ray
o Glycogen Absorptiometry (DEXA)

o Fat
o Cell Mass o Isotope Dilution
Cellular
3 o Extracellular o Bioelectrical Impedance
Level
Fluids Analysis (BIA)
o Extracellular
Solids
o Computer tomography (CT)
Functional o Magnetic Resonance
o Adipose
/Tissue- Imaging (MRI)
4 o Bone
organ o Ultrasound
o Muscle
Level o Near-Infrared Interactance
o Organ
o DEXA

o Circumferences
o Anthropometry
o Linear Dimensions
Whole- o Densitometry, Bioelectrical
5 o Lengths
body o Impedance Analysis (BIA)
o Skinfolds
o 3D Photonic Scan
o Body Volumes
o Body Surface Area

1.10.2.1 Atomic Level Multi-compartment Model

The eleven primary atomic elements are oxygen, carbon, hydrogen, nitrogen, calcium,
phosphorus, potassium, sulphur, sodium chloride, and magnesium. Each of the five

20
components is different, with no overlap between them. The discrete character of each
component minimizes misunderstanding and duplication in multi-component models.

1.10.2.2 Molecular Level Multi-compartment Model

The human body is made up of water, lipids, proteins, minerals, and carbohydrates.
Water makes roughly 60-70 percent of the body's weight depending on fat
composition. Intercellular to extracellular water ratio is a vital health parameter that
varies in diseases. The human body contains lipids. Phospholipids and sphingomyelin
are structural lipids found in cell membranes and the nervous system. Non-essential
triglycerides dominate (fat). They store energy, protect vital organs like the kidneys,
and enhance the body's appearance. A healthy adult of “normal weight” has 10% to
25% body fat in males and 15% to 35% in women. Obesity ranges from 60-70
percent.

1.10.2.3 Cellular Level Multi-compartment Model

Body composition is characterized cellularly as cell mass, extracellular fluids, and
extracellular solids. Water, proteins, and minerals make up the body's cell mass.
Extracellular fluid (plasma in the intravascular space and interstitial fluid in the
extravascular space) is 95 percent water. Extracellular solids are mostly proteins and
minerals (bone minerals and soluble minerals in the extracellular fluid). Due to its
complexity, cellular body composition is difficult to quantify.

1.10.2.4 Tissue Level Multi-compartment Model

Cells with equal functions form tissues, including muscular, connective, epithelial,
and nervous tissue. Bones are connective tissue and consist mainly of hydroxyapatite,
[ ( ) ] ( ) , bedded in a protein matrix (Thomas et al., 2000).

(1.1)

1.10.2.5 Whole-body
Whole-body measurements of body composition employ basic body factors to reveal
body composition. Formulae based on statistical connections between body
characteristics (e.g., skinfold thickness) and body composition (e.g., body fat by

21
density) can allow body composition evaluation. Body water evaluation using weight,
height, age, and gender (Shankaran et al., 2005).

1.10.3 Significance of measuring body composition

Several reasons motivate accessing the body composition. Some of the notes worthy
are as follows:

 To track the changes in body composition over time

 It helps to determine the effect of an intervention
 It aids in predicting the risk of injury that may be for instance due to low bone
mineral density
 It further helps in planning and meeting body composition goals
 It helps in accessing health risks that arise due to being overweight or
underweight(Cumberledge et al., 2018).

1.11 BODY MASS MEASUREMENTS

The body mass is usually measured in terms of the following:

1.11.1 Body Mass Index (BMI)

It is generated from a person's mass (weight) and height. The BMI is calculated by
multiplying the body mass by the square of the body height, which gives the BMI in
kg/m2. Based on tissue mass (muscle, fat, and bone) and height, the BMI classifies a
person as underweight, normal weight, overweight, or obese. There is a substantial
discussion regarding where the dividing lines between categories should be placed on
the BMI scale. Typical BMI ranges are underweight (Deurenberg et al., 1991).

1.11.2 Body Fat Percentage (BFP)

The entire quantity of fat in a human or other living entity is divided by the total mass
of the body and multiplied by 100. Where necessary type body fat is needed for life
and reproduction. Women have more significant body fat than males due to
childbearing and other hormonal activities. Body fat is adipose tissue buildup that
protects internal organs in the chest and belly. Due to childbirth and other hormonal
processes, women's essential body fat percentage is higher than men's. Storage body

22
fat is adipose tissue that protects internal organs in the chest and belly. The proportion
of body fat is used to assess fitness since it directly computes the relative body
composition of individuals without taking into account height or weight.

There are many ways to calculate body fat percentage, such as using calipers or
measuring bioelectrical impedance (Goonasegaran et al., 2012).

1.11.3 Lean Body Mass (LBM)

It is calculated by subtracting the body fat weight from the total body weight (lean
plus fat). Usually, no lean body mass % is stated. Usually, 60–90%. Rather, the
supplement measures the quantity of body fat, which is normally 10–40 percent.
Because body fat is less relevant to metabolism than lean body mass, LBM is a better
measure than total body weight for prescribing medicine and diagnosing metabolic
problems (Pavlou et al., 1985).

1.11.4 Fat-Free Mass (FFM)

Internal organs, bone, muscle, water, and connective tissue are included in this
category. Fat- free mass is different from fat mass and refers to muscle mass when
discussing weight control and body composition. The non-greasy mass includes all
bodily components except fat. It contains the body, bone, and muscle water. However,
in terms of weight loss and body composition, lean mass refers to muscle mass. Most
Americans are overweight or obese, increasing lean mass and decreasing body fat may
help improve your health (Lobo, 1987). Some writers employed BIA to assess FFM
and TBOW (TBW). Figure 1.15 illustrates the distribution of total body water (TBW),
intracellular water (ICW), extracellular water (ECW), and body cell mass (BCM)
(Kyle et al., 2003).

23
 Visceral Body cell
Protein mass

Intracellular water
(~44%)
FFM Total
Extracellular water body
(~29%) water

Bone mineral
(~29%)

FM weight (~FFM)

Figure 1.15:Schematic diagram of Fat-Free Mass (FFM) (Kyle et al., 2003).

1.11.4.1 Visceral Proteins (VPs)

VPs are used to indicate malnutrition, particularly in the elderly. Body composition
varies with VPs and age. VPs are close to FFM. Several research has examined the
association between albumins and FFM (Baumgartner et al., 1996). While (Volkert et
al., 1992) and (Bonnefoy et al., 2002) demonstrated body composition by calf
circumference and skinfold thickness techniques

The next idea is bodily water, which is part of the body's fluid makeup. Healthy Water
Balance is the word used to describe the appropriate water distribution in the body.
The optimal fluid distribution in the body is 3:2 intracellular to the extracellular fluid.

1.11.4.2 Intra Cellular Water (ICW)

It is the water inside the cells that bathes all necessary biological molecules including
the proteins and nucleic acids, consequently, the internal osmotic pressure is high and
water activity is low(Bonnefoy et al., 2002).

 Fluctuations in ICW level

24
A minor rise in ICW is not concerning. However, it is a favorable indicator of a shift
in body composition. For example, an increase in muscle mass indicates an increase in
muscle cell number and size.

Figure 1.16:Body Fluid Compartments (Clegg, 1984).

As muscles grow in bulk, they need more energy to fuel numerous cellular operations
and functions. Thus, an increased level of ICW may be related to increasing lean body
mass and is regarded as a favorable indicator that signifies the creation of healthy
body mass (Clegg, 1984). Among the factors for rising ICW are:

o Increased energy requirements

o Increased strength
o Increased immune system

1.11.4.3 Extra Cellular Fluid (ECW)

It is the extracellular fluid that makes up one-third of each multicellular organism's

bodily fluid. The interstitial fluid is the major component of extracellular fluid.
Extracellular fluid is the interior environment of all multicellular creatures, and blood

25
plasma is a component of it. The ECF is mostly composed of plasma and interstitial
fluid. The transcellular fluid is part of the ECF.

 Fluctuations in ECW

The increase in ECW in respect to ICW requires special attention. It majorly reflects
serious health risks when an excess of ECW is observed. Some of the concerns and
complications that are signaledby the increased levels of ECW are as follows:

o Inflammation

Inflammation is a process in which a part of the body or tissue gets damaged due to
some sort of injury. The human body is mechanized in such a manner that whenever
there is tissue damage, it sends additional blood supply to the inflamed area. This
results in extra water accumulation in that area. However, chronic inflammation is
observed that leads to increased ECW and long terms swelling and is usually caused
due to dysfunction or cellular stress. This further leads to several complications such
as cancer and heart disease(Clegg, 1984).

o Kidney Failure

The kidney plays an important role in filtering the blood fro m the unwanted toxins
that are produced by the body. When ECW gets accumulated due to the inability of
kidneys to filter out excess sodium in the diet, it indicates renal disease or in the long
term risk of renal failure. The swelling or the edema is the visible sign of a rise in the
extracellular water.

o Obesity

Obesity refers to the accumulation of unhealthy fat mass in the body. The excess of
body fat mass stimulates the increased production of hormones that further disrupt the
body healthy system. This further leads to the accumulation of excess ECW that
causes stress in the body’s internal organs that triggers a dangerous cyclic
effect(Falkheden and Sjogren, 1964).

26
1.11.4.4 Bone Mineral

Bone mineral is also known as bone salt. It refers to the inorganic bone phase that is
the component of bone tissue. It provides the bones the special compressive strength.
The bone mineral is composed of the following main parts

 Carbonated hydroxyapatite (Ca5 (PO 4 )3 (OH))

 Plate and globular structures as collagen fibrils of bones to form large
structures

Bone mineral is found to be a very dynamic entity of living organisms. It is found to

be continuously built, reabsorbed, and remodeled within the body. Bones are the
storehouse and reservoir of calcium that is required by the body to maintain
homeostasis (a process that balances the salt and water components within the
body)(Dalen and Olsson, 1974).

1.11.4.5 Total Body Water (TBW)

By weight, the average human adult male is approximately 60% water and the average
adult female is approximately 55%. There can be considerable variation in body water
percentage based on several factors like age, health, water intake, weight, and sex.
Various studies the adults (without age limit) and both male and female sexes, in
which the average of the adult human body is near about 65% of water. Whereas, this
can be modified based on age, sex along the amount of fat in body composition. The
amount of water in weight in this sample has obtained as 58 ±8% water for males and
48 ±6% for females(Kushner et al., 1992).

1.12 FACTORS INFLUENCING BODY COMPOSITION

Body composition is majorly affected by the following factors:

1.12.1 Genetic Composition

These are the genetic factors that govern the phenotypical appearance of an
individual. Whether an individual appears to be fatty or lean; is all determined by its
genetic constituent.

27
1.12.2 Age

This factor reflects the loss of muscle mass with the passing time or aging.However,
with regular exercise; one can overcome the creeping loss of musculature along with
bone strength.

1.12.3 Hormones

These chemical messengers play a vital role in water retention by the body that forms
the major proportion of body composition.

1.12.4 Menopause

High body fat is associated with a decrease in estrogen level in the body that in turn
reflects low energy expenditure. It is usually observed after the age of 45 years.
However, sometimes it is also related to the decrease in intense body activity or
leading a sedentary time.

1.12.5 Gender

It is observed that women exhibit more body fat as compared to men of the same age
which is justified by the nature to prepare their body for pregnancy and nursing of the
fetus.

1.12.6 Type of Physical Activity

A regular and routine that comprises an aerobic exercise or activity is found to be

associated with low body fat. However, any physical activity that is carried on for a
longer duration is found to be capable of burning calories and maintaining low body
fat. This usually involves weight resistance exercises to balance muscle mass in the
body.

1.12.7 Nutrition

This refers to the balanced nutrition of the body. For instance, when t he calories
intake is very high as compared to the calories burnt by the body then the extra
calories consumed are stored as body fat. However, balanced consumption of
macronutrients, i.e., fats, carbohydrates, proteins, and minerals is very important to
maintain healthy body composition(Spungen et al., 2003).

28
1.13 RELATIONSHIP BETWEEN OVARIAN CYCLE AND BODY
COMPOSITION

Body composition measures fat mass to lean mass. It is rich in protein, minerals, and
water. The oral cycle influences fertility and wellness. Examining reproductive health
using regular menstrual cycles Menstrual cycle length and regularity imply ovarian
steroid use. Premenopausal women's BMI fell in this study. The fat mass has gotten
less attention than excess body weight. A longer menstrual cycle increased BMI and
age of menarche, according to the study. Most active women in this study had regular
periods. Water retention and measurement discrepancies are caused by hormonal
changes.

Retention of water and measuring errors may occur during pre gnancy or menopause.
Everybody composition metric correlates with cycle length. BMI had the largest mean
cycle duration, although the relationship was nonlinear. Fat or muscle? Lack of
follicular phase estradiol is connected to a decreased risk of conception. Diet may
have altered reproductive capacity. Changing menstrual cycles increases with age.
Early ovarian cycles might be irregular for some women. Late adolescent cycles
become more regular. Women over 30 had shorter and less regular menstruation. The
loss of ovarian follicles produces a lengthier menstrual cycle. The menstrual cycle
starts now. 2nd, BMI (Body mass index). It is known to disrupt menstrual cycles.
Extremes Being underweight or overweight might affect hormones. Even little weight
changes that do not impact our BMI zone might cause hormonal changes and cyclicity
changes.

Weight changes may impact menstruation. BMI changes are associated withthe
menstrual cycle and fertility changes. Low BMI associated with irregular and heavy
menstruation Less obese ladies has more menstrual cycle troubles than normal-weight
women. Obese women are prone to PCOS.

The duration of the menstrual cycle lowered premenopausal women's BMI. The
difference in mean BMI across the three trials was statistically signific ant. Obesity
increased the probability of long cycles among college women, Harlow and
Matansoski discovered. Body fat has gotten less attention than lean body mass. In this
investigation. 2.44 percent of obese women had shorter menstrual periods than 1.83

29
percent of normal- weight women. Healthy females retain 15–20 kg of fat (25–30%) in
their hips and thighs throughout their youth and adolescence. As menstrual cycle
duration increased, so did BMI and age of menarche. More cycles with age. Changing
menstrual cycles have unclear reasons. This study found that moderately active
women had regular menstrual periods. The diverging menstrual cycle duration is
caused by increasing age and decreasing BMI.

Moderate physical activity, BMI, and age at menarche all impact the menstrual cycle.
Body mass alone does not reflect a person's health. Body composition may indicate
health. Using various methods, we may determine BF%, lean mass, fat mass, bone
mineral content, and total body water (TBW). Costs and skills vary. That's why
getting a precise BF% estimate is crucial.

It is uncertain whether physiological changes occurring during a menstrual cycle

might alter BF percent calculations. Body mass and fluid retention variations may be
connected to body pleasure. to assess nutrition and fluids An invasive diagnostic
method is used. Increased fluid retention throughout the menstrual cycle has been
connected to appearance concerns and poor body image views. Each study used BIA
with a single frequency of 50 or 100 kHz. In recent years, BIA technology has
evolved to analyze body composition utilizing frequencies up to 1000 kHz. The
reproductive cycle affects women's hydration levels more than men's, hence BIA is
useful in assessing body fat. During this period, the retaining fluid helps to change the
amount of body fat. Body composition is measured via bioelectrical impedance
analysis. Fatty tissue water slows the signal. It flies through the fat- free, water-rich
tissue-like muscle.

Usual alterations lead to overeating Weaknesses in the muscles and joints might cause
it. Exercise hormones Exercise releases hormones like beta-endorphins and
catecholamines. They may interfere with oestro gen and progesterone activities. PMS
is usual. More than 90% of menstrual women get PMS. Hormonal changes may cause
weight gain. Obesity is caused by an energy imbalance. Obesity affects women more
than men, and the risk varies by gender.

The ovaries' hormones seem to be vital. Estrogens cause female obesity. Estrogens
increase genitofemoral subcutaneous mass and decrease central AT mass in women of

30
reproductive age. Loss of estrogens improves total and lean body mass without
affecting weight.

Obesity is an epidemic. (BMI 30 is considered obese.) However, the impact of gender

on obesity is still unknown. In reality, women outnumber men. It is obvious from the
continual worldwide disparity in the prevalence of severe obesity that biological
factors, i.e. physiological sex differences, play a role Women-specific factors in the
two physiological causes of obesity, energy intake, and expenditure, have been
discovered (EE). Also, ovarian hormones, notably estrogens, influence female eating,
and EE. And ovarian hormones are linked to adipocyte metabolism. Because women
are more prone to obesity, researchers must understand how ovarian hormones affect
eating, EE, and body fat. These effects on women may be explored in numerous ways.
Women's menstrual cycles have long been examined by experts. Women with low
body fat percentages have irregular, uncomfortable, or no periods. Before beginning a
diet or training for an endurance event, find out your menstrual dysfunction threshold.
You get oestrogen from fat cells. This may cause decreased oestrogen production and
menstrual irregularities. Less body fat than low-calorie diets is considered to promote
menstrual disorder (Sun et al., 2006).

1.14 CHANGES IN BODY COMPOSITION DURING OVARIAN

CYCLE

Low body fat — females cannot menstruate below a particular body fat percentage.
Human beta-endorphins and catecholamines are released during exercise. These
hormones may influence oestrogen and progesterone action. Women have roughly 5%
more body fat than males. The female body is designed to protect her, including a
fetus. Women have more enzymes to store fat and fewer enzymes to burn fat.
Moreover, oestrogen stimulates enzymes in women's bodies, causing them to grow.

Female BIA users must be conscious of their monthly bodily cycles. Few wo men find
it beneficial to note their readings every day for a month to provide a baseline for
tracking body fat. Then take care of the monthly swings. Hydration changes due to
diet, coffee, alcohol, hard activity, stress, sickness, or taking prescribed med icines.
Women must examine weight and body fat percentage readings in the same situation
over time to track development. Your age and BMI will have a big influence on your

31
cycle. They will change our genes' cyclicity pattern. Our genetic clock dictates not
just how quickly and long we age (adolescence, maturity, and menopause), but also
our lifestyle. Habits may shorten or lengthen each stage. The mix of our genes and our
lifestyle patterns impacts our menstrual experiences on a daily, monthly, and annual
basis. We are more likely to share cycle profiles with our biological forebears,
siblings, and descendants. But it's also fun to study how the cycles alter, and if that's
due to lifestyle or new genetic combinations/mutations. That's where understanding
our present lifestyles and cyclicity about our age, BMI, and life habits come in help.
People with comparable lifestyles and ages may go through similar cycles.

The second major category is BMI (body mass index). It is known to affect
menstruation regularity. Hormonal deregulation may cause irregular menstrual cycles
in both underweight and overweight/obese women. Even minor weight changes that
do not move us into a new BMI zone might produce hormonal alterations and, as a
result, changes in our cyclicity (Borkan et al., 1983).

1.15 TECHNIQUES TO MEASURE BODY COMPOSITION

There exist several methods that can be employed for estimating the body fat
percentage at the gym, home, or with the health care practitioner. Some of the
important methods commonly used for measuring body composition are as follows:

1.15.1 Dual Energy X-ray Absorptiometry (DEXA)

Dual-energy X-ray absorptiometry (DEXA) is increasingly employed in clinical and

research settings. A DEXA scan needs a radiologist's supervision and is considered
the "Gold Standard" in body composition assessment. Total body DEXA scans
provide exact assessments of body composition, including BMC, BMD, lean tissue
mass, fat tissue mass, and fat fractional contribution. For many decades, estimating
body fatness from body density (underwater weighing) was considered the "gold
standard." Some experts say whole-body scanning (like DEXA) is the new "gold
standard." They are also evaluated against body composition measurements based on
fractional density from underwater weighing (De Lorenzo et al., 1998).

Dietary, exercise, sports, and other body composition modifying programs may all be
monitored using DEXA measures. They are also non- invasive, quick, and expose the

32
patient to fewer x-rays than a cross-country trip. For example, a DEXA test may yield
a whole body and up to 14 regional findings (Bachrach, 2000).

Figure 1.17:DXA scan (Bachrach, 2000).

1.15.2 Field Methods

They are portable and so appropriate for usage outdoors. Skinfolds, Anthropometry,
Near-Infrared Interactance, and Bioelectrical Impedance Analysis are four techniques
often used by physiologists to analyze body composition in the field. The doctor must
frequently choose an acceptable approach or prediction equation to appropriately
analyze each client's body composition. It is thus critical to assess the validity of field
methods and the predicted accuracy of equations (Blomberg et al., 1993).

1.15.2.1 Hydro-densitometry / Under-Water Weighing

The traditional method of measuring body composition is to do so underwater. The

approach, based on Archimedes' principles, produces knowledge of two
compartments: fat mass and fat- free mass. There is a live counter- force to the weight
of the displaced water. Because bone and muscle are denser than water, a person with
greater fat-free mass will weigh more. A higher percentage of fat mass in water might
make the body slimmer. Individuals are examined by submerging themselves in water
and forcing a 30 second forced expiration. This procedure is vital because retained air
in the lungs typically causes water loss. The submerged weight is recorded after the
forced decay.

33
 Figure 1.18 :Measurement of hydrostatic weight (courtesy of INCAP,
Guatemala) (Weight, 1996).

The fat body approximation and fat- free body density are used to measure these two
compartments for body composition. The fat- free mass is a heterogeneous container
that may be further split by its basic constituents: water is 73.8 percent, proteins are
19.4%, and minerals are 7.8%. Hydro densitometry is used to validate other
techniques. The graphic above shows the hydrostatic weight method used to
determine body density. Inversely proportional to the volume of a static gas at a
constant temperature, it is based on Boyle's law. Thus, under isothermal conditions,
the volume of air is reduced according to the rise in pressure (Weight, 1996).

Therefore, (1.2)

single paired situation of pressure for 1st condition

 single paired situation of pressure for 2nd condition

Voltage for 1st condition

 Voltage for 2nd condition

In contrast to isothermal conditions, under adiabatic conditions, the air is more

compressible. Poisson’s Law describes the relationship between pressure and volume
under adiabatic conditions as;

34
 (1.3)

Where, is the ratio of the specific heat of the gas at constant pressure to that at
constant volume (Cullen et al., 1990). The value of g is approximately 1.4 for air and
1.3 for CO2 (Daniels and Alberty, 1967). The system maintains the equivalency of gas
composition in the two chambers and the constancy of the g in the pressure-volume
relationship.

1.15.2.2 Air Displacement Plethysmography (ADP)

A method other than underwater weighing has been devised for assessing body
volume. Air displacement plethysmography utilizes air instead of water (ADP).
Subjects enter a sealed room that measures body volume by displacing air. Body
volume and weight (mass) are combined to calculate body density. The approach then
determines body fat percentage and lean body mass (LBM) using empirical formulae
similar to underwater weighing (for the density of fat and fat- free mass). ADP
operates based on gas laws (Ginde et al., 2005).

1.15.2.3 Body Volume Indicator (BVI)

The Body Volume Indicator (BVI) measures body form. Initially, BVI used white
light scanning equipment to assess body form. Recent developments in 3D measuring
have allowed BVI to be estimated using smartphone photos. A 3D silhouette requires
two photos. It may be used to determine body volume and fat distribution using MRI
data (Barnes, 2014).

1.15.2.4 Skin Fold Method

The skinfold test, conducted with a caliper, may also determine body composition.
Measuring the thickness of subcutaneous fat with gun-type calipers is a standard
skinfold technique. The abdominal, subscapular, arm, buttock, and thigh areas are
included. These metrics determine total body fat (Womersley and Durnin, 1977).

35
 Figure 1.19:Skin Fold Method (Womersley and Durnin, 1977).

1.15.3 MFBIA (Multi-Frequency Bioelectrical Impedance Analysis)

MFBIA is a non-invasive and low-cost way of assessing a person's body composition.

This procedure requires no special abilities, making it simple to manage. MFBIA
examines the composition of the body by calculating the impedance or resistance that
corresponds to the current flow. Fat-free conducts are better because it reduces the
barrier to current flow (Panorchan et al., 2015). Low resistance occurs in fat-free mass
due to greater electrolyte and water content. Adipose tissue is less conductive due to
high current flow resistance. High resistance is caused by low water and electrolyte
levels. The current electrode BIA analyzer is quite different from the traditional BIA
analyzer. The gel electrodes are positioned precisely using BIA analyzers. Automatic
calculation of a person's body fat and fat-free mass is done using approximated
equations. Using such equations, the impedance and information like height, mass,
gender, age, and body type are calculated (Aleman-Mateo et al., 2010).These
comparisons assessed impedance together with height, body mass, sex, age, and body
type to establish body composition.

The electric current used to compute bodily tissues is used to categorize BIA into
single or multifrequency. The single frequency analyzer penetrates the body at around
50 kHz to assess the whole body composition of segmental tissues. Unlike single-
frequency bioelectrical impedance analysis, MFBIA consumes biometric data at
diverse frequencies. The ECW and TBW may be measured using bioimpedance data

36
at low and high frequencies of 1 to 50 kHz and 100,200 or 500 kHz respectively.
Using external (red) clips, a low alternating current is fed through the body to assess
whole-body BIA. The voltage drop between the inner (black) clips is constant.
Calculate impedance (Z) using voltage-current measurements.

Figure 1.20:Multi-frequency Bioelectrical impedance measuring tool (Aleman-

Mateo et al., 2010).

The quantity of water in the body affects electric current flow. The instrument
measures how the signal is obstructed by various tissues. Tissues rich in electrolytes,
like blood, are very conductive, whereas fat and bone are not. BIA measures the
resistance to current flow through the body. Body fat is estimated by water.

1.15.3.1 Applications of MFBIA

MFBIA is a technique utilized to approximate the composition of the body. It assists

in assessing the percentage of water and fat in the body.

1. Different frequencies i.e. 0, 50, 100, 200 to 500 kHz are utilized in MFBIA for
evaluation of the content of FFM, TBW, intracellular water (ICW), and
extracellular water (ECW). MFBIA is an advantage of performing measurements
on specific sections of the body.
2. MFBIA is used to calculate the illness marker and also monitor the changes in
body fluids. The change of composition during an ovarian cycle is determined by
MFBIA. The BMI, lean mass, FM, FFM, TBW, ECF, ECS, ECM all parameters
are measured by MFBIA

37
3. To approximate the human body composition is relying upon the concept of the
electrical conductivity of the fat-free mass is much higher than that of fat.
4. To enable comparison of data from different studies and to ensure the multiple
measurements in both laboratory and clinical settings can be reliably taken.
5. The measurement of MFBIA through conventional gel electrode techniques has
effective very less or no effect. It is necessary to examine the possible effect
ovarian cycle on MFBIA measurements.
6. Multi- frequency bioelectrical impedance analysis (MFBIA) may be a relatively
simple, quick method to measure body composition, for example, fat mass and fat-
free mass, having the advantages of being portable, relatively cheap, and not
requiring specialist training for use.

The essential idea of MFBIA is that when an electrical current goes through bodily
tissues, certain extracellular components block it while intracellular components
enable it to flow freely. It is a standard technique for assessing body fat and muscle
mass. MFBIA uses a mild electric current to determine the body's impedance
(resistance). Muscle stores most of our bodily water. So, a muscular individual has
more body water, which leads to lower impedance. The approach has gained
popularity since its commercialization in the mid-1980s due to its simplicity and
mobility. It is a basic tool for assessing body fat. MFBIA measures electrical
impedance, or resistance to the passage of an electric current through human tissues,
which may be used to estimate total body water (TBW) and fat- free body mass
(FFBM) by comparing body weight and fat.

1.15.3.2 Challenges faced by MFBIA

They are simple to use, but they are less accurate than those used therapeutically or in
nutritional and medical practice. They tend to underestimate body fat. The
measurement of body fat is less shortly after eating, resulting in a day-to-day variation
of up to 4.2 percent of total body fat.

Due to lowered impedance, moderate activity before BIA measures overestimates fat-
free mass and underestimates body fat percentage. If you exercise for 90–120 minutes
before your BIA, you will gain over 12 kg in fat- free mass, but lose body fat. MFBIA
should be avoided for several hours after moderate or high- intensity exercise. MFBIA

38
has been determined to be sufficiently accurate for multi- measurement usage, and are
better suited to use to monitor changes in body composition over time for people.
Two-electrode foot-to- foot measurement is less accurate than 4-electrode (feet, hands)
and eight-electrode measurement. While certain four- and eight-electrode devices had
weak limits of agreement and systematic bias in estimating visceral fat percentage,
they had high accuracy predicting resting energy expenditure (REE) compared to
whole-body MRI and dual-energy X-ray (DEXA). Multiple frequency BIA devices
with eight electrodes had a 94% correlate with DEXA for assessing body fat
percentage. The mechanism of detecting current flow through the body (impedance) is
frequency-dependent. To measure the body composition, all multiple frequency
analyzers emit frequencies through the body ranging from 1-1000 kHz. Single-
frequency analyzers cannot discriminate between IBW and total body water at 50
kHz. The device's inbuilt computer controls the different frequencies by measuring
resistance from various body segments (Raeder et al., 2018).

1.15.4 Circumferences and other measurements

Assessment of somatic (skeletal) protein is typically determined by simple

measurements and calculations including mid-arm circumference (MAC), mid-arm
muscle circumference (MAMC), and creatinine height ratio (CHI). Creatinine height
ratio is calculated as 24- hour urine.Creatinine multiplied by 100 over the expected 24-
hour urine creatinine for height. This calculation results in a percentage that can
indicate protein depletion (McCarthy, 2006).

39
CHAP TER 2: LITERATURE REVIEW

2.1 VARIATION OF BODY COMPOSITION WITH OVARIAN

CYCLE

(Hulens et al., 2001) have evaluated the nature and size of differences in submaximal
as well as maximal exercise potential parameters among lean and obese women.
Overall, 225 healthy obese women have been selected for 18-65 years (BMI>=30 kg
/m2) and 81 non athletic women (BMI<or =26 kg/m2). Anthropometric measurements
(weight and altitude), body composition evaluation (Bioelectric impedance method),
and bicycle ergometry test have been performed for maximum maneuver testing.

(Sowers et al., 2007)had investigated the chronological age and ovarian period to
understand how this midlife can affect the size and composition of the body in
women. Nearly 543 premenopausal African-American and Caucasian women had
taken part in the study. From the experiment, it has been concluded that both
time(chronological aging), as well as ovulation, led to significant changes in aging,
body composition (fat and skeletal muscle mass), and waist circumference. These
changes have a significant impact on the formation of a metabolic environment that
can be healthy or unhealthy (Kyle et al., 2003).

(Shafe r et al., 2009) have used eight numbers of electrodes using multiple frequencies
BIA devices to determine percentage BF in adults. The classification has been
performed on the three categories named normal, overweight, and obese (Baumgartner
et al., 1996).

(Aydin et al., 2013) analyzed the influence on the composition of the body of Ethinyl
estradiol and drospirenone. Parameters of body composition were evaluated by
analysis of bioelectrical impedance. Also determined were serum androgens, lipids,
insulin resistance, and glucose metabolism. In comparison to healthy women, skinny
women with PCOS have a similar body composition.Despite no change in clinical
anthropometric measures, including weight, body mass index (BMI), and waist to hip
ratio (WHR), oral contraceptives (OC) treatment for 6 months in PCOS patients
results in an increased overall percentage and trunk fat percentage.

40
(Veasey-Rodrigueset al., 2013) examined the weight, body composition changes, and
outcome in patients treated with temsirolimus, a mTor inhibitor that has weight loss as
one of its side effects. Patients with higher-grade toxicities tended to lose more body
fat, suggesting a possible end-organ metabolic effect of temsirolimus. Sixteen patients
with advanced solid tumors treated with temsirolimus were studied; body composition
has been analyzed by utilizing computerized tomography images.

(Komarowska et al., 2013) have found one of the most common disorders named
polycystic ovary syndrome (PCOS) in women of reproductive age. The differences
observed in hormonal and metabolic profiles have also been observed in two different
groups of patients including normal and elevated Body Mass Index (BMI). The age
group with BMI<25 and BMI>25 has come under two different groups. To observe
whether psychological parameters of lean and obese patients having PCOS have
significantly different as well as there is a correlation among these features and the
concentration of different hormones.

(Saluja et al., 2014) discussed theimpact ofmenstruation, pregnancy, and menopause

on salivary flow rate, pH, and gustatory function. There was no statistically significant
difference has been observed among the groups correspond ing to salivary flow rate
but pH values have been significantly lower in post- menopausal women (p<0.05). It
has been concluded from the obtained result; the pregnant and postmenopausal
women appeared to have a reduced perception of sucrose, which can alter eating
habits, like intake of more sweet foods whereas no significant difference is observed
in taste perception of NaCl, citric acid, and quinine hydrochloride between the
subjects.

(Glintborg et al., 2014) observed whether treatment with metformin (M) or M

combined with oral contraceptive pills (OCPs) produced a better result in body
composition as compared to treatment with OCP alone. M treatment alone or in
combination with OCP was associated with weight loss and improved body
composition compared with OCP, whereas free T levels decreased during M+OCP or
OCP. Combined treatment with M+OCP should be considered as an alternative to
treatment with OCP alone to avoid weight gain in PCO.

41
(Ezeh et al., 2014) have analyzed the differences in metabolic dysfunction among
PCOS and control women related to differences in their fat to lean mass ratio. As
compared to the controls of similar BMI the women with PCOS demonstrated adverse
body composition characterized through enhanced whole-body fat relative to lean
mass that has been associated with differences in metabolic dysfunction among the
two groups.

(Van Caenegem et al., 2015) examined the grip strength (hand dynamometer),
biochemical markers of bone turnover (C-terminal telopeptides of type 1 collagen
(CTX) and procollagen 1 aminoterminal propeptide (P1NP)), total body fat and lean
mass, and areal bone mineral density (aBMD) by Dual-X-ray absorptiometry (DXA)
and fat and muscle area at the forearm and calf, bone geometry by taking the 23 trans
men (female-to-male trans persons) and 23 age- matched control women. It has
indicated that testosterone therapy in trans men induced changes in body composition
including a gain in lean body mass, muscle mass, and strength. It also hypothesized
that the increased bone turnover and discrete increases in aBMD at the total hip and
trabecular vBMD may be due to the indirect effects of testosterone therapy.

(Bubachet al., 2016) analyzed the association between age at menarche and body
composition (BMI, waist circumference, fat and fat- free mass indices, and abdominal
visceral fat thickness) in adolescence and adulthood among women. In the end, it has
to be clear from the outcome, the association between age at menarche and body
composition in adulthood is strongly related to body composition in late childhood.
Therefore, age at menarche would be a proxy of pre-pubertal body composition.

(Stachoń et al., 2016) have examined whether the tendencies and scope of changes in
different body mass, body composition along body girths across the menstrual cycle
have similar or different in women with different body build. As per the outcome of
this work, the amount of fat mass increased significantly across the menstrual cycle
considerably in case of higher hydrated around 0.66 kg as well as in case of slim
women has around 0.54 kg. The significant changes among consecutive phases of the
menstrual cycle in the waist and hip girths along with suprailiac skinfold thickness in
some age groups of women indicated the impact of the influence of fatness along with
hydration status including slenderness.

42
(Karvonen-Gutierre z et al., 2016) studied the clinical experience of the menopausal
transition through obesity status and analyze the evidence in the context of
menopausal transition and influences of reproductive hormones on the weight of the
body, composition of the body. The menopausal transition is combined with weight
gain and increased central body fat distribution, the number of experiments suggested
that changes occur in weight as per chronological aging however changes in body
composition along with fat distribution have specifically because by ovarian aging. As
per the high prevalence of obesity in midlife, several women become overweight as
they transition through menopause.

(Lukaski et al., 2017) have found a correlation between phase angle and different
nutritional statuses. It has been determined that if the phase angle is small it does not
reflect impaired nutritional status. The BIA has been used to define phase and
determined that is a simple, easy-to-use test to assess the risk of angular illness and to
evaluate the prognosis. The measurements must be implemented in strict protocols
with a phase-sensitive tool, which has been confirmed with a periodic sensitivity.
Patient results were given by BIA instruments should be trained and certified to
comply with the standardized protocols and should ask questions about non –
intermittent or clinically improper results.

(Geronikolou et al., 2017) demonstrated the differences in body composition indices

of metabolism, homeostasis including inflammation among Greek adolescent females
who suffered from PCOS and age and BMI- matched non-PCOS controls. To
accomplish the aim of this work, 13 PCOS, as well as 9 non-PCOS controls in the age
of 13-24 years, have been considered. The body composition indices have been
measured in each adolescent such as extracellular water (ECW) as inflammation
marker, complete body water as homeostasis marker, the ratio of extracellular mass to
body cell mass, fat mass (FM), fat- free mass (FFM), and intracellular water as
markers of body mass composition along with metabolism. The non- linear analysis
indicated that there have been no statistically significant differences in the features of
body composition among PCOS patients and controls.

(Chitme et al., 2017) determined the body composition of infertile women in terms of
PCOS along with analyzing the important factors to those in women without PCOS.
The authors have also observed a highly significant correlation betweenthe incidence

43
of PCOS and BMI. The prevalence of overweight, obesity, and central obesity has
been considerably higher in women with PCOS. The obtained outcome of this work
has encouraging and indicated the new ways of treating body fat in PCOS patients.
Specifically, PCOS is a very common disorder in reproductive-aged women among
25-30 years.

(Winkler et al., 2017) studied the association among body composition measures
along with a menstrual status for a large sample of young patients through a history of
anorexia nervosa along with computing the predicted probability of the resumption of
menstrual function. After that to establish whether the fat percentage is better as
compared to body mass in predicting the resumption of menses.

(Cumberledge et al., 2018)investigated the impact of the menstrual cycle as

determined by contact electrode BIA analysts, on the body composition. Forty-three
university women participated voluntarily in this study. In the following menstrual
cycle phases, subjects evaluated their body composition using four different contact
electrode BIA analyzers: menstrual, follicular, early, and late luteal. No significant
differences in body composition measurements between cycle phases have been found
irrespective of the BIA analyst used for the evaluation.

They have also demonstrated that BIA is the relatively cheapest approach to compute
the composition of the body. BIA manufacturers recommend female research, though
identifying water retention during the development of the ovaries. The key idea of this
work is to use contact electrodes to investigate the impact of the ovarian cycle on the
body's composition.

It is to be observed by the BIA that women of age 43 whose appropriate age and body
mass is taken into account as age = 21.5±1.3 years; body mass index=24± 3.8 kg/m2.
Four different BIA analysts assessed female body composition for contact electrodes.
During the menstrual phases, the analysis of body composition is categorized into
follicular, early, and late- luteal. No worthy variation in the measurements of body
composition was observed in the cycle phases, despite the various BIA analysts used
for assessment. It has to be cleared from this work; the BIA electrode for contact
electrode consumed in this research possibly has utilized at any time during the
menstrual cycle of women with modifying the composition value of the female body.

44
(Al-Bayyari, Nahla, 2018) studied the impact of Vitamin-D supplementation of the
body composition along with on the body composition as well as on the
anthropometric measurements of overweight women through PCOS and to analyze the
effect of this effect on ovaries ultrasonography and regularity of menstrual cyc le. This
study has designed a prospective randomized, double-blinded placebo-controlled
clinical trial implemented on 60 overweight women in the age group of 18-49 years
including PCOS and deficiency of Vitamin-D.

(Dmitruk et al., 2018) have determined the differences in body composition along
with distribution of fatty tissue in women through the groups discriminated against
based on their menstrual status. The status of menstrual has been determined as per
the guidelineprovided by WHO in which three groups have been determined such as;
women in the pre- menopausal period in group 1, the pre- menopausal period comes
under group 2 and in group 3 the post- menopausal period comes.The results have been
examined on the somatic built of women in distinct groups. Additionally, the
percentage of women having underweight, normal content of fatty tissue including
those with overweight as well as obesity has been computed. In the case of group 1,
the higher values of body mass, hip circumference along with most of theskin folds
have been observed, however in group 3 has characterized through a higher
percentage of body fat (PBF) and the minimum contents of lean tissue, soft tissue and
total water content in the body.

(Prado et al., 2018) examined the significant changes in decomposition of the body
between pre and post-menarche girls of the same chronological age. The presented
analysis indicated the significant changes in terms o f weight and height among pre
and post morphological studies along with classifications between the anthropometric
methods along with bioimpedance to compute the composition of the fat body.

(Di Rosaet al., 2020) collected a review about existing work of clinical trials
published between January 2014–November 2019 on people with obesity (PWO)
which observed VLCKD effects on anthropometric parameters, body
composition, satiety, lipid profile, and microbiota. The obtained outcomes from
discussed work have indicated that VLCKD could be useful to ameliorate the quality
of life and sleep of PWO. It leads to rapid weight loss and results in impro vements
in body mass index (BMI = kg/m2), waist circumference, and fat mass reduction

45
preserving lean body mass and resting metabolic rate. This eating pattern reduced the
desire to eat and increased satiety.

(Dos Santos et al., 2020) analyzed the impact of distinct kinds of exercise on
reproductive function and body composition for women with PCOS. Based ona meta-
analysis, there has been lower certainty analysis of little to no impact of only exercise
up on main reproductive outcomes. Whereas, the moderate certainty of analysis that
aerobic exercise has a positive impact on the composition of the body. Still, it has
been difficult to explore the impact of different types of exercise on health outcomes.

(Polak et al., 2020) investigated the relationship among serum androgens

concentration, insulin resistance as well as distribution of fat mass through dual-
energy X-ray absorptiometry (DXA) method in several PCOS phenotypes based onthe
Rotterdam scheme. The women datasets taken for this research havebeen divided into
a different number of women’s such as 34, 20, 20, and 15 correspondingly into
distinct phenotypes i.e. phenotype A, phenotype B, phenotype C, and phenotype D
along with 57 control subjects. The result shows that women with phenotype A have a
higher amount of visceral adipose tissue (VAT) and android/gynoid ratio (A/G) ratio
in contrast to the control groups so the metabolic disturbances must be higher
pronounced in the phenotype.

(Sare mi et al., 2020) examined and compare the BMI and physical activity level
(PAL) among women with and without infertility. The score of physical activity for
infertile women has been estimated to be low in this presented work that indicates the
sporting inactivity or very low PAL in subjects. The complete prevalence of
underweight, normal weight, overweight, and obesity in infertile subjects has been
8.7%, 26.7%, 34.7%, and 30% correspondingly. The frequency of overweight and
obesity has been reported to be significantly higher in both fertile and infertile
subjects. The physical activity score in the case of fertile women has been estimated to
be medium well as the prevalence of underweight, normal weight, overweight, and
obesity infertile subjects has been 29.8%, 40.4%, 17.2%, and 11.3% correspondingly.

(Shreffle r et al., 2020) have studied the resolution of infertility has associated with
increased life satisfaction along with self-esteem, rather than a decrease in depressive
symptoms. The considered samples for the present study including the women

46
inseveral 283 without children they satisfy the medical criteria for infertility or
perceived a fertility problem. The depressive symptoms, self-esteem along life
satisfaction have been assessed for all participants. There has not been a relationship
among infertility resolutions and change in depressive symptoms has been observed.
Involuntarilychildless women who resolved their infertility enhanced self- esteem
along with the satisfaction of life.

(Rael et al., 2020) observed the impact of female sex hormones on body fluid
regulation and metabolism homeostasis. However, it has not been clear whether
hormone fluctuations throughout the menstrual cycle (MC) along with oral
contraceptive (OC) consumption affect the body composition (BC). Therefore, the
main aim of this work has to examine the BC over the MC and OC cycle
corresponding to well-trained females. As per the obtained outcomes, the fluctuation
in sex hormones throughout the menstrual and OC cycle does not impact the BC
variables measured through bioelectrical impedance in females. So, it seems that
bioimpedance analysis can be implemented at any moment of the cycle in the case of
both i.e. eumenorrheic women along with the women using OC.

(Woodford et al., 2020) have examinedwhether phases of the MC have an impact on

common measures of body composition. The lean body tissue has approximately 73%
water fluctuations in the body because variation in women's MC may impact
measurements of body composition. However, there have been no changes in body
composition values among the three types of devices utilized to quantify body
composition. The obtained results indicated that the differences in technology
consumed to quantify body composition that may explain several results around this
study.

(Ihalainen et al., 2020) evaluated the body composition, energy availability, training
load, and menstrual status in young elite endurance running athletes (ATH) over 1-
year along with provided the investigation of how these factors differ between non-
running controls (CON), amenorrheic (AME), and eumenorreic (EUM) ATH. The
correlations to injury, illness, and performance have also been examined. The outcome
indicated that ATH has lower body mass, fat percentage, and BMI in contrast to CON
whereas, energy availability has not differed among ATH and CON.

47
(Frandsen et al., 2020) observed that an enhanced plasma estrogen concentration
together having low progesterone concentration in late-follicular (LF) leads to modest
but significant improvement in peak fat oxidation rate (PFO). The author got from the
outcome that there has not been any difference in body weight, the composition of the
body, or peak oxygen uptake among any of the menstrual phases in healthy young
eumenorrheic women has considered in this study. It has to be concluded that the
naturally occurring fluctuations in the sex hormones estrogen and progesterone have
not been effective badly for whole body PFO in young eumenorrheic women
measured during a graded exercise test.

2.2 EVALUATING TECHNIQUE FOR MEASURING BODY

COMPOSITION

(Adami et al., 1993) have investigated the bioelectrical impedance in a fairly

generous group of patients with protein malnutrition following the biliopancreatic
diversion (BPD) as an effective surgical method to treat morbid obesity. This work is
also composed of an extended distal gastrectomy with a very long Roux-en-Y that was
reconstructed 50 cm from the ileocecal valve. The main aim of the current study is to
determine if BIA measurements have been consumed to detect the changes in the
body composition of BPD patients with protein malnutrition during nutritional
support.

(Feldmar et al., 2009) described a recent approach of MFBIA for patients through
controlled ovarian hyperstimulation (COH). The MFBIA has been helpful into
estimate as well as inthe diagnosis of enormous forms of ovarian hyperstimulation
syndrome (OHSS). To perform this work, there is 53 patients have been considered
here. Impedances to the surface body were determined at 1,5,50 and 100 kHz. The
study group involved 13 patients with progression of the advanced grade III-V OHSS.

The best correlation was given by the frequency of 5 kHz (9.08 p<0.0001). In
presented work suggested that analytical values in improved OHSS ha ve enhanced as
compared to the recovery amount of oocytes (> 20 oocytes-RR 5, 71).

48
Diminishing impedance revealed an increase in OHSS development uncertainty. The
highest predictive values of OHSS were observed for impedances at 5 kHz (Miller et
al., 2016).

(Alwis et al., 2010) have presented the dual-energy X-ray absorptiometry data in
young and healthy women. The bone mineral content, bone mineral density, bone size,
lean body, and fat mass have been measured through dual-energy X-ray
absorptiometry in the total body, lumbar spine, hip, arms, and legs. The Swedish
children have similar bone mass to children in the Netherlands with higher than
children in Canada and Korea. The height, weight, bone mass, bone size, and lean
mass increased constantly from age 6 until the rapid enhancement in all traits at
puberty. The pubertal growth spurt started earlier in girls as compared to boys whereas
the spurt in boys hada higher magnitude and occurred for a longer period.

(Kaul et al., 2012) have validated a novel fully automated method in which
abdominal VF can be measured through DXA. After that, the authors have explored
the association among DXA-derived and ominal VF including different indices for
obesity. The considered obesity has; BMI, waist circumference, waist‐ to‐ hip ratio,
and DXA‐ derived total abdominal fat (AF), and SF. To execute the task in this work,
124 adult men and women of age 18-90 years represented a wide range of BMI values
measured with both DXA and CT in a fasting state within the interval of 1- hour. The
coefficient of determination (r2 ) for regression of CT on DXA values has been
obtained 0.959 for females, 0.949 for males, and combines 0.957.

(Dixon et al., 2013) used BIA techniques to analyse the food's influence on both
impedance and body fat percentage. To this end, the authors considered that 43 adults
consist of 23 females and 20 males who are between 18.5 years of age and 21.1 years
of age with a body mass index of 20.2 kg/m2 to 28 kg / m2 .Three different BIA
analyzers named Bone, Segment, and multifrequency BIA were experimented with. It
was found that the segmental BIA and multifrequency BIA were increased by 0.8 kg,
0.8 kg, and 0.7 kg respectively after 20 minutes of eating from bone to bone BIA.

(Marinangeli et al., 2013) discussed inter and intra- variation of DXA-derived body
composition measurement obtained with fan-beam DXA technology. In this current
work, the quality-control procedures have also been highlighted that must be

49
implemented before initiating the short-term single or multicenter clinical trials which
consumed DXA as a tool of body composition analysis to analyze the impacts of an
intervention on lean/fat mass loss or accretion. The authors have also provided a
discussion about considered while using DXA technology to compute the composition
of the body in the context of short-to-medium terms clinical trials that access nutrition
or exercise interventions.

(Hind, K., and B. Oldroyd, 2013) determined the in vivo precision of the Lunar
iDXA to measure the appendicular and trunk lean along with fat mass in a
heterogeneous group of adults. Two successive total body scans ha ve been performed
on each participant through re-positioning between scans. There have been excellent
agreements between consecutive scans for measurements of both fat and lean
appendicular and trunk composition. The precision for body composition corresponds
to every region has lower than 2% coefficient of variation (CV). The obtained
precision for bilateral appendicular body composition has lower than 4% and 2.5 CV
corresponds to arms and legs. It has to be clear from the obtained findings of this
work, the iDXA has been considered a valuable tool to repeat measurements of body
composition in adults.

(Carver et al., 2013) examined the precision of iDXA for total body composition and
fat distribution measurements for several obese patients. The two consecutive iDXA
scans with repositioning of the total body have been conducted for each participant.
The CV, root mean square (RMS) averages of standard deviations of repeated
measurements, the corresponding 95% least significant change, and intra-class
correlations (ICC) has been computed. The CV and RMS of the mean of the sum of
squares of differences among two measurements were also found. The obtained results
of the present work indicated that researchers may now precisely assess the total body
including some part of body composition in various obese individuals through an
iDXA scan.

(Ryo et al., 2014) demonstrated the influence of short-term calorie restriction and the
variation in VFA, determined by computed tomography (CT) and abdominal BIA has
identified. It has to be noted from the experimental analysis that the penetration of
short-term visceral fat leads to an improvement of cardiovascular risk metabolic

50
problems.The visceral fat is placed in the mesentery and the momentum delivers fat-
free acid to the liver via the portal vein.

(Mellis et al., 2014) have determined the in-vivo precision of CoreScan to measure
the VAT mass in a heterogeneous group of young elder women. To accomplish the
task, 45 adults of age around 30 ranging widely in the context of BMI has 26.0 kg/m2
received two consecutive total body scans including repositioning. The considered
samples have been divided into two small groups, normal-weight and
overweight/obese based on BMI for precision analyses. The finding of this work
indicated that the precision of DXA-VAT mass measurements enhances by BMI but
the caution must be consumed with %CV derived precision error in the case of normal
BMI subjects.

(Carver et al., 2014) evaluated the precision of GE Lunar iDXATM to assess the
BMD composed of lumbar spine L1–L4, L2–L4, the total hip, femoral neck, and total
body in several obese populations with BMI > 40 kg/m2 . There were two consecutive
iDXA scans of the total body, lumbar spine, total hip, and femoral neck that has been
calculatedto correspond to each participant.The CV, RMS average of standard
deviations of repeated measurements, and the corresponding 95% least significant
modification and ICCs have also been computed.

(Kaminsky et al., 2014) have determined the precision for both total % fat along with
all the regional % fat measures acquired through both Lunar Prodigy and Lunar iDXA
software. In current work the region of interest (ROI) has been computed
automatically through the enCORE software for total body, android, gynoid,trunk,
legs, and arm regions along with the region % fat data has consumed for analysis
purposes. Taking care of the status of body composition modifications according to
age has gained more clinical acceptance. The obtained outcome of this work added
support that DXA measurements have been consumed with a high level of confidence
for serial testing of patients.

(Tsigos et al., 2015) examined the association of body composition measurements

through a recently updated dual- frequency bioimpedance device (BIA-ACC) through
chronic stress or inflammation biomarkers along with the presence of medically
unexplained symptoms (MUS). The considered female dataset has been divided into

51
different groups in Group A contains 10,416 lean subjects with MUS, in next Group B
composed of 58, 710 lean subjects involving MUS, and at the end, 30,445
overweight/obese subjects without MUS comeunder Group C that also includes
excessive fat mass. In case of the Group B the total body extracellular water has been
higher, whereas intracellular water (ICW) has lower. The lower FM as well as higher
skeletal mass (SK) and phase angle (PA) as a contrast to Group B along with lower
circulating high sensitivity CRP levels as a contrast to Group A and Group C.

(Von Hurst et al., 2016) assessed the validation of BIA against ADP and DXA to
measure the percentage BF along with to test the reliability of such methods. In this
work, considered adult females with a wider range of BMI have been tested twice
during 5 days. After getting the outcome of this work, BIA indicated the improved
relative agreement to estimated true value but wide limits of agreement. BIA
underestimated 2% of BF across all values. DXA showed improved relative
agreement to the estimated true value along with the good absolute agreement of ADP
including wide limits of agreement as well as the under and overestimation at high and
low percentage BF levels correspondingly.

(Hicks et al., 2017) discussed several different technologies to Performa comparison

of body composition outcomes considering two phases of menstrual cycles along with
investigating whether being on hormonal birth control or not has any effect on results.
For this purpose of 39 females have been taken into account among them 15 were on
hormonal birth control (BC) and 24 were not (non-BC). Another task has been done
here which determined if body satisfaction or perception of fluid retention differed
across phases or between groups. The demographic and body satisfaction
questionnaires have been completed before testing on body composition. This testing
involves DXA, BOD POD including three separate bioelectrical impedance analyses.
There has not been a significant difference in overall body satisfaction.

(Machado et al., 2017) proposed and validated an anthropometric model for

simultaneous estimation of LST, BMC, and FM in healthy girls through a multivariate
approach of densitometry approach. From the result it has clear that it’s possible to
predict DXA body components through only four predictive measurements; body
weight (BW); supra- iliac skin fold (Sisk); horizontal abdominal skinfold (HaSk) and

52
contracted arm circumference (Caci) with high coefficients of determination and low
estimation errors.

(Ripka et al., 2017) analyzed the nutritional status of Brazilian adolescents along with
presented the skinfold thickness model (ST) to estimate body fat developed by
Brazilian samples through DXA as a reference approach. The prevalence of
overweight in adolescents has correspondingly 20.9%, and obesity 5.8%. Regression
analysis through ordinary least square method (OLS) allowed obtainment of three
equations with values of R2 = 0.935, 0.912 and 0.850, standard error estimated = 1.79,
1.78 and 1.87, and bias = 0.06, 0.20 and 0.05 correspondingly.

(Fonseca-Junior et al., 2017) have examined the validity of predictive equations

based on skinfold measurementin contrast to the DXA to estimate the body fat in
adolescent modern pentathlon athletes. The body composition of adolescent athletes
hasoften been computed scientifically along within sports through utilizing reference
equations developed through nonathlete adolescent populations.

(Silva et al., 2018)calculated the bio- impedance values obtained with MFBIA
spectroscopy (BIS, Xitron 4200) compared to a frequency of 50 kHz, measured by a
single frequency instrument (1-enkel) used by both active and elite athletes. The
experiment consists of 126 numbers of active men taking part in sports between the
ages of 20 and 39 and women athletes between the ages of 18 and 39.Bio- impedance
measurements of current single and multi- level devices were not used instead of each
other was observed. Because of the methodological as well as biological factors, there
was no consensus between devices to determine the individual values of R, X_c, and
Z which is a highly active population (Esco et al., 2015).

(Cavedon et al., 2018) have focused on twofold; such as implementing and validating
the sport-specific anthropometric predictive equation for total FM along with lean
mass components in female handball players to be utilized in the sport. Secondly,
cross-validated in female team handball players have introduced independently, the
predictive equations for BC in female athletes. The developed model has statistically
significant (P<100) for total body FM and managed R2 = 0.943, standard error to
estimate (SSE) = 1,379g. The percentage FM is managed as R2 = 0.877, SEE=2% and

53
the fat-free soft tissue mass (FFSTM) has managed as R2 = 0.834, SEE = 2,412g, the
fat-free mass (FFSTM + bone mineral content; obtained R2 = 0.829, SEE = 2,579g).

(Tinsley et al., 2018) have discussed the reliability of body volume computations
along with their usage in 4- compartment framework while employing precise
scheduling of assessments and dietary standardization. The 48 recreationally active
males and females completed two pairs of identical assessments, composed of a DXA
scan along with single- frequency bioelectrical impedance analysis. Each assessment
within a pair has been separated in 24 hours while participants have offered a
standardized die.

(Dordevic et al., 2018) have examined the variability of body composition measures
of weight-stable adults taken on the GE iDXA. The task has been performed through
the assessment of the CV% along with the least significant change (LSC) of
measurements for 3 repeated total body scans over 3- months which were the
minimum period typically desired to measure clinically significant changes in body
composition in the context of nutrition and lifestyle interventio ns. Moreover, the
precision of the same instrument through the standard protocol of two repeated scans
on the same day by repositioning of subjects. They have also hypothesized the
variability among measurements for each body segment of total body, android,
gynoid, and VAT would be enhanced as compared to those observed for the same-day
precision measures.

(Achamrah et al., 2018) have reported the small bias specifically in patients with
BMI between 16 and 18 suggested that BIA and DXA methods have interchangeable
at a certain level. Whereas the concordance among the two methods at the individual
level is lacking irrespective of BMI. The lack of concordance between BIA and DXA
methods at the individual level, irrespective of BMI has also been discussed here.
Further, this work requires to development of novel BIA-specific equations as per the
BMI class. It has also been analyzed that the patients with BMI>39 differences
between DXA and BIA varied with BMI.

(Vengerova et al., 2018) have discussed the ways to construct the academic health
physical education service in the age of 17-18-year-old females with the application of
biometric-based body composition rating tests versus ovulation or menstrual cycles.

54
The health physical education service programs have been designed with the
application of the modern active health building technologies with a special priority to
the harmonized physical development along with somatic health standards. The data
obtained from this analysis show the importance of female trainees’ body composition
tests for health-related education services being designed on the most efficient basis
with the application of modern building technologies modifiable to the ovulation or
menstrual cycles phases.

(Te wari et al., 2018) contrast the accuracy of measurement of body composition
made through DXA, analysis of CT scans, and BIA. The task performed using DXA,
CT, and BIA has been performed in 47 numbers of patients collected from two
clinical trials analyzing metabolic changes associated with major abdominal surgery
or neoadjuvant chemotherapy for esophagogastric cancer. In which DXA has been
performed the week before surgery along with before and after initiation of
neoadjuvant chemotherapy. BIA has performed during time points and utilized with
standard equations to compute FFM. However, an error understanding FFM has been
demonstrated with BIA which may be a useful modality to quantify body composition
in the clinical situation.

(Carrion et al., 2019)examined the effect of three BIA in athletic young adults. In
which total 51 numbers of participants with 26 men and 25 women have been
considered to determine fat through utilizing arm- arm bi-polar solitary frequency
device, leg-leg solitary frequency device. The measuring of PBF has been done
randomly and in equal amounts with three devices. The recurrent events ANOVA
founds to be distinct in the assessment (p <0.001) between all devices. The
measurement of correlation coefficient has been performed between arm–to–arm and
MFBIA for males and females along with the moderate relationship between leg- leg
BIA and MFBIA. This information shows a strong contract among each of the three
devices and any of them can be utilized to locate the PBF changes over time. Whereas,
distinction insignificant amount corresponds to PBF values in between devices
advised best practice for the composition of the body to be consumed constantly over
time to evaluate the valid device (Deurenberg et al., 1988).

(Beato et al., 2019) researched the effects of bariatric surgery on women's body
composition (BC). Body mass between 42 kg / m2 is subjected to gastric bypass

55
surgery for experimental data of 20 Class III obese females aged 24 to 34 years. The
experiment was conducted in body 230 and DLW before six months and after six
months of surgery. The accuracy was investigated in terms of square root mean error.
The mean square error of -1.40 to 0.06 kg has been observed (Dixon et al., 2008).

(Grzegorczyk et al., 2019) depicted the correlations among the selected parameters of
body composition assessed through DXA and anthropometric methods. This work has
been performed on 50 women aged 51-85. The adipose mass and fat-free mass have
been assessed through the DXA method along the nutritional status of the participants
has been analyzed through anthropometric methods. The considered parameters
analyzed through DXA have been significantly correlated with selected parameters
assessed with anthropometric methods. The Waist to height ratio (WtHR) parameter
has significantly correlated with DXA parameter, VAT mass, gynoid region % fat,
and android region % fat.

(Marra et al., 2019) provided a useful insight into the utilization of BC methods with
the aim of prevention and clinical practice. Whereas, the total body DXA scans have
been utilized to derive a 3-compartment BC model included with BMC, FM, and
FFM. Such models have possessed some drawbacks; the accuracy of BIA
measurements has degraded when particular predictive equations along with
standardized measurement protocols have not been consumed. However, the
limitations of DXA havethe safety of repeated measurements, cost, and technical
expertise. In which the bioelectrical phase angle and Bioelectrical Impedance Vector
Analysis (BIVA) has correspond to the clinical approach to body composition, free
from prediction equations- inherent errors and assumptions, although quantities of
body compartments have not been measured.

(Ripka et al., 2020) aimed to represent the reference data for Southern Brazilian
adolescents composed of percentile curves, of whole body LM, LMI, ALM, and FM.
This has been a cross-sectional study of adolescents aged 12–17 years having body
composition assessed through DXA. Percentile values along with reference curves
employing the Lambda, Mu, and Sigma method (LMS) have been computed for LM,
LMI (lean mass/height2), ALM, and FM. The data on 541 adolescents (68.6% boys)
has been considered for comparison purposes. The sex differences in growth
trajectories have been observed for absolute along with adjusted LM, with boys

56
producing higher LM quantity with advancing ages as compared to girls. There has
been observed from the outcome that 66.9% and 17.4% difference between the ages of
12 and 17 for boys and girls correspondingly.

(De Macêdo Cesário et al., 2020) have validated the association between the body adiposity
index (BAI) and dual‐ energy X‐ ray absorptiometry (DXA) methods to analyze the body
composition and the validity of BAI to verify the percentage of fat in children and
adolescents. The BAI and DXA estimates of %F has strongly correlated (boys: r = .71, P <
.0001; girls: r = .72, P < .0001). The linear regression analyses showed that BAI has
significance to estimate the %F in the total sample (P < .0001). For boys, the %F analyzes
performed by BAI and DXA has not presented any differences when compared (P = .2).

(Damasceno et al., 2020) developed and designed the prediction equation for body
composition assessment through utilizing the anthropometric measures of elder
women. This is a cross-sectional co-relational study with 243 older women ± 64.5
years old and BMI ± 28.70 kg/m². For the development of the equation, it has utilized
the method of hold-out sample validation. The participants have randomly divided
into an equation development group (96 elderly women) and a group for validation
(147 elderly women). Total body mass, height, waist and hip circumferences, ratio
waist-hip ratio, and BMI has been measured. The whole-body DXA assessed body
composition i.e. percentage of body fat, fat mass, and fat-free mass.

(Barnas et al., 2020) developed a DXA criterion (DC) equation to the DXA
corresponds to a collected dataset of women. The obtained DC equation for men has
been validated on a separate sample of men from which it was created (Ball et al.,
2014). Results showed excellent agreement between the DXA (18.0 ± 5.9%) and the
DC equation for men (19.1 ± 6.3%), moreover to a low SEE (2.72%) and high
correlation (r = .934).

57
CHAP TER 3: AIMS AND OBJECTIVES

After a review of the literature, it has been observed that the length of the menstrual
cycle and BMI is increased. Since the changes in BMI is due to the changes in the
amount of fat and not because of variation in density of tissue that makes up the
FFM.Overall change in BM is influenced by the effect of the fat content of the body
and no study has been done to see the changes in body fat and its relation with BMI as
well as ovarian cycle length. Further research is needed to be able to develop the
technique and to evaluate the variability of body composition at different age
groups.BIA is used to evaluate different compositions. The menstrual cycle is a highly
regulated, physiological process that makes conception and pregnancy possible. From
the start of the ovarian cycle (menarche) to its cessation (menopause), monthly
ovarian bleeding (menses) is regulated by hypothalamic and pituitary hormones. Even
the smallest changes in hormone levels can result in menstrual abnormalities. A
variety of conditions and factors (e.g., medication, stress)can cause such hormonal
changes, which are not necessarily pathological. Abnormal menstrual patterns are
identified based on changes in frequency, intensity, and onset of bleeding. A common
manifestation of menstrual cycle abnormalities includes amenorrhea, dysmenorrhea,
and menorrhagia.Discomfort before the onset of menstruation that is accompanied by
psychiatric, gastrointestinal, neurological symptoms is referred to as a premenstrual
syndrome.Amenorrhoea is the absence of menstrual periods. Women who are elite
athletes or who exercise a lot on regular basis are at risk of developing athletic
amenorrhoea.Exercise related hormones and low levels of body fat are thought to
affect how the sex hormones(estragon and progesterone)work.

Body composition measurement tool: Several aspects of body composition, in

particular the amount and distribution of body fat and the amount and composition of
lean mass, are now understood to be important health outcomes in infants and
children. Their measurement is increasingly considered in clinical practice; however,
pediatricians are often unsure as to which techniques are appropriate and suitable for
application in specific contexts. The gold standard for body composition analysis is
cadaver analysis, so no in vivo technique may be considered to meet the highest
criteria of accuracy. Only multi-component frameworks are now taken into account
that are sufficient for accuracy to serve as reference or methods of criterion in the

58
molecular approach up to the composition of body measurement (make a distinction
among fat and fat-free masses) against which different methods needs to be solved.

Several techniques are available, varying in complexity and ease of use, and each
makes assumptions that may affect its suitability for different conditions. A single
technique is unlikely to be optimal in all circumstances. A further important issue is
that of the difficulty of validating techniques in humans. Invivo techniques do not
measure body composition directly, but rather predict it from measure ments of
body properties. Thus all techniques suffer from two types of error: methodological
error when collecting raw data, and error in the assumptions by which raw data are
converted to final values. The relative magnitude of these errors varies between
techniques. Despite these limitations, more sophisticated approaches to BIA have the
potential both to improve accuracy and to increase the specificity of outcomes, and
such progress is desirable given the ease with which measurements can be made in
most age groups. Multifrequency measurements provide additional information about
water distribution. These are active areas of research but knowledge is currently
insufficient to justify the routine clinical application. The measuring tool is less
available in the field(Earthman, 2015).

3.1 AIM OF THE STUDY

Keeping all the above- mentioned problems in mind and minimal research has been
conducted on the influence of the menstrual cycle and its consequences on body
composition. The key objective of this work is to analyze the impact of the ovarian
cycle on the composition of the body observed through a 4 contacts electrode multi-
frequency bioelectrical impedance analyzer.

3.2 OBJECTIVES

The purpose of this study is to measure the variables of body composition at different
age groups of women throughout the cause of the ovarian cycle. The objective of the
present study was to:

 To evaluate the changes in the pattern of ovarian cycle during different age
groups.

59
 To evaluate the changes in parameters of changes in body composition during
the various age groups of ovarian cycle.

 Compare the physiological changes during the different age groups of the
ovarian cycle.

60
CHAP TER 4: MATERIALS AND METHODS

The chapter summarizes the materials such as machines and the software involved
during experimentation is discussed followed by the discussion of the steps involved
in the proposed work.

4.1 MATERIALS

The materialistic requirements of the study are summarized as follows:

 Subjects

 Instrumentation (Body stat Quad Scan 4000)

 Impedance disposable electrode

 MFBIA Calibrator

 Bluetooth

 Intel i5 Processor

 GB RAM

 Weighing Machine

 Height Scale.

 Sanitizer sprays.

 Inch tape(Height, Waist, Hip size measurement)

 Patient Bed

 Desktop with body stat quad scan software.

61
4.2 SUBJECTS

The body composition parameters were evaluated on 44 premenopausal females by

using MFBIA in the different age groups(17-25),(26-30),(31-40), (41-50), (51-
59).Between 17 to 60 years residing in state territory of Jammu and Kashmir,India.All
of the study participants were healthy and did not suffer from any gynecological
complications. All the subjects had not attended menopause experiencing normal
menstrual cycle. The experimental groups of all the females were divided into five
subgroups viz., based on BMI menstrual cycle length and ovulation duration. The key
criterion for selecting these age groups is that a female's ovarian cycle begins around
the age of 14 or 15. However, since this is not the case for all girls, we have chosen 17
as the beginning age. The menstrual cycle will be closed after a particular age is
reached, therefore the finishing age is 60. For experimental analysis of this work, few
female volunteers have participated. Women's body composition has been examined
throughout all phases of the ovarian cycle, i.e. phase of ovarian, follicular, early, and
luteal.

4.2.1 Exclusion Criterion

 Missing limbs.

 Pacemaker or any implanted assistive device.

 No pathological problem in previous or present records.

 Ensure that the individual had not gone through any surgical process.

 No Alcohol consumption is permitted within 48 hours of the test.

 After performing the seven days of test continuously, there are no diuretic
medications are allowed.

4.2.2 Inclusion Criterion:

 In this 44 middle-aged volunteers were taking part.

62
 Every volunteer visited the laboratory 4 times to test that distinguished by
time of exactly one week, along with each visit lasting less than 30
minutes.

 In all compositions, measurements, female volunteers had been suggested

to wear a T-shirt and shorts. The tool walls–mounted stadiometer along
with a digital scale has been consumed to measure Age (17-25), (26-30) ,
(31-40), (41-50), (51-59). Height (158.08±8.11) cm, Weight (55.11±10.8)
kg, Body mass (21.13±4.7)kg/m2. Length of ovarian cycles (28.0±3.5)
days.

 The name of the subject and all information associated with participation
in the study were kept confidential and have not been used in any written
portion of the final research outcome.

 The main information such as age, height waist, and hip circumferences
was recorded as per guidelines of Emily A, et al.2018.Women volunteers
were asked to follow the approved guidelines.

 Volunteers were asked to follow the approved guidelines. Before 2-hours

of the test, volunteers are not permitted to eat or drink. Before 30 minutes
commence of test the bladder must be emptied.

 Each female volunteer completed a two- month calendar to report two

preceding ovarian cycles starting and ending. These details were used to
determine the average length of the cycle and the specifications for all test
visits.

 The testing procedure had been conducted while the ovarian (cycle days1-
7),

 Follicular (cycle days 8-14), and late luteal (cycle days 15-21) (cycle days
22-28). Each female volunteer completed a questionnaire on-screen to
conclude the use of contraceptives before the test.

63
4.3 INSTRUMENT USED (BODY STAT QUAD SCAN 4000)

The quad scan 4000 non- invasively measures the flow of current through the body at
four different frequencies 5, 50, 100, & 200kHz.Low frequencies have difficulty in
penetrating the cell wall and pass predominately through the extracellular spaces,
while higher frequencies can penetrate the cellular membrane and pass through both
intracellular and extracellular spaces. By applying Body stat's own researched
equations, the system quickly determines values for body composition, hydration
status, and cellular health. It is well documented that in a disease state, extracellular
fluid space is a proven indicator of improving or declining cellular status.

Body statuses the multi- frequency bioimpedance raw data measurement values it
obtains for extracellular water and total body water to determine each patient’s unique
prediction marker an increase in this value shows further deterioration in health status
while a decrease shows cellular improvement.Diuretics,accurate fluid monitoring, and
investable decrease of lean muscle mass are all concerns for patients. In addition,
Quad scan4000 is used to detect malnutrition in patients with normal or high body fat.
Body cell mass can easily be obscured by an expansion of extracellular fluid which
will not be detected by looking at an overall increase in total body weight.

Figure 4.1:Body stat Quad Scan 4000

Immediately following measurement, an impedance graph will be displayed. The

impedance graph was viewed to ensure that there are no bumps and does not look
smooth; we have the option to reject the test and repeat the measurement immediately

64
with no inconvenience to the patient. Phase angle BIVA and the prediction marker are
obtained directly from resistance, reactance, or impedance and evidence in the
literature indicated that they could be used in prognostic or nutrition markers.

The Bodystat Quadscan 4000 was the device of choice for those wishing to find an
affordable reliable, trusted, and an accurate machine capable of giving immediate and
accurate results of body composition and hydration, which was also portable and easy
to use. The Quadscan 4000 measures cellular health, nutritional status, and fluid shifts
in and out of the cells. This device is used extensively within ICU, Oncology, diabetes
&obesity, and renal clinics. It had been successful in the assessment of dry weight for
dialysis patients(Orea-Tejeda et al., 2008).

4.3.1 Bodystat Quadscan 4000 measurements

The instrument accesses the following components for measuring body composition:

 ICW% &ICW volume

 ECW% &ECW volume

 TBW% &TBW

 Third space water

 Body cell mass

 ECW/TBW nutrition index

 Plus, Normal %levels

 Body fat% & Fat Weight

 Body lean mass

 Dry lean mass

 Basal Metabolic Rate

 BMR/Bodyweight

65
  Average daily calorie requirement

 Waist/Hip ratio

 Body Mass Index

 Prediction Marker

 Impedance Values at 5, 50, 100,200kHz

 Resistance 50 kHz

 Reactance 50 kHz

 Phase Angle 50 kHz

4.3.2 Body stat Quadscan 4000 Window software:

Body composition data was instantly downloaded directly from the Quadscan4000
using Bluetooth technology. It saved time and eliminates the risk of error. The Body
Stat Quad scan 4000 included the platinum software with the device.

The program features are:

 Body composition analysis report: includes printout on the subject’s body

composition results and additional trending history reports, allowing the user
to track lean/fat ratios, water, and impedance values over some time

 Hydration report: includes a one-page report of an individual’s hydration

results and additional trending history reports allowing the user to track water,
ECW, ICW,TBW, etc. over some time

 Weight loss report: Allowed a user to develop a series of exercises to

determine the number of kilocalories these will burn. These adjustment mats
then are incorporated into a user’s diet program. Goal weight and projected
targets from a wide selection of activities may be set.

66
  Health report: Assessment of health status relating lifestyle with facility related
to lifestyle changes on-screen factors include blood pressure, cholesterol
&diabetes.

 Advanced: Allowed the user to print the BIVA graphs& raw data values for
impedance, resistance, reactance, and phase angle and prediction marker

4.3.3 Protocol used with Bodystat Quadscan 4000

In this study, a group of girls is chosen and their menstrual cycle is followed for a
length of time. Many characteristics are chosen for this, including their age, weight,
height, hydration level, and other body components. Quadscan 4000 has been chosen
to monitor all of these bodystats.

4.3.4 Features of Bodystat Quad scan 4000:

 Bodystat Quadscan 4000 Bioimpedance unit

 Comprehensive hardware users guide on CD

 Cable leads

 120 disposable long electrodes

 Calibrator

 Duracell batteries AA (6)

 Body stat Quadscan 4000 software program with E-mail software support

 Bluetooth module fitted to download stored test data into a PC

 Customized padded shoulder bag (Achamrah et al., 2018).

4.3.5 The principle involved in the study with Bodystat Quad scan 4000:

The basic principle involved in the working of Bodystat Quad scan is that t he
frequency used affects the mechanism of detecting current flow through the body
(impedance). This is because the current cannot penetrate the cellular membrane at

67
low frequencies, it passes mostly via the extracellular space. Based on this principle
the measurements are made in the present study.

4.4 IMPEDANCE DISPOSABLE ELECTRODE

The subjects were recorded within a minute of electrode placement, whilst ensuring
that they did not make contact with any metal objects. A change in subject position
was well within a time frame of 2 min. Measurements were taken using an
ImpediMed Inc tetrapolar bioimpedance spectroscopy unit (Pinkenba, Qld, Australia).
This device, which applies a constant current, scans 256 frequencies between 4 kHz
and 1000 kHz, repeating this procedure six times with a 1-sec interval. Using this
approach, any slight movement artifacts or changes in the resistance and reactance
values due to cable movement, change in the stance, body, or electrode movement
were minimized.

Figure 4.2:Applications of Electrodes for Impedance Measurements

The subjects were in a standing position with equal weight on both legs, and in the
case of Human # 1 & 3, recordings were also made in a sitting position with both legs
on the floor. In Human #1, recordings were additionally made in a sitting position
without foot contact to the floor, and in a lying (supine) position.

Electrode choice may affect the measurements by creating a capacitance between the
electrode surface and the skin. In Human #1, to assess electrode-type effects on
mfBIA recordings, two types of Ag/AgCl electrodes were also used (Ambu – Blue
Sensor, DK-2750, Ballerup, Denmark). The one, N-type with integrated gel, was
44 mm × 22 mm, and the other, a SU-type with gel removed to expose the electrode,
was 49 mm × 33 mm. The SU-type electrode surface was attached to the skin with the
same conductance paste (Ten20) as used previously with the platinum electrodes. The

68
positions of the Ag/AgCl electrodes were the same as those for the platinum
electrodes. Recordings were taken within a 3 min time frame of each other(Olson et
al., 1979).

4.5 BLUETOOTH

MFBIA device has inbuilt Bluetooth and is used to transfer the measured data from
the device to the computer and can be used for diagnosing and analyzing the
physiological status of individual Bluetooth internal modules the analyzer can be
connected via a standard wireless communication port(Sami, 2016).

4.6 WEIGHING MACHINE

The principle of weight measurement is used in the weighing machine. When the body
is moved to a weighing machine, it is pulled towards the center of the earth that exerts
the force on the weighting machine which is used to calibrate the body mass in terms
of numbers(Del Castillo et al., 2017). Therefore, in BIA measurement a weighting
machine measures the mass of the body exerted on the machine as illustrated in Figure
4.3.

Figure 4.3:Weighing Machine

69
4.7 HEIGHT MEASUREMENT SCALE

Figure 4.4:Height Measurement by Healthcare Practitioner

Height measurement is mandatory to make the record to person’s height in respect to

its weight. For this purpose, inch tape or the measuring scale is used for height
measurement and this data is recorded by the healthcare professional in the system as
illustrated in the Figure. The ideal height to weight variation is also shown in the
Table for reference during experimentation(Freitag et al., 2010).

Table 4.1: Ideal Height to Weight Variations

Height Weight

Inches Centimetres Kilograms

4’6” 137 28.5 – 34.9

4’7” 140 30.8 – 37.6

4’8” 142 32.6 – 39.9

4’9” 145 34.9 – 42.6

4’10” 147 36.4 – 44.9

70
 4’11” 150 39 – 47.6

5’0” 152 40.8 – 49.9

5’1” 155 43.1 – 52.6

5’2” 157 44.9 – 54.9

5’3” 160 47.2 – 57.6

5’4” 163 49 – 59.9

5’5” 165 51.2 – 62.6

5’6” 168 53 – 64.8

5’7” 170 55.3 – 67.6

5’8” 173 57.1 – 69.8

5’9” 175 59.4 – 72.6

5’10” 178 61.2 – 74.8

5’11” 180 63.5 – 77.5

6’0” 183 65.3 – 79.8

4.8 MFBIA CALIBRATOR

It is used from time to time to calibrate the instrument and to make the instrument
result accurate. It controls the device's output current and frequency and helps
maintain the system's sensitivity (Colls et al., 2000).

4.9 METHODOLOGY

The overview of the methodologyused which is involved in the present work is given
in Figure.

71
 Figure 4.5 :Methodology used

Steps involved in the measurement of BIA using the methodology used are as follows:

1. Every volunteer visited the laboratory for testing four times, separated by an
exact time of one week and each visit lasted less than 30 minutes.
2. Women volunteers were advised to wear a t-shirt and a short stadiometer for
all body composition measurements, and the digital scale was used to measure
the height (cm) and weight (kg).
3. BIA measurements were carried out with subjects lying in the supine position.
Nonconductive tetra polar technique the Quad Scan 4000 unit with four
electrodes.

72
 Figure 4.6: Electrode placement

4. Two electrodes were placed on the right ankle with one just proximal to the
third metatarsophalangeal joint (positive) and one between the medial and
lateral malleoli (negative).
5. Multifrequency (5, 50,100 & 200 kHz) are used for the evaluation of fat-free
mass (FFM), total body water (TBW), intracellular water (ICW), and
extracellular water content (ECW).
6. In this study, we also evaluated the results of the ovarian cycle on the
composition of the body using MFBIA.
7. Experiments were carried out on 44 women at universities of appropriate age
and body weight Age (17-25), (26-30), (31-40), (41-50), (51-59) years; body
weight index = 24.4 ± 3.8 kg / m2 ).
8. Women's body composition is evaluated using MFBIA analysts at all phases of
the ovarian cycle, namely ovarian, follicular, early, and late luteal phases. The
research study indicated that the result is to assess the variability of the
composition of the body in different age groups.

During the ovarian (cycle days 8-14), finally luteal (cycle days 15-21), and late luteal
(cycle days 22-28) the testing process was carried out. Each female volunteer
completed a pre-screening questionnaire to conclude the use of contraceptives before
the test.

The subject was weight in standardized lightweight clothes without shoes on platform
manual scale balance. BIA measurements are carried out on subjects lying at a supine
position on a flat non-conductive bed. Two electrodes were placed on the right wrist
with one just proximal to the third metacarpo-phalangeal joint(Positive) and one on

73
the wrist next to the ulnar head(Negative),Two electrodes were placed at the right
ankle with one just proximal to the third metatarsophalangeal joint(Positive) and one
between the medial and lateral malleoli(Negative),ECW, ICW, and TBW were
calculated by using manufacturer software.

4.9.1 Estimation of Body Hydration using MFBIA

The QuadScan 4000 units are battery operated and easy to use requiring no specialist
skills.The unit has been electronically precision-engineered to the highest quality
standards offering the user a safe and efficient means of measurement.

The basic principle of the method is that lean tissue,which consists essentially of
electrolyte-containing water,conducts the electric current,whereas the fat acts as an
insulator,the impedance of the body is therefore determined largely by the low–
impedance lean tissue.Regression equations are then derived which relate impedance
to FFM or TBW measured by independent techniques.

At 50 kHz, a proportion of the applied current is unable to penetrate the cell

membranes and therefore passes only through the extracellular space.At this
frequency, BIA is only able to predict TBW measured by independent techniques.

Furthermore,the measurement of TBW alone is only of limited value in the nutritional

or functional assessment of seriously ill patients.Patients with ongoing infections have
been shown to retain fluid response to nutritional support and weight gain is due to
expansion of extracellular water(ECW) space.It has been emphasized that weight gain
cannot be viewed as an improvement in nutritional status as it does not reflect a n
improvement in protein storage.There is also evidence that surgical patients who
respond to nutritional support with an increase in ECW have increased postoperative
complication rates compared with patients who lose water, and that may benefit from
longer courses of nutritional support.

Hence a measure of the distribution of TBW between extracellular and intracellular

spaces may provide a useful index of the well- being response to feeding of a critically
ill patient(prediction marker).

74
Similarly,it may be necessary to measure the extra and intracellular fluid levels in
competitive athletes to fully assess their hydrational status and measure performance
levels at various levels of hydration intracellularly.

4.9.2 Principles of used parameters

In this study some parameters were chosen as they influence the ovarian cycle. The
principles of chosen parameters include the following:

 Age

Women's menstrual periods definitely vary as they get older! Various women's
menstrual periods are heavy and irregular over the first few years after menarche,
which is common in some cultures. The cycles grow more regular in the late teen and
early twenties. Women in their 30s had the shortest menstrual periods and the least
amount of unpredictability. With the loss of ovarian follicles that occurs when women
reach their forties, the duration of the menstrual cycle lengthens. Women enter
menopause when all of their ovarian follicles have been drained, and their monthly
cycle comes to a stop. Based on this fact, various age groups are included in the study.

 Weight

Menstrual patterns might be affected by changes in body weight. Extremes in BMI are
linked to alterations in menstrual cycle and reproductive patterns, according to
research. To start and continue menstrual periods, a certain percentage of body weight
is required. As a result, women who lose too much weight or are underweight may
have changes in their menstruation, such as cessation of menstruation or extended
periods of amenorrhea. Excess body fat and weight are also linked to irregular and
heavy menstrual periods. Obese women have a greater risk of menstrual cycle
abnormalities than women with a normal body mass index. Obese women are more
likely to have conditions like polycystic ovarian syndrome. This change in the body
weight is to some extent linked with the women reproductive health and forms the
basic principle that govern the women health to some extent. The fact and figures are
further analyzed to have the real time interpretation and conclusion.

75
  Height

Girls go through a growth spurt one to two years before they start menstruating.
Puberty strikes most females between the ages of 8 and 13, while the growth spurt
comes between the ages of 10 and 14. In the year or two following their first period,
they only gain 1 to 2 inches in height. This is the time when they achieve adult height.
By the age of 14 or 15, the majority of females have reached adult height. Depending
on when a female first receives her period, this age might be lower.

 Hydration levels

The water plays most important role in human life. During your period, being
dehydrated may cause cramping and discomfort. Therefore, itis one of the principle
necessities of women especially during reproductive age. The hormonal imbalances
and a bloated stomach are common during periods. Your body retains more water
when your oestrogen and progesterone levels decline. As a result, several
physiological changes occur, which may worsen as people age.

 Body fat

Body fat is known to have an effect on the period's regularity. Due to hormonal
pathway deregulation, both extremes — underweight and overweight/obese — may
cause abnormalities in the menstrual cycle. Smaller weight changes that do not put us
in a different BMI zone, on the other hand, may be significant enough to trigger
hormonal alterations and, as a result, changes in thecycle.

76
CHAP TER 5: RESULTS

The chapter provides detailed information about the number of participants involved
in the evaluation of the presented research study. This section is followed by the
parametric evaluation of body composition concerning different parameters,
frequencies, and ovarian phases used for analyzing the variations observed in the
body measurements. The section also shows that the females of appropriate age,
height, weight, body mass, and phase of the ovarian cycle were considered for
experimentation.

5.1 SUBJECT DISTRIBUTION

In this study, 44 middle-aged females were considered to study ovarian cycles with
the help of the MFBIA analysis tool to analyze the body composition of women
during various phases of the ovarian cycle.(Table 5.1) lists the distribution of subjects
concerning different age groups.

Table 5.1: Participants’ Distribution

Age Groups < 25 Yrs 25-30 Yrs 31-40 Yrs 41-50 Yrs >60Yrs TOTAL

(21.06 ± (28.0 ± (36 ± (55.6 ± (31.67 ±

(Mean ±SD) (45 ± 2.82)
2.5) 1.6) 3.03) 3.51) 10.96)
Number of
16 8 11 6 3 44
Participants

6
< 25 Yrs
16
25-30 Yrs
31-40 Yrs
41-50 Yrs
11
>60Yrs
8

Figure 5.1: Participants’ Distribution

77
The graphical representation of the table shown in the form of a pie chart in Figure
(5.1) shows that the highest number of volunteer females belongs to the teenage, i.e.,
16 females participated in the research whose age was below 25 years. The next
highest number of female volunteers belongs to the age group that covers subjects
with ages between 31 years and 40 years. However, there were only 3 participants
with age more than 60 years of age. All the observations made during the study are
demonstrated as Mean± Standard Deviation.

5.2 PARAMETERS USED FOR ANALYSIS OF BODY

COMPOSITION

The body composition studies are usually based on the basic assumption that the
human body is composed of two distinct chemically separated components that are
labeled as fat mass and fat- free mass. In this context, the following parameters are
computed to evaluate the performed study.

5.2.1 Body Mass Index (BMI)

It is a widely used term to express body composition in terms of an index of relative

weight while taking into consideration the age and gender of the subject.

(5.1)

Where, represents the weight of the subject and

represents the height of the subject under study. BMI is an index that is globally used
for defining the nutrition status and practiced as the predictive parameter of clinical
importance that is widely used to determine the risk of diabetes.

5.2.2 Body Fat (BF)

The percentage of body fat is a much more accurate parameter than BMI to predict
the exact amount of fat present in the body. A healthy female human body comprises
18 to 28% BF as compared to 10% to 20% BF observed for the male population.

5.2.3 Fat Mass (FM)

(5.2)

78
 (5.3)

Where, is the weight of the participant and represents the fat-

free mass of the participant

5.2.4 Fat-Free Mass (FFM)

This type of body mass is nearly constant and comprises bones, muscles, water,
internal organs, and connective tissues present in the body.

(5.4)

Where is total body water and is the hydration constant

5.2.5 Body Water

The total amount of body water present in the human body also termed as Total Body
Water (TBW) is further divided into two groups as follows:

5.2.5.1 Extra Cellular Water(ECW)

This is the water compartment that surrounds the cells and tissues and is always
present outside the cells.

5.2.5.2 Intra Cellular Water (ICW)

This refers to the water that is present within the cells and is required to carry on the
intracellular activities of the cell.

The maintenance of healthy water level in the body is essential to lead a healthy life.
These parameters symbol any alarming change that can be addressed to overcome any
health risk.

5.2.6 Body Cell Mass (BCM)

It is the total mass of the metabolically active components of the body that comprises
internal organs, muscles, and blood as connective tissue, etc. It is used to access the
nutritional status of the body.

79
5.2.7 Extra Cellular Mass (ECM)

It comprises the extracellular fluid along with the solid mass that comprises bones and
cartilage present in the body. The major role of this mass is to provide solid support to
the body in addition to transporting wastes, nutrients, and fluids within the body.

5.3 BODY COMPOSITION ANALYSIS USING MFBIA IN

OVARIAN PHASE

In the body mass assessment,a large number of parameters are determined such as
body mass (kg), body fat (percent), lean body mass, FM, ICW, ECW, and TBW using
MFBIA in the ovarian phase. The parametric values are along with mean and standard
deviation are summarised in (Table 5.2). The weight and height are observed as the
basic requirements of the body composition analysis study with an average weight of
and height of .

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 Table 5.2:Body composition (MFBIA) in Ovarian Phase

Age Groups < 25 Yrs 25-30 Yrs 31-40 Yrs 41-50 Yrs >60Yrs TOTAL

Weight 51.6 ± 9.8 46..0 ± 2.67 54.63 ± 3.82 65.33 ± 4.27 79.33 ± 5.50 55.11± 10.84

(Kg) (39 – 68) (42 – 50) (50 – 61) (61 – 72) (74 – 85) (39-85)

Height 160.9 ± 8.0 161.2 ± 7.3 155.0 ± 5.3 151.8 ± 8.9 157.3 ± 11.01 158.0± 8.11

(cm) (149 – 172) (147 – 170) (149 – 165) (144 – 168) (146– 167) (144-172)

19.3 ± 3.3 25.2 ± 2.9 24.5 ± 3.8 26.3 ± 5.0 22.2 ± 7.8 22.8 ± 4.7
BMI
(15.8 –
(21.1 – 29.1) (20.6 – 30.1) (19.6 – 31.9) (15.9 – 31.0) (15.8-31.0)
25.7)

19.86± 3.1 23.7 ± 4.1 28.0 ± 4.09 30.08± 2.55 32.33 ±2.96 24.93± 11.5
BF%
(16.2-26.8) (19.1-29.2) (22.5-35.1) (28.2-34.8) (29.2-35.1) (16.2-35.1))

17.93 ± 8.9 28.7 ± 5.30 28.25 ± 6.51 40.5 ± 5.36 43.96 ± 6.3 27.33 ± 11.5
% FM
(11.01– (21.01–
(21.6– 35.0) (40.1– 53.9) (39.9– 50.28) (11.01-54.3)
38.4) 38.09)

50.6 ± 5.57 47.7± 6.42 47.6 ± 5.51 46.04± 5.03 45.3 ± 5.2 48.3 ± 5.6
% TBW
(43.1– (42..0 –
(41.8 – 54.3) (40.16 – 53.9) (34.9-50.28 ) (34.9-58.0)
58.0) 56.0)

.910 ± .040 .955 ± .439 0.719 ± 0.397 .981 ± 0.646 .912 ± 0.604 .899± .381
ECW/ICW
(.910–
(.902–1.045) (.890 – 1.035) (.908– 1.002) (.863– 980) (.863-1.055)
1.055)

8.26± 5.38 17.3 ± 6.40 22.1 ± 7.94 27.8 ± 7.9 34.8± 13.3 17.8± 10.9
FM (Kg)
(3.2– 20.6) (8.9– 25.2) (10.8– 30.4) (16.9 – 35.6) (19.6– 44.6) (3.2-44.6)

31.5 ± 8.7 35.3 ± 5.9 37.6 ± 2.28 42.6 ± 2.4 39.3 ± 3.8 35.7± 7.4
FFM (Kg)
(22..0 –
(28.0 –43.2) (34.0–40.1) (39.6-45.8) (35.3-42.9) (22.0-46.1)
46.1)
22.38 ±
19.01 ± 1.72 20.68 ± 1.25 20.65 ± 2.67 22.8± 4.06 21.1± 4.7
7.20
BCM (Kg)
(14.7–
(16.3-21.8) (18.3– 22.2) (17.4–24.9) (18.2-25.6) (14.7-21.8)
36.5)

14.7 ± 5.20 18.06 ± 1.04 17.9 ± 1.19 17.9 ± 0.67 18.16 ± 2.09 16.7± 3.5
ECM (Kg)
(6.9– 22.4) (16.1 – 19.5) (16.0 – 19.0) (16.9 – 19.0) (15.8 – 19.8) (6.9-22.4)

27.7±2.2 29.2±3.2 32.4±2.2 31.6±1.86 31.0±0 29.9±2.9

Length of
Cycle
(30-24) (33-24) (35-29_ (35-30) (31-31)

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Figure 5.2: SD and mean range values of different body composition in females.

The mean and SD for various parametric values used in the study of body mass
analysis are plotted in (Figure 5.2).The study further reveals that with the increase in
the age of subjects from a considerable increase in the %FM
from FFM from 39 3 3 8 , BCM
from2 38 7 20 to 22 8 4 06, and ECM to18 16 2 09 is observed.
The increase in the overall body compositions values with the decrease in ECM with
age shows that there is a strong correlation between impedance and body composition.
It is also observed that it reflects the body composition in terms of TBW, ECW/ICW
decreases with the rising age of the female group.

5.4 Impedance Analysis at different frequencies

In the evaluation study, impedance analysis for legs, arms, and the trunk at 50 kHz,
100 kHz, and 200 kHz were determined. The observations for impedance analysis are
summarized in (Table 5.3) showing variation in terms of mean and standard deviation.

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 Table 5.3: Measurements driven from (MFBIA) impedance and phase angle
(Females) at different frequencies

Age Groups
< 25 Yrs 25-30 Yrs 31-40 Yrs 41-50 Yrs >60Yrs TOTAL
(Mean ±SD)

757 ± 85.2
Impedance 804 ± 68 776 ± 75 735 ± 94 702 ± 52 649 ± 50
(581 –
(50 khz) (622 – 1004) (632 – 963) (636 – 862) (638 – 784) (581 – 739)
1004)

Impedance 754 ± 60 735 ± 65 691 ± 88 659 ± 56 603 ± 43 711 ± 79

(100 khz) (581 – 938) 598 –921 (590 – 829) (601 – 731) (546 – 669) (546 – 938)

Impedance 765 ± 65 740 ± 61 673 ± 95 616 ± 52 605 ± 45 706 ± 90

(200 khz) (581 – 938) (577 – 928) (556 – 829) (605 – 724) (527 – 685) (556 – 938)

Figure 5.3:Variation in impedance with age in females

(Figure 5.3)represents the variation in the impedance concerning different age groups
of females. It is observed that with the increase in age of female’s impedance
decreases with minor variations in each age group. The decrease in impedance value

83
with increasing age suggests that impedance is an indicator of function and general
health. It is not only an indicator of the body composition but also shows the
comparison of the values with the overall mean. The calculated average value of
impedance is computed to be at the frequency of 50 KHz, at
100 KHz, and at 200 KHz, respectively. Table 5.3 depicts the mean and
standard deviation. The lesser the standard de viation the best the model. Here if
observed, the female of age group greater than 60 have low impedance values even at
50 khz, 100khz, and 200 khz. And with the average values, the impedance of 100 kHz
has a low standard deviation. This reveals that the female of age group greater than 60
should take utmost care regarding their general health.

5.5 PHASE ANGLE ANALYSIS AT DIFFERENT FREQUENCIES

It is further observed that the variation in phase angle and impedance is highly
affected with sex, age, BMI, FM%. Therefore, in this section, the measurements were
drawn concerning changes in the phase angle and age groups of subjects. The
observations are summarized in (Table 5.4) with phase angles varied from 50 khz to
200 khz.

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 Table 5.4: Measurements driven from MFBIA phase angle at different
frequencies

Age Groups
< 25 Yrs 25-30 Yrs 31-40 Yrs 41-50 Yrs >60Yrs TOTAL
(Mean ±SD)

Phase 6.93 ± 0.59 6.91 ± 0.54 6.90 ± 0.73

6.89± 0.64 6.84 ± 0.28 6.89 ± 0.54
Angle
(6.20– (6.19 – (6.12 –
(5.50 – 8.39) (6.40 – 7.13) (5.50 – 8.22)
(50 khz) 8.22) 7.69) 7.92)

Phase 8.9 ± 0.42 8.80 ± 0.49

9.18 ± 0.52 8.45 ± 0.23 8.35 ± 0.52 8.87 ± 0.53
Angle
(7.89 – (7.99 –
(8.04 –10.25) (8.09 – 8.45) (7.64 –8.94) (7.64–10.25)
(100 khz) 9.89) 9.52)

11.43 ±
Phase 9.67 ± 1.77 9.21 ± 2.09 8.59 ± 0.96
9.8 ± 1.89 1.81 10.02 ± 1.96
Angle
(8.32 – (8.11 – (7.69 –
(8.06 –16.24) (8.39– (7.69– 16.24)
(200 khz) 14.16) 13.12) 10.82)
13.17)

Figure 5.4:Variation in phase angle with females age group

85
The observed significance level of the study summarized in the above table is found to
be for all analyses. It is observed that impedance for phase age of 50 khz is
found to be for the age group of females with ageless than 20 years that
increases to for the age group that covers females of age more than 60
years. Similarly, the impedance varies from for phase
angle measurement at 100 kHz and for phase angle
measurement at 200 kHz. Figure 5.4 shows the variation in impedance analysis along
with different age groups of females concerning different age groups. It is observed
that the least impedance is observed for phase angle of 50 khz followed by 100 khz
and 200 kHz with an average value of
This also illustrates that the highest impedance is observed when a phase angle
of 200 kHz is used for analysis.

The distribution of impedance and phase angle was fairly normal in the above data.
The overall mean phase angle and impedance in all categories, there is a significant
decrease in phase angle and impedance with increasing age in females.

5.6 BODY MEASUREMENTS VARIATION WITH PHASES OF

OVARIAN CYCLE

The women's body composition was also assessed during different ovarian cycle
phases using MFBIA analysts. The different phases used in the analysis are described
in (Table 5.5).

Table 5.5 :Phase Distribution for Analysis

Sr. No. Phase Cycle (days)

1 Ovarian 1–7

2 Follicular 8 – 14

3 Early Luteal 15 – 21

4 Late Luteal 22-28

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The first phase used for the analysis consists of day 1 to day 7 days of the ovarian
cycle that is also named the ovarian phase of the cycle. The follicular phase is
comprised of day 8 to day 14 of the cycle. However, the luteal phase is categorized
into two sub-categories, namely the early luteal phase that comprises day 15 to day 21
of the cycle with the late luteal phase that covers day 22 to day 28 of the cycle.

5.6.1 %FM variation with Ovarian Cycle

(Table 5.6) summarizes the body compositions of females that are examined through
MFBIA analyzers during different phases of the ovarian cycle. Thenumber of
variables including fat mass in terms of percentiles was determined at different
frequencies was also determined.

Table 5.6:% FM variation with Ovarian Cycle

AGE
OVARIAN FOLLICULAR EARLY LUTEAL LATE LUTEAL
GROUP

<25 YRS 17.93 ± 8.9 17.8 ± 8.8 18.03 ± 9.01 18.03 ± 9.01

25-30 Yrs 28.7 ± 5.30 27.6 ± 5.20 29.6± 5.50 29.6± 5.50

31-40 Yrs 28.25 ± 6.51 28.01 ± 6.41 29.03 ± 7.41 29.03 ± 7.41

41-50Yrs 40.5 ±5.36 40.3 ± 5.04 40.7 ± 5.55 40.7 ± 5.55

>60Yrs 43.96 ± 6.3 43.78 ± 6.09 44.10 ± 6.5 44.10 ± 6.5

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 Figure 5.5:Correlation of ovarian cycle phases with female age group & FM%

The graphical illustration of the variation in %FM is shown in (Figure 5.5). It is

observed that %FM is more dependent on the age of the subject as compared to the
phase of the ovarian cycle. For instance, it is observed that %FM for subjects with age
group less than 25 years is found to be 17 93 8 9 during the days of ovarian phase,
17 8 8 8 during days of the follicular phase, 18 03 9 01 during the days of early
luteal phase and 18 03 9 01 during the late luteal phase. This shows that the %FM
almost remains unchanged during various phases of the cycle. However, a
considerable variation that is found in different age groups is also due to differences in
the physique of the population under study.

5.6.2 FFM variation with Ovarian Cycle

(Table 5.7) shows the variation observed for FFM measurements of the subjects in
different phases ofthe ovarian cycle as described in (Table 5.5). The graphical
representation of the observations summarized here is given in (Figure 5.6).

88
 Table 5.7:FFM variation with Ovarian Cycle

AGE
OVARIAN FOLLICULAR EARLY LUTEAL LATE LUTEAL
GROUP

<25 YRS 31.5 ± 8.7 31.5 ± 8.6 31.7 ± 8.8 31.7 ± 8.9

25-30 Yrs 35.3 ± 5.9 35.5 ± 5.9 35.7± 6.0 35.7 ± 6.1

31-40 Yrs 37.6 ± 2.28 37.8 ± 2.28 37.6 ± 2.38 37.6 ± 2.41

41-50 Yrs 42.6 ± 2.4 42.8 ± 2.4 42.8± 2.5 42.9 ± 2.6

>60Yrs 39.3 ± 3.8 39.6 ± 3.9 39.6 ± 3.9 39.7 ± 4.01

It is observed that the highest FFM is found during the luteal phase as compared to the
ovarian and follicular phase irrespective of the age group under study. For instance,
FFM observed in the ovarian phase is 35 3 5 9, follicular phase is 35 5 5 9, the
early luteal phase is 35 7 6 0 , and late luteal phase is 35 7 6 1 for subjects
belonging to the age group of 25 years to 30 years. Similar observations are also
found for subjects of age more than 60 years of age with the highest FFM exhibited in
the luteal phase 39 6 3 9 and 39 7 4 01 as compared to 39 3 3 8 in the
ovarian phase and 39 6 3 9 in the follicular phase. Overall it is observed that the
highest FFM is exhibited in the luteal phase irrespective of the age of the subjects.

Figure 5.6:Correlation of ovarian cycle phases with female age group & FFM
(Kg)

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5.6.3 FM (kg) variation with Ovarian Cycle

Fat mass is also computed to the weight of the subject. The variations observed in FM
for different age groups used in the study forthe ovarian cycle are summarized (Table
5.8).

Table 5.8:FM (kg) variation with Ovarian Cycle

AGE LATE
OVARIAN FOLLICULAR EARLY LUTEAL
GROUP LUTEAL
<25 YRS 8.26± 5.38 8.18± 5.28 8.28± 5.49 8.28± 5.49

25-30 Yrs 17.3 ± 6.40 17.4 ± 6.40 17.6 ± 7.50 17.6 ± 7.50

31-40 Yrs 22.1 ± 7.94 21.9 ± 7.7 22.0 ± 8.02 22.0 ± 8.02

41-50 Yrs 27.8 ± 7.9 27.6 ± 7.7 27.8 ± 7.9 27.5 ± 7.5

>60Yrs 34.8± 13.3 34.6± 13.2 34.5 ± 13.2 34.7 ± 13.1

Figure 5.7:Correlation of ovarian cycle phases with female age group &
impedance

The correlation of ovarian cycle with FM is illustrated in (Figure 5.7) for the different
age groups of females. It is observed that FM increases with the increase in the age of

90
the subjects which means that higher FM is observed for older females tha n the young
teenagers. However, minor variation is observed for FM to phases of the ovarian
cycle. For example, as illustrated by the observations, females having age group of 25
years to 30 years exhibits FM of in the ovarian phase, in the
follicular phase, early luteal phase, and and late luteal
phase. Similar, trends for FM have also been observed in different age groups. This
shows that FM does not demonstrate large variation with different phases of the
ovarian cycle however, FM is found to be highest in luteal phases of the ovarian
cycle.

5.6.4 BMI variation with Ovarian Cycle

Body mass index is a very common parameter used for body measurement analysis.
The observed BMI for the female subjects of different age groups is summarized in
(Table 5.9) to different phases of the ovarian cycle. The table reflects that due to
change in physique with the rising age, BMI also varies in different age groups.

Table 5.9:BMI variation with Ovarian Cycle

AGE FOLLICULA EARLY LATE

OVARIAN
GROUP R LUTEAL LUTEAL
<25 YRS 19.3 ± 3.3 19.2 ± 3.2 19.5 ± 3.3 19.6 ± 3.3

25-30 Yrs 25.2 ± 2.9 25.1 ± 2.8 25.3 ± 3.1 25.2 ± 3.1

31-40 Yrs 24.5 ± 3.8 24.4 ± 3.7 24.6 ± 3.9 24.4 ± 3.9

41-50 Yrs 26.3 ± 5.0 26.2 ± 4.8 26.3 ± 4.9 26.3 ± 4.9

>60Yrs 22.2 ± 7.8 22.1 ± 7.7 22.3 ± 7.7 22.3 ± 7.7

(Figure 5.8) represents the graphical representation of the observations made for BMI
summarized in the above table. It is observed that higher BMI is observed in the luteal
phases of the ovarian cycle as compared to the ovarian or follicular phase irrespective
of the age of the subjects. For example, the subjects belonging to the age group of 31
years to 40 years exhibits 24 5 3 8 BMI in ovarian phase, 24 4 3 7 in follicular

91
phase, 24 6 3 9 in early luteal phase and 24 4 3 9 in late luteal phase. This
shows that BMI is higher in luteal phase of ovarian cycle as compared to other phases.

30

25

20 BMI <25 YRS

BMI 25-30 Yrs
15
BMI 31-40 Yrs
BMI 41-50 Yrs
10
BMI >60Yrs

0
OVARIAN FOLLICULAR EARLY LUTEAL LATE LUTEAL

Figure 5.8: BMI variation with Ovarian Cycle

The study reveals that there is co-relation of body compositions with ovarian cycle
phases when compared each phase such as ovarian, follicular, luteal and late luteal
with female age group of 44 females, which is already divided into five groups. The
change in body compositions shows that with the increase in age group of females
there is change in each phase of ovarian cycle and for each cycle mean and SD values
has been set up which shows if there is slight change in mean ad SD values this shows
the body composition changes with female age group and if these values change there
will be change in body composition and MFBIA values shows variation which
reflects the ovulation cycle and composition value will change or show some
variations in FM%,FFM,BMI and impedance will show a great change. These
variations of compositional values and change in body fluids will show change in
cycle of ovarian phase and thus causes change in hormones or hormonal imbalance,
and effects fertility of females and pregnancy. This graphical study also proves if
there is decrease in value of impedance, fat mass will increase and shows BMI of
obese female.

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5.7 STATISTICAL ANALYSIS

The data were measured using version 13.0 of Sigma Plot. Each value is shown as
the; mean ± standard deviation. For each female volunteer, each of the four phases of
the ovarian cycles is considered as the variables of body composition. A repeated
measure ANOVA was used to determine the result in the variables for the various
phases of the ovarian cycle. The meaning level for all analyses was p ≤ 0.05.

In this cross-sectional study we assessed age related pattern of changes in body fat
tissue and FFM in females. The group of interest consisted of female’s individual
younger than 60 years. It is aimed to define body composition, estimated fro m
MFBIA and to evaluate changes that occur with aging. Body fat mass is most variable
component of the human body. In women age related changes and regional obesity
have been observed mainly after menopause. Women the body mass index is
significantly increases at the age of 31-40 and 50- 60 age groups proceeding age
groups. Moreover, if the FFM decreases body weight increases slightly. It may be
assumed that the continuation of this trend is decreasing FFM and increasing BCM
mean FFM was greatest between 41-50 years old females and decreased thereafter.
FFM increased slightly but not significantly in younger women than 30 years old
compared with women 31-40 and decreased slightly in women older in 60 years.

The analysis corresponds to composition of body is done during ovarian phases such
as follicular, early, and late leutal. From the study it was concluded that contact
electrodes used in the research can be consumed at every time while processing
ovarian cycle without change in the value of composition (Lukaski et al., 2017).
MFBIA helped to predict and to diagnose severe form of ovarian cycle (Genton et al.,
2018). Significant differences between body fat group estradiol profile across whole
ovarian cycle no such relation found negative (Sowers et al., 2007, Kushner, 1992 and
Shafer et al., 2009).Human body composition is based upon the principle of electrical
conductivity of the fat free tissue mass (Ellis et al., 1999).

The outcome of this study shows the analysis of MFBIA in the body in which 44
middle-aged women are considered as ovarian cycles using the MFBIA analytics tool.
In Body it is used to analyse women's body composition during various phases of the
ovarian cycle. We considered the females with the appropria te number, age, height,

93
and weight, body mass and ovarian length.ANOVA tests were conducted during this
study, and it was observed that variables of body composition were evaluated for four
phases of ovarian cycles. Significance level for all analyses was p≤0.05.

The factors, which could be responsible in variation of ovarian cycle, are understood.
The outcome of the present study shows that females with less age group who are
physically active experiences regular ovarian cycles. Increased age of menarche and
decrease in body mass index shows, increase in duration of ovarian cycles from public
health perspective understanding the effects of ovarian cycle of moderate, levels of
physical activity, body mass index and age at menarche is most relevant. It is almost
importance to identify the cause and deviation of ovarian cycles to enable normal
reproductive health of females.

Reference values were developed parameters of body composition derived from

MFBIA.These parameters can be used to evaluate individual measurement results
compared to healthy female of higher age group. Normal values for TBW & ECW are
innovative may allow evaluating normal hydration.

94
CHAP TER 6: DISCUSSION

The analysis of body composition is the clinical assessment of tissue and fluid
compartments in the human body in terms of the parameter such a; FM, FFM, BCM,
ECM, TBW, ICW and ECW. Nutritional intake, losses, and needs are reflected in
body composition throughout time. Malnutrition, defined as a decrease of FFM has
been linked to worse survival, poorer clinical outcomes, and quality of life in cancer
patients, as well as higher drug toxicity. In a variety of clinical situations, including
sarcopenic obesity and chronic diseases, measuring body composition with available
methods such as dual X-ray absorptiometry, computed tomography, and bioelectrical
impedance analysis quantifies FFM loss, whereas body weight loss and body mass
index only FFM loss are insignificant (Kin et al., 1991). The ability to document the
success of nutritional support, change the choice of disease-specific and nutritional
treatments, and evaluate their effectiveness and suspected toxicity is all made possible
by measuring body composition. Malnutrition causes a loss of FFM of varying
intensity, which is linked to a loss of fat mass whose importance grows with the
length of malnutrition. Its prevalence is especially high among the elderly, people
with chronic conditions, and patients in hospitals, and it is anticipated to rise over the
next decade as the deleterious impact of malnutrition on clinical outcomes grows.
Even in older sitting participants with pre-existing sarcopenia or patients with chronic
diseases, advances in medical technology and treatments have been shown to extend
survival. As a result, the percentage of patients with low FFM will rise, lowering their
general health, functional abilities, and enjoyment of life. Indeed, in cancer patients,
FFM loss is unequivocally linked to decreased survival, unfavorable clinical
outcomes, such as an increased rate of infection, complications, hospitalizations,
length of stay and recovery, and toxicity to treatments, all of which raise health-care
costs.

In clinical practice, the primary goal of body composition assessments is to assess

nutritional health by measuring FFM and FM. In present research, these are used to
access the female health status during ovarian cycle. In patients admitted to the
hospital or outpatients those are nutritionally vulnerable, a clinical analysis of
nutritional status is commonly indicated. The BMI and the percentage of weight loss
provide little insight into the respective contributions of FFM and FM in changes in

95
body mass in various chronic diseases. The total body impedance may be used to
estimate total body water, and by assuming that total body water is constant, FFM,
which contains body water, may be calculated (Thibault et al., 2002). In this work it
has observed that, there is no fixed trend found to analyse the parameters such as
BF%, BMI, %FM, FFM BMC, ECM, TBW, ECW and ICW in terms of varying age
of women. Although the variation occurs in such parameters i.e. BMI varies from
BF% from , %FM fro m
, %TBW from , FFM from
, BCM from , ECM from
for the population of female volunteers with age
varying from less than 25 years to more than 60 years.

Overweight and obesity have become more common in different countries. The
composition of a population's gender, age, ethnicity groupings, and numerous
environmental and lifestyle factors all play a role in these variances. Due to these
factors, changes in fat mass (FM) and lean mass (LM) can differ between populations.
As a result, extrapolations from one population with a certain gender, age, and ethnic
composition to another group with a different factors are unknown. Understanding
changes in body composition with increasing age in relation to sarcopenic obesity and
the importance of changes in FM and FFM in relation to the development of general
metabolic diseases within a population are just a few of the clinical implications of
body composition reference data (Maheshwari et al., 2007). The changes in the
composition of the ageing human body are both gerontological and geriatric in nature,
reflecting genetic variables as well as environmental influences like physical activity,
nutrition, and disease, as well as normal ageing processes. Long-term patterns in body
subjects are significant in the study of normal ageing, or ageing in general. Body
composition is also important in the research of obesity, malignancies, clinical
pharmacology, and biochemistry in the field of clinical geriatrics. Changes in body
mass, body fat, and body water are common in both illness and ageing. The
pharmacokinetic and pharmacodynamic properties of medications taken in old age
may be altered as a result of these changes in body subjects (Steen, B., 1988). The
difference in BMI from the youngest to the oldest age groups has been
correspondingly for men and women. The findings of presented
work indicates that reference data on body composition in Swedish populations. BMI

96
and FM were higher among older age groups compared with the younger ones. BMI,
calculated as weight/ height is also widely used as an index of relative weight, often
expressed as standard deviation score (SDS) to take into account age and sex (Wells
and Fewtrell, 2006). After the age of 20, and up to the age of 50, an increase in body
weight was frequent, which was primarily due to an increase in adipose tissue. The
growth in adipose tissue that occurs with age is distributed in a predictable manner,
with the majority of the gain occurring in the central abdominal areas and
subcutaneous fat loss occurring in the limbs (Bembenet al., 1998).

Because they can be read as absolute values or by defining individuals or groups of

individuals as normal or abnormal, FFMI and BFMI are used to follow the impact of
illness, treatment, or ageing in individuals and groups (low and high). Lower FFM can
also be determined using FFMI. This is due to changes in body composition or
reduced lengths in older subjects as they get older. Individuals with high muscle mass
but no extra BF are also identified by FFMI and BFMI. At the group or population
level, these changes are significant (Kyleet al., 2003). The traditional terminology of
body fat and fat distribution cannot be used to assess health risks in the elderly. In
comparison to younger adults, the elderly have decreased muscle and bone mass,
greater fluid volumes outside the cells, and a lower body mass. The elderly have a
more difficult time judging their body composition than younger people. The elderly
have a more difficult time judging their body composition than younger people.
Muscle mass and function, bone mineral and body fluid distribution, as well as body
fat and fat distribution, all require non-invasive procedures. Diseases, disability, and
health status all have varied associations with different patterns of "disordered body
composition”. Using traditional anthropometric metrics like BMI, waist/hip ratio, or
mid-arm muscle area, it's difficult to spot these patterns (Baumgartner, R. N., 2000).
The necessity to explain shortages or excesses of a component that are suspected or
known to be related with health hazards frequently prompts the analysis of body
composition in humans. Body fat and bone mineral density (BMD) values provide
clinical diagnoses in illnesses including obesity and osteoporosis, with implications
for the design of relevant therapies. During infancy and childhood, nutritional
assessment based on body composition can help guide appropriate nutrition and
nutrition management. The measurement methods available range from simple to
complicated, with each approach having its own set of limits and measuring errors.

97
Before picking a measurement method, consider the clinical significance of the body
space to be examined, as more advanced procedures are less accessible and more
expensive. Body composition measurements are used in many fields of b iology and
medicine to gain a better knowledge of nutritional and growth status assessment in
disease states and population treatment (Khosla et al. 1996).Although bone mass and
body weight normally correlate well in adults and children, the ratio may not apply to
obese youngsters. Obese prepubertal and early puberty children are said to have a
higher or equivalent bone mineral density, although their bone mineral content may
be lower than non- fats. In regard to the prospective repercussions of dietary
regulation, as appropriate in obese children, bone mineral status is especially
significant in childhood, which is a vital time for bone mineral accumulation (Lee and
Gallagher, 2008). In our work, the body mass assessment large number of parameters
are determined such as body mass (kg), body fat (percent), lean body mass, FM, ICW,
ECW, and TBW using MFBIA in the ovarian phase. The parametric values are along
with mean and standard deviation are summarised in Table 5.2. The weight and height
are observed as the basic requirements of the body composition analysis study with an
average weight of and height of .

The content of bone minerals diminishes as people age; this is termed as 'osteopenia'
as opposed to the associated pathological process in which the architecture of the bone
alters, resulting in 'osteoporosis'. During the ninth decade, bone mineral density drops
by 30% on average, compared to the peak in the third and fourth decades. After
menopause, bone mineral loss accelerates rapidly in women. Reduced anabolic
hormone levels have been linked to musculoskeletal atrophy and impaired function in
older women. This change in bone mineralization with age is not linked to any
apparent changes in vitamin D nutrients, as measured by circulating vitamin- D levels.
The ageing of the cartilaginous drug replacement causes changes in the body's joints,
which is worsened by the degenerative effects of cumulative use over time
(Andersenet al., 1997). In peritoneal dialysis (PD) patients, malnutrition is a
substantial predictor of mortality. Extracellular mass (ECM) is made up of
metabolically dormant tissues, whereas BCM is made up of metabolically active
tissues. The ratio of ECM to BCM is a particularly sensitive indicator of malnutrition.
Bone, cartilage, ligaments, and non- metabolically active tissues, as well as
extracellular water, make up ECM. The assessment of body tissues, organs, and their

98
distribution in living people is possible because of the body composition
measurements. As a result of cellular metabolic and respiratory activities, the body
mass is responsible for practically all of the body's basic energy usage [Avramet al.,
2010].

The optimal percentage of bodily water varies. Different elements have an impact on
it. This percentage is influenced by gender, age, and body composition. Women have
a lower body water percentage than men in general. Because women have more tissue
than men, this is the case. Adult women's optimal proportion will be between 45 and
60%, while adult men's optimal percentage will be between 50 and 65% of their entire
body weight. Even 5% extra body water than the typical adult is advised for the
correct athletic body types. The total body water (TBW) volume is given as a function
of age, but much of the published data is average and ranges from 20 to over 50 years
old. The availability of current longitudinal data from healthy persons is required for
the evaluation of clinical levels of TBW and trends with age [Chumlea et al., 1999].
The graphical representation of the table shown in the form of pie chart in Figure 5.1
shows that the highest number of volunteer females belongs to the teen age, i.e., 16
females participated in the research whose age was below 25 years. Next highest
number of female volunteers belongs to the age group that covers subjects with age
between 31 years and 40 years. In women age related changes and regional obesity
have been observed mainly after menopause. Women the body mass index is
significantly increases at the age of 31-40 and 50- 60 age groups proceeding age
groups.

Multifrequency bioelectrical impedance analysis (MF-BIA) can discriminate between

intracellular and external components quickly and accurately. The MF-BIA readings
for total body water (TBW), intracellular water (ICW), and extracellular water (ECW)
content show a strong association with the isotopic dilution methodology and double
energy X-ray absorptiometry data. The increase in ECW in respect to ICW requires
special attention. It majorly reflects serious health risk when excess of ECW is
observed in current study. The hydration component of the body composition
approach is 73.3% of lean body mass (LBM), while fat content is determined by
subtracting LBM from body weight. Age, gender, and physical size all influence total
fluid volume. In healthy subjects, ICW content may be linked to the muscular
component in particular. The TBW content o f men remains generally steady during

99
early adulthood, and then declines at a rate of 0.3 kg/yr until it hits a low point
between 70 and 80 years. The TBW content in women was shown to stay constant
between young adulthood and middle age; however, at the a ge of 70 years, the TBW
content dropped dramatically by 0.7 kg each year (Ohashi et al., 2018). The mean and
SD for various parametric values used in the study of body mass analysis are plotted
in Figure 5.2. The study further revels that with the increase in the age of subjects
from a considerable increase in the %FM from
FFM from , BCM from
, and ECM is observed. The
increase in the overall body compositions values with decrease in ECM with age
shows that there is strong correlation between impedance and body composition. It is
also observed that it reflects the body composition in terms of TBW, ECW/ICW
decreases with the rising age of female group.

The impedance or resistance to the flow of an electric current through body fluids,
primarily in lean tissue, is measured by bioelectrical impedance analysis (BIA). Due
to the presence of intracellular fluid and electrolytes, as well as a large amount of
adipose tissue, lean tissue has a low impedance. As a result, impedance is proportional
to the entire volume of bodily water. Because muscle has the highest water content of
all body tissue, BIA uses an electric current to estimate muscle mass based on the
current's conduction through water. The use of BIA to determine body composition is
a safe and cost-effective technology that eliminates the use of radiation. It's based on
the differential in resistance between fat and lean body parts when an electric current
is sent through them. It may calculate the total body fat content using the total body
water content as a starting point (Goswami and Munna, 2007). In the evaluation study,
impedance analysis for legs, arms and the trunk at 50 kHz, 100 kHz, and 200 kHz
were determined. Presented work indicates the variation in the impedance with
reference to different age groups of female. It is observed that with the increase in age
of female’s impedance decreases with minor variations in each a ge group. The
decrease in impedance value with increasing age suggests that impedance is an
indicator of function and general health. It is not only an indicator of the body
composition but also shows the comparison of the values with overall mean. The
calculated average value of impedance is computed to be at frequency of
50 KHz, at 100 KHz and at 200 KHz, respectively.

100
The phase angle varies depending on gender, age, BMI, and fat proportion. Phase
angle's utility in clinical and epidemiological circumstances has been limited due to a
lack of reference values. Such data are required in epidemiological studies to correctly
quantify individual deviations from the population mean and to investigate the impact
of phase angle on various outcomes. The phase angle is a measurement derived from
the reactance and resistance of BIA. Resistance and reactance measurements can also
provide a BIA with a phase angle estimate. Although BIA body composition
projections are frequently based on population-specific comparisons, the phase angle
is calculated directly without any conversion to individual body sections. The idea of
phase angle is based on resistance and reactance variations while an alternating
current passes through the examined tissues (Gonzalez et al., 2016). It is further
observed that the variation in phase angle and impedance is highly affected with sex,
age, BMI, FM%. The observed significance level of the study summarized in above
table is found to be for all analysis. It is observed that impedance for phase
age of 50 khz is found to be for the age group of females with age less
thn 20 years that increases to for the age group that covers females of
age more than 60 years. Similarly, impedance varies from
for phase angle measurement at 100 khz and from
for phase angle measurement at 200 khz. Figure 5.4 shows the variation in
impedance analysis along different age group of females with respect to d ifferent age
groups. It is observed that least impedance is observed with respect to phase angle of
50 kHz followed by 100 kHz and 200 kHz with an average value of
This also illustrates that highest impedance is
observed when phase angle of 200 khz is used for analysis. This distribution of
impedance and phase angle proved to be fairly normal in the present study. The
overall mean phase angle and impedance in all categories, there is significant decrease
in phase angle and impedance with increasing age in females.

Body mass alone does not provide an encompassing view of an individual's health. It
is ideal to establish body composition in order to better assess whether or not an
individual is in good health. Depending on the method utilized, we can estimate
several variables such as percentage BF%, lean mass, fat mass, bone mineral content,
and TBW. The various approaches differ in terms of cost and the degree of experience
required by the tester to complete the assessment successfully. Many different

101
subgroups of people can benefit from the BF outcomes. BF% is a useful metric for
tracking progress in a fitness or weight loss plan in the general population. The
importance of having the most precise estimation of BF% is due to this use of body
composition calculations. Body composition equipment manufacturers have
established various test methodologies to achieve the most valid and trustworthy
results. Participants are often advised to test when they are fast and adequately
hydrated in order to receive the most precise estimates about methods. In addition,
before being tested, participants are requested to exercise to confirm that there is no
increase in body temperature or dehydration owing to sweat loss. It has bee n
questioned if menstruation can impact the consistency of these assessments due to
possible fluid retention, weight increase, and hormonal variations that occur during
the menstrual cycle. Whether uncertain number of women should be checked during
their menstrual cycle is still up in the air. The women's body composition was also
assessed during different ovarian cycle phases named as; (a) Ovarian, (b) Follicular
(c) Early Luteal and (d) Late Luteal. The variation of body composition with phases
of ovarian cycle has also been observed in this study. Ovarian cycle has an important
bearing on fertility and health of women. Irregular menstrual cycles have especially
been used as an indicator of reproductive health. The length and regularity of
menstrual cycle is an indicator of cumulative exposure of ovarian steroids. Among the
premenopausal women in the present study there is decrease in BMI.

i. The first phase (Ovarian) used for the analysis consists of day 1 to day 7 days
of the ovarian cycle that is also named as ovarian phase of the cycle.
ii. The second or, follicular phase is comprising of day 8 to day 14 of the cycle.
iii. Third or Luteal phase is categorised into two sub categories, namely early
luteal phase that comprises of day 15 to day 21 of the cycle (refer table 5.5)
iv. Finally, last phase of ovarian cycle i.e. Late luteal phase that covers day 22 to
day 28 of the cycle.

Hormonal changes due to pregnancy or menopause may also cause water retention
and variations in measuring. There is a significant positive association with cycle
length for each body composition measure. The relationship between each body
composition measure and cycle length was nonlinear with the longest mean cycle
length occurring with greater BMI. Body fat mass or body lean mass small reduction

102
in the levels of estradiol during the follicular phase, even not related to significant
changes in the length of menstrual cycle is associated with decreased probability of
conception. A regulatory role of nutritional status on reproductive ability was
hypothesized. There is greater probability in menstrual cycle change with women age
with menarche. The variation of body composition with ovarian cycle in terms of
%FM, by varying FFM also the variation of FFM in Kg, BMI, for female age group
including graphical representation is given in figure 5.5, figure, 5.6, figure 5.7 and
figure 5.8 correspondingly. Table 5.6 summarizes the body compositions of females
that are examined through MFBIA analysers during different phases such as; ovarian,
follicular, early luteal and late luteal of ovarian cycle in different age group i.e. <25
Yrs, 25-30 Yrs, 31-40 Yrs, 41-50Yrs and >60Yrs. It is observed that %FM for
subjects with age group less than 25 years is found to be during ovarian
phase, during follicular phase, during early luteal phse
and during late luteal phase.

In the context of FFM, its higher value is analysed during luteal phase as compared to
ovarian and follicular phase irrespective of the age group under study. For instance,
FFM observed in the ovarian phase is , follicular phase is ,
early luteal phase is , and late luteal phase is for subjects
belonging to age group of 25 years to 30 years. Similar observations are also found
for subjects of age more than 60 years of age with highest FFM exhibited in luteal
phase as compared to in ovarian phase and
in follicular phase.

The correlation of ovarian cycle with FM (Kg) for different age group of females. It is
observed that FM increases with the increase in the age of the subjects which means
that higher FM is observed for older females that the young teenagers. However,
minor variation is observed for FM with respect to phases of ovarian cycle. For
example, as illustrated by the observations, females having age group of 25 years to
30 years exhibits FM of in ovarian phase, in follicular
phase, early luteal phase and and late luteal phase.

It is observed that higher BMI is observed in the luteal phases of the ovarian cycle as
compared to ovarian or follicular phase irrespective of the age of the subjects. For
example, the subjects belonging to age group of 31 years to 40 years exhibits

103
 BMI in ovarian phase, in follicular phase, in early luteal
phase and in late luteal phase. This shows that BMI is higher in luteal
phase of ovarian cycle as compared to other phases.

Final analysis indicates that the %FM almost remains unchanged during various
phases of cycle. However, a considerable variation that is found in different age
groups is also due to difference in the physique of the population under study. Overall
it is observed that highest FFM is exhibited in luteal phase irrespective of the age of
the subjects. Similar, trends for FM have also been observed in different age groups.
This shows that FM does not demonstrate large variation with different phases of
ovarian cycle however, FM is found to be highest in luteal phases of the ovarian
cycle. In terms of BMI, its highest value is found in luteal phase of ovarian cycle as
compared to other phases. The study reveals that there is co-relation of body
compositions with ovarian cycle phases when compared each phase such as ovarian,
follicular, luteal and late luteal with female age group of 44 females. In existing work
(Michels et al., 2017) indicates that the factors which may be responsible for ovarian
cycle variance have been identified. According to the findings of the current study,
women in lower age groups who are physically active had more frequent ovarian
cycles. Increased age of menarche and decrease in BMI indicates that enhancement in
duration of ovarian cycles from the perspective of public health. To understand the
impacts of ovarian cycle of moderate, different levels of physical activity, BMI along
with age at menarche is important. It has also observed relevant for recognizing the
cause and deviation of ovarian cycle to make it enable normal reproductive female
health.

The change in body compositions shows that with the increase in age group of
females there is change in each phase of ovarian cycle and for each cycle mean and
SD values has been set up which shows if there is slight change in mean ad SD values
this shows the body composition changes with female age group and if these values
change there will be change in body composition and MFBIA values shows variation
which reflects the ovulation cycle and composition value will change or show some
variations in FM%, FFM, BMI and impedance will show a great change. These
variations of compositional values and change in body fluids will show change in
cycle of ovarian phase and thus causes change in hormones or hormonal imbalance,

104
and effects fertility of females and pregnancy. This graphical study also proves if
there is decrease in value of impedance, fat mass will increase and shows BMI of
obese female. In Body it is used to analyse women's body composition during various
phases of the ovarian cycle. We considered the females with the appropriate number,
age, height, and weight, body mass and ovarian length.ANOVA tests were conducted
during this study, and it was observed that variables of body composition were
evaluated for four phases of ovarian cycles. The significance level for all analyses has
been p≤0.

105
CHAP TER 7: SUMMARY AND CONCLUSION

The present study reviews the most commonly used methods for essential part of
making sexual reproduction possible. Ovarian cycle is essential for the production of
eggs, and for preparation of uterus for pregnancy. In humans the length of
menstruation varies greatly among women with 28 days (Heyward and Wagner, 2004)
(Lukasi, 2000). Each cycle can be divided into three phases based on events in the
ovary. Endocrine changes block the ovary during ovarian cycle (Vanovski et al., 1996)
(Nunez et al., 1997). Ovarian cycle is the series of changes that female goes to a
possible pregnancy in human females it occurs repeatly at the age of menarche. The
ovarian cycle consists of the phases which are follicular phase, ovulatory and luteal
phase were as uterine is divided in ovarian prolifera tory and secreatory phase
(Heymsfield et al., 2005).

The level of follicle stimulating increases at lesser degree ovulatory lasts 16 to 32

hours. Once the ovulation occurs the follicle that contained the egg transforms into
something called corpus luteum and begins to produce progesterone and estrogens
Enumenorrhea denotes normal regular menstruation or ovarian cycle 3 to 4 days.
Presence of two different estrogens receptors in the hypothalamus estrogens receptor
alpha is responsible for negative feedback (Heyward and Wagner, 2004, Buchholz et
al., 2004 and Deurenberg et al., 1988). Progesterone is an indigenous steroid hormone
in an ovarian cycle. Another type of estrogens called estetrol (E4) is produced during
premarin.Premarin a commonly prescribed estrogenic during produce firm urine of
pregnant mares contains the steroidal estrogens, equine, and equilenin.Progestrone
steroid hormone involves in ovarian cycle (Dixon et al., 2008, Gomez et al., 1993,
Mitchell et al., 1993 and Damavandi, 2008).

Ovarian disease is conditions that happen to young women and can affect their
reproductive system and health (Teilmann et al., 2006) (Chumlea et al., 1987).
MFBIA is used to define phase and easy to use to access the risk of angular illness
(Gleichauf and Roe, 1989) (Gonzalez-Correa and Caicedo-Eraso, 2012). The
chronological age and ovarian period is to consider how this midlife can affect size
and composition of the body in women(Cumberledge et al., 2018) (Feldmar et al.,
2009). The classification has been performed on the categories named as normal,

106
overweight, and obese (168,169) (Silva et al., 2018).

Considers hoe these could influence body size and composition in midlife(Beato et al.,
2019).Menopause did not affect the energy intake. These techniques allow the
measurement of FFM, Bone mineral content, ECW, TBW, EW, total adipose and
ectopic fat depots (Kyle and Bosaeuset, 2004). Ovarian hormone levels and nutritional
status hormone levels come from studies on women anorexia nervosa. Low body fat
mass and low level of estrodiol inhibition of ovarian cycle (Janssen et al.,
2000).Hormonal level can influence nutritional status higher level of follicular
estradiaol has been observed in cycles. It is impossible to determine if observed
changes in the level of reproductive hormones by nutritional status (Kushner et al.,
1996, Barbosa et al., 2005, Kyle et al., 2004 and Barbosa and Barros, 2005). However,
the association of full range body composition measure cyclicity is not determined.
Body contain many substances more neutral fat does not bind water, nitrogen and
electrolyte and result of tissue analysis on fat free basis (Cornish et al., 1993) (Segal et
al., 1991). We determined body fat mass, BMI of lower abdomen value of females we
derived equation between body fat mass, age, BMI circumferences for female
percentage fat mass is also varies with age.

7.1 CONCLUSION

A number of physiological changes occur during puberty to menarche including rapid

increase in physical size, hormonal fluctuations and marked changes in body
composition and timing of marked changes in body compositions. Furthermore,
adipose tissue as a major component of body composition is involved in hormonal
interaction, growth and maturation factors. The majority of measured various body
component (TBW, ECM, FFM) can lead to better understanding including obesity,
age related changes, ovarian phases change with increase in age, in addition this
knowledge allows predictions to be made about subsequent health outcomes and helps
to identify timing for more effective.MFBIA is better easy to handle, portable, takes
less time, and does not cause any harm to humans.MFBIA is a repetitive, easy and
efficient method for evaluating fat free mass in p hysiological and pathological
conditions. It is advisable to use scientifically certified analyzer to ensure reliable
measurements to place correct electrodes and respect standard procedures. The major
observations of the present study are summarized under three sections to fulfill the

107
designed objectives of the study.

7.1.1 Parametric Analysis of Body Composition for different Age Groups

 The variation in the length of cycle is observed from 27 to 31 days with

considerably lengthy cycles found in elderly females.
 There is no fixed trend found for the measurement of BF%, BMI, %FM, FFM
BMC, ECM, TBW, ECW and ICW with respect to age of subjects. However,
BMI varies from BF% from
, %FM from , %TBW
from , FFM from ,
BCM from , ECM from
for the population of female volunteers with
age varying from less than 25 years to more than 60 years.

7.1.2 Impedance and Phase Angle Analysis at Different Frequencies

 The experimentation in terms of impedance analysis shows that highest

average impedance of is observed at 50 khz followed by a n
average impedance of at 100 khz and at 200 khz
 Similar measurements are also performed for variation in phase angle from 50
kHz to 200 kHz. It is observed that least impedance is observed with respect to
phase angle of 50 kHz followed by 100 kHz and 200 kHz with an average
value of This also illustrates
that the highest impedance is observed when phase angle of 200 kHz is used
for analysis.

This distribution of impedance and phase angle proved to be fairly normal in the
present study. The overall mean phase angle and impedance in all categories, there is
significant decrease in phase angle and impedance with increasing age in females.

7.1.3 Body Composition variation with Phases of Ovarian Cycle

The major aim of the study is to analyse variation in body composition in reference to
different phases of the ovarian cycle.

108
 It was observed that %FM distribution remained almost unchanged in different
phases of the ovarian cycle. However, a considerable variation that is observed
with respect to different age groups is also due to difference in the physique of
the population under study.
 The FFM analysis shows that highest FFM is exhibited in luteal phase
irrespective of the age of the subject used in the study.
 In case of FM measurements, it is observed that it does not demonstrate a very
large variation with different phases of ovarian cycle. In fact, FM exhibits to
be the highest in luteal phases of the ovarian cycle.
 BMI analysis shows that highest BMI is observed in the luteal phases of the
ovarian cycle as compared to ovarian or follicular phases irrespective of the
age of the subjects.

109
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