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Medical Biochemistry for

Physiotherapy Students
Medical Biochemistry for
Physiotherapy Students

Harpreet Kaur
Assistant Professor
Department of Biochemistry
Gian Sagar Medical College and Hospital
Ram Nagar, Banur
Dist. Patiala (Punjab), India

Jagmohan Singh
Principal and Professor
Gian Sagar College of Physiotherapy
Ram Nagar, Banur
Dist. Patiala (Punjab), India

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Medical Biochemistry for Physiotherapy Students


© 2008, Harpreet Kaur, Jagmohan Singh
All rights reserved. No part of this publication should be reproduced, stored in a retrieval system, or transmitted in any form or
by any means: electronic, mechanical, photocopying, recording, or otherwise, without the prior written permission of the
authors and the publisher.

This book has been published in good faith that the material provided by authors is original. Every effort is made to ensure
accuracy of material, but the publisher, printer and authors will not be held responsible for any inadvertent error(s). In case
of any dispute, all legal matters are to be settled under Delhi jurisdiction only.

First Edition : 2008


ISBN 978-81-8448-379-6
Typeset at JPBMP typesetting unit
Printed at Gopsons Papers Ltd., A-14, Sector 60, Noida
Dedicated to
Our Parents, Teachers,
Friends and Students
Foreword

In modern days of advanced medical care, Biochemistry has attained a key position in the
field of diagnosis and prognosis of diseases. Physiotherapy is a highly upcoming branch of
medical sciences. There is a great role of Biochemistry in Physiotherapy. I appreciate the
meritorious work done by Harpreet Kaur and Jagmohan Singh in writing this book Medical
Biochemistry for Physiotherapy Students. This book will definitely help the students of
Physiotherapy to study the subject of Biochemistry better.
I congratulate both of them for successful completion of this work and wish this book a
great success.

Dr Kamaljit Singh
Professor and Head
Department of Biochemistry
Gian Sagar Medical College and Hospital
Banur, Dist. Patiala

Chief Executive Officer


Gian Sagar Educational and Charitable Trust
HO: SCO 109-110, Sector 43-B
Chandigarh
Foreword

It gives me immense pleasure to write foreword for the book titled Medical Biochemistry for
Physiotherapy Students authored by Harpreet Kaur and Jagmohan Singh. This book is a much
needed work in the absence of a comprehensive explanatory textbook written for degree
level Physiotherapy students.
In my opinion, Medical Biochemistry for Physiotherapy Students lives up to the promise of its
title, and I am delighted that the authors found the time and dedication needed to write this
book which will enable them to share their experience as teachers of Physiotherapy.
I appreciate both of them for successful completion of the book.

Dr MS Sohal
Professor
Department of Sports Sciences
Punjabi University
Patiala
Preface

The book titled Medical Biochemistry for Physiotherapy Students has been designed to cater to
the long felt needs of students of physiotherapy. The book is also useful for professionals,
teachers, doctors, rehabilitation professionals, other paramedics and public in general.
The book is developed in a simple and concise format that is easily and readily
understandable. The clear, readable type and bold headings make the text easy to comprehend
and follow. The book is adequately illustrated with diagrams and tables to assist the students
in readily grasping the essential points.
The concepts in this book are presented in a simple and straight-forward style. Illustrations
and examples have been frequently used to clarify the topics in detail and are further explained
as a concept.
We hope that the book will provide complete guidance to the students. The book is also
useful for teachers, nursing students and students of other allied courses.

Harpreet Kaur
Jagmohan Singh
Acknowledgements

Compilation of such a comprehensive work cannot be done single handedly. The following
persons have also been an invaluable source of opinions and comments. Thanks goes to all of
them for their encouragement and support:
• Dr MS Sohal, Professor, Dept. of Sports Sciences, Punjabi University, Patiala.
• Dr Kamaljit Singh, Professor and Head, Dept. of Biochemistry, Gian Sagar Medical College
and Hospital, Banur, Patiala.
• Dr Minni Verma, Professor, Dept. of Biochemistry, Gian Sagar Medical College and
Hospital, Banur, Patiala.
• Dr Sukhdev Singh, Vice Principal, Desh Bhagat Dental College, Muktsar.
• Dr Amrit Kaur, Professor and Head, Dept. of Biochemistry, Adesh Institute of Medical
Sciences and Research, Bathinda.
• Dr RL Mattoo, Professor, Dept. of Biochemistry, Adesh Institute of Medical Sciences and
Research, Bathinda.
• Dr Paramvir Singh, Lecturer, Dept. of Sports Sciences, Punjabi University, Patiala.
In preparing a textbook like this, I have utilized the knowledge of a number of stalwarts
in my profession and consulted many books and journals. I wish to express my appreciation
and gratitude to all of them including the related authors and publishers.
My special thanks to Shri JP Vij, CMD, M/s Jaypee Brothers Medical Publishers (P) Ltd.
and his whole team for publishing this book.
Contents

1. Introduction ......................................................................................................................... 1
2. Water and Electrolytes ...................................................................................................... 6
3. Chemistry of Carbohydrates ......................................................................................... 12
4. Chemistry of Lipids ......................................................................................................... 21
5. Chemistry of Amino Acids and Proteins ................................................................... 28
6. Chemistry of Nucleotides and Nucleic Acids .......................................................... 37
7. Enzymes ............................................................................................................................. 43
8. Vitamins ............................................................................................................................. 53
9. Bioenergetics ..................................................................................................................... 69
10. Digestion and Absorption .............................................................................................. 74
11. Metabolism of Carbohydrates ...................................................................................... 82
12. Metabolism of Lipids .................................................................................................... 100
13. Metabolism of Proteins ................................................................................................ 109
14. Minerals ........................................................................................................................... 117
15. Fluid and Electrolyte Balance and Imbalance ........................................................ 124
16. Biophysics ........................................................................................................................ 128
17. Metabolism of Specialized Tissues .......................................................................... 135
18. Hormones ......................................................................................................................... 143
19. Diet and Nutrition ......................................................................................................... 158
20. Radioactivity and Radioisotopes ............................................................................... 165
21. Normal and Abnormal Constituents of Urine ........................................................ 170

Index ................................................................................................................................... 177


Chapter 1

Introduction

INTRODUCTION TO BIOCHEMISTRY Biochemistry can be divided into four


main branches:
Biochemistry is the science which deals with 1. Medical Biochemistry
the chemistry of living organisms, both plants 2. Animal Biochemistry
and animals. 3. Plant Biochemistry
Cell is the structural unit of all forms of 4. Biochemistry of Micro-organisms.
life. Although protoplasm of cell is different Medical Biochemistry deals with the
in each kind of animal and plant cell but the following with which the medical students
chemical composition, organization and are mainly concerned. i.e. chemistry of tissues
processes occurring in each type is similar in and foods, digestion and absorption, respi-
many respects. ration, blood, cell membrane and physical
The relationship of the living beings to chemistry, tissue metabolism, glands, excre-
their environment; the processes by which an tion, biochemical disorders in various
exchange of chemical substances takes place diseases etc.
between the living organism and its environ- STRUCTURE AND FUNCTION OF
ment through digestion, absorption and CELL CONSTITUENTS
excretion; the processes by which the
Cells are the smallest structural and functional
absorbed materials are utilized for synthetic
units of all living organisms. The cells of the
reactions leading to growth and development
living kingdom are divided into two cate-
of tissues; the metabolic breakdown of the gories:
materials to supply energy; the mechanisms 1. Prokaryotic cells
which regulate with precision all these 2. Eukaryotic cells
processes by means of hormonal and neuro- Prokaryotic cells lack a well defined
regulatory stimuli – all these come under the nucleus and possess simple structure. These
purview of biochemistry. include various bacteria.
2 Medical Biochemistry for Physiotherapy Students
Eukaryotic cells possess a well defined Cells
nucleus and are more complex in their
structure and function. The higher organisms
(animals and plants) are composed of Tissues
eukaryotic cells. We are discussing eukaryotic
cells in detail.
Cells when grouped together forms tissues Organs
which perform specialized functions. Tissues
when grouped together forms organs and
different organs forms organ systems e.g. Organ System
digestive system which is responsible for
Fig. 1.1: Cell and organ system
taking in, digesting and absorbing food and
involves a number of organs including the
stomach and intestines (Fig. 1.1). They include:
A cell consists of plasma membrane inside 1. Nucleus
which there are a number of organelles 2. Mitochondria
floating in a watery fluid called cytosol (Fig. 3. Ribosomes
1.2). Organelles are small structures with 4. Endoplasmic reticulum (ER)
highly specialized functions. 5. Golgi apparatus

Fig. 1.2: Structure of a cell


Introduction 3
6. Lysosome The membrane proteins perform several
7. Centrioles functions:
8. Microfilaments • They give the cell its immunological
9. Microtubules identity.
• They can act as specific receptors for
1. Plasma Membrane hormones and other chemical messengers.
The boundary of every cell has a thin mem- • Some are enzymes.
brane of thickness 75Å units. This is known • Some are involved in transport across the
as unit membrane or plasma membrane. membrane.
The most important function of biologic
The plasma membrane consists of two
membranes is to restrict the exchange of
layers of phospholipids (fatty substance) with
substances among various compartments.
some proteins embedded in them (Fig. 1.3).
Thus, different types of body fluids are
Those that extend all the way through the
separated from each other by different
membrane may provide channels that allow
membranes.
the passage for example electrolytes, non-lipid
soluble substances, etc. 2. Nucleus
The phospholipids molecules have a head It is a spherical or oval organelle. Within the
which is hydrophilic (attracting water) and a nucleus, hereditary units of the cell called
tail which is hydrophobic (water repelling). genes are present which control cellular struc-
The phospholipids bilayer is arranged like a ture and direct many cellular activities. The
sandwich with the hydrophilic heads aligned nuclear genes are arranged in a single file
on the outer surfaces of the membrane and along structures termed chromosomes.
the hydrophobic tails forming a central water Human body cells have 46 chromosomes.
repelling layer. These differences influence Each body cell contains a nucleus except red
the transfer of substances across the mem- blood cells.
brane. The nucleus is contained in a membrane
similar to the plasma membrane but it has
tiny pores through which some substances can
pass between it and the cytoplasm i.e. the cell
excluding the nucleus.
Nucleoli are present inside the nucleus.
They are clusters of proteins, DNA, RNA that
are not enclosed by a membrane. Nucleoli are
the sites of assembly of ribosomes and contain
a type of RNA called ribosomal RNA (Fig.
1.4).

3. Mitochondria
These are the largest components in cyto-
Fig. 1.3: Structure of plasma membrane plasm. These are described as the power
4 Medical Biochemistry for Physiotherapy Students
of ATP is most efficient in the final stages of
cellular respiration. This is called oxidative
phosphorylation, requiring the presence of
oxygen.

4. Ribosomes
These are tiny granules composed of RNA
Fig. 1.4: Structure of nucleus and proteins. They are also found on the outer
surface of rough endoplasmic reticulum
house of the cell and each cell may contain (RER). Bacterial ribosomes consists of two
from 50 to 2500 mitochondria depending on subunits of unequal size, one of sedimentation
the respiratory activity of the cell. The cells coefficient of 50S and other of 30S. In
of skeletal muscle, kidney and liver contain eukaryotes, 80S ribosome is found (60S and
large amount of mitochondria while those of 40S). They synthesize proteins from amino
heart muscle contain less. acids using RNA as the template.
It has two membranes – the outer mem-
brane and the inner membrane. The outer 5. Endoplasmic Reticulum
membrane is smooth while the inner mem- These are a series of membranous canals in
brane is folded to form ridges or cristae which the cytoplasm. They are of two types:
extend into the matrix of the mitochondria. a. smooth endoplasmic reticulum (SER)
Elaborate folds of the cristae provide an b. rough endoplasmic reticulum (RER)
enormous surface area for a series of chemical Smooth endoplasmic reticulum synthesizes
reactions called cellular respiration. Two lipids and steroid hormones and is also
spaces – intercristae space and the matrix associated with detoxification of some drugs.
space are thereby developed. The matrix Rough endoplasmic reticulum is site of
space is rich in enzymes (Fig. 1.5). synthesis of proteins that are extruded from
cells i.e. enzymes and hormones that pass out
of their parent cell to be used by other cells
in the body.

6. Golgi Apparatus
Fig. 1.5: Structure of mitochondria This consists of stacks of closely folded
flattened membranous sacs. It is present in
Mitochondria are involved in cellular all cells but is larger in those cells that
respiration, the process by which chemical synthesize and export proteins. The proteins
energy is made available in the cell. This is in move from the ER to the golgi apparatus
the form of adenosine triphosphate (ATP). where they are packaged into secretory
ATP is a high energy compound. Synthesis vesicles, sometimes called secretory granules.
Introduction 5
The vesicles are stored and when needed sible for equal distribution of the characters
they move to the plasma membrane, through in the offspring.
which the proteins are exported.
9. Microfilaments and Microtubules
7. Lysosomes
These are contractile structures in the cyto-
They are oval or spherical membrane bound plasm involved in the cell and of organelles
bodies formed by the golgi apparatus. They within the cell, the movement of cilia (small
contain a variety of enzymes involved in projections from the free surface of some cells)
breaking down fragments of organelles and and the organization of proteins in the plasma
large molecules (e.g. RNA, DNA, carbo- membrane. They also maintain the charac-
hydrates, proteins) inside the cell into smaller teristic shape of the cell.
particles that are extruded from the cell as
waste materials. Lysosomes in WBC contain 10. Cytoplasm
enzymes that digest foreign materials such The cytoplasm is the structure less material
as microbes. filling the rest of the cell (aqueous matrix) in
which all the cell components float. It is a
8. Centrioles
colloidal solution of proteins containing nearly
These are two short, cylindrical structures 70% of water besides both organic and
known to exist on either side of the nucleus inorganic substances such as glucose, potas-
at right angles to each other. They are not sium, magnesium etc. The cytoplasm also
bound by any membrane. They help in the contains a small amount of RNA and the
equal division of the chromosomes by taking enzymes for glycolysis, gluconeogenesis and
them apart and they are, therefore, respon- HMP shunt.
Chapter 2

Water and Electrolytes

WATER Properties

Water is the most abundant of all chemical 1. Water is colourless, odourless but in larger
compounds and it exists in the form of solid, masses has a blue tinge due to the presence
liquid and gas. of the finely divided material.
2. Water at sea level freezes at 0ºC or 32ºF
Sources and boils at 100ºC or 212ºF.
3. On freezing, water rapidly expands and
1. It occurs in the form of ice – vast areas of
increases its volume by nearly 100%. It
the colder region.
becomes less dense and floats on water.
2. It occurs in the liquid state – 5/7th of
4. The volume of steam is produced about
earth’s surface.
1700 times larger than the original volume
3. Soil contains water which is necessary for
of water. Confining this steam under
plant life.
pressure in an autoclave raises the boiling
4. Body is 2/3rd water.
point of water and hence increases the
5. It occurs in food in varying quantities.
temperature.
Necessity of Water 5. It is an excellent universal solvent.

Life cannot exist without water, e.g. the Chemical Properties


human body without water – its muscles will 1. It is very stable compound and even when
refuse to function, food would not be heated to 2500ºC, only a small percentage
digested, elimination of waste and the of water molecules dissociate into hydro-
building of new tissue would stop and gen and oxygen and these elements reunite
regulation of body temperature will be so on as the temperature is lowered but it
disturbed. can be decomposed by an electric current.
Water and Electrolytes 7
2. Water reacts with many compounds to Hard Water
break them into simpler substances by the
The water containing the bicarbonates and
process called hydrolysis e.g. digestion of
sulphates of calcium and magnesium is called
carbohydrates, fats and proteins.
as hard water.
3. Water combines directly with many
Hardness is of two types:
compounds to form crystalline substances
1. Temporary hard water
called hydrates. Plaster of Paris when
2. Permanent hard water
mixed with water sets to a hard mass and
it is used in making surgical casts. Temporary Hard Water
This water contains the bicarbonates of
calcium and magnesium and it can be softened
by boiling because heat changes the soluble
bicarbonates into insoluble carbonates of
calcium and magnesium.
The water which assists in formation of
crystals is called water of crystallization or
water of hydration. Some crystalline sub-
stances contain no water of hydration. These
are called as anhydrous compounds e.g. sugar,
salt. When a compound takes up atmospheric
water, it is called deliquescent e.g. calcium
chloride. Permanent Hard Water
This contains chlorides and sulphates of
Impurities of Water
sodium and magnesium. It can be softened
Naturally occurring water contains more or by adding caustic soda, slaked lime and by
less foreign matter derived from the soil. special processes.
This foreign matter is either mineral or
organic matter and it may be either dissolved
or suspended in the water.
Depending upon the mineral matter found
in water it is classified as:
1. Soft water
2. Hard water
Purification of Water
Soft Water
Water contains organic impurities such as
The water which contains little or no mineral sewage, refuse and inorganic impurities like
matter and lathers easily is called as soft water. salts, bicarbonates, chlorides and sulphates
8 Medical Biochemistry for Physiotherapy Students
of calcium, magnesium, iron, lead etc. and also 3. Water keeps various surfaces moist such
has dissolved gases like nitrogen and carbon as the mucous membrane of the nose, eyes
dioxide. Pathogenic bacteria may also be and throat and prevents discomfort. It also
present. moistens the joint and act as the solvent
in the large intestine helping in the elimi-
Methods of Purification nation of faeces.
4. Water acts as a temperature regulator by
1. Distillation: Boiling water and condensing
equalizing temperature of various parts of
the resulting steam in a different container
the body through the circulating fluid. The
is called as distillation. It is the most
regulation of the body temperature is
effective method of purifying water.
accomplished largely by the skin – by
2. Boiling: Water may be made fit for
controlling the amount of heat lost from
drinking purposes by boiling. This process
the body through altering the amount of
does not remove any dissolved solid
blood sent to the surface and the amount
matter. But kills the disease producing
of perspiration. The major portion of heat
micro-organisms.
lost from the body is removed through
3. Filtration: The suspended matter in water the evaporation of water from skin.
such as clay and undissolved organic 5. It helps in digestion. It is the principal
matter may be removed by filtration. It is constituent of the secretion of glands
a very economical method of purifying (digestive juices). It is essential in rapid
water for drinking purposes. chemical action and the absorption of
4. Aeration: When exposed to the air, water digested foods. It is one of the reacting
in time purifies itself. The oxygen of the substances in many chemical changes
air dissolves in the water, acts chemically (hydrolysis). Practically, all the chemical
on the bacteria and destroys them. processes which take place during diges-
tion and many of these which occur in the
Functions/Physiologic tissues are of this nature i.e. breaking
Importance of Water down of complex substances into simple
1. It is a component of protoplasm and substances by means of chemical reaction
tissues. It keeps most of the constituents with water. Enzymes speed up these
of the cell contents (protoplasm) in solu- reactions.
tion so that chemical changes essential to
maintain cell function can take place. It ELECTROLYTES
maintains the proper degree of dilution The substances which conduct the electric
in the tissue fluids which bathe the cells, a current in solution are called electrolytes e.g.
condition necessary for their functioning. HCl, NaCl and the substances which fail to
2. It provides fluid medium for transporting pass the current are called as non electrolytes
food materials away from the tissues. It is e.g. sugar.
necessary for the proper excretion of So, electrolytes include acids, bases and
waste material many of which are excreted salts and non electrolytes include pure water,
in urine. alcohol and sugar.
Water and Electrolytes 9
The body contains electrolytes and is a 2. Action on indicators: Many coloured
conductor of electricity. Its advantage is taken compounds changes colour in the presence
by using the electrocardiograph, an instru- of acids. So these are used as an indicators
ment which measures the variation in the for detecting acids e.g. blue litmus dye
feeble current released during the contraction which changes to a red colour in the pre-
and relaxation of the heart. sence of an acid.
The common acids HCl, HNO3, H2SO4 and 3. Action on metals: When active metals such
the bases NaOH and KOH are ionized into as sodium, magnesium or zinc are placed
in acid solutions, they react chemically and
water, so they are called strong electrolytes.
hydrogen gas is usually liberated. Because
Other acids and bases such as acetic acid and
of this corrosive action, the acids are kept
ammonium hydroxide show only a small
in glass containers.
degree of ionization, so they are called as
4. Action on oxides and hydroxides: Many acids
weak electrolytes.
react with the oxides and hydroxide of
metals. Neutralization is a useful reaction
Acids
for counteracting the action of an acid or
Acids give hydrogen ions (H+) in solution. a base.
Many of our food contain acids and they are 5. Action on carbonates and bicarbonates:
constantly forming in the body. The gastric Whenever an acid solution comes into
juice and the urine are acid fluids. contact with a carbonate or bicarbonate,
effervescence comes out due to the
Classification of Acids evolution of CO2 gas.
6. Action on tissues: The physiological action
1. Organic acids of strong, concentrated acids upon the
2. Inorganic acids tissue is to destroy the tissues through
All of the organic acids contain carbon and corrosive action. But much diluted acids
they are often derived from plant and animal only abstract water from the tissues. So in
sources. Organic acids are weak. Its examples fevers, mild acids may be used to diminish
are acetic acid C2H4O2, citric acid C6H8O7, the thirst because they stimulate the flow
lactic acid C3H6O3, tartaric acid C4H6O6, etc. of saliva.
The important inorganic acids are known as
mineral acids. Its examples are hydrochloric Precautions While Handling Acids
acid HCl, nitric acid HNO3, sulphuric acid and Bases
H 2 SO 4 , boric acid H 3 BO 3 , carbonic acid Acids and bases are designated as corrosive
H2CO3 etc. chemicals. Corrosive chemicals are those
which when ingested, inhaled or allowed to
Properties of Acids come in contact with skin can destroy living
1. Sour taste: When acid is dissolved in water, tissue. E.g. acids like sulphuric acid (H2SO4),
it gives sour taste to the water e.g. sour nitric acid (HNO3) and bases like sodium
taste of lemons, oranges, grapes is due to hydroxide (NaOH), potassium hydroxide
citric acid. (KOH).
10 Medical Biochemistry for Physiotherapy Students
Storage 4. Lactic acid: It checks growth of bacteria in
large intestine.
These should be stored at low level to avoid
5. Hypochlorous acid: It acts as a disinfectant,
injury which could be caused if these chemi-
antiseptic and bleaching agent.
cals were accidently knocked off a shelf. Do
6. Boric acid: It acts as an antiseptic for
not store KOH or NaOH in a bottle having a
inflamed mucous membranes in the eyes,
ground glass stopper because these chemicals
nose or mouth.
absorb carbon dioxide from the air forming
carbonates, which can cement the stopper in Bases
the bottle.
A base is a compound which gives hydroxyl
Safe Use ions (OH–) in water solution.
1. Never mouth pipette with a corrosive Properties
chemical. The accidental swallowing of it
1. Soapy feeling: Whenever strong base is
can cause severe internal injury.
dissolved in water, the solution feels
2. Always pour a corrosive chemical at below
the eye level, slowly and with a great care slippery or soapy.
to avoid splashing. 2. Bitter taste: They have bitter taste.
3. Wear suitable protective gloves when 3. Action on indicators: Bases reverse the
opening a container of a corrosive chemi- colour change which acids produce with
cal and when pouring it. Place a cloth over indicators. Bases turn red litmus paper
the neck and cap of the container when blue while acids turn blue litmus to red.
opening it. 4. Reaction with acids: They react with acids
4. Dissolve a solid corrosive chemical such to form salts. This is very important
as NaOH in water with great care, mixing function in the body for maintaining the
in small amounts at a time to dissipate the alkaline reaction in the blood.
heat produced. 5. They react with fats to form soaps. Strong
5. When diluting concentrated acids parti- bases destroy tissues by attracting the
cularly H 2SO 4, always add acid to the water, dissolving the albumin and reacting
water. Never add water to the acid. with the fats.
Adding water to acid can produce 6. They are used for removing certain stains.
sufficient heat to break a glass container.
Some Bases of Importance
6. Always use a strong carrier to transport
bottles of corrosive chemicals. 1. Sodium hydroxide: It is also known as
caustic soda. It is used as a household
Some Acids of Importance article as a drastic cleanser because it
1. Hydrochloric acid: It helps in gastric reacts with grease to make soap. Commer-
digestion of proteins. It also acts as an cially it is used in the manufacture of hard
antiseptic by destroying fermentation of soaps.
bacteria in the intestinal tract. 2. Calcium hydroxide: It is also known as
2. Nitric acid: It acts as a coagulant in testing slaked lime. Lime water is used in
for albumin in the urine. medicine to overcome high acidity in the
3. Boric acid: It acts as an antiseptic for stomach. Lime water is used in diet for
inflamed mucous membranes in the eyes, building bones and teeth. It serves as a
nose or mouth. special antidote for oxalic acid poisoning.
Water and Electrolytes 11
3. Ammonium hydroxide: In medicine, 4. Essential for life: Salts do not furnish heat
ammonium hydroxide is used as a heart or energy, but they are very essential to
respiratory stimulant. life because
4. Magnesium hydroxide: In medicine, a thick a. They make up the protoplasm, tissue and
suspension known as milk of magnesia is bone structure.
used as an antacid. Magnesium compound b. They keep up the elasticity and irrita-
is a laxative in its physiological action. bility of the muscles.
c. They maintain the neutrality of slightly
Salts acid or alkaline condition of the body
Salts are formed by a reaction between an secretions, blood and other fluids.
acid and a base where a salt and water are d. They assist in maintaining normal osmo-
formed. tic pressure conditions.

NaOH + HCl ————


> NaCl + H2O
Medical Salts
Importance of Salts in the Body 1. Sodium chloride: Its common name is
common salt. It is used as a saline solution.
1. Formation of salts in the body: Acids are
2. Mercuric chloride: It is used as an antiseptic.
constantly being formed in the body. The
3. Calcium sulphate: Its common name is
body fluids with the exception of the
Plaster of Paris. It is used as casts for
gastric juices and urine are alkaline or
broken bone.
neutral. As a result of neutralization, many
Acids, bases and salts are called electro-
salts are produced and excreted in the
lytes when dissolved in water because they
urine, sweat and faeces.
ionize into two oppositely charged part called
2. Neutralization in blood: Any change in the
ions.
normal acidity or alkalinity of the different
parts of the human body affects markedly
Theory of Arrehenius Ionization
every cell and organ for instance. If the
alkalinity of the blood is too low (acidosis) 1. The molecules of electrolytes break up in
or too high (alkalosis), the heart will be water solution into definite component
affected so that it will not pump uniformly. parts called ions. This type of dissociation
3. Buffer action in the body: The process of is called ionization. Non electrolytes do
digestion and cell activity produce nume- not ionize.
rous substances which enter the blood 2. Some ions are positively charged and
stream. These substances may be acidic, others are negatively charged. The
some alkaline and neutral. The blood conductance of electricity by solutions of
maintains a constant pH which ranges electrolytes is accomplished by the ions.
normally from 7.35 to 7.45. This constancy 3. The abnormal effect of electrolytes as
of hydrogen ion concentration is possible compared to the non electrolytes in the
because of the presence in the blood of same concentration are due to an increase
certain molecules and related ions which in the total number of particles as a
acts as buffers. consequence of ionization.
Chapter 3

Chemistry of Carbohydrates

Carbohydrates are composed of the elements 1. Monosaccharides


carbon, hydrogen and oxygen. The name 2. Disaccharides
carbohydrates literally mean ‘hydrates of 3. Oligosaccharides
carbon’. The empirical formula is (CH2O)n. 4. Polysaccharides
Carbohydrates are defined chemically as
aldehyde or ketone derivatives of higher Monosaccharides
polyhydric alcohols or compounds which These are also known as simple sugars.
yield these derivatives on hydrolysis. Monosaccharides are those which cannot be
further hydrolyzed into simpler forms.
FUNCTIONS OF CARBOHYDRATES
Empirical formula is:
1. Carbohydrates are the dietary sources of CnH2nOn
energy (4 C/g) for all organisms.
They can further be subdivided as follows:
2. Carbohydrates are precursors for many
a. Depending on the number of carbon atoms
organic compounds (fats, amino acids).
it contains like trioses if 3 carbon atoms
3. Carbohydrates (as glycoprotein and
are present, tetroses if 4 carbon atoms, etc.
glycolipids) participate in the structure of
b. Depending on the presence of aldehyde
cell membrane and cellular functions.
or ketone groups
4. These are storage form of energy for
energy in the form of glycogen General Aldosugars Ketosugars
Formula
5. These are structural components of many
Trioses (C3H6O3) Glyceraldehyde Dihydroxyacetone
organisms like the cell walls of bacteria,
Tetroses (C4H8O4) Erythrose Erythrulose
fibrous cellulose of plants.
Pentoses (C 5H10 O5) Ribose Ribulose
Hexoses (C 6H12 O6) Glucose Fructose
Classification of Carbohydrates
Heptoses(C 7H 14O 7) Glucoheptose Sedoheptulose
Carbohydrates are divided into four main
groups depending upon the number of Thus, trioses with aldehyde group are
monomer units present in the molecule: called aldotrioses and trioses with ketone
Chemistry of Carbohydrates 13
group are ketotrioses. Similarly, tetroses with
aldehyde group are aldotetroses and tetroses
with ketone group are ketotetroses and so
on.

D-Series and L-Series


The D- and L-isomers are mirror images of
each other. The orientation of the H and OH
groups around the carbon atom just adjacent
to the terminal primary alcohol carbon e.g.
C-atom 5 in glucose determine the series. If
this OH group is on right side, the sugar is of
General Properties D-Series and if it is on left side, it belongs to
the L-Series.
Isomerism
Naturally occurring monosaccharides in
Compounds that have the same chemical the mammalian tissues are mostly of D-form.
formula are called isomers. For example
fructose, glucose, mannose and galactose are
all isomers of each other, having the same
chemical formulae. Therefore, isomerism is
exhibited by organic compounds having same
molecular formulae. Due to this property,
although these compounds have same
molecular formulae, they have different
physical and chemical properties.
The presence of asymmetric carbon in the
compound results in the formation of isomers
of that compound.
Optical Activity of Sugars
Asymmetric Carbon Atom
All the compounds having asymmetric carbon
A carbon atom to which four different atoms
atoms can rotate the beam of plane polarized
or group of atoms is attached is said to be
light and are said to be optically active. An
asymmetric. The number of isomers of a
compound depends on the number of isomer which can rotate the plane of polarized
asymmetric carbon atoms and is given by 2n light to the right is called as dextrorotatory
where n indicates the number of asymmetric and is designated as (d) or (+) while the
carbon atoms in that compound. Glucose isomer which rotates the plane of polarized
contains 4 asymmetric carbon atoms and thus light to the left is called as laevorotatory and
has 16 isomers. is designated as (l) or (-).
14 Medical Biochemistry for Physiotherapy Students
When equal amounts of dextrorotatory
and levorotatory isomers are present in a
mixture, the resulting mixture is optically
inactive and is said to be a racemic mixture.

α – and β – Anomers of Glucose


The cyclic structures of glucose is retained in
solution, but isomerism takes place about
position 1. This is formed by optical rotation,
known as mutarotation, by which the posi-
tions of –H and –OH groups are changed PROPERTIES OF MONOSACCHARIDES
around carbon 1.
Example: When D- Glucose is dissolved Physical Properties
in water, its specific rotation is +111º. Gra- They are colourless, crystalline compounds,
dually, it decreases and remains constant at readily soluble in water and sweetish in taste.
+52.5º. These two forms are referred to as α- Their solutions are optically active and exhibit
and β- glucose respectively. Blood sugar is the phenomenon of mutarotation.
α-β-D-glucose.
Chemical Properties
Reactions due to aldehyde or ketone groups:
1. Osazone formation: The reducing sugars
form characteristic crystals. These are
obtained by adding a mixture of phenyl
hydrazine hydrochloride and sodium
acetate to the sugar solution and then
heating it in a boiling water bath for 30 to
45 minutes and allowing to cool slowly.
Precipitates are separated and spread on
slide for examination under low power of
microscope. These compounds have
characteristic crystal structures, melting
Epimers points and precipitation times.
Isomers formed as a result of interchange of Glucosazone crystals are fine, yellow
the –OH and –H on carbon atoms 2, 3 and 4 needles in fan shaped aggregates, descri-
of glucose are known as epimers. Mannose bed as “bundle of hay”. Glucose, maltose,
and galactose are formed by epimerization mannose form the same osazones.
of carbons 2 and 4 respectively. In the body, Lactosazone crystals are irregular
of epimerization takes place by the enzyme, clusters of fine needles and look like a
epimerase. “powder puff”.
Chemistry of Carbohydrates 15
Maltosazone crystals are star shaped and HNO3
compared to sunflower petals. D-Glucose ———> D-Glucosaccharic acid
c. Uronic Acids: When an aldo sugar is
oxidized in such a way that the primary
alcohol group is converted to -COOH
group, without oxidation of aldehyde
group, a uronic acid is formed.
D-Glucose ——> D-Glucuronic acids
D-Galactose ——> D-Galactouronic acid
This is not possible under laboratory
conditions, but readily occurs in living
Osazone formation is a useful means of tissues by enzyme action.
preparing crystalline derivatives of sugars
and are valuable in identification of sugars. Biological Importance
2. Reduction of sugars to form sugar alco-
Uronic acids are components of structural
hols: The monosaccharides may be redu-
material like chondroitin and mucoitin sul-
ced to their corresponding alcohols by phuric acids and glycoproteins. Certain toxic
reducing agents such as sodium amalgam substances are conjugated with glucuronic
or with hydrogen under pressure. Simi- acid in the liver and excreted as less toxic sub-
larly, ketoses may also be reduced to form stances. Bile pigments and steroid hormones
keto alcohol. For example, that are relatively insoluble, are also conju-
2H gated with glucuronic acid to render them
D-Glucose —————> D-Sorbitol more soluble and readily excretable in urine.
D-Galactose —————> D-Dulcitol 1. Interconversion of sugars: Glucose, fructose
D-Mannose —————> D-Mannitol and mannose are interconvertible in weak
D-Fructose —————> D-Mannitol + alkaline solutions such as Ba(OH) 2 or
D-Sorbitol Ca(OH) 2 . It is because weak alkaline
3. Oxidation of sugars to produce sugar solutions of sugars undergo molecular
acids change known as “tautomerization” where
a. Mild oxidizing agents like bromine ‘H’ atoms migrate from one carbon to
water oxidize the -CHO to a -COOH to another to form “enediol” compounds. All
form sugar acids. sugars give the same enediol which
Br 2 tautomerizes to all three sugars. This
D-Glucose —————> D-Gluconic acid interconversion of related sugars by the
action of dilute alkali is referred to as
b. Strong oxidizing agents like HNO 3 , “Lobry de bruyn-Van Ekenstein reaction”.
Oxidize the -CHO group as well as the 2. Reducing action of sugars: Monosaccharides
primary alcoholic group to produce by virtue of their free aldehyde or ketone
sugar acids called “saccharic acids” or group in their structure reduce free
“aldaric acids”. metallic cations such as Cu 2+ ions in
16 Medical Biochemistry for Physiotherapy Students
alkaline solution at high temperature. All Indican = Carbohydrate + Indoxyl
the monosaccharides and the two disaccha- (aglycone)
rides, maltose and lactose reduce cupric 2. Acetylation or ester formation: The alcohol
ions present in Benedict’s reagent and also group of sugars may react with acids to
in Fehling’s reagent. Lactose and maltose form esters. Phosphoric acid esters of
have free reducing groups hence these are sugars are important as intermediate
capable of reducing metallic ions but since products during metabolism. Examples
sucrose does not have a free reducing are: Glucose-1 -phosphate, Glucose-6-
group, it is a non reducing sugar. phosphate, Fructose-6-phosphate, Galac-
tose-1-phosphate, etc.
Reactions due to the Alcoholic Groups 3. Dehydration by strong acids: When sugars
1. Glycosides: Glycosides are compounds are treated with concentrated H2SO2 in
formed by the condensation reaction cold or on heating with a strong solution
between a sugar and alcohol. The carbo- of HCl, pentoses form cyclic aldehyde
hydrate residue is attached by an acetal called “furfural” whereas hexoses form
linkage of carbon-1 to the -OH group of “hydroxy methyl furfural”.
non carbohydrate residue called aglycone. This reaction forms the basis of quanti-
The aglycones may be methyl alcohol, tative determination of carbohydrates.
glycerol, phenol, hydroquinones, sterols Examples are: Molisch test, Seliwanoff’s
and anthraquinones. test, Bial-orcinol test.
The glycosides are named according to the
SUGAR DERIVATIVES OF
carbohydrate they contain. If it contains
BIOLOGICAL IMPORTANCE
glucose, forms glucoside. If galactose, it
forms galactoside and so on.
A simple example is the methyl glucoside Amino Sugars (Hexosamines)
formed when a solution of glucose in Sugars containing -NH 2 group are called
boiling methyl alcohol is treated with 0.5% amino sugars.
HCl as a catalyst. a. Glucosamine (2-amino D-glucose): It is a
Aglycone: The non carbohydrate portion constituent of mucopolysaccharides and
of the glycoside is called the aglycone. mucoproteins such as hyaluronic acid,
Glycosides do not reduce alkaline copper heparin and blood group substances. It is
sulphate because sugar group is combined, present in the cell walls of fungi and in
i.e. aldehyde group is converted to an the shells of crustaceans (lobster, crab, etc)
acetal group. as chitin. Hence glucosamine is called
Examples “chitosamine”.
Cardiac glycosides = Carbohydrate + b. Galactosamine (2-amino D-galactose): It
Digoxin (aglycone) occurs in sulphated mucopolysaccharides
Chemistry of Carbohydrates 17
as chondroitin sulphates which are present Examples are:
in cartilages, bones, tendons and heart 1. Maltose – It yields two molecules of
valves, hence galactosamine is also called glucose on hydrolysis.
as chondrosamine. Maltose ———> glucose + glucose
2. Lactose – It yields one molecule of glucose
and one molecule of galactose on hydro-
lysis.
Lactose ———> glucose + galactose
3. Sucrose – It yields one molecule of glucose
and one molecule of fructose on hydrolysis
Sucrose ———> glucose + fructose

The disaccharides are of two types:


1. Reducing disaccharides which have free
c. Certain antibiotics such as erythromycin
aldehyde or keto group e.g. maltose,
contain amino sugars responsible for
lactose.
antibiotic activity.
2. Non-reducing disaccharides which have
Deoxy Sugars no free aldehyde or keto group e.g.
sucrose.
Deoxy sugars are those in which a hydroxyl
group attached to the ring structure has been Maltose
replaced by a hydrogen atom. They are
obtained on hydrolysis of certain substances Maltose, also known as malt sugar is
that are important in biologic processes. An composed of two α- D- glucose units held
example is the deoxyribose occurring in together by α (1→4) glycosidic bond. The free
nucleic acids (DNA). aldehyde group present on C 1 of second
Also found as a carbohydrate of glyco- glucose answers the reducing reactions,
proteins is L-fucose and of importance as an besides the osazone formations.
inhibitor of glucose metabolism is 2-deoxy-
glucose.

Disaccharides
These are those sugars which yield two
molecules of monosaccharides on hydrolysis;
they may be same or different.
The two units of monosaccharides are It is produced during the course of
joined by a glycosidic bond. Empirical formula digestion of starch by the enzyme amylase.
is: It is hydrolyzed by the intestinal enzyme
Cn(H2O)n-1 maltase to two molecules of glucose.
18 Medical Biochemistry for Physiotherapy Students
Sucrose heldtogether by α (1→4) glycosidic bond. The
anomeric carbon of C1 glucose is free; hence
Sucrose, also known as cane sugar is made
lactose exhibits reducing properties and
up of α-D-glucose and β-D-fructose. The two
forms osazones.
monosaccharides are held together by a
glycosidic bond (α 1 →β 2 ) between C 1 of
α-glucose and C2 of β- fructose. The reducing
groups of glucose and fructose are involved
in glycosidic bond, hence sucrose is a non
reducing sugar and it cannot form osazones.

Lactose or milk sugar is an animal


disaccharide and is present to the extent of
5% in milk only. It is synthesized in mammary
glands and during lactation may appear in
urine. It is hydrolyzed by the intestinal
enzyme lactase to glucose and galactose.
Sucrose or cane sugar is a plant disaccharide
and is present in high concentration in sugar Oligosaccharides
cane and sugar beet. Sucrose is used for
sweetening purpose. It is hydrolyzed by the These are those sugars which yield 3–10
intestinal enzyme sucrase to glucose and monosaccharides on hydrolysis. Examples
fructose. include
Raffinose – fructose + galactose + glucose
Stachyose – 2 molecules of galactose +
Invert Sugar
glucose +fructose
Sucrose is dextrorotatory (+62.5 o ) but its
hydrolytic products are laevorotatory because Polysaccharides
fructose has a greater specific laevo-rotation
These are those sugars which yield more than
(-92 o ) than the dextro-rotation of glucose
ten molecules of monosaccharides on hydro-
(+52.5o). As the hydrolytic products invert the
lysis. Empirical formula is:
rotation, the resulting mixtures of glucose and
(C6H10O5)n
fructose (hydrolytic products) is called as invert
These are usually tasteless and form
sugar and the process is called as inversion.
colloid with water. Polysaacharides are of
Honey is an example of invert sugar and
two types:
the presence of fructose accounts for the
1. Homopolysaccharides
greater sweetness of honey.
2. Heteropolysacchraides
Homopolysaccharides are also known as
Lactose
homoglycans. These are polymers of same
Lactose, also known as milk sugar is made monosaccharide units. Examples are starch,
up of β-D-galactose and β-D-glucose glycogen, inulin, cellulose, dextrins, etc.
Chemistry of Carbohydrates 19
Heteropolysaccharides are also known as 3. Inulin
heteroglycans. These are polymers of diffe-
Inulin is found in tubers and roots of dahlias,
rent monosaccharide units. Examples are
artichokes and dandelions. It is hydrolysable
mucopolysaccharides like heparin, chondriotin
to fructose and hence it is a fructosan. No
sulphate etc.
colour is given when iodine solution is added
1. Starch to inulin solutions. This starch is easily soluble
in warm water. It is used in physiologic
Starch is stored as a reserve food in cereals
investigation for determination of the rate of
and tubers of plants. A starch molecule
glomerular filtration.
contains two polysaccharides; amylase and
amylopectin. Amylase consists of D-glucose 4. Dextrins
units linked by α-1, 4-glycosidic linkages in
the form of a straight chain. Amylopectin is Dextrins are substances formed in the course
similar to amylase but also contains α-1, of the hydrolytic breakdown of starch. The
6-glycosidic bonds to form a branched struc- partially digested starches are amorphous.
ture. Branching occurs after every 24-30 Limit dextrins that give a red colour when
glucose units. tested with iodine are first formed as
hydrolysis reaches a certain degree of
2. Glycogen branching. These are called erythrodextrins.
Glycogen is also called animal starch as it is As hydrolysis proceeds, the iodine colour is
stored in animals in their liver and muscle. It no longer produced. These are the so-called
has a structure similar to amylopectin with achroodextrins. Finally, only reducing sugars
straight chain of α-1, 4-linked glucose units will appear.
and α-1, 6-branched points. Branching occurs
after every 12- 18 glucose units making it a 5. Cellulose
highly branched molecule with a tree like Cellulose is the chief constituent of the
structure. Liver glycogen is readily available framework of plants. It gives no colour with
source of glucose during starvation. Glycogen iodine and is not soluble in ordinary solvents.
is non-reducing and gives a red colour with It consists of long, straight chains of β-D-
iodine. glucopyranose units linked by β (1 ——> 4)
Differences between starch and glycogen:
bonds. It is not subject to attack by the
a. Starch is of plant origin whereas glycogen
digestive enzymes of humans because of the
is of animal origin.
β-linkage. Thus, it is an important source of
b. Glycogen is much more branched than the
bulk in the diet.
starch. In starch, the branching is after
every 24 to 30 glucose units, whereas in
6. Chitin
glycogen, the branching is after every 8
to 10 glucose units. It is present in the exoskeletons of inver-
c. Starch gives blue colour with iodine tebrates like crab shell or insect wings, it is
solution whereas glycogen gives red made up of acetylated glucosamine (chito-
colour. samine) units.
20 Medical Biochemistry for Physiotherapy Students
Heteropolysaccharides mainly in cartilage, bone, cornea, skin, arterial
wall, etc.
Heteropolysaacharides or heteroglycans or
mucopolysaccharides or glycosaminoglycans
Dermatan Sulphate
Mucopolysaccharides are composed of
acetylated glycosamine (aminosugar) and These are made of D-glucuronic acid and L-
uronic acid units; some are made up of ioduronic acid. They occur in the skin, aorta,
aminosugar and monosaccharide units cardiac valves, tendons, arterial walls.
without the presence of uronic acid. They are
essential components of tissues present either Keratan Sulphate
in free form or in combination with proteins, These are composed of N-acetyl glucosamine
known as “mucoproteins or glycoproteins”. and D-galactose. They occur in cornea,
Some important mucopolysaccharides are as cartilage, intervertebral discs and loose
follows: connective tissues.
Hyaluronic Acid
Heparin
It is made of repeating units of N-acetyl
It is a polymer of D-glucosamine and either
glucosamine and D-glucuronic acid. It is
of the two uronic acids—D-glucuronic acid
present in skin, connective tissue, vitreous
or L-ioduronic acid. It is present in mast cells
humour, synovial fluid and cartilage and acts
in lungs, liver, skin and intestinal mucosa. It
as a ground substance or connecting sub-
acts as an anticoagulant.
stance and acts as a lubricant and shock absor-
bent.
Blood Group Antigens
Chondroitin Sulphate These contain peptides or amino acids and
These are made of N-acetyl galactosamine carbohydrates. They contain galactosamine,
and D-glucuronic acid. They are present glucosamine, fucose, etc.
Chapter 4

Chemistry of Lipids

Lipids are a heterogeneous group of com- 5. Lipids are required for the absorption of
pounds related, either actually or potentially, fat-soluble vitamins.
to the fatty acids. 6. Lipoproteins and glycolipids are essential
Chemically, lipids are defined as esters of for maintaining cellular integrity.
fatty acids with various alcohols. They have 7. Lipids form important constituent of
the common property of being: nervous tissue.
1. Relatively insoluble in water.
2. Soluble in non polar solvents such as ether, Classification of Lipids
chloroform, acetone and benzene. Bloor has proposed the following classifi-
Because of their insolubility in aqueous cation of lipids:
solutions, body lipids are generally found A. Simple Lipids. These are esters of fatty
compartmentalized as in the case of mem- acids with various alcohols.
brane associated lipids and droplets of triacyl- B. Compound Lipids
glycerol in adipocytes or transported by These are esters of fatty acids with alco-
plasma in association with protein as lipo- hols but in addition they also contain
protein particles. other substituents like carbohydrate,
protein, sulphate, phosphoric acid etc.
Importance of Lipids C. Derived Lipids
1. Lipids are important dietary constituents. These are obtained by hydrolysis of
These give 9 C/g. simple and compound lipids.
2. Lipids are important constituents of the
Simple Lipids
cell membrane along with the proteins.
3. Lipids serve as a thermal insulator in the These are esters of fatty acids with various
subcutaneous tissues and around certain alcohols and are further divided into two sub
organs. classes – neutral fats and waxes.
4. Lipids supply so-called essential fatty acids
which cannot be synthesized by the body Neutral Fats
and are essential in the diet for normal These are esters of fatty acids with glycerol
health and growth. and are also known as triglycerides or
22 Medical Biochemistry for Physiotherapy Students
triacylglycerol. Triacylglycerol contains three Chemical Properties
molecules of fatty acids esterified with one
1. Hydrolysis: Triglycerides are hydrolysed
molecule of glycerol. Similarly, monoacyl-
to produce free fatty acids and glycerol
glycerol and diacylglycerol are also other
by the lipases at alkaline pH and in the
examples of neutral fats, they contain one,
presence of water. The hydrolysis takes
and two molecules of fatty acids esterified
place in a stepwise manner. Diglycerides
with one molecule of glycerol respectively.
are formed first, followed by mono-
Out of these three, triglycerides are most
glycerides and finally the monoglycerides
important.
are split to free glycerol and fatty acids.
Triglycerides can be simple or mixed type.
Simple type of triglycerides contain same type
of fatty acid residue at all the three carbons
e.g. tristearin or tristeroyl glycerol.

2. Hydrogenation: Unsaturated fats can be


hydrogenated by the addition of hydro-
gen across the double bonds of the fatty
Mixed type of triglycerides contains two acids in the presence of nickel as catalyst
or three different types of fatty acid residues to give fully saturated fats. The above
e.g. 2- stearo- 1,3- oleopalmitin and 1- palmito process is called hardening of oils where-
– 2- stearo- palmitin. by vegetable oils are hydrogenated to
produce commercial cooking fats.
3. Saponification: The hydrolysis of trigly-
cerides by alcoholic potassium hydroxide
or by sodium hydroxide is called saponi-
fication. The resultant products are
glycerol and potassium or sodium salts of
Physical Properties of Fats fatty acids known as soaps.
1. They are insoluble in water, but readily
soluble in organic solvents like ether,
chloroform, benzene etc.
2. They are readily soluble in hot alcohol but
slightly soluble in cold.
3. They are tasteless, odourless, colourless
and neutral in reaction. 4. Rancidity: The term rancidity is used to
4. Their melting points are low. represent the deterioration of fats and oils
5. The specific gravity of fats is lower than in an unpleasant taste. Rancidity occurs
water, so, it floats on water. when fats and oils are exposed to air,
Chemistry of Lipids 23
moisture, light, bacteria, etc. This occurs This classification is based on the type of
due to the formation of peroxides at the alcohol present. In glycerophosphatides,
double bonds of unsaturated fatty acids. glycerol is the alcohol group. In phospho-
Therefore, fats containing unsaturated fatty inositides, inositol is the alcohol group. In
acids are more susceptible to rancidity. phospho-sphingosides, sphingol is the alcohol
Rancidity can be prevented by the use of group.
antioxidants. Antioxidants like tocopherols, The phospholipids include the following:
1. Phosphatidic acid and Phosphatidyl
hydroquinone, gallic acid and α-naphthol
glycerols
are added to the commercial preparations
Phosphatidic acid is important as an
of fats and oils to prevent rancidity.
intermediate in the synthesis of triacyl-
glycerols and phospholipids but is not
Waxes
found in greater quantity in tissues.
If the fatty acid is esterified with a mono- Cardiolipin is a phospholipid that is found
hydric alcohol of high molecular weight in membranes of mitochondria. It is
instead of glycerol, the resulting compound formed from phosphatidyl glycerol.
is called a wax. These resemble fats and are 2. Phosphatidyl choline (Lecithin)
usually solid. In the human body, the The lecithins contain glycerol and fatty
commonest waxes are esters of cholesterol. acids as in simple fats, but they also contain
True waxes are esters of higher fatty acids phosphoric acid and choline. The lecithins
with cetyl alcohol (C16H33OH) or other higher are widely distributed in the cells of the
straight chain alcohols. These are formed as body, having both metabolic and struc-
tural functions in membranes.
secretions and are mostly protective in
Lecithin on hydrolysis give glycerol, fatty
function, by many animals.
acid, phosphoric acid and choline.
Dipalmityl lecithin (DPL) is a very effective
Compound Lipids
surface effective agent, preventing adhe-
These are esters of fatty acids with alcohols rence, due to surface tension, of the inner
but in addition, they also contain other surfaces of the lungs.
substituents like carbohydrate, protein,
sulphate, phosphoric acid, etc.

Phospholipids
Phospholipids are included in a class of
compound lipids. These are the lipids
containing, in addition to fatty acids and an
Phosphatidyl Ethanolamine (Cephalin)
alcohol, a phosphoric acid residue. They also
have nitrogen containing bases and other The cephalins differ from lecithins only in that
subsituents. ethanolamine replaces choline. If the base is
They are divided into three classes: ethanolamine, then it is called phosphatidyl
1. Glycerophosphatides ethanolamine or ethanol amine cephalin. If
2. Phospho-inositides the base is amino acid serine, then it is called
3. Phospho-sphingosides phosphatidyl serine or serine cephalin.
24 Medical Biochemistry for Physiotherapy Students
Cephalins on hydrolysis yield glycerol, Glycolipids
fatty acids, ethanolamine or serine.
Lipids containing carbohydrate moiety are
called glycolipids. They contain a special
Phosphatidylinositol
alcohol called sphingosine or sphingol and
They are found in brain, liver, heart and nitrogenous base in addition to fatty acids
soyabean. They contain no base but have but do not contain phosphoric acid or
inositol in its place. glycerol. These are of two types:
a. Cerebrosides
Cardiolipin b. Gangliosides
It is an important phospholipids of mito-
chondrial membrane. It is a diphosphatidyl Cerebrosides
glycerol in which two phosphatidic acids are They are present in greater concentration in
joined by a molecule of glycerol. These white matter of brain and myelin sheath of
phosphatidic acids are particularly rich in the nerve. They comprise of a molecule of fatty
polyunsaturated fatty acids especially linoleic acid, an amino alcohol sphingosine and a
acid. sugar usually galactose.

Sphingomyelins Types
Phospholipids containing sphingosine are Four types of cerebrosides are found
called sphingomyelins. They contain a depending upon the type of fatty acid present:
complex base sphingosine in addition to a. Kerasin – contains lignoceric acid as the
phosphoryl choline. A fatty acid is attached fatty acid.
to the amino group of the sphingosine. No b. Cerebron - contains hydroxy lignoceric
glycerol is present. Sphingomyelins are acid as the fatty acid.
present in all tissues especially in brain and c. Nervon - contains nervonic acid as the
other nervous tissues. Sphingomyelins on fatty acid.
hydrolysis yield sphingosine, fatty acid, d. Oxynervon - contains hydroxy nervonic
phosphoric acid and choline. acid as the fatty acid.
Functions of Phospholipids
Clinical Aspect
1. They form structures of membrane, matrix
Gaucher’s Disease
of cell wall, myelin sheath, microsomes
and mitochondria. This disease occurs due to the deficiency of
2. They help in transport of cholesterol. enzyme β–glucosidase. Normally this enzyme
3. They help in formation of lipoproteins. hydrolyzes glucocerebrosides to form
4. They play a role in synthesis of compounds ceramide and glucose. In the absence of the
called eicosanoids including prostaglan- enzyme glucocerebroside cannot be converted
dins, leukotrienes. to ceramide due to which tissue gluco-
Chemistry of Lipids 25
cerebroside levels increase. Clinical features Apolipoproteins are divided by structure
include hepatomegaly, splenomegaly, pig- and function into classes A to H, with most
mentation of skin, etc. classes having subclasses, for example apo-
A1 and apo- CII.
Gangliosides
They are present in greater concentration in Functions of Lipoproteins
gray matter of brain. They comprise of a 1. They act as transport vehicles for lipids in
molecule of fatty acid, an amino alcohol the lipid plasma.
sphingosine, a sugar usually galactose or 2. They deliver the blood components
glucose, one molecule of N–acetyl galacto- (cholesterol, triacylglycerol etc.) to various
samine and upto three molecules of N-acetyl tissues for utilization.
neuraminic acid (NANA or sialic acid).
Types of gangliosides are GM1, GM2, GM3 DERIVED LIPIDS
and GD3.
Fatty Acids
Sulpholipids Fatty acids are obtained by hydrolysis of fats.
Lipids possessing sulphate groups are called A fatty acid consists of a hydrocarbon chain
sulpholipids. These are largely found in white with a terminal carboxyl group. They are
matter of brain and to a less extent in liver, divided into saturated and unsaturated fatty
kidney, salivary gland, testis etc. acids.
Saturated fatty acids are those which do
Lipoproteins not contain any double bond in their
structure. General formula for saturated fatty
Lipoproteins are complexes of lipids and
acids is CnH2n+1. Examples of saturated fatty
proteins. The lipid part consists of fats,
acids include acetic acid, propionic acid,
phospholipids, cholesterol and its ester and
butyric acid, capric acid, palmitic acid, stearic
free fatty acids. These are found in cell
acid, etc.
membranes, milk, egg yolk etc.
These are molecular complexes of lipids
and specific proteins called apolipoproteins.
These are in constant state of synthesis,
degradation and removal from the plasma.
The lipoprotein particles include: chylo-
microns, very low density lipoproteins
(VLDL), low density lipoproteins (LDL) and
high density lipoproteins (HDL). Unsaturated fatty acids are those which
Apolipoproteins associated with lipo- contain one or more double bonds in its
proteins serve as structural components of the structure. General formula for unsaturated
particles, provide recognition sites for cell fatty acids is:
surface receptors and serve as activators or CnH2n-1COOH
coenzymes for enzymes involved in lipo- They can be monosaturated or polyun-
protein metabolism. saturated.
26 Medical Biochemistry for Physiotherapy Students
Monounsaturated fatty acids (MUFA) are sources. Linoleic is the most important
those fatty acids which contain one double because it is the precursor from which the
bond in its structure, e.g. oleic acid and other two essential fatty acids (linolenic acid
palmitoleic acid. These monounsaturated and arachidonic acid) can be synthesized in
fatty acids are present in nearly all fats. the body.
Biologically, arachidonic acid is the most
important because it is the precursor from
which eicosanoids like prostaglandins and
leukotrienes are formed.

Properties
Polyunsaturated fatty acids (PUFA) are
those fatty acids which contain more than one 1. Saturated fatty acids are relatively
resistant to oxidation outside the body
double bond in its structure. Examples include
whereas unsaturated fatty acids are
linoleic acid, linolenic acid, arachidonic acid.
slowly but spontaneously oxidized in the
Sources and structure is given below:
presence of air leading to rancidification.
Name of Structure Sources 2. In general, increasing the chain length
fatty acid increases the melting temperature of a fatty
Linoleic acid 18:2(9,12) Corn, peanut, acid but addition of a double bond
cottonseed, decreases the melting temperature.
soyabean oil
Linolenic acid 18:3(9,12,15) Found frequently
Therefore, the presence in membrane
with linoleic acid lipids of fatty acids that contain double
but particularly in bonds helps maintain the fluid nature of
linseed oil those lipids
Arachidonic acid 20:4(5,8,11,14) Found in small
quantities with
linoleic and Functions of Essential Fatty Acids
linolenic acids but They are required for:
particularly in
Peanut oil. 1. membrane structure and function.
2. transport of cholesterol.
PUFA are also known as essential fatty 3. formation of lipoproteins.
acids because they cannot be synthesized in 4. prevention of fatty liver.
the body and have to be provided from natural 5. synthesis of eicosanoids.
Chemistry of Lipids 27
The common names and structures of some group at C-3 is classified as sterols. Choles-
fatty acids are given below: terol is the major sterol in the body.
The structure of cholesterol consists of four
Common name Structure fused rings A,B,C and D with the carbons
Formic acid 1 numbered in sequence and an eight mem-
Acetic acid 2:0 bered, branched hydrocarbon chain attached
Propionic acid 3:0
Butyric acid 4:0
to the D ring. This structure is known as
Capric acid 10:0 cyclopentanoperhydrophenanthrene nucleus.
Palmitic acid 16:0 Its molecular formula is C27H45OH.
Palmitolic acid 16:1(9)
Stearic acid 18:0
Oleic acid 18:1(9)
Linoleic acid 18:2(9,12)
Liolenic acid 18:3(9,12,15)
Arachidonic acid 20:4(5,8,11,14)
Lignoceric acid 24:0
Nervonic acid 24:1(15)

The numbers in the table indicate the


number of carbons in the chain and the
number and position of double bonds. For Cholesterol has 27 carbon atoms, a
example, acetic acid, 2:0 indicates it has 2 hydroxyl group, a double bond, 2 methyl
carbon atoms and no double bond i.e. it is groups and a side chain at C-17. Cholesterol
saturated fatty acid. Similarly, arachidonic is synthesized by virtually all tissues but liver,
acid, 20:4 (5,8,11,15) indicates it has 20 carbon intestine, adrenal cortex and gonads make the
long chain and 4 double bonds which are largest contributions. Cholesterol is a
between carbons 5-6, 8-9, 11-12 and 14-15. component of all cell membranes and is a
precursor of bile acids, steroid hormones and
Alcohols vitamin D.
Cholesterol is found in largest amounts in
Alcohols found in lipid molecules include
normal human adults as:
glycerol, cholesterol and higher alcohols like
• Brain and nervous tissue- 2%
cetyl alcohol usually found in waxes. The
• Liver – 0.3%
unsaturated alcohols are important pigments.
• Skin – 0.3%
Phytyl alcohol is a constituent of chlorophyll
• Adrenal cortex – 10% or more.
and lycophyll, which is a polyunsaturated
alcohol and occurs in tomatoes as a purple Cholesterol occurs in two forms in the
pigment. body i.e. free form and ester form. In ester
form, it is esterified with fatty acids at –OH
Steroids and Sterols at C-3 position. In brain and nervous tissue,
free form predominates, while in adrenal
Steroids are often found in association with cortex, it occurs mainly in esterified form.
fat. Steroids with 8- 10 carbon atoms in the Much of the plasma cholesterol is in esterified
side chain at C-17 and an alcohol hydroxyl form.
Chapter 5

Chemistry of Amino Acids and Proteins

CHEMISTRY OF AMINO ACIDS carboxylic acid moiety attached to the same


α- carbon atom.
Amino acids contain the carboxyl group
At physiologic pH, (approx. pH, 7.4), the
(COOH), an amino group (NH2) and a side
carboxyl group is dissociated; forming the
chain (R group) bonded to the α- carbon atom.
negatively charged carboxylate ion (COO–)
Alpha-amino acids have both an amino and a
and the amino group is protonated (NH3+).
Although about 300 amino acids occur in
nature, less than one-tenth of these occur in
proteins. Complete acid-, base- or enzyme
catalyzed hydrolysis of proteins produces the
20 L-amino acids. Proteins from all forms of
life- plant, animal or microbial contain the
same 20 amino acids.
Classification of Amino Acids
Amino acids are classified on the basis of
acidic, basic or neutral groups as follows:
They are basically of three main types:
1. Neutral amino acids
2. Acidic amino acids
3. Basic amino acids
Neutral Amino Acids
Neutral amino acids are those which contain
one amino group and one carboxyl group. So,
these are called as monoamino mono carboxy-
lic acids.
These are further classified into four types:
1. Aliphatic amino acids
2. Aromatic amino acids
3. Heterocyclic amino acids
4. Sulphur containing amino acids
Chemistry of Amino Acids and Proteins 29
Aliphatic amino acids include Glycine, Acidic Amino Acids
Alanine, Valine, Leucine, Isoleucine, Serine Acidic amino acids are those which contain
and Threonine. one amino group and two acid or carboxylic
groups. So, these are known as monoamino
dicarboxylic acids.
Acidic amino acids include Aspartic acid
and Glutamic acid.
Aromatic amino acids include Phenyla-
lanine and Tyrosine.

Basic Amino Acids


Heterocyclic amino acids include Tryptophan Basic amino acids contain two amino groups
and Histidine. and one carboxylic acid. So, these are known
as diamino monocarboxylic acids.
Basic amino acids include Arginine, Lysine
and Histidine.

S- containing amino acids include Cysteine,


Cystine and Methionine.
Functions of Amino Acids
1. Amino acids are monomeric constituents
of proteins.
2. Amino acids are the important dietary
sources of nitrogen.
3. Some amino acids give rise to specialized
products like
- Tryptophan synthesizes B-complex
vitamin niacin.
- Tyrosine forms thyroid hormones.
- Glycine and cysteine help in synthesis
of bile salts.
30 Medical Biochemistry for Physiotherapy Students
4. Some amino acids are glucogenic amino from essential amino acids. Examples include
acids, i.e. they are converted to carbohy- rest of the 10 amino acids like glycine, alanine,
drates. So, such type of amino acids after serine, cysteine, tyrosine, aspartic acid,
their conversion to carbohydrates are used glutamic acid, asparagine, glutamine and
as source of energy proline.
5. Amino acids like glycine and cysteine are The amounts and proportions of the
used in detoxification reactions. constituent amino acids, particularly the
6. Methionine acts as active methionine by proportion of essential amino acids determine
transferring its methyl (CH 3) group to the nutritional quality (biological value) of the
various substances by a process known as protein. A protein which has the right quantity
transmethylation. of each essential amino acid for growth and
maintenance has high biological value e.g.
Nutritional Classification of Amino Acids milk proteins and egg albumin. On the other
Essential Amino Acids hand, a protein with poor biological value has
a limited supply of one or more essential
Essential amino acids are the amino acids amino acids e.g. proteins from cereals and
which are not synthesized by the body and pulses. Biological value is also influenced by
therefore must be provided through the diet. the digestibility of the protein.
These are eight in number.
1. Phenylalanine PEPTIDE BOND
2. Methionine In the process of formation of peptide bond,
3. Tryptophan the COOH group of one amino acid is joined
4. Threonine to the NH2 group of another amino acid by a
5. Valine covalent bond. In this process, a molecule of
6. Isoleucine water is eliminated.
7. Leucine
8. Lysine

Semi Essential Amino Acids


Semi essential amino acids are those amino
acids which are not synthesized by the body
in sufficient quantity during growth. So, these
are known as growth promoting factors.
These become essential in growing children,
pregnant women and lactating mothers. These
include arginine and histidine.
For example, Glycine and glycine can
Non-Essential Amino Acids
combine to form the dipeptide glycyl-glycine.
Non-essential amino acids are those amino Valine and alanine can form the dipeptide
acids which can be synthesized by the body valylalanine through the formation of a
Chemistry of Amino Acids and Proteins 31
peptide bond. Peptide bonds are not broken 3. Oxytocin: It is a nonapeptide (9 amino
by conditions that denature proteins such as acids) secreted by posterior pituitary
heating or high concentrations of urea. gland. It acts on uterine muscles before
parturition and in the ejection of milk.
4. Vasopressin: It is also known as ADH
(antidiuretic hormone) and is secreted by
posterior pituitary gland. It is a nonapep-
tide (9 amino acids). It influences reabsorp-
tion of water from distal and collecting
tubules of the kidney.
5. TRH (Thyrotropic hormone): It is a
tripeptide (3 amino acids) and is secreted
by hypothalamus. It stimulates pituitary
gland to release TSH.
Three amino acids can be joined by two
6. Angiotensins: Angiotensin I is a decapetide
peptide bonds to form a tripeptide; similarly,
(10 amino acids) which is converted to
amino acids can be linked to form tetrapep-
Angiotensin II (8 amino acids). The latter
tides and pentapeptides. When a few amino
has more hypertensive effect. It stimulates
acids are joined in this manner, the structure
thirst, dilatation of blood vessels of
is called an oligopeptide. When many amino
voluntary muscle and brain and increases
acids are joined, the product formed is called
aldosterone secretion from adrenal gland.
polypeptide.
7. Gastrointestinal hormones: Gastrin, secretin
Peptides of Biological Importance and pancreozymin are the gastrointestinal
peptides which acts as hormones which
1. Glutathione: It is a tripeptide (3 amino stimulates secretion of bile and other
acids) and is composed of glutamic acid, enzymes of digestive juices.
cysteine and glycine. In humans and other
animals, glutathione is required for the
General Characteristics of Amino Acids
action of several enzymes and of insulin.
Glutathione and the enzyme glutathione 1. Optical activity: Amino acids that have an
reductase function either in insulin asymmetric carbon atom exist as D- and
degradation or in formation of correct L- forms that are mirror images of each
disulfide bonds in insulin. other. The two forms in each pair are
2. Bradykinin and kallidin: Bradykinin is a termed as stereo-isomers or optical
nonapeptide (9 amino acids) and kallidin isomers. Glycine is an exception because
is a decapeptide (10 amino acids). These its α-carbon has two hydrogen substi-
are smooth muscle hypotensive agents tuents and therefore, is optically inactive.
liberated from specific plasma proteins All the amino acids found in proteins are
exposed to snake venom or trypsin. of the L-configuration.
32 Medical Biochemistry for Physiotherapy Students
Chemistry of Proteins

Proteins are high molecular weight polymers


of a group of low molecular weight mono-
mers called amino acids. The amino acids are
joined by peptide bond.

2. Zwitter ion and isoelectric pH: Amino acids


Biological Importance of Proteins
contain both acidic (COOH) as well as
basic (NH2) groups. They can donate a 1. Proteins form an essential part for the
proton or accept a proton hence they are particular structures in the body (structural
regarded as ampholytes. Thus, depending proteins) e.g. collagen of bones and
upon pH of the solution an amino acid can cartilages or keratins of hair and nails.
act either as a cation or anion. This 2. Proteins give energy to body when
property of the charged particles such as carbohydrates are not present in adequate
amino acids is called as amphoteric nature amounts. They provide about 4 C/g.
of the molecule. 3. Many proteins act as catalysts, thus
enhancing the rate of chemical reactions
to such extents as needed by the living
cells.
4. Most of the enzymes are hormones e.g.
insulin.
3. Isoelectric pH: The isoelectric pH (pI) is the 5. Some proteins serve as carrier for the
pH at which an amino acid is electrically transport of various substances e.g.
neutral i.e. where the sum of the positive
haemoglobin and caeruloplasmin.
charges equals the sum of negative
6. Some proteins bind to certain substances
charges. At physiologic pH, all amino acids
and store them in different tissues, thus
have both a negatively charged group and
acting as storage proteins e.g. ferritin.
a positively charged group. They are,
7. Some proteins like gamma globulins act
therefore, dipolar ions or zwitter ions.
as antibodies and provide immunity to the
body.
Physical Properties of Amino Acids
1. Amino acids are soluble in polar solvents CLASSIFICATION OF PROTEINS
such as water and ethanol but they are
insoluble in non polar solvents such as Proteins are classified according to the
benzene or ether. solubility and physical properties as
2. Their melting point is above 200oC. 1. Simple proteins.
3. The aromatic amino acids tryptophan and 2. Conjugated/Compound proteins.
histidine absorb ultraviolet light. 3. Derived proteins.
Chemistry of Amino Acids and Proteins 33
Simple Proteins proteins, blood group substances, hor-
mones like chorionic gonadotrophin, TSH
Simple proteins are further subdivided into
are glycoproteins.
different classes which are as follows:
iii. Phosphoproteins – In phosphoproteins,
i. Albumins – Albumins are present in serum phosphoric acid residue is attached. Casein
as serum albumin, in egg as ovoalbumin of milk and vitellin of egg yolk are
and in milk as lactalbumin. These are examples.
soluble in water and dilute salt solutions. iv. Metalloproteins – Fe, Co, Mn, Zn, Cu, and
ii. Globulins - Globulins are present in serum Mg are some of the metallic ions asso-
as serum globulin, in egg as ovoglobulin ciated with proteins. The proteins are
and in milk as lactoglobulin. These are usually enzymes and require the metallic
soluble in neutral and dilute salt solutions. ions as activators.
iii. Prolamines – Gliadin of wheat and zein of v. Lipoproteins – These are formed in combi-
maize are the prolamines. These are nations with lipids as their prosthetic
insoluble in water but soluble in 60 – 80% group. Phospholipids protein complex is
alcohol. also called lecithoprotein. They are found
iv. Glutelins – The proteins of wheat (glutelin), in milk, blood cell nuclei, egg yolk mem-
rice (oryzenin) and other cereals belong to brane etc.
this group. These are soluble in dilute acids vi. Chromoproteins – The prosthetic group is
and alkalies. a coloured compound in chromoproteins.
v. Scleroproteins – They are animal proteins For example haemoglobin, flavoprotein,
etc.
present in hair, horn, nails, cartilage and
bone. Collagen of bone, keratin of hair are Derived Proteins
few examples. These are insoluble in water, Derived proteins are formed from simple and
dilute acids and alkalies. conjugated proteins by physical and chemical
vi. Histones – They are basic proteins rich in means.
arginine and histidine. They are soluble The products of partial hydrolysis of pro-
in water, dilute acids and salt solutions. teins are often classified as derived proteins.
Nucleohistones, globin of haemoglobin i. The primary derived proteins are the
are few examples. denatured or coagulated or first hydro-
lysed products of protein. Examples
Conjugated Proteins include coagulated proteins like cooked
Conjugated proteins are simple proteins proteins, coagulated albumin, proteans like
which are complexed or conjugated with the fibrin from fibrinogen.
non protein part i.e. prosthetic group. ii. The secondary derived proteins are the
i. Nucleoproteins – These are compounds degraded (due to breakdown of peptide
made up of simple basic proteins such as bonds) products of proteins. Examples
histone with nucleic acids as the prosthetic include proteoses (hydrolytic products of
group e.g ribonucleoproteins. proteins), peptones (hydrolytic products
ii. Glycoproteins – They have oligosaccharide of simpler structure than the proteoses),
chains attached. Most of the plasma polypeptides and peptides.
34 Medical Biochemistry for Physiotherapy Students

CLASSIFICATION OF PROTEINS BASED STRUCTURE OF PROTEINS


ON FUNCTIONAL PROPERTIES
There are 4 levels of protein structure:
1. Structural proteins: Keratin of hair and 1. Primary structure – Primary structure of
nails, collagen of bone. a protein refers to the order and sequence
2. Contractile proteins: Actin, myosin. of amino acids in a polypeptide chain in
3. Defence proteins: Immunoglobulins. which different amino acids are linked
4. Transport proteins: Haemoglobin, serum through the peptide linkages.
albumin. 2. Secondary structure – Secondary structure
5. Enzymes or catalytic proteins: Pepsin, of protein is formed by folding or twisting
hexokinase. the polypeptide chain possessing primary
6. Storage proteins: Ovoalbumin, glutelin. structure. Two main types of bonds are
7. Hormonal proteins: Insulin, growth involved for the formation of secondary level
hormone. of protein structure. The foldings are the
results of linking of the carboxyl and amino
CLASSIFICATION OF PROTEINS BASED ON groups of the peptide chains by means of
THE SHAPE OF THE PROTEIN MOLECULE hydrogen and disulphide bonds.
Hydrogen bonds: Hydrogen bonds are weak
On the basis of shape of protein molecule, bonds. They result from the sharing of
the proteins are categorized into two classes hydrogen atoms between the nitrogen and
which are as follows: the carbonyl oxygen of different peptide
1. Globular or corpuscular proteins. bonds in the same or different polypeptide
2. Fibrous or fibrillar proteins. chains.

Globular or Corpuscular Proteins


These have an axial ratio (length: width) of
less than 10 and hence possess a relatively
spherical or ovoid shape. Examples include
some enzymes, blood transport proteins and
antibodies.

Fibrous or Fibrillar Proteins


These have axial ratio greater than 10 and
hence resemble long ribbons or fibres in Disulphide bonds: The disulphide bond
shape. Examples include collagens, elastins interconnects two portions of polypeptide
and keratins. chains through cysteine residues.
Chemistry of Amino Acids and Proteins 35
c. β- bends: They reverse the direction of a
polypeptide chain, helping it to form a
compact, globular shape. These are usually
found on the surface of protein molecules.
3. Tertiary structure – Tertiary structure of
proteins is formed when the polypeptide
chain having secondary structure is folded
and twisted upon itself. The forces or
This is of following types: bonds which give stability to the tertiary
a. α- helix: This is the most common type of structure are vanderwaal forces, ionic
secondary structure, it is spiral structure. bonds and hydrogen bonds.
It is stabilized by extensive hydrogen
bonding and it consists of 3- 6 amino acids
per turn.

4. Quaternary structure – Many proteins


consist of a single polypeptide chain; these
are monomeric proteins. But many other
consist of two or more polypeptide chains
b. β- sheet: In this surface appears pleated. that may be structurally identical or totally
So, it is also known as β- pleated sheet. unrelated. The arrangement of these poly-
The two or more chains may be parallel peptide subunits is called the quaternary
structure of the protein. If there are two
or anti-parallel.
subunits, the protein is called dimeric, if
three subunits trimeric and if several
subunits then multimeric. Subunits are held
together by non-covalent interactions like
hydrophobic interactions, hydrogen
bonds, ionic bonds as in haemoglobin.
Quaternary structure is observed in
several oligomeric proteins such as
haemoglobin (a tetramer) and several
isoenzymes like lactic dehydrogenase (a
tetramer) or creatine phosphokinase (a
dimer). Disintegration of the monomeric
subunits results in loss of the biological
activity of protein.
36 Medical Biochemistry for Physiotherapy Students
consists of a series of changes in the protein
molecule brought about by the various
physical and chemical agents and these
changes often affect the viscosity, particle size,
solubility etc. Denaturating agents include
heat, organic solvents, mechanical mixing,
strong acids or bases, detergents and ions of
heavy metals such as lead and mercury.

DENATURATION OF PROTEINS

The comparatively weak forces responsible


for maintaining secondary, tertiary and
quaternary structures of proteins are readily
disrupted by a variety of manipulations with Denaturation may, in rare cases, be rever-
a resulting loss of enzymatic activity. Protein sible, in which case the protein refolds into
denaturation results in the unfolding and its original native structure when the dena-
disorganization of the protein’s structure turing agent is removed. However, most
which are not accompanied by hydrolysis of proteins, once denatured, remain perma-
peptide bonds. In general, denaturation nently disordered.
Chapter 6

Chemistry of Nucleotides
and Nucleic Acids

Nucleotides are energy rich compounds that


drive metabolism (primarily biosynthesis) in
all cells. They serve as chemical signals. They
are structural components of a number of
enzyme cofactors and metabolic inter-
mediates.
Nucleic acids are biopolymers of high
molecular weight with mononucleotide as
their repeating units, just as amino acids are
Nitrogenous Bases
the repeating units of proteins. The nucleic
acids are DNA (deoxyribonucleic acid) and The nitrogenous bases are derivatives of two
RNA (ribonucleic acid). DNA is found mainly parent compounds, pyrimidine and purine.
on the chromatin of the cell nucleus whereas Free purines and pyrimidines are weakly
most of the RNA (90%) is present in the cell basic compounds and are thus called bases.
The base is joined covalently (at N -1 of
cytoplasm and a little (10%) in the nucleolus.
pyrimidines and N - 9 of purines) in an N–
The nucleic acids DNA and RNA are the
glycosyl linkage to the 1 carbon of the pentose
molecular repositories for genetic infor- and the phosphate is esterified to the 5
mation. carbon.
Nucleotides have three components:
a. A nitrogenous base Pyrimidine Bases
b. A pentose (sugar) These are all derived from their parent
c. A phosphoric acid heterocyclic compound pyrimidine.
38 Medical Biochemistry for Physiotherapy Students
bases i.e. cytosine occurs in both DNA and
RNA, thymine occurs in DNA only whereas
uracil occurs in RNA only.

Pyrimidine bases are three in number:


1. Cytosine (C)
2. Thymine (T)
3. Uracil (U)

Sugars (Pentoses)
Two kinds of pentoses are found in nucleic
acids. The recurring deoxyribonucleotides
units of DNA contain 2–deoxy–D–ribose and
the ribonucleotide units of RNA contain D–
ribose.
In nucleotides, both types of pentoses are
in the form of their β–furanose structure
Purine Bases (closed five membered ring structure).

Purine ring is more complex than the


pyrimidine ring. It can be considered as the
product of fusion of a pyrimidine ring with
an imidazole ring.

Phosphoric Acid
The molecular formula of phosphoric acid is
H3PO4. It contains 3 monovalent hydroxyl
groups and a divalent oxygen atom, all linked
to the pentavalent phosphorous atom.
Purine bases are two in number:
1. Adenine (A)
2. Guanine (G)
Purine bases, i.e. adenine and guanine
occur in both DNA and RNA. Pyrimidine
Chemistry of Nucleotides and Nucleic Acids 39
Nucleoside 2. They carry chemical energy in cells.
Nucleotides may have 1, 2 or 3 phosphate
A molecule composed of sugar (ribose or
groups covalently linked at the 5’ – OH of
deoxyribose) and a base (purine or pyrimi-
ribose. These are referred to as nucleoside
dine) without a phosphoric acid group. So,
monophosphate if they contain one phos-
from this, nucleotides can be defined as
phate group (NMP), nucleoside diphos-
phosphorylated nucleosides.
phate if they contain two phosphate groups
Example of nucleoside is adenosine
(NDP) and nucleoside triphosphate if they
present in RNA which contains purine base contain three phosphate groups (NTP).
adenine and pentose sugar ribose. Another
example is deoxyadenosine present in DNA
which contains purine base adenine and
pentose sugar deoxyribose. Similarly, other
examples include guanosine, deoxyguanosine
etc.
Examples of nucleotides are AMP, dAMP,
GMP, dGMP, CMP, dCMP, UMP and dTMP.
AMP (adenosine monophosphate) contains
purine base adenine, pentose sugar ribose and
one phosphate group. Nucleosides and
nucleotides will be more clear from the table
given below. 3. Nucleoside triphosphates are used as a
source of chemical energy to drive a wide
Base Nucleoside Nucleotide Nucleic acid variety of biochemical reactions.
Adenine Adenosine AMP RNA ATP is by far the most widely used, but
Deoxyadenoside dAMP DNA UTP, GTP and CTP are used in specific
Guanine Guanosine GMP RNA
reactions.
Deoxyguanosine dGMP DNA
Cytosine Cytidine CMP RNA The hydrolysis of ATP and the other
Deoxycytidine dCMP DNA nucleoside triphosphates is an energy
Uracil Uridine UMP RNA yielding reaction because of the chemistry
Thymine Deoxythymidine dTMP DNA of triphosphate structure.
The bond between the ribose and the
So it is clear from the above table that,
phosphate is an ester linkage. The α–β and
DNA consists of adenine, guanine,
β-α linkages are phosphoric acid
cytosine, thymine, deoxy ribose sugar and
anhydrides. Hydrolysis of the ester
phosphate and RNA consists of adenine,
linkage yields about 14 KJ/mol, whereas
guanine, cytosine, uracil, ribose sugar and hydrolysis of each of the anhydride bonds
phosphate. yields about 30 KJ/mol.
4. Nucleotides are components of many
Functions of Nucleotides
enzyme cofactors. A variety of enzyme
1. They are the subunits of nucleic acids. cofactors serving a wide range of chemical
40 Medical Biochemistry for Physiotherapy Students
functions includes adenosine as a part of 2. The base composition of DNA in a given
their structure. In these cofactors, the species does not change with the orga-
adenosine portion does not play part nism’s age, nutritional state or change of
directly in the primary function, but environment.
removal of adenosine from these struc- 3. Sum of purine residues equals the sum of
tures generally results in drastic reduction pyrimidine residues.
of their activities. A+G=T+C
5. Some nucleotides are the intermediates in
cellular communication. Cells respond to Structure of DNA
their environment by taking cues from DNA is a double helix. DNA contains recur-
hormones or other chemical signals in the ring units of adenine, guanine, cytosine,
surrounding medium. The interaction of thymine, deoxyribose sugar and phosphate.
these extra cellular chemical signals (first It is a polynucleotide.
messengers) with receptors on the cell DNA consists of two helical DNA chains
surface lead to the production of second coiled around the same axis to form a right–
messengers inside the cell, which in turn handed double helix. The two helices are
lead to adaptive changes in the cell interior.
Second messenger is a nucleotide. One of
the most common is the nucleotide 3’, 5’ –
cyclic adenosine monophosphate (3’, 5’ -
cAMP).

STRUCTURE OF NUCLEIC ACID


1. Primary structure: The primary structure of
a nucleic acid is its covalent structure and
the nucleotide sequence.
2. Secondary structure: Any regular, stable
structure taken up by some or all of the
nucleotides in a nucleic acid can be referred
to as secondary structure.
3. Tertiary structure: The complex folding of
large chromosomes within the bacterial
nucleoid and eukaryotic chromatin is
considered as tertiary structure.

Chargaff’s Rule
The base composition of DNA generally
varies from one species to another.
1. DNA isolated from different tissues of the
same species has the same base compo-
sition.
Chemistry of Nucleotides and Nucleic Acids 41
wound in such a way so as to produce 2 defined by Watson and Crick. Different forms
interchain spacings or grooves, a major (or are:
wide) groove and a minor groove (or narrow) 1. B – DNA
groove. 2. A – DNA
The two chains/strands of the helix are 3. Z – DNA
antiparallel, i.e. one strand runs in the 5’ to 3’ The Watson- Crick structure is also
direction and the other in the 3’ to 5’ direction. referred to as B–form of DNA. The B–form
The diameter of the helix is 20Å. The bases is the most stable structure. A–DNA is right
are 3.4Å apart along the helix axis and are handed helix. Z–DNA is left handed helix.
related by a rotation of 36º. Therefore, the In A – DNA, rise per base pair is 0.23 nm
helical structure repeats after 10 residues on and number of base pair per helical turn is 11.
each chain i.e. at intervals of 34Å. In other In Z – DNA, rise per base pair is 0.38 nm and
words, each turn of the helix contains 10 number of base pair per helical turn is 12.
nucleotide sequences.
The backbones of alternating deoxyribose Denaturation of DNA
and negatively charged phosphate groups are Solutions of carefully, isolated native DNA
on the outside of the double helix and the are highly viscous at pH 7.0 and room tempe-
bases (purines and pyrimidines) of both rature (20-25ºC). When such a solution is
strands are stacked inside the double helix. subjected to extremes of pH or to tempera-
The planes of the bases are perpendicular to tures above 80-90ºC, its viscosity decreases
the helix axis. The planes of the sugars are sharply, indicating that DNA has undergone
almost at right angles to those of the bases. a physical change. This involves disruption
The two chains are held together by of hydrogen bonds between the paired bases
hydrogen bonds between pairs of bases. and the interactions between the stacked
Adenine always pairs with thymine by 2 bases. As a result, double helix unwinds to
hydrogen bonds and guanine with cytosine form to single strands. No covalent bonds in
with 3 hydrogen bonds. This specific position- the DNA are broken.
ing of the bases is called base complimentarity. Renaturation of DNA is a rapid one step
The DNA double helix or duplex is held process. When the temperature or pH is
together by two sets of forces: returned to the biological range, the unwound
1. Hydrogen bonding between complemen- segments of the two strands spontaneously
tary base pairs. rewind or anneal to yield the intact duplex.
2. Base-stacking interactions. However, if the two strands are completely
separated, renaturation occurs in two steps.
Different Structural Forms of DNA
The first step is relatively slow, because the
Many significant deviations from the Watson two strands must first find each other by
– Crick structure of DNA are found in cellular random collisions and form a short segment
DNA and these may play important roles in of complementary double helix. The second
DNA metabolism. These structural variations step is much faster: the remaining unpaired
do not affect the key properties of DNA bases successively come into register as base
42 Medical Biochemistry for Physiotherapy Students
pairs, and the two strands zipper themselves Genetic Code
together to form the double helix.
The genetic code is dictionary that gives the
Each species of DNA has a characteristic
correspondence between a sequence of
denaturation temperature or melting point: nucleotide bases and a sequence of amino
the higher its content of G≡C base pairs, the acids. Each individual word in the code is
higher the melting point of the DNA. This is composed of three nucleotide bases. These
because G≡C base pairs, with three hydrogen genetic words are called codons.
bonds are more stable and require more heat The four nucleotide bases are used to
energy to dissociate than A=T base pairs. produce the three base codons. There are
Circular DNA therefore, 64 different combinations of bases
taken three at a time (43).
In certain micro-organisms, such as bacteria
and bacteriophages, the DNA molecule does Characteristics of Genetic Code
not possess free 3’ and 5’ ends but the two 1. Specificity: The genetic code is specific –
ends of the molecule are joined together that is, a specific codon always code for
forming a circular DNA. the same amino acid.
2. Universality: The genetic code is universal
CELLULAR FUNCTIONS OF DNA (with a few minor exceptions) – i.e. the
specificity of the genetic code has been
DNA controls every aspect of cellular
conserved from very early stages of
function. evolution, with only slight differences in
1. It makes copies of itself as a cell divides the manner the code is translated.
and thus transfers genetic information from 3. Redundancy: The genetic code is redundant
one generation to the next known as (sometimes called degenerate). Although
replication. each codon corresponds to a single amino
2. DNA also determines the properties of a acid, a given amino acid may have more
living cell and regulates biological than one triplet coding for it. For example,
information by control of protein syn- arginine is specified by six different
thesis. This flow of biological information codons.
from one class of nucleic acid to another, 4. Nonoverlapping and commaless: The genetic
i.e. from DNA to RNA is known as code is nonoverlapping and commaless,
transcription and from there to protein is i.e. the code is read from a fixed starting
called translation. point as a continuous sequence of bases,
DNA molecule thus serves as a template, taken three at a time. For example,
both for the transcription of information ABCDEFGHIJKL….is read as ABC/DEF/
from RNA as well as for the replication of GHI/JKL….without any punctuation
information into daughter DNA. between the codons.
Chapter 7

Enzymes

Enzymes are defined as biocatalysts, protein 9. Abnormal conditions can also be caused
in nature, specific in actions which are not by the excessive activity of the specific
themselves used up in the reaction. enzymes.
Substances acted upon by an enzyme are
called its substrates while those produced by Chemical Nature of Enzymes
its action are called its products. Most of the enzymes are proteins with a few
exceptions. Their catalytic activity depends
Properties of Enzymes
upon the integrity of their native protein
1. Enzymes are proteins. conformation. If an enzyme is denatured or
2. Enzymes are the reaction catalysts of dissociated into subunits, catalytic activity is
biological systems. usually lost. If an enzyme is broken down
3. Enzymes remain unchanged in mass and into its component amino acids, its catalytic
form on completion of the reaction. So, activity is also destroyed.
very small amounts of an enzyme may be Few enzymes are simple proteins while
repeatedly used to change large amounts some are conjugated proteins. In such
of its substrates. enzymes, the non-protein part is called
4. Enzymes have extraordinary catalytic prosthetic group and the protein part is called
power. as apo-enzyme. The complete structure, i.e.
5. They have a high degree of specificity for Apo-enzyme + prosthetic group =
their substrates. holoenzyme.
6. They accelerate specific chemical reactions. Certain enzymes with only one polypep-
7. They act by lowering the activation energy tide chain are called as monomeric enzymes,
required for forming or cleaving covalent e.g. ribonuclease. Several enzymes possess
bonds. more than one polypeptide chain and are
8. Deficiency of enzymes result in genetic called as oligomeric enzymes, e.g LDH (lactate
disorders. dehydrogenase), hexokinase, etc.
44 Medical Biochemistry for Physiotherapy Students
Ribozymes 3. The coenzymes are also attached to the
protein at a different but adjacent site so
A few enzymes are made up of RNA and
as to be in a position to accept the atoms
called ribozymes, e.g. the RNA splicing
or groups which are removed from the
ribozyme of the protozoan Tetrahymena
substrate.
thermophilia and the RNase P of bacteria
4. NAD and NADP coenzymes function as
Escherichia coli. RNase P is actually made of hydrogen acceptors in dehydrogenation
both RNA and protein, but its RNA subunit reactions.
itself is the catalytic subunit and can catalyze 5. The chief function of CoA is to carry acyl
the cleavage of the tRNAs of E.coli even when groups and they are used in the oxidative
freed from the protein subunit. decarboxylation of pyruvic acid and
synthesis of fatty acid and acetylation.
Coenzymes 6. The function of TPP is to carry active
Many reactions of substrates are catalyzed aldehyde group.
by enzymes only in the presence of a specific 7. The tetrahydrofolic acid is expressed as a
non protein organic molecule called the carrier of formate and it is used in the
coenzyme. synthesis of purines and pyrimidines.
Coenzymes are defined as heat stable, 8. The pyridoxal phosphate is involved in
transamination reactions.
dialyzable, non-protein, organic molecules.
Many coenzymes are derived as the
Classification of Enzymes
physiologically active forms from the consti-
tuents of vitamin B complex such as Based on the type of chemical reactions and
mechanism of reaction, IUB (International
Vitamin Coenzyme Union of Biochemistry) divided enzymes into
Thaimine (B1) TPP six major classes that are as follows:
Riboflavin (B2) FMN 1. Oxidoreductases
Niacin NAD+, NADP+ 2. Transferases
Pyridoxine (B6) Pyridoxal phosphate 3. Hydrolases
Pantothenic acid CoA 4. Lyases
5. Isomerases
Biotin Active Biotin
6. Ligases
Folic acid Tetrahydrofolic acid
Cyanocobalamin (B12) Cobamide
Lipoic acid Lipoic acid Oxidoreductases
Enzymes which catalyze oxidations reduc-
Functions of Coenzymes
tions between two substrates A and B are
1. Their function is usually to accept atoms known as oxidoreductases.
or groups from a substrate and to transfer Areduced + Boxidized –––––
>Aoxidized + Breduced
them to other molecules.
2. They are less specific than are enzymes For example, alcohol dehydrogenase, lactate
and the same enzyme can act as such in a dehydrogenase, xanthine oxidase, glutathione
number of different reactions. reductase.
Enzymes 45
Transferases For example, DNA ligases, glutamine
synthetase, succinate thiokinase.
These enzymes catalyze the transfer of some
group or radical X from one molecule A, to
ROLE OF METAL IONS IN ENZYMES
another molecule, B.
A.X + B ———> A + B.X The activity of many enzymes depends on
For example, hexokinase, phosphogluco- the presence of certain metal ions (Table 7.1).
mutase, aspartate transaminase, alanine In some cases, the requirement is specific for
transaminase, ornithine carbamoyl trans- a particular metal. Carbonic anhydrase shows
ferase, choline acetyl transferase. no activity upon removal of zinc and no other
metal is known to replace zinc in this enzyme.
Hydrolases In other cases, more than one metal is able to
These enzymes catalyze hydrolysis by bring about activation; for example Mg2+,
addition of water. Mn2+ or Zn2+ activates enolase (2-phospho-
glycerate → phosphoenolpyruvate + water).
A – B + H2O ———> AH + BOH
In a few cases it appears that two metal ions
For example, pepsin, trypsin, esterases, may be required by the enzyme; for example
glucose -6-phosphatase. pyruvate phosphokinase requires both Mg2+ and
K+ (pyruvate + ATP ———> phosphopyruvate
Lyases + ADP).
These are those enzymes that facilitate
Table 7.1: Representative metalloenzymes (The enzymes
removal of small molecule from a large either contain the metal or are activated by it)
substrate. Metal Enzyme
A – B + X – Y———> AX – BY Zn2+ Carbonic anhydrase
Lactic dehydrogenase
For example, fumarase, histidine decarboxy- Mg2+ Peptidases
lase, argininosuccinase. Hexokinases
Mn2+ Arginase
Phosphoglucomutase
Isomerases Fe2+ Cytochrome oxidase
Catalase
Enzymes catalyzing interconversion of Cu2+ Tyrosinase
optical, geometric or positional isomers are Phenolase
known as isomerases.
A ———> A’ Factors Affecting Enzyme Action
1. Effect of pH: The pH of the reaction medium
For example, retinal isomerase, racemases,
affects reaction velocity. The relationship
triose phosphate isomerases.
of enzyme activity to pH is also
represented by a bell shaped curve which
Ligases
has its peak at the optimum pH. This pH
Enzymes involved in joining together two is characteristic for each enzyme and is an
substrates are known as ligases. H+ concentration at which the enzyme
A + B ———> A – B reacts at maximum speed. The optimum
46 Medical Biochemistry for Physiotherapy Students
pH for most of the intracellular enzymes
is in the neutral range i.e. around 7.0. The
optimum pH for pepsin is about 2.0 while
for enzymes of the pancreatic juice the
optimum pH is nearly 8.0.
The optimum pH of an enzyme depends
upon the ionization of the enzyme as well
as of the substrate. Change in pH affects
ionization of the protein molecule thereby
decreasing the number of active sites. At
an extremely high or low pH, enzyme gets
denatured.
3. Effect of enzyme concentration and substrate
concentration: If the concentration of
substrate is lower than the concentration
of the enzyme, then all the active groups
of the enzyme will not be utilized for the
formation of enzyme substrate complex.
The rate of reaction will be less.
When the concentration of substrate
increases, more and more active sites will
be used for the formation of ES complex
then the rate of reaction will increase.
2. Effect of temperature: The rate of an enzyme 4. Effect of product concentration: Products
catalyzed reaction generally increases with formed because of chemical reaction may
temperature, within the temperature accumulate and this excess of product may
range in which the enzyme is stable and lower the reaction.
retains its full or maximum activity. 5. Effect of oxidation: The sulfhydryl (SH)
Enzyme catalyzed reactions have an groups of many enzymes mainly the
optimum temperature at which the oxidoreductases are essential for enzyme
reaction is most rapid. For most of the activity. Many oxidizing agents including
enzymes, the optimum temperature is near the oxygen of air oxidize these –SH groups
the body temperature i.e. between 37-40ºC. giving rise to the formation of disulphide
Above the optimum temperature, the (S-S) linkage. This results in loss of
reaction rate decreases as enzymes being enzymatic activity.
protein in nature are denatured by heat 6. Effect of activators: Many ions and molecules
and becomes inactive. have the capacity to activate some
Enzymes 47
enzymes. Metal ions are activators of a The ES complex soon dissociates into the
number of enzymes. Pepsin (as proenzyme unchanged enzyme and the product (P), i.e.
pepsinogen) is activated by H+ to form the E + S ——> ES complex ——> E + P
active enzyme. Enzymes themselves This is known as Michaelis- Menten theory
activate other enzymes or proenzymes, of enzyme action.
i.e. enterokinase activates trypsinogen to The active site of an enzyme is constituted
form active trypsin. Trypsinogen, chymo- by several amino acid residues, located far
trypsinogen are known as zymogens. apart from each other in the peptide chain
7. Effect of radiation: Enzymes are highly but closed up by the folding of the latter. So,
sensitive to short wavelength radiations the active site has a complex 3- dimensional
such as ultra violet light, X- ray, β- rays or form with a predominantly non polar cleft or
γ-rays. The enzymes may get oxidized by crevice which receives and binds the sub-
the peroxides formed by high-energy strate. Inside this cleft, the side chains of
radiation. specific amino acid residues bind with specific
8. Effect of ionic strength: The activity of many groups of the substrate while some others
enzymes is affected by the concentration participate in changing the latter. Any change
of ions in the solution. The sensitivity of in the 3-dimensional form of the active site
enzymes to the concentration of ions may alters its substrate binding and catalytic
be due to the presence of a diffuse ionic activities.
atmosphere around each enzyme protein Koshland’s induced fit model proposes
molecule, which attracts ions of opposite that in the absence of the substrate, the active
sign. This produces a double layer, the site does not possess the fully active and
thickness of which is inversely complementary conformation for the latter.
proportional to the ionic strength of the But either on the approach of the substrate,
or during its binding with the enzyme, or
solution.
following an initial superficial binding of the
MECHANISM OF ENZYME ACTION two, the flexible active site changes its
conformation into the final catalytic form.
(HOW ENZYMES WORK)
This model explains complex saturation
Substrates bind with specific active or catalytic kinetics, competitive inhibition and allosteric
sites of the enzyme molecule to form highly modulation of enzymes.
reactive enzyme-substrate complexes (ES).
Following types of bonds may link specific Specificity
groups of the substrate with specific amino An enzyme can bind only such substrates as
acid side chains at the active site of the have groups and steric conformations fitting
enzyme: with its active site. So, it can act specifically
1. hydrogen bonds on only a limited number of substrates
2. electrostatic bonds fulfilling such specifications. Many enzymes
3. van der Waals forces act only on a specific type of covalent bond
4. hydrophobic interactions associated with specific groups or residues
5. covalent bonds in the substrate (group specificity), e.g. trypsin
48 Medical Biochemistry for Physiotherapy Students
hydrolyzes peptide bonds connected to the S = substrate concentration
α-COOH groups of basic amino acids. Many Km = Michaelis constant
enzymes like fumarase act specifically on either
the cis or the trans isomer of the substrate 1. When [S] is very much less than Km, adding
but not on both the isomers (cis-trans [S] to Km in the denominator, change its
specificity). Some enzymes such as L- amino
value very little. Hence, [S] term can be
acid oxidase and D- amino acid oxidase act upon
dropped from the denominator.
either the L or the D isomer (D- L stereo
specificity). Stereo specificity results from a
Vmax [S] Vmax [S]
three point bonding between the enzyme and Vi = =
the substrate. K m + [S] Km
Vmax
ENZYME KINETICS Since = Constant (K)
Km
In enzyme catalyzed reactions, if the concen- Vi = K [S]
tration of substrate [S] is increased, while all
other conditions are kept constant, the initial In other words, when the substrate con-
velocity V1 (the velocity measured when very centration is considerably below that
little substrate has reacted), increases to a required to produce half-maximum velo-
maximum value, Vmax and no further. city (Km), the initial velocity Vi depends
upon the substrate concentration [S].

2 When [S] is very much greater than Km,


Km can be dropped from the denominator.

This states that when the substrate


Michaelis-Menten Equation
concentration [S] far exceeds the Km value,
The substrate concentration that produces the initial velocity Vi is maximum , i.e. Vmax
half maximum is termed the K m value or 3 When [S] = Km
Michaelis constant. The Michaelis- Menten
equation is expressed as follows:
Vmax [S]
Vi = —————
Km + [S]
Where,
Vi = Initial velocity
Vmax = Maximum velocity
Enzymes 49
This states that when the substrate
concentration is equal to the Km value,
the initial velocity Vi is half-maximum.

Enzyme Inhibition
Enzyme inhibitor is defined as a substance
which binds with the enzyme and brings
about a decrease in its catalytic activity. The
enzyme inhibitor may be organic or inorganic
in nature. Following are the various types of
enzyme inhibitions.
1. Reversible inhibition
2. Irreversible inhibition
3. Feedback inhibition

Reversible Inhibition Vmax . However, the [S] at which Vi = ½ V max,


The inhibitor binds non covalently with the the K m , will increase in the presence of
enzyme. The enzyme inhibition can be inhibitor.
reversed if the inhibition is removed. The Example:
reversible inhibition can be Enzyme: Succinate dehydrogenase (SDH)
a. Competitive Substrate: Succinic acid
b. Non-competitive Competitive inhibitor: Malonic acid.
The competitive inhibitor compete with
Competitive Inhibition succinic acid for binding at the active site of
A competitive inhibitor competes with the SDH.
substrate for the active site of an enzyme, but
a reaction usually does not occur once the Non-competitive Inhibition
inhibitor (I) is bound. While the inhibitor A non-competitive inhibitor is one that binds
occupies the active site, it prevents binding to a site distinct form that which binds the
by the substrate. substrate. Inhibitor binding does not block
substrate binding (or vice versa). The enzyme
Competitive inhibitors are often compounds is inactivated when inhibitor is bound,
that often resemble the substrate and combine whether or not substrate is also present. The
with the enzyme to form an EI complex. When inhibitor effectively lowers the concentration
enough substrate is present the probability of active enzyme and hence lowers the
that an inhibitor molecule will bind is apparent Vmax. There is often little or no effect
minimized, and the reaction exhibits normal on Km.
50 Medical Biochemistry for Physiotherapy Students
reaction. When a substrate (A) is converted
to product (P) through various intermediates
(B,C,D etc.), the end product of the reaction
inhibits the first enzyme of the pathway. In
this type of inhibition, the accumulation of
the end product slows down the whole
reaction sequence and as the end product is
consumed the synthesis continues. Since it is
partially competitive both Vmax and Km are
altered.

Example: There are many enzymes which Enzymes in Clinical Diagnosis


require free sulfhydryl groups (SH) for
activity, are non-competitively inhibited by Plasma enzymes can be classified into two
heavy metal ions like Hg2+, Pb2+, Ag2+ etc. major groups. First, a relatively small group
of enzymes are actively secreted into the
Urease is an example of an enzyme which
plasma by certain organs. For example, the
experiences heavy metal inhibition.
liver secretes zymogens (inactive precursors)
of the enzymes involved in blood coagulation.
Irreversible Inhibition Second, a large number of enzyme species
The inhibitors bind covalently with the are released from cells during normal cell
enzymes and inactivate them. The inhibition turnover. These enzymes are normally
of enzyme activity is irreversible. intracellular and have no physiologic function
in the plasma.
Examples: In healthy individuals, the levels of these
- Iodoacetate is an irreversible inhibitor of enzymes are fairly constant and represent a
enzyme such as glyceraldehyde-3-phosphate steady state in which the rate of release from
dehydrogenase. Iodoacetate combines with cells into the plasma is balanced by an equal
sulfhydryl (SH) groups at the active site rate of removal from the plasma. The
of these enzymes and makes them inactive. presence of elevated enzyme activity in the
- Diisopropyl fluorophosphates (DFP) plasma may indicate tissue damage accom-
irreversibly binds with enzymes con- panied by increased release of intracellular
taining serine proteases, acetylcholine esterases enzymes.
etc. Changes of Plasma Enzyme Levels
in Disease States
Feedback Inhibition
Many diseases that cause tissue damage result
Feedback inhibition refers to the inhibition in an increased release of intracellular
of the enzyme by the end product of the enzymes into the plasma. The activities of
Enzymes 51
many of these enzymes are routinely deter- For example
mined for diagnostic purposes in diseases of Lactate dehydrogenase (LDH)
the heart, liver, skeletal muscle and other It occurs in five isoenzyme forms. Each
tissues. The level of specific enzyme activity form consists of 4 subunits (polypeptide chain)
in the plasma frequently correlates with the in the following combination:
extent of tissue damage. Thus, the degree of Isoenzyme form Subunits Location
evaluation of a particular enzyme activity in LDH 1 HHHH Heart, RBC
plasma is often useful in evaluating the LDH 2 HHHM RBC, Heart
LDH 3 HHMM Liver, Lung and spleen
prognosis for the patient. LDH 4 HMMM Liver, Lung and spleen
LDH 5 MMMM Skeletal muscle
Diagnostic Importance of Enzymes
Some enzymes show relatively high activity Creatine Phosphokinase or
in only one or a few tissues. The presence of Creatine Kinase (CPK or CK)
increased levels of these enzymes in plasma
It is a high energy compound and acts as a
thus reflects damage to the corresponding
ready source of energy in the muscles. It has
tissue. For example the enzyme alanine
three isoenzyme forms, each having 2 sub-
transaminase, i.e. ALT also called as glutamate
units.
pyruvate transaminase, i.e. GPT is abundant in
Isoenzyme form Subunits Location
the liver. The appearance of elevated levels
CPK 1 BB Brain
of ALT in plasma signals possible damage to
CPK 2 MB Heart muscle
hepatic tissue. Increases in plasma levels of
CPK 3 MM Heart and skeletal muscle
enzymes with a wide distribution provide a
less specific indication of the site of cellular
injury. This lack of tissue specificity limits the Serum Enzyme in Muscle Diseases
diagnostic value of many plasma enzymes. 1. CPK
2. SGOT/SGPT
Isoenzymes 3. LDH
Isoenzymes are physically distinct forms of a
same enzyme having same catalytic activity. CPK (Creatine phosphokinase).
Thus, they catalyze the same reaction. It is also known as CK (creatine kinase). It
Although they possess identical or very catalyses the following reaction:
similar activity, the enzymes from different
sources are not identical proteins since they
may differ in physical, biochemical and
immunological properties. Due to the It is a high energy compound and acts as
difference in their molecular weights and a ready source of energy in the muscles. It is
sizes, their rates of migration in electro- found in high concentration in skeletal muscle,
phoresis are found to be different. myocardium and brain but not found in liver
52 Medical Biochemistry for Physiotherapy Students
and kidney. It appears to be a sensitive SGPT (Serum glutamate pyruvate transaminase):
measure of myocardial infarction, but remains It is also known as ALT (Alanine transaminase).
normal in patients with liver diseases. Liver is the major source. It is increased in all
CPK increases in muscular dystrophy types of liver diseases like viral hepatitis,
injuries of skeletal muscle particularly CPK- cirrhosis, liver cancer and drug induced
MM. CPK is increased in MI and other heart jaundice.
diseases. In this case, CPK- MB increases LDH (Lactate dehydrogenase)
markedly. CPK also increases in head injury
It catalyses the following reaction:
and diseases of the brain, it is CPK- BB form
of CPK that is increased.
Normal value is 4 – 60 IU/L.

SGOT (Serum glutamate


The enzyme is present in almost all the
oxaloacetate transaminase)
tissues, so its increase in the serum is
It is also known as AST (aspartate amino- relatively non-specific.
transferase). Heart, liver, skeletal muscle and LDH level mainly increases in the following
RBC’s are the major sources of SGOT. It is condition:
specific for heart. It is increased in: – MI (LDH 1 and LDH 2 increases)
– Skeletal muscle diseases (LDH 5 increases)
1. Heart muscle diseases – MI
– Liver diseases (LDH 3 and LDH 4 increases)
2. Liver diseases – mainly viral hepatitis, – Cancer of lung, liver and many other organ
jaundice and cirrhosis of liver diseases (LDH 3 and LDH 4 increases)
3. Muscle diseases –muscular dystrophy. Normal value is 60- 250 IU/L.
Chapter 8

Vitamins

Vitamins may be defined as potent organic 5. Synthesis in the body: Some vitamins are
compounds which are found in foods in synthesized in the body, e.g. vitamin A
variable and minute quantity and synthesized from provitamin carotene and vitamin D
in body and required in minute amounts for from ultraviolet irradiation of vitamin D
normal growth and reproduction etc. precursors. Some members of vitamin B
The general characteristics of the vitamins complex are synthesized by the micro-
are: organisms in the intestinal tract. Vitamin
C is also synthesized in some animals, e.g.
1. Distribution: The vitamins are widely
rats.
distributed in nature, both in the animal
6. Destruction: Vitamins are not destroyed in
and vegetable kingdoms. All vitamins are the digestive process and are absorbed as
manufactured in plants. The animals can such. Hence, all vitamins are effective
manufacture a few only but can store all when administered orally.
to some extent.
2. Store: Vitamins can be stored in the body Classification
to some extent, e.g. fat soluble vitamins are
All vitamins are broadly divided into two
stored in the liver and the subcutaneous
groups according to solubility:
tissue, vitamin C in adrenal cortex, etc.
1. Fat soluble vitamins
3. Daily requirement: Vitamins can perform
- Vitamin A
their work in very low concentrations. - Vitamin D
Hence, the total daily requirement is - Vitamin E
usually very small. The daily need of any - Vitamin K
vitamin for any individual is not a fixed 2. Water soluble vitamins
quantity. It varies according to the rate of - Vitamin C
metabolism. - Vitamin B complex
4. Fate: Vitamins are partly destroyed and Vitamin B complex further include
partly excreted. - Vitamin B1
54 Medical Biochemistry for Physiotherapy Students
- Vitamin B2 β-carotene – Plant foods contain β –
- Niacin carotene which on cleavage gives two vitamin
- Vitamin B6 A molecules whereas α and γ-carotenes give
- Pantothenic acid only one molecule of vitamin A. β-carotene
- Lipoic acid is an antioxidant and because of this property
- Biotin it will account for its possible anticancer
- Folic acid activity.
- Vitamin B12
Sources
VITAMIN A: ANTIXEROPHTHALMIC FACTOR
Animal sources include cod liver oil, milk,
Chemistry butter, egg, fishes, etc. Vegetable sources
Vitamin A is often used as a collective term include vegetable oils which are rich sources
for several biologically active molecules. The of carotene, e.g. carrots, spinach, green leafy
term retinoids includes both natural and vegetables, yellow fruits such as mangoes,
synthetic forms of vitamin A. tomatoes, etc.
Four important forms of vitamin A are:
Retinol (OH) or vitamin A alcohol – A Daily Requirement
primary alcohol containing an (β-ionone ring The RDA for adults is 1000 retinol equivalents
with an unsaturated side chain). (RE) for males and 800 RE for females. One
Retinal (CHO) or vitamin A aldehyde – It RE = 1 μg of retinol, 6 μg of β–carotene or 12
is derived from the oxidation of retinol. μg of other carotenoids.
Retinoic acid (COOH) or vitamin A acid –
It is derived from the oxidation of retinal. Absorption and Storage
In the small intestine, the vitamin A esters of
food are hydrolysed to fatty acids and free
vitamin. Vitamin A and carotene are absorbed
through the lymphatics and appear in the
blood plasma as ester. Presence of fats and
bile salts helps the process.
Storage occurs chiefly in the liver (95%)
as esters. When needed, retinol is released
from the liver and transported to extrahepatic
tissues by the retinol binding protein (RBP).

Functions
1. Role in vision: The visual mechanism by
which vitamin A functions in the vision is
known as Wald’s visual cycle or Rhodopsin
cycle.
Retina contains two types of receptor cells:
cones and rods. Cones are specialized for
Vitamins 55
colour and detail vision in bright light. be converted to cis-form. Third, all-trans-
Rods are specialized for visual activity in retinol from blood may be first isomerized
dim light. When light waves strike these to 11-cis-retinol which can then be
receptors, chemical changes are produced converted to 11-cis-retinal by retinol
which give rise to nerve impulses. Rod cells dehydrogenase.
contain photosensitive pigment rhodopsin 11-cis-retinal then combines with opsin in
or visual purple. It contains opsin as its the dark to form rhodopsin. As the rods
apoprotein and retinine as prosthetic are meant for vision in dim light,
group which is present as 11-cis retinal. rhodopsin should be adequately available
Light falling on rhodopsin converts it into for night vision.
opsin and all-trans-retinal in a series of 2. Growth – Animals deprived of vitamin A
events. Bathorhodopsin is formed first, lose their appetites and fail to grow. Bone
growth is slow and fails to keep pace with
then lumirhodopsin, then metarhodopsin
growth of the nervous system, leading to
I and II. Finally metarhodopsin gets split
central nervous system damage.
into opsin and all-trans-retinal. All-trans-
3. Epithelial tissue – Vitamin A is essential for
retinal has to be converted to 11-cis-retinal
normal differentiation of epithelial tissues
to be used again. This can happen by three
and mucus secretion. The epithelium of a
ways. First, all-trans-retinal may be wide variety of organs in the body under-
isomerized to 11-cis-retinal by retinal goes changes in vitamin A deficiency. The
isomerase. Second, the all-trans retinol primary cause involves atrophy of the
from blood may be converted to all-trans- epithelium ad the formation of a stratified
retinal by retinene reductase which may epithelium.
4. Reproduction - Retinol and retinal are
essential for normal reproduction, sup-
porting spermatogenesis in the male and
preventing foetal resorption in the female.
5. Bones and Teeth – Bone growth is markedly
impaired in vitamin A deficiency.
Cessation of growth in parts of the body
as well as abnormal growths are encoun-
tered. Poor development of the
ameloblasts and odontoblasts of the teeth
with a consequent lack of dentine or
abnormal formation of it. Both abnormal
bone and tooth formation are reversed by
vitamin A administration.

Deficiency Signs
1. On general growth – Failure of growth.
2. On eye – Night blindness (Nyctalopia: the
individuals have difficulty to see in dim
56 Medical Biochemistry for Physiotherapy Students
light since the dark adoptation time is
affected) degeneration of lacrimal glands,
redness, abnormally dry and lusterless
condition of the eye-ball (xeropthalmia)
with consequent keratinisation and
degeneration of cornea (keratomalacia).
3. On epithelial tissues – The epithelium of the
respiratory passages becomes stratified
and degenerates.
4. On bones – abnormal bone growth occurs
in certain parts of vertebral column and
skull.

Hypervitaminosis A
In growing rats, large doses (4,000 units/day)
cause loss of weight, atrophy of skin, loss of cholesterol in plants and animals respectively.
hair, ulceration in the eyes, haemorrhage, In plants, the irradiation of ergosterol leads
decalcification of bones with spontaneous to the production of ergocalciferol (vitamin
fracture and lowering of plasma prothrombin. D 2 ). In animals, it is converted to chole-
In man, effects include drowsiness, calciferol (vitamin D3) by irradiation of skin.
sluggishness, severe headache, vomiting and
Sources
peeling of skin of mouth.
Rich sources are liver and viscera of fish and
VITAMIN D the liver of animals. Good sources are eggs
and butter. In body, it is also obtained by the
Chemistry photolysis of 7-dehydrocholesterol in skin.
Vitamin D is a group of sterol compounds
that occur in nature chiefly in animals but also Daily Requirement
in plants and yeasts. It includes vitamin D2 The RDA for adults is about 400 I.U. One
and vitamin D3. international unit is defined as the biologic
The D vitamins are generated from the activity of 0.025 μg of pure crystalline vitamin
provitamin ergosterol and 7-dehydro- D3.

Active form
Calcitriol (1,25-DHCL) calcidiol 25-hydroxylase
Cholecalciferol 25 HCC (Calcidiol)
CD3, calcidiol α-hydroxylase (kidney)

1,25 DHCC
calcitriol
Vitamins 57
Absorption and Storage 50-60 percent in growing children and 30-40
percent in adults, decreases to a lower value
The vitamin D is readily absorbed in small
in the case of rickets.
intestines. After absorption, the vitamin is
stored largely in the liver, kidneys, intestines, Osteomalacia
adrenal glands and bones.
In this, the action of bones becomes softer
Functions than the rachitic bones. The loss of calcium is
greater than that of phosphorous. The disease
1. Vitamin D binds to the chromatin of target
is prevalent in such countries particularly in
tissue and expresses the genes for calcium
women because of the custom that keeps them
binding protein as well as Ca2+ ATPase in
intestinal cells. This increases the Ca 2+ indoor and also prevents them from exposure
absorption by actively transporting Ca2+ to sunlight. In both the conditions, there is a
across the plasma membrane against decrease in serum calcium level alongwith the
electrochemical gradients. increased urinary excretion of phosphorous.
2. It increases the reabsorption of calcium Besides there is also an increase in serum
and excretion of phosphorous in kidneys. alkaline phosphatase in these two conditions.
3. It promotes bone resorption and calcium
Hypervitaminosis D
mobilization to raise the levels of Ca and
P in blood in association with PTH. Extremely large doses of vitamin D have been
4. It lowers the pH in certain parts of the reported to cause
gut such as colon and produces increase - loss of weight
in urinary pH. - reduced excretion of calcium and phos-
phate
Deficiency - increased blood calcium
- nausea
The deficiency of vitamin D leads to rickets
- vomiting
in children and osteomalacia in adults.
- headache
- drowsiness
Rickets
- diarrhoea
It is primarily a disease of growing bones. In Extensive deposits of calcium are found
this, the deposition of inorganic materials on in the soft regions which are not normally
the matrix of bones fails to occur, although calcified such as kidney, heart, artery etc.
matrix formation continues. As a result, bones
may become soft and flexible leading to VITAMIN E
enlarged skulls, swollen joints, knock-knees
or bow legs, beaded ribs and protruding Chemistry
chest (pigeon breast). Dentition becomes Vitamin E refers to a group of compounds
delayed in rachitic children. The first tooth known as tocopherols. Four unsaturated
in such babies appears between 6th and 9th alcohols, i.e. α, β, γ and δ- tocopherols occur
month at which time it has appeared in half in nature. These tocopherols differ slightly
of the normal babies. The product of Ca and in structure in their side chain; α- tocopherol
P of normal blood which is usually around is most potent of them.
58 Medical Biochemistry for Physiotherapy Students

Sources 3. It protects RBC from haemolysis by oxidi-


zing agents (e.g. hydrogen peroxide).
Tocopherols are present in small quantities
4. It is closely associated with reproductive
in green leafy vegetables (spinach and lettuce),
functions and prevents sterility. It pres-
milk and milk products and egg yolk. Oils
erves and maintains germinal epithelium
such as corn oil, cotton seed oil and sunflower
of gonads for proper reproductive func-
oil are the richest sources.
tion.
5. It is required for cellular respiration-
Daily Requirement
through electron transport chain.
Vitamin E intake is related to the intake of 6. It prevents rancidity.
polyunsaturated fatty acids, i.e. 0.4 mg/g of
polyunsaturated fatty acids. Deficiency Disease

Absorption Deficiency of vitamin E can be seen in children


with fat malabsorption. Sprue, creatinuria,
Vitamin E is absorbed along with fat in the peptic ulceration, abnormal red cell haemo-
small intestine. Bile salts are necessary for the lysis and their diminished life span are some
absorption. In the liver, it is incorporated into of the findings in vitamin E deficiency in
lipoproteins (VLDL and LDL) and trans- humans.
ported. Vitamin E is stored in adipose tissue, The characteristic symptoms of experi-
liver and muscle. The normal plasma level of mentally induced vitamin E deficiency vary
tocopherol is less than 1 mg/dl. from animal to animal. In mature female rats,
sterility develops because of resorption of
Functions foetus after conception while in males the
1. Tocopherols act as powerful antioxidants germinal epithelium of the testes degenerates
by preventing the auto oxidation of and the spermatozoa becomes non-motile.
vitamin A and carotenes.
2. It protects the lipids of biological mem- VITAMIN K
branes against oxygen by acting as anti-
Chemistry
oxidants (i.e. prevent the peroxidation of
polyunsaturated fatty acids that occur in The name vitamin K has been derived from
membranes throughout the body). the Danish word Koagulation because of its
Vitamins 59
important role in blood clotting. Various 2. It is involved in oxidative processes taking
compounds related to 2- methyl-1, 4- naptho- place in photosynthesis of plant kingdom.
quinone have been described to have vitamin 3. It is involved in electron transport chain
K activity. and is involved in oxidative phosphory-
The most important form which occurs lation.
naturally is vitamin K1 (Phylloquinone). It 4. Vitamin K is also required for carbo-
was isolated from plants and found to have xylation of glutamic acid residues of
phytyl side chain. The other forms include osteocalcin, a calcium binding protein
vitamin K2 (menaquinone) which is synthe- present in the bone.
sized by the intestinal flora and is found in
animal tissues. It has a side chain with 7 Deficiency Diseases
isoprenoid units. Vitamin K3 (menadione) is
a synthetic compound available commercially 1. The deficiency of vitamin K causes loss of
for therapeutic uses. blood clotting power. The infants may
show signs of vitamin K deficiency by
Sources developing haemorrhage.
2. Vitamin K deficiency causes haemorragic
Cabbage, cauliflower, tomatoes, alfa alfa,
disease of the new born.
spinach and other green leafy vegetables are
3. Vitamin K deficiency is caused by fat
good sources. It is also present in egg yolk,
malabsorption which may be associated
meat, liver, cheese and dairy products. It is
with pancreatic dysfunction, biliary disease
also synthesized by intestinal flora.
and atrophy of the intestinal mucosa.
Daily Requirement 4. Sterilization of the large intestine by
antibiotics can result in deficiency when
Dietary requirement of vitamin K is not of dietary intake is limited.
much importance under normal circumstances 5. Short circuiting of the bowel as a result of
as it is provided by intestinal bacteria in surgery may also cause deficiency which
adequate quantities. The suggested RDA may not respond even to large doses of
(Recommended Dietary Allowance) for an
vitamin K. Water soluble form of vitamin
adult is 70-140 μg/day.
K, i.e. K3 is alone useful in such cases.
Absorption and Storage
Hypervitaminosis K
Vitamin K is taken in the diet or synthesized
The parenteral administration of too large
by the intestinal bacteria. Its absorption takes
doses of vitamin K (e.g. 30 mg/ day for 3 days)
place along with fat (chylomicrons) and is
to infants has been shown to produce hyper-
dependent on bile salts. Vitamin K is transpor-
bilirubinaemia, i.e. jaundice in some cases. This
ted along with LDL and is stored mainly in
liver and to a lesser extent in other tissues. is due to increased breakdown of RBC.

Functions VITAMIN C

1. Vitamin K helps in the synthesis of Chemistry


prothrombin. It is concerned with blood Vitamin C, which is hydrophilic, acts as an
clotting factor. antioxidant in solution. Vitamin C is a powerful
60 Medical Biochemistry for Physiotherapy Students
reducing agent and is oxidized to dehydro- 2. It is required for functional activities of
ascorbic acid. Both forms are biologically fibroblasts and osteoblasts and conse-
active. It is stable in acidic solution at low quently for formation of Mucopoly-
temperatures but undergoes destruction in saccharides (MPS) of connective tissues,
alkaline solution when in contact with air. osteoid tissues, dentine and intercellular
cement substance of capillaries.
Sources 3. It has a role in the conversion of folic acid
to a physiologically active form, tetra-
It is widely distributed in citrus fruits, green
hydrofolic acid.
chillies, guava and tomatoes which are good 4. It helps in the absorption of Fe by conver-
sources whereas amla is the richest source of ting the inorganic ferric iron to the ferrous
vitamin C. form.
5. It is capable of both activating and
Daily Requirement
inhibiting different groups of enzymes.
About 60-70 mg vitamin C per day will meet Arginase and papain are activated whereas
the requirement. Additional intakes (20- 40% urease is inhibited.
increase) are recommended for women 6. It is required as a co-factor for hydroxy-
during pregnancy and lactation. lation of tryptophan to form 5- OH deri-
vative, in the pathway of biosynthesis of
Biosynthesis and Metabolism serotonin.
7. It is involved in electron transport in the
Many animals can synthesize ascorbic acid
microsomal fraction.
from glucose via uronic acid pathway.
However, man, other primates, guinea pigs Deficiency Diseases
and bats cannot synthesize ascorbic acid due The prolonged deficiency of vitamin C causes
to the deficiency of a single enzyme namely scurvy which is characterized by multiple
L-gulonolactone oxidase. haemorrhages. Early symptoms include
Vitamin C is rapidly absorbed from the loosening of teeth and joints pain. In severe
intestine. It is not stored in the body to a cases, there may be bleeding from the nose,
significant extent. Ascorbic acid is excreted GIT or genitourinary tract. Wound healing is
in urine as such or as its metabolites – delayed due to deficient formation of
diketogulonic acid and oxalic acid. collagen. Gums are swollen and becomes
spongy and bleeds on slightest pressure.
Functions Osteoid of bone is poorly laid and minerali-
1. The main function of vitamin C is the zation of bone is poor. The bones are weak
formation of tissue collagen or intracellular and readily fractures.
cement substance. This is because vitamin Scurvy may also be observed in infants
C is essential to the activity of the enzyme and called as infantile scurvy. Infants lose
collagen proline hydroxylase, which appetite and weight with painful tenderness
catalyzes the conversion of proline to of the extremities, bleeding from gums and
hydroxyproline. Hydroxyproline is found mucous membrane. Long bones show cessa-
exclusively in collagen and is vital in tion of osteogenesis. Because of the role of
maintaining the tertiary structure of this vitamin C in absorption, transport and
major vertebrate protein, i.e. collagen. storage of iron its deficiency also leads to
Vitamins 61
anaemia which is of hypochromic microcytic Functions
type.
Thiamine pyrophosphate (TPP) is used mainly
as a coenzyme in the carbohydrate metabolism.
THIAMINE 1. In oxidative decarboxylation of α-keto-
It is also known as Vitamin B1, anti-beriberi acids: Pyruvic acid and α-ketoglutaric acid
factor. which are intermediates in the carbo-
hydrate metabolism are oxidatively
Chemistry decarboxylated to acetyl CoA and succinyl
Thiamine consists of a substituted pyrimidine CoA respectively.
ring joined by a methylene bridge to substi- Pyruvic acid ———> Acetyl CoA
α-Ketoglutaric acid ———> Succinyl CoA
tuted thiazole ring.
2. In transketolation reaction: An inter-
Thiamine is water soluble. It is thermo-
mediate step in Hexose monophosphate
labile, destroyed in alkaline medium but shunt pathway where transfer of
thermostable in acidic medium. glycolaldehyde group from D-xylulose-5
phosphate and glyceraldehyde.
D-xylulose-5-PO4 ———> D-Sedoheptu-
lose-PO,
+ +
D-Ribose-5-PO4 ———> D-glyceraldehyde-
Thiamine occurs in the cells largely as its PO4
active coenzyme form, i.e. thiamine pyro-
3. In yeast: In yeast TPP acts as coenzyme
phosphate (TPP), also called cocarboxylase.
for nonoxidative decarboxylation of α-
TPP is the biologically active form, formed
Keto acids, i.e. pyruvate to CO 2 and
by the transfer of pyrophosphate group from
acetaldehyde.
ATP to thiamine.
Sources Deficiency
Cereals, pulses, oil seeds, nuts and yeast are Deficiency gives rise to a disease known as
good sources. Thiamine is mostly concentrated beriberi. It is found in areas where polished
in the outer layer (bran) of cereals. Polishing rice is the major component of the diet.
of rice removes about 80% of thiamine. Vitamin It is a disease of the nervous system and
B1 is also present in animal foods like pork, is characterized by polyneuritis (degeneration
liver, heart, kidney, milk, etc. of the peripheral nerves), muscular atrophy,
cardiovascular changes and gastrointestinal
Daily Requirement disorders. At first, there is weakness and
The daily requirement of thiamine depends fatigue. This is followed by symptoms like
on the intake of carbohydrate. A dietary anorexia, insomnia, anaesthesia and tachy-
supply of 1-1.5 mg/day is recommended for cardia. Finally the major symptoms may
adults. For children the RDA is 0.7-1.2 mg/ follow one of the following three courses (and
day. The requirement marginally increases accordingly beriberi is of three types):
in pregnancy, lactation, old age and - symptoms involving nervous system (dry
alcoholism. beriberi).
62 Medical Biochemistry for Physiotherapy Students
- symptoms associated with oedema and riboflavin is flavin adenine dinucleotide
effusions (wet beriberi). (FAD) which is a dinucleotide of FMN and
- symptoms involving heart (acute perni- AMP.
cious anaemia). The two coenzymes form an integral part
of flavoprotein containing enzymes and are
RIBOFLAVIN important in oxidation-reduction reactions
It is also known as vitamin B2. or lactoflavin. requiring oxidoreductases. Isoalloxazine ring
serves as the acceptor of two hydrogens and
Chemistry the flavoproteins inturn get reduced.

Riboflavin consists of a heterotricyclic


structure (isoalloxazine) to which is attached
ribitol. The ring structure is conjugated; thus,
riboflavin is a coloured and fluorescent
pigment. It is relatively heat stable but
sensitive to irreversible decomposition upon FMN is an important constituent of
exposure to visible light. cytochrome-C-reductase and L-amino acid
oxidases. FAD is used as a coenzyme with
glycine oxidase, xanthine oxidase and D-amino
acid oxidases. FAD also forms an integral part
of the acyl CoA dehydrogenase and is thus
important for the oxidation of fatty acids.

Deficiency Symptoms
Deficiency of riboflavin is not common but is
associated with the deficiency of iron and
with pellagra. Deficiency symptoms include
the lesions of lips, cheilosis (fissures of the
Sources angles of the mouth), glossitis (the tongue
Milk, liver, kidney and heart are rich sources. appearing smooth and purplish), localised
Green leafy vegetables are good sources. dermatitis of face besides functional and
organic disorders of the eyes such as photo-
Daily Requirement phobia, burning and itching of the eyes.
The average daily requirement is about 1-2
mg/day. NIACIN
Functions Niacin is also known as “Pellagra Preventive
Riboflavin with phosphoric acid forms flavin Factor” because of its role in the prevention
mononucleotide (FMN) which is a coenzyme of pellagra, a disease mainly affecting the
of the vitamin. The other coenzyme of skin.
Vitamins 63
Chemistry 3. The conversion of 3-OH butyrate to
acetataoacetate is NAD dependent.
Niacin is the generic name given to the two 4. NADP + is used as a coenzyme with
compounds, i.e. nicotinic acid and its amide isocitrate dehydrogenase in the conver-
called nicotinamide. Both these compounds sion of isocitrate to oxalosuccinate.
contain a pyridine ring system. Niacin and 5. NADP+ is used as a coenzyme with glu-
niacinamide are white crystalline, water cose-6-phosphate dehydrogenase which
soluble compounds. converts glucose-6-phosphate to 6-phos-
phogluconolactone.

Deficiency
Deficiency occurs mainly in areas where
maize is the chief constituent of the diet
because niacin in maize is in an unusable form.
Deficiency causes pellagra. Pellagra develops
Sources within 6 to 8 weeks of severe deficiency. It is
characterized by 3D’s which are dermatitis,
Niacin is found in liver, cheese, whole cereals, diarrhoea and dementia:
eggs, fish are best sources whereas wheat,
vegetables and rice bran are moderate Clinical Feature
sources. The body can synthesize it from the - Redness of the skin in parts exposed to
amino acid trytophan. light, especially in the neck.
- Anorexia, nausea, dysphagia and infla-
Daily Requirement mmation of the lining of the mouth.
- Delirium, mental disturbance and demen-
The daily requirement is about 12 to 17 mg.
tia.
Functions PYRIDOXINE
NAD and NADP are the coenzyme form of It is also known as vitamin B6.
niacin. These are involved in a variety of
Chemistry
oxidation-reduction reactions.
Pyridoxine, pyridoxal and pyridoxamine are
1. It acts as a coenzyme for the enzyme
all interconvertible in the body and can act
lactate dehydrogenase (LDH) which
as vitamin B6.
converts lactate to pyruvate.

2. It acts as a coenzyme for the enzyme pyru- These all are water soluble. They are also
vate dehydrogenase complex. This soluble in alcohol and slightly soluble in
enzyme converts pyruvate to acetyl CoA. certain fat solvents.
64 Medical Biochemistry for Physiotherapy Students
Sources
Egg yolk, fish, meat, milk are the richest
sources. Vegetables like cabbage and legumes
and foods such as whole grains, etc. contain
moderate amounts of this vitamin.

Daily Requirement 5. It is required as a coenzyme for activation


The daily requirement is about 1.2 to 1.4 mg. of serine which is required for synthesis
of sphingomyelin.
Functions 6. It is required for active transport of amino
acids through cell membrane and intestinal
Pyridoxal phosphate acts as a coenzyme – it absorption of amino acids.
is mainly involved with metabolism of amino
acids. Deficiency
1. It acts as a coenzyme for the enzyme
transaminases in transamination reactions. In many species, pyridoxine deficiency causes
2. It acts as a coenzyme for the enzyme a hypochromic microcytic anaemia and
decarboxylases in decarboxylation reac- retardation of growth.
tion. Amino acids are decarboxylated to In man, the deficiency of pyridoxine gives
form corresponding amines, e.g. conver- rise to certain symptoms gradually. The first
sion of histidine to histamine and gluta- effect is a fall in haemoglobin level and
mate to GABA. anaemia followed by lethargy, depression
and mental confusion. Unlike adults, the
deficiency symptoms appear quickly in
infants, when pyridoxine is lacking in their
milk. General irritability, vomiting, diarrhoea
and convulsions are observed.
The symptoms of pyridoxine deficiency
have been observed in patients receiving
isonicotinic acid hydrazide (INH) in the
treatment of tuberculosis. This is due to the
3. In tryptophan metabolism, pyridoxal-P antagonistic action of INH to pyridoxine as
acts as a coenzyme for the enzyme they are structural analogues.
kynureninase which converts 3-OH
kyunurenine to 3-OH anthranillic acid PANTOTHENIC ACID
which ultimately forms nicotinic acid.
Chemistry
Thus, in vitamin B 6 deficiency, niacin
synthesis from tryptophan does not take Pantothenic acid is an amide of pantoic acid
place. and β- alanine. It is a yellow viscous oil. It is
4. It is required for the synthesis of δ-amino destroyed by heat in dry conditions. It is
levulinic acid, the precursor for heme soluble in water and when food is cooked
synthesis. and the water discarded, some vitamin is lost.
Vitamins 65
d. The oxidation of fatty acids requires
coenzyme A at the initial step of activation
of fatty acyl CoA.
e. CoA is also required for the activation of
succinic acid, used in the biosynthesis of
heme.
Sources f. It also forms the prosthetic group of acyl
carrier protein which participates in fatty
Pantothenic acid occurs in all animal and plant acid synthesis.
tissues. Rich sources are yeast, liver, eggs,
potatoes, whole cereals and legumes. Deficiency

Daily Requirement No definite lesions due to the deficiency of


panthothenic acid had been observed in man.
The daily requirement is about 3-7mg. As pantothenic acid is universally present in
almost all types of foodstuffs, deficiency
Functions diseases associated with the lack of this vitamin
Pantothenic acid functions in a number of are not manifested. Pantothenic acid deficiency
biochemical reactions in the form of its symptoms includes headache, fatigue, impai-
red motor activity, muscle cramps and
coenzyme, i.e. coenzyme (CoA).
gastrointestinal disturbances. It has also been
a. It is involved in the conversion of pyruvic
used in the burning feet syndrome and in the
acid to acetyl CoA (active acetate) which
healing of ulcers and bed sores.
is used in the synthesis of citrate and
enters citric acid cycle. BIOTIN

Chemistry
It consists of a fused imidazole and thiophene
ring with a fatty acid chain. Two forms of
biotin exist, allobiotin and epibiotin. Biotin is
This acetyl CoA is used in the synthesis of optically active. It is soluble in water and ethyl
acetylcholine, a chemical transmitter at the alcohol but is insoluble in chloroform and
nerve synapse. ether. It is heat stable and is resistant to both
acids and alkalies.

b. It is involved in the conversion of α-keto


glutaric acid to succinyl CoA.
c. Acetyl CoA is also a precursor of choles-
terol and steroid biosynthesis.
66 Medical Biochemistry for Physiotherapy Students
Sources Deficiency
Biotin is widely distributed in both animal In most animals including man, intestinal
and plant sources. Animal foods like liver, bacteria synthesise appreciable amounts of
kidney and milk are very good sources of biotin. It is because of this reason that biotin
this vitamin. Vegetables contains moderate deficiency in human beings, fed on biotin free
amounts, tomatoes and yeast being good diets, cannot be produced. However, biotin
sources of this vitamin. deficiency may be induced by sterilization of
intestine and by feeding with raw egg white.
Daily Requirement Avidin, the egg white protein, inactivates
The average daily requirement is about 3 to biotin by eliminating it from an otherwise
7 mg. complete diet. Such a deficiency in man leads
to dermatitis, loss of hair, decrease in weight
Functions and oedema. The lesions on skin appear with
Biotin serves as a carrier of CO2 in carboxy- changes in posture and gait.
lation reactions.
1. The conversion of pyruvate to oxalo- FOLIC ACID
acetate by biotin dependent pyruvate
carboxylase is essential for the synthesis Chemistry
of glucose from many non carbohydrate It is also known as pteroyl glutamic acid. It is
sources.
composed of pterin ring attached to p-
2. The first reaction for the synthesis of fatty
aminobenzoic acid (PABA) and conjugated
acids, i.e. the conversion of acetyl CoA to
with one or more glutamic acid residues.
malonyl CoA requires biotin as a co-
Humans cannot synthesize PABA or attach
enzyme.
the first glutamic acid. Folic acid is a yellow
3. Propionyl CoA is produced in the meta-
bolism of certain amino acids (valine, substance, slightly soluble in water but
isoleucine, threonine, etc.) and degrada- insoluble in fat solvents.
tion of odd chain fatty acids. Its further
metabolism is dependent on biotin. Sources
4. In the transcarboxylation reactions, in the It is found in liver, kidney, fresh leafy vege-
catabolism of branched chain amino acids. tables and yeast. It is also synthesized by
bacteria in the large intestine.

Daily Requirement
The daily requirement is about 200 μg.
Vitamins 67
Functions Deficiency
Folic acid is reduced to give its coenzyme Folic acid deficiency is not generally observed
form, i.e. tetrahydrofolic acid (FH4). as it is synthesized by the intestinal flora but
may occur in pregnancy, intestinal mal-
absorption or on antibiotics therapy. Its
deficiency leads to megaloblastic anaemia,
glossitis and gastrointestinal disorders.

CYANOCOBALAMIN

Chemistry
It is also known as vitamin B 12. or anti-
Folic acid is then converted to N 5 – pernicious anaemia factor. It is also called as
formyltetrahydrofolic acid (N5-CHO. FH4) or cyanocobalamin due to the presence of a
folinic acid in the liver and is thus used in the cyano group and a cobalt atom. The central
transfer of one carbon groups (one carbon ring structure of vitamin B12 is a corrin ring
moiety) from a donor to an acceptor molecule. system similar to porphyrins but with a
Besides formyl (-CHO), one carbon moiety central cobalt atom linked to a cyano group
may be in the form of (-HCOO-), methenyl and to a nitrogen of the imidazole group of
(=CH-), methylene (-CH2-), methyl (-CH3),
the nucleotide which is esterified through
hydroxymethyl (-CH 2 OH) or formimino
aminopropanol and propionic acid to the ring
(-CH = NH-) groups. All the forms are
IV of the corrin ring.
interconvertible by the NADP + or NAD +
The cyano group may be replaced by
dependent dehydrogenases.
nitrate (-NO2) or hydroxyl (-OH) group to
The sources of one carbon moiety may be
give nitritocobalamin or hydroxocobalamin
histidine releasing formimino group,
which are haemopoetically active.
β-carbon of serine giving hydroxymethyl
Vitamin B12 is absorbed from the gastro-
group or glycine and methionine donating a
intestinal tract in the presence of a constituent
methyl group.
of gastric juice (intrinsic factor) and is stored
One carbon moiety carried by FH 4 is
utilised in many important reactions, such as in the liver.
a source of carbon at positions 2 and 8 in the
Sources
biosynthesis of a purine ring, in the conver-
sion of glycine to serine, uracil to thymine Cyanocobalamin is almost absent in plant
homocysteine to methionine and in the materials. Animal tissues are good sources
synthesis of choline. In some of these of this vitamin. Egg yolk and milk contain
reactions, vitamin B12 is also required. adequate amounts.
68 Medical Biochemistry for Physiotherapy Students
Daily Requirement Deficiency
The daily requirement is about 3 μg. A nutritional deficiency of this vitamin is
usually not observed on account of its
Functions
widespread nature in foodstuffs. However,
Methylcobalamin and deoxyadenosylco- deficiency may be observed in individuals
balamin, both act as vitamin B12 coenzymes who abstain from all animal products. The
and are called cobamide coenzymes. These deficiency disease, pernicious anaemia, is
have several important roles in different
characterized by RBC’s becoming abnormally
metabolic processes in the body.
large and fewer in number (1-3 million/cu.mm
a. Conversion of methylmalonyl CoA to
instead of the normal 4-5 million/cu.mm). The
succinyl CoA.
b. Methylation of homocysteine to form patient weakens, loses weight and the
methionine. nervous system is also gradually affected
c. Isomerization of dicarboxylic acids, e.g. because there occurs demyelinization of the
glutamic acid into β-methyl-aspartic acid. large nerve fibres of the spinal cord.
Chapter 9

Bioenergetics

Bioenergetics describes the transfer and between the reactants and products. It is
utilization of energy in biologic systems. It represented as ΔG.
makes use of a concept of free energy change. Free energy is related to heat content (H)
Changes in free energy (ΔG) provide a and entropy (S) by the following equation:
measure of the energetic feasibility of a ΔG = ΔH - TΔS.
chemical reaction and can therefore allow Enthalpy (H) which is a measure of the
prediction of whether a reaction will take heat content represents the entire chemical
place. The free energy concept is essential for energy. Entropy (S) represents the disorder
understanding the unique role of adenosine or state of randomness in the molecule. Thus,
triphosphate (ATP) plays in transferring the free energy is not the entire chemical
energy from energy yielding catabolic energy contained, but the chemical energy
processes to energy requiring reactions. less the entropy which is influenced by
temperature (T).
Concept of Energy The change in free energy, ΔG can be used
Energy is defined as the capacity to do work. to predict the direction of a reaction at constant
Energy exists in a variety of forms and these temperature and pressure. If G is negative,
forms are inter-convertible. there is a net loss of energy. The reaction is
said to be exergonic. If G is positive, there is a
Free Energy net gain of energy. The reaction is said to be
Any reaction will be spontaneous and endergonic and energy must be added to the
exergonic, if it is accompanied by a decrease system to make the reaction go on.
of free energy. Free energy is the actual
available energy, i.e. the chemical energy ATP-energy Rich Compound
capable of causing chemical reactions. It is ATP consists of a molecule of adenosine to
represented as difference in free energy which three phosphate groups are attached.
70 Medical Biochemistry for Physiotherapy Students
If one phosphate is removed, ADP is Most biological oxidations are only biolo-
produced; if two phosphates are removed, gical dehydrogenations in which hydrogen
AMP results. The free energy of hydrolysis is removed from the metabolites.
of ATP is approx –7300 cal/mol for each of Biological oxidations are brought about
the two terminal phosphate groups. Because with different enzymes, coenzymes and
of this large negative ΔG, ATP is called a high electron carriers which takes place in the
energy phosphate compound. mitochondria of the cells.
The different enzymes associated are
BIOLOGICAL OXIDATIONS a. Oxidases
Biological oxidations are oxidations taking b. Aerobic dehydrogenases
place in the living systems. During biological c. Anaerobic dehydrogenases
oxidations, the reacting chemical systems d. Hydroperoxidases
move from a higher energy level to a lower e. Oxygenases
one and hence there is liberation of energy.
The oxidations are therefore exergonic. The The different coenzymes involved are
energy liberated as heat is converted to a. NAD+
chemical energy by the formation of ATP
b. NADP+
which is one of the important energy-rich
c. FMN
compounds. The formation of ATP from ADP
and Pi is termed phosphorylation and as it is d. FAD
concurrent with oxidation, the process is e. ETF (electron transferring flavoprotein)
known as oxidative phosphorylation. The f. CoQ (coenzyme Q)
energy of oxidations trapped in ATP is used g. Lipoate
for synthetic reactions, muscular contraction,
nerve conduction, active transport and other The electron carriers involved are
processes that require energy (endergonic a. cytochrome b
processes). b. cyt c1
Oxidation is defined as addition of c. cyt c
oxygen, removal of hydrogen or removal of d. cyt a
electrons. e. non-heme iron (NHI)

Oxidases
Oxidases are enzymes that catalyze the
removal of hydrogen from a substrate but
use only oxygen as a hydrogen acceptor. They
form water as a reaction product. Uricase and
monoamine oxidase enzymes are exceptions
which form H2O2 as a product.
Bioenergetics 71

Hydroperoxidases
Aerobic Dehydrogenases Hydroperoxidase are enzymes that uses
hydrogen peroxide as a substrate. Examples
Aerobic dehydrogenases are enzymes that
include peroxidase which is found in milk,
catalyze removal of hydrogen from a
plants, leukocytes and erythrocytes and
substrate. The hydrogen can be given to
catalase which is found in animals and plants.
molecular oxygen or artificial acceptors like
methylene blue. Hydrogen peroxide is
formed as a product.

Oxygenases
Oxygenases are enzymes which catalyze the
Examples include L-amino acid dehydro- incorporation of oxygen into a substrate
genase, glucose oxidase, xanthine dehydro- molecule. These have two subclasses.
genase, etc. 1. Dioxygenases: These catalyse the incorpo-
ration of 2-atoms of oxygen into the
Anaerobic Dehydrogenases substrate.
A + O2 ———>AO2
Anaerobic dehydrogenases are enzymes Examples includes tryptophan dioxygenase,
which catalyze the removal of hydrogen from homogentisate dioxygenase, etc.
a substrate but not able to use oxygen as
hydrogen accepter. These are classified as:
a. Pyridine nucleotides: Under this group
comes coenzyme NAD+ and NADP+. The 2. Monoxygenases: These catalyse the incor-
effective part which participates in the poration of only 1- atom of oxygen into
reaction is nicotinamide. the substrate. The other oxygen atom is
b. Flavonucleotides: These are FMN and FAD. removed as water. A cosubstrate is
The effective part which takes part in the required for this purpose.
reaction is riboflavin. A–H + O2 + ZH2 ———> A–OH + H2O + Z
c. Cytochromes: The cytochromes are iron- Monoxygenases
containing hemoproteins and include cyt. Examples include phenylalanine hydroxy-
b, c1, c, a etc. lase, tyrosinase, etc.
72 Medical Biochemistry for Physiotherapy Students
ELECTRON TRANSPORT CHAIN proteins and cytochromes, to molecular
oxygen on other.
Energy rich molecules like glucose or fatty Not all substrates are linked to the
acids are metabolized by a series of oxidation respiratory chain through NAD + linked
reactions ultimately yielding CO2 and water.
dehyrogenases. Some are linked directly to
The metabolic intermediates of these reactions
flavoprotein dehydrogenases which are then
donate electrons to specialized coenzymes
linked to cytochromes.
NAD+ and FAD to form energy rich reduced
coenzymes NADH and FADH 2 . These
reduced coenzymes can in turn donate a pair
of electrons to a specialized set of electron
carriers collectively called the electron
transport chain or respiratory chain. As
electrons are passed down the electron
transport chain, they lose much of their free
energy. Part of this energy can be captured
and stored by the production of ATP from
ADP and inorganic phosphate. The remainder
β-hydroxy acyl CoA, β-hydroxy butyrate,
of the free energy not trapped as ATP is
malate, isocitrate, glutamate, 3-phospho-
released as heat.
glyceraldehyde dehydrogenases are all
NAD+ dependent. The hydrogen atoms are
Location
directly taken up by NAD+. Pyruvate and α-
The electron transport chain is present in the ketoglutarate dehydrogenases are oxidsed by
inner mitochondrial membrane and is the final lipoate, Fp combination and subsequently
common pathway by which electrons derived NAD+ followed by flavin system.
from different fuels of the body flow to On the other hand, succinate and α-
oxygen. Electron transport and ATP synthesis glycerophosphate do not depend on NAD+ .
by oxidative phosphorylation proceed conti- They are oxidised by flavoprotein system
nuously in all cells of the body that contain which contains non heme iron and shunted
mitochondria. on to CoQ system. While acyl CoA also does
not require NAD +, it requires in addition
Reactions
to flavoprotein (FAD) system, another
The main respiratory chain in mitochondria flavoprotein, i.e. electron transferring
proceeds from the NAD+ linked dehydro- flavoprotein (ETF) and the relay is continued
genase systems on one hand, through flavo- by CoQ.
Bioenergetics 73
OXIDATIVE PHOSPHORYLATION

The term oxidative phosphorylation is a


descriptive blend of the two processes
operating simultaneously, i.e. the oxidation
of a metabolite by oxygen via electron
transport chain and the phosphorylation of
ADP. The necessary energy is trapped gene-
rated by the flow of electrons from a
substrate molecule undergoing oxidation. The
chain functions within the cell mitochondria
and liberates energy, a large amount of which
is used to form ATP from ADP and phos-
CoQ is the collecting point in the respi- phate. This is the major route for ATP pro-
ratory chain for reducing equivalents derived duction.
from other substrates. Electrons flow from
Q through the series of cytochromes (cyt b,
P:O Ratio
c1, c, a, a3) and molecular oxygen at the last
stage to form water. The P:O ratio refers to the atoms of phosphate
utilized to the atoms of oxygen consumed in
Inhibitors of the ETC oxidation or the number of molecules of ATP
The inhibitors of the respiratory chain act at synthesized per pair of electrons carried
sites I, II and III of the chain. through ETC.
Site I: (NAD ——> Fp step) is inhibited by The mitochondrial oxidation of NADH
barbiturates, piercidin A (antibiotic), rotenone has a P:O ratio of 3 whereas oxidation of
(fish poison), some steroid drugs and mercu- FADH2 is 2.
rials.
Site II: (cyt b ——> cyt c1 step) is inhibited Sites of Oxidative Phosphorylation
by dimercaprol and antimycin A.
Site III: (at cyt a3 step): is inhibited by CN, The P:O ratio of 3 for NADH oxidation
CO, H2S and azide. indicates that there are three reactions in the
ETC that are exergonic to result in the
synthesis of 3 ATP molecules. The three sites
are
a. Oxidation of FMNH2 by CoQ
b. Oxidation of cyt b by cyt c1
c. cyt oxidase reaction
There are only two sites for oxidation of
FADH2 since the first site is bypassed.
Chapter 10

Digestion and Absorption

The term digestion may be defined as the DIGESTION AND ABSORPTION OF


process of biochemical transformation of CARBOHYDRATES
complex and larger food particles in the gut
enzymatically into a simple form suitable for Different forms of carbohydrates which are
generally included in the diet are:
absorption and assimilation, the complex and
1. Polysaccharides
higher molecules being unsuitable for
2. Oligosaccharides
absorption.
3. Disaccharides (Maltose, lactose, sucrose)
The digestive system consists of gastro-
4. Monosaccharides (glucose, fructose).
intestinal tract (GIT) and accessory organs.
The GIT consists of mouth, pharynx, oeso-
Monosaccharides need no further diges-
phagus, stomach, small intestine (duodenum,
tion because all carbohydrates are absorbed
jejunum, and ileum), large intestine (caecum,
in the form of monosaccharides.
ascending colon, transverse colon, des-
Cellulose cannot be appreciably digested
cending colon, and sigmoid colon), rectum
in the human alimentary canal. Digestion of
and anal canal. Accessory organs include polysaccharides and disaccharides starts in
tongue, teeth, liver, gall bladder, pancreas, the saliva and is completed in the succus
etc. entericus, i.e. intestinal juice.
Digestion of disaccharides chiefly takes
Functions of the Digestive System place in the intestinal juice but may take place
1. Ingestion of food. to a slight extent by other digestive juices.
2. Digestion of food.
In the Saliva (mouth)
3. Secretion of various digestive juices.
4. Absorption of water, salts, vitamins and Saliva contains
end products of food digestion. a. Chiefly salivary amylase or ptyalin
5. Excretion of heavy metals, toxins etc. b. Traces of maltase
Digestion and Absorption 75
Salivary amylase acts on boiled starch only. It Sucrase (invertase)
Sucrose —————————> fructose + glucose
cannot penetrate the intact cellulose covering
of the unboiled starch particle. Strong acid lactase
Lactose —————> glucose + galactose
destroys ptyalin. Effects of salts such as
maltase
chlorides are necessary for ptyalin action. Maltose —————> glucose + glucose
Ptyalin digests starch upto the maltose stage oligo –α-1,6 – glucosidase
Isomaltose ———————————> glucose + glucose
only. Ptyalin hydrolyses only α–1, 4 linkages
but not the α–1, 6 linkages.
Intestinal juice may contain traces of
In the Gastric Juice amylase. It is supposed to act on that little
quantity of starch and dextrin that might have
When the food gets mixed with the gastric escaped pancreatic digestion.
juice, the action of amylase ceases due to high
acidity. Gastric juice does not possess any ABSORPTION
carbohydrate splitting enzyme, but gastric
HCl can carry on some hydrolysis of sucrose. Absorption is a process by which the end
products of digestion pass through the
In the Pancreatic Juice intestinal epithelium and enter the blood
stream. Most absorption occurs in the small
The pancreatic juice contains two enzymes intestine where there is large surface area
acting on carbohydrates. which is provided by intestinal villi and
a. pancreatic amylase – acting on starch and microvilli.
dextrin.
b. maltase in traces – acting on maltose. General Principles of Absorption
1. Diffusion: This depends on relative
Pancreatic amylase can act on both boiled and
concentration of the substance on either
unboiled starch. Its action is much more rapid
side of a permeable membrane. The
than ptyalin. Most of the starch is converted
concentration of the end products of
into maltose within a few minutes. Salts and
digestion is much higher in the intestine
chlorides ions are essential for its action. than in blood in health. The diffusion
Pancreatic amylase is not present in the indicates diffusion currents from the lumen
pancreatic juice of infants up to the age of of the intestine into the blood stream and
about 6 months. Hence, during this period, thus helps absorption.
the baby should not be given any starchy 2. Hydrostatic pressure: The amount of
food. absorption is directly proportional to the
hydrostatic pressure in the intestine. The
pancreatic amylase
Starch and glycogen ————————>maltose + dextrin intestinal contents at the height of diges-
+ isomaltose tion exert considerable pressure which is
all the more enhanced by the tone and
In the Intestinal Juice movement of the intestine. This higher
It contains oligo-α–1,6 – glucosidase which acts pressure causes filtration of water and
on maltose, dextrin, isomaltose to form dissolved substances across intestinal
glucose. epithelium.
76 Medical Biochemistry for Physiotherapy Students
3. Osmotic pressure: The colloidal osmotic The absorption of sugar takes place at
pressure in the capillary loop of the villus different rates from the intestine indicating
is fairly high and is nearly two to three that absorption is an active process.
times higher than the blood pressure Galactose > glucose > fructose > mannose.
existing in it. This high osmotic pressure If the absorption coefficient of glucose is
helps in drawing the fluid from the lumen taken as 100, that of other monosaccharides
into blood. is as follows:
4. Adsorption: This process is also involved D – Galactose 110
in the absorption of certain substances. It D – Glucose 100
is a process by which a particular substance D – Fructose 43
while being absorbed from the intestine D – Mannose 19
can carry another substance along with it.
5. Hydrotropy: This is a process by means of Galactose and glucose with a higher rate
which substances insoluble in water are of absorption are said to be actively absor-
converted into soluble forms. In this way bed. They are transported across mucosal cell
they are brought into a suitable state for membrane by an energy dependent process.
absorption. This action is effected by bile Mannose is absorbed passively by simple
salts upon fatty acids and similar products. diffusion.
6. Passive and active transport: In the passive Rate of absorption of proximal jejunum is
transport mechanism, special physical
three times greater than that of distal ileum.
force is not required. As for instance if
No carbohydrate higher than the mono-
any particular food substance remains in
saccharide can be absorbed directly into the
higher concentration in the lumen of the
bloodstream in normal health and if adminis-
intestine, the energy for movement is
tered parenterally, they are eliminated as
derived from the higher concentration of
foreign bodies.
the food substance. In the active transport
mechanism, there is a carrier mechanism
Active Transport of Glucose
which helps in movement of the food
substance against the electric forces. ATP Active transport of glucose is by a carrier
supplies energy to this carrier system. protein sodium-dependent glucose trans-
porter (SGLT 1) which is present in the brush
ABSORPTION OF CARBOHYDRATES border of intestinal epithelial cells. The carrier
Carbohydrate digestion is complete when protein has the following characteristics—
food materials reach small intestine and all • Two binding sites, one for sodium and one
complex dietary carbohydrates like starch for glucose
and glycogen and the disaccharides are • Carrier protein is specific
ultimately converted to simpler monosaccha- • It is mobile
rides. All monosaccharides, i.e. products of • It is sodium dependent and energy
digestion of dietary carbohydrates are dependent.
practically completely absorbed almost A sodium dependent glucose transporter
entirely from the small intestine. binds both glucose and sodium at separate
Digestion and Absorption 77
sites and transports them both through the 9. Inherited deficiencies of enzymes like
plasma membrane and the intestinal cell. Both sucrase and lactase can interfere with
glucose and sodium are released into the hydrolysis of corresponding disaccharides
cytosol. The sodium is transported down its and their absorption.
concentration gradient and at the same time
causes the transporter to carry glucose against DEFECT IN DIGESTION AND ABSORPTION
its concentration gradient. The free energy OF CARBOHYDRATES
required for this active transport is obtained
The overall process of carbohydrate digestion
from the hydrolysis of ATP linked to Na+
and absorption is so efficient in healthy
pump that expels Na + from the cell in
individuals that ordinarily all digestible
exchange of K+.
dietary carbohydrate is absorbed by the time
Factors Controlling Active the ingested material reaches the lower
Transport of Glucose jejunum. However, because predominantly
monosaccharides are absorbed, any defect in
1. Active transport of sugar is depressed by a specific disaccharidase activity of the
agents like cyanide, malonate, fluoro- intestinal mucosa causes the passage of
acetate. undigested carbohydrate into the large
2. Plorihizin which interacts with the intestine. Therefore, because of the presence
membrane site at which sugar enters, of this osmotically active substance, osmotic
inhibits intestinal absorption of glucose diarrhoea results as water is drawn from the
and galactose. mucosa into the large intestine. This is
3. Ouabain (cardiac glycoside), an inhibitor reinforced by the bacterial fermentation of
of the sodium pump, inhibits active the remaining carbohydrate to two- and three-
transport of glucose. carbon compounds plus large volumes of CO2
4. State of mucous membrane and length of and H2 gas causing flatulence.
time of contact also affects active transport
of glucose. If mucous membrane is not DIGESTION AND ABSORPTION OF LIPIDS
healthy, absorption will suffer. In hurried
bowel, length of contact is less and as such Different forms of lipids taken in the diet are:
absorption will be less. 1. Neutral fats
5. Thyroid hormones increase absorption of 2. Phospholipids
hexoses and act directly on intestinal cells. 3. Cholesterol
6. Deficiency of adrenal cortex hormones 4. Fatty acids and glycerol
decreases absorption due to decreased Fatty acids and glycerol do not require
concentration of sodium in body fluids. any digestion because they are absorbed as
7. Insulin has no effect on absorption of such.
glucose.
8. Absorption is decreased in states of Dietary Sources of Lipids
deficiency of B – vitamins namely thiamine, 1. Animal sources: Dairy products like milk,
pyridoxine and pantothenic acid. butter, ghee, meat, fish.
78 Medical Biochemistry for Physiotherapy Students
2. Vegetable sources: Various cooking oils from The pancreatic lipase is the most important
various seeds namely sunflower oil, which hydrolyses triglycerides containing
groundnut oil, cottonseed oil, mustard oil short chain fatty acids as well as long chain
etc. and fats from other sources. fatty acids. Other two enzymes are required
Digestion of fat starts in the stomach and for phospholipids and cholesterol respec-
ends in the succus entericus (intestinal juice). tively.
Role of bile salts in pancreatic lipase
In the Stomach activity: Bile salts are required for proper
The digestion of lipids begins in the stomach, functioning of the enzyme.
catalyzed by an acid-stable lipase, most of which 1. Bile salts help in combination of lipase with
is thought to originate from glands at the back co–lipase (small protein) in the intestinal
of the tongue (lingual lipase). However, the lumen.
rate of hydrolysis is very slow because the 2. Bile salts help in the emulsification of fats.
lipid is not yet emulsified. As water–soluble lipases can act only on
the surface of water–insoluble fat droplets
Gastric lipase (lipolytic enzyme): It acts best in
so, bile salts help in lipase activity by
slightly acid medium but is destroyed by
emulsifying fats into fine droplets by
0.02% HCl in 15 minutes. Hence, its action
takes place only in the initial stages of gastric lowering the surface tension and thus
secretion when the acidity is not high. It acts increasing the surface area of these
best on emulsified fat, i.e. egg–yolk, milk etc. droplets for lipase activity.
Due to these characteristics, gastric lipase is Triglyceride→1,2-diglyceride→2-monoglyceride
not of much importance.
In the Intestinal Juice
In the Pancreatic Juice
Intestinal juice also contains another lipase.
Fat digestion is mainly carried out in the Since under normal conditions, fat digestion
duodenum by pancreatic lipase (or sometimes is almost completed by pancreatic juice, this
called as steapsin) and bile salts acts as an lipase is of little importance. It is needed only
effective emulsifying agent for fats. Pancreatic
to deal with that little quantity of which may
juice and bile enter the upper small intestine,
have accidentally escaped pancreatic diges-
the duodenum.
tion.
Secretion of pancreatic juice is stimulated
by the
ABSORPTION OF FATS
1. passage of an acid gastric contents (acid
chyme). The end products of lipid digestion are mainly
2. by secretion of gastrointestinal hormones. glycerol, fatty acids and also the partially split
Pancreatic juice contains a number of products such as monoglycerides and digly-
lipolytic enzymes: cerides. With the aid of the bile salts, these
1. Pancreatic lipase (steapsin) products of fat digestion enter the mucosal
2. Phospholipase A2 cells of the small intestine where digestion of
3. Cholesterol esterase. fats may be completed through the action of
Digestion and Absorption 79
the intestinal lipase. The resultant products are fistula. Feeding triacylglycerols in which the
glycerol and fatty acids. Resynthesis of fatty acids are of medium chain length (less
triglycerides now occurs, by the utilization than 12 carbons) in place of dietary fat results
of partial glycerides and the liberated free in a disappearance of chyluria.
acids. The resynthesized triglycerides then
pass into the lymphatics of the abdominal DIGESTION AND ABSORPTION
cavity and through the thoracic duct the fats OF PROTEINS
appear in blood as 1 μm diameter particles
Proteins which we take in our diet are either
called chylomicrons. Structurally the
from animal source or vegetable source.
chylomicrons consist mainly of a triglyceride
Animal sources are milk and dairy products,
core covered with phospholipids and protein.
meat, fish, liver, eggs. Vegetable sources are
The bile salts are carried by the portal blood
cereals, pulses, peas and beans, nuts.
to the liver and excreted in the bile back to
Different forms in which proteins are taken
the intestine.
in diet are as follows:
Phospholipids obtained from the diet may
1. Various types of albumins and globulins.
be absorbed as such because of its hydrophilic
This is the chief form of food protein.
nature. These may travel by the way of the
2. Nucleoprotein
portal blood directly to the liver. Phospho-
3. Caseinogen (of milk)
lipids synthesis and turnover may increase
4. Collagen and gelatin
in the intestinal wall during the absorption
5. Mucin
of fat.
6. Elastin
The absorption of cholesterol is facilitated
Of these proteins, elastin cannot be
by esterification with fatty acids which is
digested. All the other varieties are broken
catalyzed by cholesterol esterase.
down upto the stage of amino acids by
DEFECT IN DIGESTION AND endopeptidases, e.g. pepsin, trypsin, chymotrypsin
ABSORPTION OF LIPIDS and by the exopeptidases in the lower part of
the intestinal tract.
Lipid Malabsorption Protein digestion starts in the gastric juice
(stomach) and is completed in the intestinal
It can occur due to obstruction to flow in bile
juice (succus entericus) due to the absence of
due to biliary obstruction, diseases of the
the proteolytic enzyme in saliva.
pancreas and tropical sprue which leads to
increased fat content of faeces causing
In the Gastric Juice
steatorrhoea.
Pepsin is the proteolytic enzyme of gastric
Chyluria juice. It acts with the help of HCl and converts
It is an abnormality in which the patient all digestible proteins upto the peptone stage.
excretes milky urine because of the presence Proteolytic enzymes other than pepsin (e.g.
of an abnormal connection between the cathepsin, parapepsin, gastricism) are also
urinary tract and the lymphatic drainage present in gastric juice. 10-15% of protein is
system of the intestine, a so called chylous broken down to amino acids in stomach.
80 Medical Biochemistry for Physiotherapy Students
Properties of Pepsin activates chymotrypsinogen into chymotrypsin.
Chymotrypsin coagulates milk. It also hydro-
1. Pepsin is protein in nature.
lyses casein and gelatin. Both trypsin and
2. It remains as pepsinogen in the peptic cells
chymotrypsin are endopeptidases.
of the stomach.
Further breakdown of proteins is carried
3. HCl of gastric juice converts it into active
out by the exopeptidases for example:
pepsin.
a. Aminopeptidases
4. Optimum pH is 2.0, i.e. highly acidic. In
b. Carboxypeptidases
alkaline or neutral pH, it is inactive.
c. Tripeptidases
5. It attacks the peptide linkages of the
d. Dipeptidases
protein molecules and breaks them at
Aminopeptidase splits off those amino acids
these linkages.
from the protein molecule which possess a
6. It acts on proteins and digests them upto
free amino group. Similarly, carboxypeptidase
peptone. After complete peptic digestion
takes away those ones which possess a free
about 1/10th of the total number of
carboxyl group. Tripeptidase and dipeptidase
peptide linkages are broken down.
split off tri- and di- peptides to amino acids.
7. Its action on living gastric mucous mem-
brane is prevented by the presence of
In the Intestinal Juice
antipepsin in the gastric mucosa.
8. Pepsin in the presence of HCl digests Digestion of protein in the intestinal juice
protein to peptone through the following depends upon the tryptic activity as the
stages – Protein→acid metaprotein → proteolytic enzyme present in the intestinal
primary proteoses→secondary proteoses juice is unable to hydrolyze the protein as such.
→peptone. Intestinal juice contains enzyme erepsin.
Erepsin consists of a mixture of enzymes
In the Pancreatic Juice including aminopeptidases and dipeptidases.
Erepsin cannot act on native proteins. It acts
Trypsin is the proteolytic enzyme of pancreatic
upon lower peptides and converts them
juice. Trypsin is secreted as inactive trypsino-
completely into amino acids.
gen. It is activated by the action of entero-
peptidase or enterokinase present in the intestinal ABSORPTION OF PROTEINS
juice. Activation can also be brought about
by the calcium salts. Once the trypsin is formed Dietary proteins under physiological condi-
from trypsinogen, it auto catalytically activates tions are absorbed as amino acids which are
rest of the inactive trypsinogen. the end products of digestion. Small amounts
Trypsin can act upon native proteins as well of peptides may also be absorbed from
as the products of protein digestion, such as alimentary canal.
metaprotein, proteose, peptones, polypep- In infants, the cellular lining of the
tides, etc. and converts them to lower alimentary canal, being delicate is more
peptides, i.e. di - and tri-peptides. permeable to these higher forms. Moreover,
Inactive pancreatic juice also contains in unhealthy conditions of the alimentary
chymotrypsinogen which is inactive. Trypsin tract such as diarrhoea, dysentery, etc. where
Digestion and Absorption 81
the mucous membrane is ulcerated – these digestion, along with many other compounds
foreign proteins may be absorbed in greater such as vitamins, water and mineral salts are
amounts and may cause various disturbances absorbed. When the contents reach the small
in the body. intestine, the absorption process with the
However, under physiological conditions, exception of water is normally completed.
proteins are absorbed in the form of amino Here more water and NaCl are absorbed and
acids only. the remaining material leaves the body as
Absorption of amino acids may be either faeces.
an active or a passive process. D-isomers The water content of the faeces is usually
which are less rapidly absorbed are carried from 60 to 70% by weight. The 20 to 30% dry
by passive diffusion whereas the more matter is composed primarily of undigested
rapidly absorbed, L-isomers are actively dietary constituents such as cellulose material,
transported. Amino acids are absorbed from fatty material, mineral matter and bacteria.
ileum and distal jejunum. Oligopeptides like The undigested food protein, carbohydrate
di- and tri- peptides are absorbed from
and fat amount to very little, since the
duodenum and proximal jejunum.
digestion and absorption of these substances
DEFECT IN DIGESTION AND is normally 95 to 98% complete. Practically
ABSORPTION OF PROTEINS all the nitrogen present is of bacterial origin.
The normal dark brown colour of the
In individuals with a deficiency in pancreatic faeces is due to bile pigment derivatives. The
secretion (for example, due to chronic pan- primary pigment is stercobilin arising from
creatitis, cystic fibrosis or surgical removal
oxidation of the precursor stercobilinogen.
of the pancreas), the digestion and absorption
Small amounts of bilirubin and biliverdin are
of fat and protein is incomplete. This results
sometimes present.
in the abnormal appearance of lipids called
The large intestine is the site of bacterial
steatorrhea and undigested protein in the
multiplication. Many billions of bacteria may
faeces.
be excreted daily in the faeces. E. coli is
Formation and Composition of Faeces ordinarily the predominating organism,
During the passage of intestinal contents although many others are frequently found
through the small intestine, the products of in the faeces.
Chapter 11

Metabolism of Carbohydrates

METABOLISM CARBOHYDRATE METABOLISM

Metabolism refers to all the chemical reactions Carbohydrate metabolism is essentially the
of the body. They can be anabolic (synthetic) metabolism of glucose and of substances
or catabolic (breakdown). related to glucose in their metabolic pro-
cesses. During digestion, polysaccharides and
Anabolism disaccharides are hydrolysed into the
monosaccharides glucose (about 80%), fruc-
Chemical reactions that combine simple tose and galactose. Some fructose is conver-
substances into more complex molecules are ted into glucose as it is absorbed through the
collectively known as anabolism, e.g. forma- intestinal epithelial cells.
tion of peptide bonds between amino acids The three monosaccharides are absorbed
thereby building amino acids into proteins. into the capillaries of the villi of the small
Also fatty acid can be built into phospholipids intestine and then are converted through the
that form the plasma membrane and glucose hepatic portal vein to the liver. Liver cells
into glycogen through anabolic reactions. convert very much of the remaining fructose
and practically all the galactose to glucose.
Catabolism Fate of Carbohydrates
The chemical reactions that break down 1. ATP production: If the cells require imme-
complex organic compounds into simple ones diate energy, glucose is oxidized by the
are known as catabolism, e.g. chemical cells. Each gram of carbohydrates pro-
digestion in which breaking of bonds of food duces about 4 kilocalories (Kcal). Glucose
molecules releases energy. These are not needed for immediate energy (ATP)
generally hydrolysis reactions that release the production can enter one of the several
chemical energy in organic metabolism. metabolic pathways.
Metabolism of Carbohydrates 83
2. Amino acid synthesis: Glucose can be used pyruvic acid (aerobic glycolysis) or during the
to form amino acids which then can be lack of oxygen such as during exercise to lactic
incorporated into proteins. acid (anaerobic glycolysis). Glycolytic
3. Glycogenesis: The liver can store a small enzymes are present in the extramito-
amount of excess glucose by converting it chondrial compartment of the cell. While
into glycogen (glycogenesis). Later, when aerobic glycolysis occurs in all the tissues like
blood glucose start to decrease, hepatic liver, kidney and erythrocytes, anaerobic
cells can convert glycogen back to glucose glycolysis takes place only in muscle.
(glycogenolysis) and release it into the In certain mammalian tissues and cell
blood. types (erythrocytes, renal medulla, brain etc.),
4. Lipogenesis: If the glycogen storage areas glucose is the sole source of metabolic energy
are filled up, liver cells and fat cells can through glycolysis. All the sugar derivatives
transform the glucose to glycerol and fatty that occur in the glycolytic pathway are the
acids that can be used for synthesis of D-isomers.
triglycerides (lipogenesis). Triglycerides
(TGs) are then deposited in adipose tissue Steps of Glycolysis
which has virtually unlimited capacity. 1. Glucose is irreversibly activated to
When glucose enters cells, glucose is phos- Glucose-6-phosphate by hexokinase in the
phorylated. It combines with a phosphate presence of ATP and Mg2+. Glucokinase
group produced by the breakdown of ATP also catalyses similar reaction in liver in
to form glucose-6-phosphate. Enzymes that the fed state. Glucose- 6- phosphate is also
catalyse phosphorylations are kinases. Phos- produced via, Glucose- 1- phosphate from
phorylation traps glucose in the cell so that it glycogen in the presence of inorganic
cannot move back out. Liver cells, kidney phosphate by phosphorylase.
tubule cells and intestinal epithelial cells have 2. Glucose- 6- phosphate is later isomerised
the necessary enzyme (phosphatase) to remove to fructose- 6- phosphate by phosphohexose
the phosphate group which enables glucose isomerase.
to diffuse out of the cell and into the blood 3. Fructose-6-phosphate is then phospho-
stream. rylated by phosphofructokinase to form
fructose 1, 6-diphosphate. This is an
GLYCOLYSIS irreversible reaction and requires ATP.
It is also known as EMP pathway, i.e. 4. Thereafter, fructose-1, 6-diphosphate (the
Embden-Meyerhof Parnas pathway. In hexose molecule) is cleaved by aldolase to
glycolysis, a molecule of glucose is degraded 3-phosphoglyceraldehyde (glyceralde-
in a series of enzyme catalyzed reactions to hyde-3-phosphate) and dihydroxy acetone
yield two molecules of pyruvate. During the phosphate (2 triose molecules).
sequential reactions of glycolysis, some of the 5. Dihydroxyacetone phosphate is also
free energy liberated from glucose is converted to 3-phosphoglyceraldehyde by
conserved in the form of ATP. phosphotriose isomerase.
This process of catabolism of glucose 6. Subsequently, 3-phosphoglyceraldehyde
occurs either in the presence of oxygen to enters the pathway. In the presence of
84 Medical Biochemistry for Physiotherapy Students
NAD+ and inorganic phosphate, glyceral- 7. In the next step, 1, 3-biphosphoglyceric
dehyde-3-phosphate dehydrogenase oxidizes 3- acid is converted to 3-phosphoglyceric
phosphoglyceraldehyde to 1, 3-biphos- acid. This reaction is catalysed by phospho-
phoglycerate (1, 3-biphosphoglyceric glycerokinase and generates one ATP. It is
acid). called as substrate level production of
ATP. Arsenic acid, if present, results in
uncoupling of oxidation and phosphory-
lation and inhibits ATP production.
8. Thereafter, 3-phosphoglyceric acid is
converted to 2-phosphoglyceric acid by
phosphoglucomutase. During this conversion
2,3-biphosphoglyceric acid is formed as an
intermediate which also acts as a coenzyme
for this reaction.
9. Enolase then converts 2-phosphoglyceric
acid to a high energy compound i.e. phos-
phoenolpyruvate. The enzyme requires
Mg 2+ or Mn 2+ and is inhibited by the
presence of fluoride.
10. Later phosphoenol pyruvate is converted
to pyruvate, irreversibly. The reaction is
catalysed by pyruvate kinase and generates
one ATP.
11. Pyruvate is converted to lactate under
anaerobic conditions by lactate dehydro-
genase and NADH+H+.

Importance of Glycolysis
1. Glycolysis is meant for energy.
2. Glycolysis produces ATP in the absence
of oxygen because this allows skeletal
muscles to perform efficiently even when
aerobic oxidation becomes insufficient and
it allows tissue with significant glycolytic
activity.
3. The deficient activity of pyruvate kinase, the
enzyme of glycolysis, produces the disease
haemolytic anaemia.

Regulation of Glycolysis
The three irreversible steps catalysed by the
Reactions of glycolysis various kinases, i.e. hexokinase, phosphofructo-
Metabolism of Carbohydrates 85
kinase and pyruvate kinase are the sites of 3. CoA-SH
regulation of glycolysis. Insulin stimulates 4. FAD
these enzymes and thereby increasing the 5. NAD+
utilization of glucose into glycolysis. On the 6. Mg2+
other hand, glucagon inhibits the process.

Bioenergetics/Energy Production
Reaction Energy Number of
consumed Oxidative Decarboxylation of Pyruvic Acid
produced as ATPs
Glucose to glucose-6- ATP –1 In this oxidative decarboxylation reaction,
phosphate
one NADH + H + is produced, which on
Fructose-6-phosphate to
fructose-1, 6-diphosphate ATP –1 entering respiratory chain will give 3
Glyceraldehyde-3-P- 2 NADH + H +
2×3= +6 molecules of ATP. However, because 2
dehydrogenase pyruvate will give 2 acetyl CoA molecules,
Phosphoglycerate kinase 2 ATP +2 so total molecules of ATP are 6 in this reaction.
Pyruvate kinase 2 ATP +2
Under Anaerobic Conditions
Net ATP yield (ATP produced – ATP Under anaerobic conditions, pyruvate is
consumed): 10 – 2 = 8. reduced to lactate. The reaction is catalysed
by lactate dehydrogenase (LDH) which requires
Fate of Pyruvate NADH + H+
Under Aerobic Conditions
Under aerobic conditions, pyruvate is trans-
ported into mitochondria via pyruvate trans- Formation of Lactate
porter. In the mitochondria, pyruvate is
oxidatively decarboxylated to acetyl CoA by TRICARBOXYLIC ACID CYCLE (TCA CYCLE)
pyruvate dehydrogenase complex. Because
reaction involves both oxidation and loss of It is also known as citric acid cycle, Kreb’s
CO 2 (decarboxylation), it is termed as citric acid cycle, Kreb’s cycle. TCA cycle
oxidative decarboxylation reaction. consists of a series of reactions in mitochondria
Pyruvate dehydrogenase complex consists of which catalyses the oxidation of acetyl CoA
29 molecules of pyruvate dehydrogenase (PDH), to CO2 and H2O in aerobic conditions giving
8 molecules of flavoprotein containing dihydro- out energy. It is the final common pathway
lipoyl dehyrogenase and 1 molecule of dihydro- for metabolism of carbohydrates, fats and
lipoyl transacetylase. proteins through formation of 2 carbon
The enzyme complex requires six co- compound acetyl CoA. It is the mechanism
enzymes/cofactors. by which much of the free energy liberated
1. TPP during the oxidation of carbohydrate, lipids
2. Lipoic acid and amino acids is made available.
86 Medical Biochemistry for Physiotherapy Students
These reactions occur in a cyclic manner of citrate to isocitrate is inhibited by
and generate large amounts of ATP since the fluoroacetate.
enzymes of the cycle are located in the 3. In the presence of isocitrate dehydrogenase,
mitochondria facilitating the transfer of isocitrate is first converted to oxalo-
reducing equivalents from the Kreb’s cycle succinate which is later decarboxylated to
to the respiratory chain, the enzymes of which form α-ketoglutarate.
are also located in the inner mitochondrial 4. Similar to the conversion of pyruvate to
membrane. acetyl CoA, α-ketoglutarate also under-
goes oxidative decarboxylation and is
Steps of Citric Acid Cycle converted to succinyl CoA. The reaction
1. In the first step of the Kreb’s cycle, acetyl requires α-ketoglutarate dehydrogenase
CoA (as formed from pyruvate under complex and the five coenzymes (TPP,
aerobic conditions) combines with oxalo- NAD+, FAD, coenzyme A and lipoic acid).
acetate and forms citric acid (a tricar- 5. Succinyl CoA is then changed to succinate
boxylic acid). The reaction is catalysed by by succinate thiokinase. During this reaction,
citrate synthetase (a condensing enzyme). a molecule of GTP is formed. This is known
2. Citrate is then rearranged to form cis- as substrate level production of ATP since
aconitic acid which is subsequently a high energy molecule is formed at the
changed to isocitrate. Both the steps are substrate level without the involvement
carried out by aconitase. The conversion of the respiratory chain.

TCA Cycle
Metabolism of Carbohydrates 87
6. Succinate is converted to fumarate by FAD Energy yield during complete oxidation
containing enzyme succinate dehydrogenase. of one molecule of glucose gives 38 molecules
7. With the addition of water by fumarase, of ATPs.
fumarate changes to malate. (Glycolysis = 8, Pyruvate to acetyl CoA =
8. Finally, malate dehydrogenase, in the presence 6 and TCA cycle = 24)
of NAD+, converts malate to oxaloacetate
which re-enters the cycle. Biological Significance of Citric Acid Cycle
The TCA Cycle Serves Citric acid cycle plays a dual role and is
Five Major Functions important both in the oxidation as well as
1. It produces most of the carbon dioxide synthetic processes. Due to its amphibolic
made in human tissues. nature, it is catabolic for the oxidation of
2. It is the source of much of the reduced carbohydrates, lipids and proteins whereby
coenzymes that drive the respiratory chain these substances are completely oxidized to
to produce ATP. CO2 and H2O and release a large amount of
3. It converts excess energy and inter- energy. It is also important for the anabolic
mediates the synthesis of proteins and reactions as various intermediates of the cycle
fatty acids. are used for the biosynthesis of nonessential
4. It provides some of the precursors used in amino acids and glucose.
the synthesis of proteins and nucleic acids.
5. Its components control directly (product Yeast Fermentation
precursor) or indirectly (allosteric) other
The fermentation of yeast by monosaccharides
enzyme systems.
involves exactly the same chemical reactions
as glycolysis in animal tissues down to the
Energy Production during
production of pyruvic acid. Yeast then decar-
Citric Acid Cycle
boxylates the pyruvic acid to acetaldehyde
Reaction Energy Number and reduces the latter to ethyl alcohol.
produced of ATPs
as
HMP SHUNT
Isocitrate to oxalosuccinate NADH+H+ 3
α-ketoglutarate to succinyl NADH+H+ 3 Hexose monophosphate shunt (HMP shunt)
CoA
Succinyl CoA to succinate GTP 1 or pentose phosphate pathway is the second
(Substrate level major pathway for the metabolism of glucose.
phosphorylation)
Succinate to fumarate FADH 2 2
The enzymes for this pathway are present in
Malate to oxaloacetate NADH+H+ 3 the extra mitochondrial fraction of the cell.
Total 12 This pathway is operative in many tissues
such as liver, erythrocytes, lactating mammary
Since 2 molecules of pyruvate are pro- glands and adipose tissue.
duced during glycolysis and it yields 2
Steps of HMP Shunt
molecules of acetyl CoA which in one cycle
yields 12 ATPs, so 2 acetyl CoA in two cycles 1. In the first step, glucose is converted to
will give 24 molecules of ATPs. glucose-6-phosphate by the enzyme
88 Medical Biochemistry for Physiotherapy Students
hexokinase. Thereafter, glucose-6-phosphate 3. In the second stage of the shunt, ribulose-
by glucose-6-phosphate dehydrogenase is 5-phosphate is utilised in a multicyclic
converted to 6-phosphogluconolactone process. Ribulose-5-phosphate is changed
and subsequently to 6-phosphogluconate. to ribose-5-phosphate by ribose-5-phosphate
During these processes, NADP+ is reduced ketoisomerase. Ribulose-5-phosphate is also
to NADPH + H+. converted to xylulose-5-phosphate by
2. In the next reaction, 6-phosphogluconate ribulose-5-phosphate epimerase.
is oxidized by 6-phosphogluconate dehydro-
4. In the next reaction, carbon skeletons of the
genase to form an intermediate 3-keto-6-
pentoses are rearranged. A transketolase
phosphogluconate. This reaction also
generates NADPH + H+. Subsequently the transfers a ketol group (the first 2 carbon
first carbon atom is removed as CO2 and atoms) from xylulose-5-phosphate onto
3-keto-6-phosphogluconate is decarboxy- ribose-5-phosphate forming sedoheptulose-7-
lated to ribulose-5- phosphate. phosphate and glyceraldehyde-3-phosphate.

HMP Shunt
Metabolism of Carbohydrates 89
Subsequently, an aldol group (3-carbon 4. Microsomal cytochrome P450 system (in
moiety) is transferred by transaldolase from liver) brings about the detoxification of
sedoheptulose-7-phosphate onto glyceral- drugs and foreign compounds by hydroxy-
dehyde-3-phosphate forming fructose-6- lation reactions involving NADPH.
phosphate and erythrose-4-phosphate. 5. Phagocytosis requires the supply of
NADPH. It is the engulfment of foreign
Metabolic Significance of HMP Shunt particles including micro-organisms
HMP shunt is unique in generating two carried out by white blood cells.
important products, pentoses and NADPH, 6. NADPH which is produced in erythro-
needed for the biosynthetic reactions and cytes maintains the concentration of
other functions. reduced glutathione which is essentially
required to preserve the integrity of RBC
Importance of Pentoses membrane.
7. NADPH is necessary to keep the ferrous
In the HMP shunt, hexose are converted into iron of haemoglobin in the reduced state
pentoses, the most important being ribose- 5 so that accumulation of methemoglobin is
– phosphate. This pentose or its derivatives prevented.
are useful for the synthesis of nucleic acids
(RNA and DNA) and many nucleotides such GLYCOGEN METABOLISM
as ATP, NAD+/FAD and CoA.
Skeletal muscle is capable of synthesizing The synthesis (glycogenesis) and degradation
pentoses although only the first enzymes of of glycogen (glycogenolysis) are not simply
HMP shunt are active. It, therefore, appears the reversal of one series of reactions. Instead,
that the complete pathway of HMP shunt may each process is an entirely separate metabolic
not be required for the synthesis of pentoses. pathway catalyzed by a different set of
enzymes. The formation of glycogen occurs
Importance of NADPH mainly in liver and muscle. The liver contains
1. NADPH is required for the reductive about 4- 6% of glycogen as per its weight i.e.
biosynthesis of fatty acids and steroids; 72-108 g and the muscle contains about 0.7%
hence, HMP shunt is more active in the of its weight, i.e. about 245 g.
tissues concerned with lipogenesis, e.g. The function of muscle glycogen is to act
adipose tissue, liver, etc. as a readily available source of hexose units
2. NADPH is used in the synthesis of certain for glycolysis within the muscle itself. Liver
amino acids involving the enzyme gluta- glycogen is largely concerned with export of
mate dehydrogenase. hexose units for maintenance of the blood
3. There is production of H2O2 in the living glucose, particularly between meals.
cells which can chemically damage unsatu- If glucose is not needed immediately for
rated lipids, proteins and DNA. This is ATP production, it is combined with many
prevented through antioxidant reactions other molecules of glucose to form a long
involving NADPH. chain molecule called glycogen.
90 Medical Biochemistry for Physiotherapy Students
GLYCOGENESIS

Glycogenesis is defined as a process of


formation of glycogen from glucose mole-
cules.

Steps of Glycogenesis
1. Glucose is phosphorylated to glucose-6-
phosphate by enzyme hexokinase. This
reaction is common to the first reaction in
the pathway of glycolysis from glucose.
2. In the next step, glucose-6-phosphate is
then converted to glucose-1-phosphate in
a reaction catalyzed by the enzyme
phosphoglucomutase with Mg 2+ ions as
cofactor.
3. Next, glucose-1-phosphate reacts with
uridine triphosphate (UTP) to form the
active nucleotide uridine diphosphate Glycogenesis
glucose (UDPG) catalyzed by the enzyme
UDPG pyrophosphorylase with the release
of inorganic pyrophosphate.
6 glucose residues) to a neighbouring
4. In the next step, by the action of the
chain to form a α-1, 6-linkage, thus
enzyme glycogen synthetase, the C1 of the
establishing a branch point in the mole-
activated glucose of UDPG forms a
cule. The branches grow by further
glycosidic bond with the C4 of a terminal
additions of α-1, 4- glucosyl units and
glucose residue of glycogen, liberating
further branching.
uridine diphosphate (UDP). A preexisting
Thus, under the combined action of
glycogen molecule or primer must be
glycogen synthetase and branching enzyme, the
present to initiate the reaction.
glycogen molecule is formed.
5. The addition of a glucose residue to a pre-
existing glycogen chain or ‘primer’ occurs GLYCOGENOLYSIS
at the non-reducing, outer end of the
molecule so that the branches of the Breakdown of glycogen to glucose is called
glycogen tree become elongated as glycogenolysis. It involves a debranching
successive α– 1,4- linkages occur. mechanism of phosphoryltic cleavage of α-l,
6. When the chain has been lengthened to 4-linkage to yield glucose-1-phosphate. It is
between 6 and 11 glucose residues, a catalyzed by phosphorylase, which is a rate-
second enzyme, the branching enzyme acts limiting step in glycogenolysis.
on the glycogen. This enzyme transfers a Liver and muscle, both contain phospho-
part of the α-1, 4-chain (minimum length rylase. In both, the enzyme can be in “active”
Metabolism of Carbohydrates 91

Type Name Enzyme deficiency Organs affected Clinical features


I Von Gierke’s Glucose-6-phosphatase Liver, kidney Hypoglycaemia,
disease and intestine ketosis,enlarged liver,
gouty arthritis.
II Pompe’s disease α-1,4-glucosidase Liver, heart, striated Cardiomegaly,
(acid maltase) and smooth muscle Muscle hypotonia
leading to muscle
weakness.
III Forbe’s disease Debranching enzyme Liver, muscle Moderate
and heart hypoglycaemia,
acidosis, enlarged
liver.
IV Anderson’s disease Branching enzyme Most tissues Moderate hypoglycaemia,
cirrhosis of liver, splenomegaly
V Mc Ardle’s disease Muscle phosphorylase Skeletal Splenomegaly,
muscle Muscle cramps on exercise,
weakness and stiffness of muscle.
VI Her’s disease Liver phosphorylase Liver Mild hypoglycaemia, enlarged
liver, acidosis.

enzyme. The combined action of phosphorylase


and other enzymes leads to the complete
breakdown of glycogen molecule.

Glycogen Storage Diseases


These are a group of inherited disorders
associated with glycogen metabolism in which
abnormal type or quantity of glycogen is
deposited in different tissues. According to
Glycogenolysis
the deficiency of the enzyme involved, there
are six types of the glycogen storage diseases.
and “inactive” forms and both are inter-
convertible. Pyridoxal phosphate is required GLUCONEOGENESIS
for the activity of muscle phosphorylase. The synthesis of glucose or glycogen from
The terminal glucose residues from the noncarbohydrate compounds is known as
outermost chains of the glycogen molecule gluconeogenesis. The major substrates or
are removed sequentially until approximately precursors for gluconeogenesis are lactate,
four glucose residues remain on either side pyruvate, glucogenic amino acids, propionate
of α-l, 6- branch. Then another enzyme and glycerol.
α-1,4- α 1, 4 glucan transferase, transfers a Gluconeogenesis occurs mainly in the
trisaccharide unit from one branch to the cytosol, although some precursors are pro-
other, exposing the α-l, 6 branch point. The duced in the mitochondria. Gluconeogenesis
hydrolytic splitting of the α-l, 6-linkages mostly takes place in liver and, to some
requires the action of a specific debranching extent, in kidney.
92 Medical Biochemistry for Physiotherapy Students
Importance of Gluconeogenesis to phosphoenolpyruvate. High energy
Brain and central nervous system, erythro- phosphate in the form of GTP or ITP is
cytes, testes and kidney medulla are depen- required in this reaction and CO2 is liberated.
dent on glucose for continuous supply of
energy. Glucose is the only source of supply-
ing energy to the skeletal muscle under
anaerobic conditions.

Reactions of Gluconeogenesis
Gluconeogenesis closely resembles the
reversed pathway of glycolysis, although it Pyruvate to Phosphoenolpyruvate Conversion
is not the complete reversal of glycolysis.
Mainly 3 reactions of glycolysis are irrever- Fructose-1,6-biphosphate to Fructose-6-
sible. The rest of the reactions are common phosphate Conversion
for both glycolysis and gluconeogenesis .The
This reaction is catalyzed by a specific enzyme
three irreversible steps of glycolysis are
fructose-1,6-biphosphatase. This enzyme is
catalysed by the enzymes, namely hexokinase,
present in liver, kidney and striated muscle.
phosphofructokinase and pyruvate kinase.

Gluconeogenesis from Pyruvate


1. Pyruvate to phosphoenolpyruvate conver-
sion. Fructose-1, 6-Biphosphate to Fructose-6-
2. Fructose-1,6-biphosphate to fructose-6- Phosphate Conversion
phosphate conversion.
3. Glucose-6-phosphate to glucose conver- Glucose-6-phosphate to
sion. Glucose Conversion
The conversion of glucose-6-phosphate is
Pyruvate to Phosphoenolpyruvate catalyzed by another specific phosphatase,
Conversion glucose-6-phosphatase. It is present in intestine,
liver and kidney. It is absent from muscle and
This reaction occurs in two parts. First,
adipose tissue.
enzyme pyruvate carboxylase converts pyruvate
to oxaloacetate in the presence of biotin, ATP
and CO2. However, because gluconeogenesis
occurs in cytosol and this reaction takes place
in mitochondria and oxaloacetate is imper- Glucose-6-Phosphate to Glucose Conversion
meable, so oxaloacetate is first converted to
malate which is permeable. Oxaloacetate is
Gluconeogenesis from Amino Acids
regenerated from malate. In the cytosol,
second enzyme phosphoenolpyruvate carboxy- After transamination or deamination,
kinase catalyzes the conversion of oxaloacetate glucogenic amino acids form either pyruvate
Metabolism of Carbohydrates 93

The Pathway of Gluconeogenesis


94 Medical Biochemistry for Physiotherapy Students
or members of the TCA cycle. Therefore, the phosphate (DHAP) by enzyme glycerol-3-
reactions described above account for the phosphate dehydrogenase. This DHAP is an
conversion of glucogenic amino acids to intermediate in glycolysis which can be used
glucose. for glucose production.

Gluconeogenesis from Propionate FRUCTOSE METABOLISM


Propionate enters the main glucogenic Fructose is obtained by the body either
pathway via the TCA cycle after its conver- directly from diet or because of the hydrolysis
sion to succinyl CoA. Propionate with other of sucrose in the intestine. It is converted to
fatty acids is activated to propionyl CoA in glucose for its utilisation by the body by
the presence of thiokinase, ATP and CoA. various routes.
Propionyl CoA carboxylase acts on this in 1. Fructose is changed to fructose-1-phos-
presence of ATP and biotin and converts to phate by the specific enzyme fructokinase
methyl malonyl CoA which is then converted present in liver, muscle, intestine and
to succinyl CoA in presence of vitamin B12 kidney.
coenzyme. Fructokinase deficiency results in fructo-
suria, as a result of which fructose appears
Gluconeogenesis from Glycerol in urine, after a high fructose diet is
Glycerol is a product of metabolism of adipose ingested.
tissue. The enzyme glycerokinase converts Fructose-1-phosphate is cleaved by the
glycerol to glycerol-3-phosphate which is enzyme aldolase B (which is different from
further converted to dihydroxyacetone aldolase, also called aldolase A, which acts

Metabolism of Fructose
Metabolism of Carbohydrates 95
on fructose-1,6-bisphosphate). Aldolase B
splits fructose-1-phosphate into dihy-
droxyacetone phosphate and D-glyceral-
dehyde. The individuals with aldolase B
deficiency suffer from hereditary fructose
intolerance.
Subsequently, glyceraldehyde is phos-
phorylated by ATP in the presence of triose
kinase and is changed to 3-phospho-
glyceraldehyde. Dihydroxyacetone phos-
phate along with glyceraldehyde- 3-phos-
phate conjugates to form glucose-6- phos-
phate by the reversal of glycolytic reactions.
2. Alternatively, fructose-l-phosphate is
phosphorylated to form fructose-l,6-
bisphosphate by the enzyme 1-phospho-
Metabolism of Galactose
fructokinase and enters glycolytic path-
way.
glucose (UDP-glucose) in the presence of
3. Fructose may also change to fructose-6-
the enzyme galactose-1-phosphate uridyl
phosphate by hexokinase. It is then
transferase forming glucose-1-phosphate
isomerised to glucose-6- phosphate and
and UDP-galactose. Glucose-1-phosphate
either directly enters glycolysis or is
is changed to UDP-glucose by the enzyme
hydrolysed to produce free glucose.
UDP-glucose pyrophosphorylase and can be
GALACTOSE METABOLISM used in glycogenesis.
In liver, UDP-galactose is also changed to
Galactose is released into the blood stream UDP-glucose by the enzyme UDP-galactose-
by the hydrolysis of lactose due to the action 4-epimerase. Subsequently, UDP-glucose is
of lactase in the intestine. It can be utilised in also used in glycogen synthesis.
the body in the liver, the brain and the lacta- 2. In the brain and the nervous tissue, UDP-
ting mammary glands. Most of the dietary galactose reacts with sphingosine in the
galactose is changed to glucose in the liver presence of the enzyme galactose-sphingosine
and this ability of the liver to utilise galactose transferase and is changed to galactosyl-
is important in the assessment of the liver sphingosine for use in the synthesis of
function test, after a galactose load (galactose gangliosides and cerebrosides.
tolerance test). 3. In the lactating mammary glands, UDP-
1. In liver, galactose is phosphorylated by galactose combines with glucose in the
the enzyme galactokinase forming galactose- presence of the enzyme lactose synthetase
1-phosphate, more rapidly in children and forms lactose for its secretion into
than in adults. Subsequently, galactose-1- milk. Hereditary deficiency of the enzyme
phosphate reacts with uridine diphosphate uridyl transferase (galactose-1-phosphate uridyl
96 Medical Biochemistry for Physiotherapy Students
transferase) results in galactosemia, i.e. the Insulin
accumulation of galactose in blood. In
Insulin is released into blood stream under
patients with galactosemia increased levels
the direct influence of hyperglycemia. It is a
of galactose in the blood also results in
polypeptide, containing 2 chains and is
galactosuria (increased excretion of
produced by the β-cells of the islets of
galactose in urine), mental retardation and
langerhans of the pancreas. Insulin lowers
formation of cataract.
blood glucose concentration by increasing the
uptake of glucose by the extrahepatic tissues
REGULATION OF BLOOD GLUCOSE
for its oxidation. Insulin also promotes
The blood glucose level is maintained within glycolysis and glycogenesis, both in liver and
the normal physiological range of 60-100 mg/ muscle. At the same time, it suppresses
100 ml in the post-absorptive state. Mainte- glycogenolysis in liver and kidneys. Besides,
nance of the blood glucose level is regulated insulin also suppresses gluconeogenesis.
by a number of factors such as a balance bet- Hormones which have hyperglycemic
ween different metabolic pathways involving action include glucagon, epinephrine, gluco-
various tissues, several hormones and thres- corticoids, growth hormone and thyroxine.
hold of the kidneys.
Glucagon
By Metabolic Processes It is produced by the α-cells of the islets of
Various metabolic pathways regulate blood langerhans of the pancreas. Its synthesis is
glucose. Glycolysis, HMP shunt and glyco- stimulated under the influence of hypo-
genesis lower blood glucose while glyco- glycemia. It acts only in the liver and promo-
genolysis and gluconeogenesis increase blood tes glycogenolysis. It also promotes gluco-
glucose concentration. neogenesis from amino acids and lactate.
In the fasting state or on a low carbo-
hydrate diet when blood glucose level is Epinephrine
reduced, gluconeogenesis and glycogenolysis Epinephrine also called adrenaline is secreted
are stimulated in the liver and add glucose by adrenal medulla. It is regarded as a
to the blood stream. hormone in the first line of defence against
In the fed state, when blood glucose hypoglycemia. It promotes glycogenolysis in
concentration increases, its cellular transport both liver and muscle. Epinephrine also
in the liver and utilisation by the glycolytic promotes gluconeogenesis in liver. The
reactions is increased. Glycogenesis is also hormone inhibits the release of insulin from
stimulated in the liver and muscle. pancreas, thereby, decreasing the transport
and utilisation of glucose in different tissues.
By Hormones
Glucocorticoids
Several hormones regulate blood glucose
concentration. Insulin is the hormone which Glucocorticoids are the steroids secreted by
lowers blood glucose, various other hor- adrenal cortex and are antagonists to insulin.
mones increase blood glucose level. Cortisol is the major glucocorticoid present
Metabolism of Carbohydrates 97
in blood. Glucocorticoids promote gluco- insulin. There are two major clinical classes
neogenesis in liver and glucose oxidation by of the disease, i.e. insulin-dependent and non-
liver. Glucocorticoids also facilitate the action insulin-dependent diabetes mellitus.
of other hyperglycemic hormones (permissive
effect) such as glucagon, epinephrine and Insulin-dependent Diabetes Mellitus
growth hormone which further inhibit the Insulin-dependent diabetes mellitus (IDDM)
uptake and utilisation of glucose by the or type I diabetes also called juvenile-onset
peripheral tissues. diabetes because it usually appears in
childhood or in younger age group (less than
Growth Hormone
40 years of age). There is an absolute defi-
Growth hormone is secreted by anterior ciency of insulin due to a gradual depletion
pituitary. Its secretion is stimulated under the of the β-cells of the pancreas. The β-cells are
influence of hypoglycemia. Growth hormone destroyed by an autoimmune process.
also decreases glucose uptake and its Patients with IDDM can usually be
utilization by the muscle and promotes recognized by the abrupt appearance of
lipolysis in the adipose tissue. polyuria (frequent urination), polydypsia
(excessive thirst), and polyphagia (excessive
Thyroxine hunger), often triggered by stress or an
Thyroxine is secreted by the thyroid gland. illness. These symptoms are usually accom-
It stimulates the intestinal absorption of panied by fatigue, weight loss and weakness.
glucose and promotes glycogenolysis and The diagnosis is confirmed by a fasting
gluconeogenesis in liver. Thyroxine also blood glucose (greater than 140 mg/dl)
causes the destruction of insulin. Hyperthy- commonly accompanied by ketoacidosis.
roidism is generally associated with hyper- Blood glucose concentration rises to more
glycemia and mild diabetes. than 200 mg/dl following the oral adminis-
tration of 75-100 g of glucose. Glucose also
By Kidneys appears in the urine at this stage. In contrast,
normal individuals show fasting blood
Kidney also regulates blood glucose that is glucose level of 70-90 mg/dl and a rise to only
continuously filtered by the glomeruli and is about 130-140 mg/dl, after a glucose load.
completely reabsorbed by the renal tubules.
However, when blood glucose concentration Non-insulin-Dependent Diabetes Mellitus
exceeds 170- 180 mg/dl, tubules fail to
reabsorb all the filtered glucose and it starts Majority of the diabetic patients (over 80%)
appearing in urine (glycosuria). suffer from non-insulin-dependent diabetes
mellitus (NIDDM) or type II diabetes, also
DIABETES MELLITUS called maturity-onset diabetes since this
usually occurs in middle-aged people (more
Diabetes mellitus is defined as a state of than 40 years old), who are obese. The
chronic hyperglycemia caused by a relative occurrence of the type II disease is almost
or absolute deficiency of pancreatic hormone completely determined by genetic factors.
98 Medical Biochemistry for Physiotherapy Students
NIDDM develops gradually without obvious the GTT is performed following starvation
symptoms. The metabolic alterations are or a diet low in carbohydrate, glucose
milder than those for the IDDM. tolerance will be reduced which will make
NIDDM is characterized by hyperglycemia the results of the test difficult to interpret.
often with hypertriglyceridemia. Inspite of 2. Ideally, the test should be performed in
high levels of insulin, glucose levels are poorly the morning. The patient should be
controlled because of the lack of normal instructed not to eat, drink, or smoke for
response to insulin. Blood glucose concen- 10-16 hours before the test.
tration is greater than 140 mg/dl. 3. Patient should not be under fear or anxiety
about the possibility of being a diabetic if
GLUCOSE TOLERANCE TEST (GTT) so it leads to false positive results. There-
fore, it is the duty of the technician to
A GTT measures the ability of the body to prepare the patient psychologically or
tolerate, or cope with, a standard dose of mentally and convince the patients.
glucose. The degree of tolerance to the 4. The patient should not have excess amount
glucose is mainly dependent on the rate of of exercise.
glucose absorption and on the insulin dose. 5. If the patient is not well, the test should
As the glucose is absorbed, the level of be postponed.
glucose in the blood rises and the normal 6. The patient should not receive any drugs
response is for insulin to be released from at least 3 days before the test.
the pancreas to lower the glucose level.
Tolerance is reduced when insulin is insuffi- Method
cient.
The test is usually carried out in the early
Indications of GTT morning after overnight fast. Fasting blood
sample and urine is also collected. 100 g of
1. To know the family history of diabetes glucose dissolved in about 150-200 ml of
mellitus. water is given to drink.
2. Symptoms and signs comparable with Venous blood for the estimation of blood
diabetic without any complications.
glucose is collected at ½ hourly intervals for
3. Glucosuric patients with normal fasting
2 ½ hours or hourly intervals for 3 hours after
blood sugar.
the ingestion of glucose. Urine specimens are
4. Border line of glucose in PPBS.
also collected at the same time.
5. Reactive hypoglycemia for 3 hour or
Blood glucose is estimated in each samples
longer period after food intake.
6. In pregnancy conditions with history of and the urine is tested for the presence of the
abortions, stillbirth and large baby. sugar.

Interpretation
Preparation of the Patient
Normal Glucose Tolerance Curve (GTC)
1. Before the test, the patient should be on a
diet containing not less than 150 g of Fasting blood glucose level should be bet-
carbohydrate per day for at least 3 days. If ween 60-100 mg%. The highest peak value is
Metabolism of Carbohydrates 99
reached within 60 min and does not exceed Conditions Associated with
the renal threshold, i.e. 170-180 mg%. Urine Diminished Glucose Tolerance
sugar is negative in all the samples. A typical 1. Due to lack of insulin, there will be
response is shown on page 84. decreased tissue utilisation of glucose,
which is seen in diabetes mellitus.
Fasting ½ 1 1½ 2 2½ 2. Increased glycogenolysis and gluconeo-
Blood glucose 75 130 150 100 65 76 genesis. This is seen in glucocorticoid
Urine sugar — — — — — — excess and hyperthyroidism.
3. Increased rate of absorption which is seen
Diabetic Type of GTC in thyrotoxicosis.
4. Decreased glycogen storage which is seen
Fasting blood glucose level is definitely in severe hepatic diseases and glycogen
raised- 110 mg% or more. The highest peak storage.
value is usually reached after 60- 90 min and
exceeds the normal renal threshold (see page Increased Glucose Tolerance
84). Increased glucose tolerance curve is charac-
For moderate diabetes mellitus terized by a flat response. Conditions asso-
ciated with increased tolerance:
Fasting ½ 1 1½ 2 2½
1. Hypothyroidism
Blood glucose 130 200 280 260 220 170
2. Hypoadrenalism
Urine sugar — ++ ++ ++ ++ +–
3. Hypopituitarism
4. Malabsorption from the GIT
For severe diabetes mellitus 5. Renal glycosuria
6. Hyperinsulinism or insulinoma.
Fasting ½ 1 1½ 2 2½
This is also characterized by
Blood glucose 230 300 345 365 350 330
i. Fasting hypoglycemia.
Urine sugar ++ +++ +++ ++++ +++ +++
ii. Slight increase in blood glucose follow-
ing glucose ingestion.

Intravenous Glucose Tolerance


Preparation of Patient
Poor absorption of orally given glucose may
result in a flat tolerance curve. Some patients
are unable to tolerate a large amount of
carbohydrate load. In these patients, an
intravenous glucose tolerance test may be
performed to eliminate the factors related to
the rate of the glucose absorption. The
preparation of patient is as same as that of
oral GTT. The dose of glucose is 0.5 g/kg
Glucose Tolerance Curve body weight given as 25 g/dl solution.
Chapter 12

Metabolism of Lipids

The first stage in the utilization of triglycerides β – oxidation, in which two-carbon fragments
for energy is hydrolysis of these compounds are successively removed from the carboxyl
into fatty acids and glycerol and their transport end of the fatty acyl CoA, producing acetyl
to the active tissues where they will be oxidized CoA.
to give energy. Almost all cells can use fatty
acids as a source of energy. Glycerol after Transport of Fatty Acids into the
conversion into glycerol-3-phosphate, enters Mitochondria
glycolytic pathway for glucose breakdown and After the fatty acid is taken up by a cell, it is
in this way, it is used for energy. converted to the CoA derivative by fatty acyl
Triglycerides which form 85–90% of total CoA synthetase (thiokinase) in the cytosol.
lipids are stored in the adipose tissue and
serve as energy reserve of the body.
Phospholipids, glycolipids and cholesterol are
major components of cell membranes.
Transport of lipids is in the form of lipo-
proteins. Chylomicrons, VLDL, LDL, HDL,
albumin–free fatty acid complex are the
different lipoprotein complexes that transport Activation of Fatty Acid

lipids in the blood stream.


The mobilization is initiated by hydrolysis Because β–oxidation occurs in the mito-
of triglycerides into fatty acids and glycerol chondrial matrix, the fatty acid must be
(by the enzyme hormone sensitive lipase). transported across the mitochondrial inner
membrane which is generally impermeable
β–OXIDATION OF FATTY ACIDS to bulky, polar molecules such as coenzyme
A. Therefore, a specialized carrier in this
The major pathway of catabolism of saturated membrane transports the acyl group from the
fatty acids is a mitochondrial pathway called cytosol into the mitochondrial matrix. This
Metabolism of Lipids 101
carrier is carnitine and the transport process 1. Acyl CoA undergoes dehydrogenation by
is called carnitine shuttle. First, an acyl group enzyme acyl CoA dehydrogenase to form Δ2
is transferred from the cytosolic coenzyme A trans-enoyl CoA.
to carnitine by enzyme carnitine acyltransferase 2. Enoyl CoA hydratase brings about hydration
I, forming acylcarnitine. The enzyme is to form the product β-hydroxyacyl CoA.
located on the outer surface of the inner 3. β-hydroxyacyl CoA dehydrogenase catalyses
mitochondrial membrane. Second, the the second oxidation and generates
acylcarnitine group is transported across the NADH. The product formed is β-ketoacyl
membrane to the mitochondrial matrix, where CoA.
it is transferred to another molecule of 4. The final reaction is the liberation of two
coenzyme A by enzyme carnitine acyltransferase carbon compound, acetyl CoA by enzyme
II on the inner surface of the inner mitochon- thiolase.
drial membrane.

Carnitine Shuttle

β –oxidation Proper
The one cycle of β–oxidation consists of a
sequence of four reactions that result in the
shortening of the fatty acid chain by two
carbons. The steps include an oxidation that β -oxidation Proper
produces FADH 2 , hydration, a second
oxidation that produces NADH and a thiolytic
Energy Yield from α -oxidation
cleavage that releases a molecule of acetyl
CoA. These four steps are repeated for The oxidation of a molecule of palmitic acid
saturated fatty acids of even numbered (C15H31COOH) to CO2 and water yields the
carbon chains, each cycle producing an acetyl following number of ATPs:
group plus one NADH and one FADH2. The The complete oxidation of palmitic acid
final thiolytic cleavage produces two acetyl undergoes 7 cycles and gives 8 acetyl CoA
groups. molecules. In one cycle 1 NADH + H + , 1
102 Medical Biochemistry for Physiotherapy Students
FADH2 are produced. Therefore, in 7 cycles
7 NADH + H+, 7 FADH2 are produced.
7 NADH + H+, each provide 3 ATP when
oxidized by the ETC 21
7 FADH 2, each provide 2 ATP when
oxidized by the ETC 14
1 acetyl CoA molecule when converted by
TCA cycle to water, carbon dioxide produces
12 ATPs. Therefore, 8 molecules produce
8 × 12 = 96.
Energy production from one molecule of
palmitic acid is 21 + 14 + 96 = 131 Oxidation of Fatty Acids with
Odd Number of C-Atoms
Net yield is 131 – 2 = 129 ATPs because 2
molecules are used at the first step, i.e.
activation reaction.
or pyruvate oxidation into ketone bodies. The
Oxidation of Fatty Acids with compounds categorized as ketone bodies are
acetoacetate, 3-hydroxybutyrate (β-hydroxy-
Odd Number of Carbons
butyrate), and acetone. They are transported
The β–oxidation of a saturated fatty acid with in the blood to the peripheral tissues, where
an odd number of carbon atoms proceeds by they can be reconverted to acetyl CoA and
the same reaction steps as that of a fatty acid oxidized by the TCA cycle.
with an even number of carbon atoms until Ketone bodies are important sources of
the final three carbons are reached. This energy for the peripheral tissues because
compound called propionyl CoA is metabo- 1. They are soluble in aqueous solution, and
lized by a two step pathway. therefore do not need to be incorporated
First, propionyl CoA is carboxylated in lipoproteins or carried by albumin as
forming methylmalonyl CoA. The enzyme do the other lipids;
propionyl CoA carboxylase and coenzyme biotin 2. They are produced in the liver during
are required for the reaction to occur. Next, periods when the amount of acetyl CoA
the carbons of methylmalonyl are rearranged present exceeds the oxidative capacity of
forming succinyl CoA which can enter the the liver;
TCA cycle. The enzyme methylmalonyl CoA 3. They are used in extrahepatic tissues such
as the skeletal and cardiac muscle and
mutase requires a coenzyme form of vitamin
renal cortex, in proportion to their
B12 (deoxyadenosylcobalamin) for its action.
concentration in the blood. Even the brain
KETONE BODIES can utilize ketone bodies for fuel if the
level rises sufficiently. (Note: This is
Liver mitochondria have the capacity to divert important during prolonged periods of
any excess acetyl CoA derived from fatty acid fasting).
Metabolism of Lipids 103
Synthesis of Ketone Bodies or CoA, and therefore cannot itself use them as
Ketogenesis fuels. However, extrahepatic tissues, inclu-
ding the brain but excluding cells lacking
The first step is the formation of acetoacetyl
mitochondria (for example, red blood cells),
CoA. Then, a molecule of acetyl CoA com-
efficiently oxidize acetoacetate and 3-
bines with acetoacetyl CoA to produce 3-
hydroxybutyrate. 3-hydroxybutyrate is oxi-
hydroxy-3-methylglutaryl CoA (HMG CoA)
dized to acetoacetate by 3-hydroxybutyrate
by the enzyme HMG CoA synthase which is
dehydrogenase, producing NADH. Acetoacetate
the rate-limiting step in the synthesis of
receives a coenzyme A molecule from succinyl
ketone bodies and is present in significant
CoA. This reaction is reversible, but the
quantities only in the liver. HMG CoA is
product, acetoacetyl CoA, is actively removed
cleaved to produce acetoacetate and acetyl
by its conversion to two acetyl CoA. The liver
CoA. Acetoacetate can be reduced to form 3-
lacks succinyl CoA: acetoacetate CoA
hydroxybutyrate with NADH as the hydro-
transferase (thiophorase), and therefore is
gen donor, or it can be spontaneously
unable to use acetoacetate as a fuel.
decarboxylated to form acetone.

Ketolysis

Excessive Production of Ketone


Bodies in Diabetes Mellitus
When the rate of formation of ketone bodies
Ketogenesis is greater than the rate of their use, their levels
begin to rise in the blood (ketonaemia) and
Utilization of Ketone Bodies
eventually in the urine (ketonuria). These two
Although the liver constantly produces low conditions are seen most often in cases of
levels of ketone bodies, their production starvation or severe diabetes mellitus. In
becomes much more significant during diabetic individuals with severe ketosis,
starvation, when ketone bodies are needed urinary excretion of ketone bodies may be as
to provide energy to the peripheral tissues. high as 5000 mg/24 hr, and the blood
The liver actively produces ketone bodies, but concentration may reach 90 mg/dL (versus
it cannot reconvert acetoacetate to acetoacetyl less than 3 mg/dL in normal individuals.
104 Medical Biochemistry for Physiotherapy Students
BIOSYNTHESIS OF FATTY ACIDS

There are three systems for fatty acid


synthesis
1. Extramitochondrial system or De novo
synthesis of fatty acids (lipogenesis).
2. Mitrochondrial chain elongation 1. Fatty acid synthesis starts with the
3. Michrosomal chain elongation. transfer of acetyl CoA to the cysteinyl-SH
group of ACP with the help of acetyl
De Novo Synthesis of Fatty Acids
transacylase subunit of the fatty acid
The main site of synthesis of fatty acids inside sythetase complex and forms acetyl-S-
the cells is outside the mitochondria i.e., in synthetase. Malonyl transacylase transfers
cytosol. All the enzymes required for it are malonyl CoA onto the pantotheinyl-SH
present in cytosol. Acetyl CoA is the starting group of ACP to form malonyl-S-ACP.
material and palmitic acid is the end product.
Acetyl CoA is produced in mitochondria from
different sources such as pyruvate obtained
from carbohydrates (aerobic glycolysis) and
glucogenic amino acids, or directly from lipids
(β-oxidation) and ketogenic amino acids. Since
mitochondrial membrane is impermeable to
acetyl CoA, it condenses with oxaloacetate
and forms citrate. Citrate enters the cytoplasm
2. The condensing enzyme (β-ketoacyl-ACP
and is degraded to acetyl CoA and oxaloace-
synthetase) causes condensation of acetyl-
tate by ATP citrate lyase.
S-synthetase with malonyl-S- ACP and
In cytoplasm acetyl CoA is first converted
forms acetoacetyl-S-ACP (acetoacetyl
to malonyl CoA by biotin containing enzyme,
enzyme or β-ketoacyl enzyme).
acetyl CoA carboxylase, irreversibly. This
reaction occurs in the presence of NADPH,
ATP and CO2.
Biosynthesis of fatty acids requires fatty
acid synthetase which is a multienzyme
complex containing 6 enzymes and an acyl
carrier protein (ACP). Six subunits of the
3. β-Ketoacyl-ACP reductase with NADPH
multienzyme complex are acetyl transacylase,
malonyl transacylase, β-ketoacyl-ACP +H+ reduces acetoacetyl-S-ACP to form β-
synthetase, β-ketoacyl-ACP reductase, β- hydroxybutyryl-S-ACP (β-hydroxyacyl-S-
hydroxyacyl-ACP dehydratase and enoyl- ACP).
ACP reductase.
The ACP has two –SH groups called
pantotheinyl-SH group and the cysteinyl-SH
group.
Metabolism of Lipids 105
4. β-hydroxyacyl-ACP dehydratase removes rounds, chain elongation continues and
H2O from β−hydroxyaceyl-S-ACP forming finally palmityl CoA is synthesized.
crotonyl-S-ACP. Deacylase removes CoA and forms
palmitate.
The reducing equivalents (NADPH + H+)
used in the biosynthesis of fatty acids are
derived from HMP shunt reactions.
5. Crotonyl-S-ACP is reduced by Enoyl-ACP
reductase utilizing NADPH+ H+and is Mitochondrial Chain Elongation
converted to butyryl-S-ACP. De novo synthesis forms palmitic acid which
is used as a starting material for the synthesis
of higher fatty acids in mitochondria.
Chain elongation of palmitic acid in
mitochondria takes place by the successive
Butyryl-S-ACP again accepts a molecule additions of acetyl CoA. Condensation of
of malonyl CoA and with the repeated palmitic CoA with acetyl CoA forms β-
ketostearyl CoA which utilises NADPH + H+
and is reduced to β-hydroxystearyl CoA.
Removal of a molecule of water converts β-
hydroxystearyl CoA to α, β-unsaturated
stearyl CoA which is reduced to stearyl CoA
by using NADPH + H+.

Microsomal Chain Elongation


The process of chain elongation in microsomes
is similar to fatty acid biosynthesis (De novo
synthesis) in the cytoplasm. Although it also
utilises malonyl CoA but uses CoA instead
of ACP as the acyl carrier. Medium chain
saturated fatty acids and monounsaturated
fatty acids with C18 (oleic acid) are used for
chain elongation.

CHOLESTEROL METABOLISM

Virtually all tissues in humans synthesize


cholesterol, although liver, intestine, adrenal
cortex and reproductive tissues including
ovaries, testes and placenta make the largest
contributions to the body’s cholesterol pool.
Cholesterol is the most abundant sterol in
humans. The liver plays a central role in the
Biosynthesis of fatty acids regulation of the body’s cholesterol balance.
106 Medical Biochemistry for Physiotherapy Students
Synthesis
All the carbon atoms in cholesterol are
provided by acetate and NADPH provides
the reducing equivalents.
1. First two acetyl CoA molecules condense
to give a molecule of acetoacetyl CoA by
the action of enzyme thiolase.
2. Next, a third molecule of acetyl CoA is
added to form HMG-CoA (3-hydroxy-3-
methylglutaryl CoA). This reaction is
catalysed by the enzyme HMG-CoA
synthetase.
3. HMG-CoA is converted to mevalonic acid
by the enzyme HMG-CoA reductase. This
reaction uses two molecules of NADPH
as the reducing agent and releases CoA.
This is the rate-limiting step in cholesterol
synthesis.
4. Mevalonic acid is converted to isoprene
unit, i.e. isopentenyl pyrophosphate. This
reaction requires ATP and Mg2+ ions as
cofactor.
5. Two isopentenyl phosphate molecules
condense to give geranyl pyrophosphate
releasing pyrophosphate.
6. Geranyl pyrophosphate condenses with
one molecule of isoprene unit to give
farnesyl pyrophosphate.
7. Two molecules of 15-carbon farnesyl pyro-
phosphate combine releasing pyro-
phosphate and are reduced forming the
30-carbon compound squalene.
8. Squalene is converted to 27-carbon com-
pound cholesterol by ring closure and
removal of three methyl groups.
Biosynthesis of cholesterol

Fate of Cholesterol
The sterol structure of cholesterol cannot be b. Secretion of cholesterol into the bile which
metabolized to CO 2 and H 2O in humans. transports it to the intestine for elimina-
Sterol ring is eliminated from the body by: tion.
a. Conversion to bile acids which are excreted c. Conversion to vitamin D.
in the faeces. It occurs in liver. d. Conversion to steroid hormones.
Metabolism of Lipids 107
Blood Cholesterol and remnants after releasing both Apo-A and
Cardiovascular Disease Apo-C from it. The fatty acids released are
Persons with increased levels of blood taken up by the tissues, while the chylomicron
cholesterol have a high incidence of athero- remnants are taken up by the liver. The
sclerosis, a chronic disease characterized by enzyme lipoprotein lipase is present on the
accumulation of deposits of cholesterol, walls of blood capillaries in many tissues like
cholesteryl esters and cellular debris on the lungs, adipose tissue, heart, spleen and
inner surfaces of the large and medium sized lactating mammary gland, etc. and attached
arteries. As the disease progresses, the to the capillary membrane with the proteo-
deposits reduce or even, stop the flow of glycan chains of heparin sulphate. If heparin
blood causing coronary artery disease. The is injected into the body, there will be rapid
cells normally irrigated by the affected artery release of lipoprotein lipase into the circu-
are deprived of oxygen and nutrients, and lation. Both Apo-CII and some phospholipids
die rapidly. If this interruption of blood flow are co-factors for lipoprotein lipase. Insulin
occurs in an artery of the heart, a myocardial also activates lipoprotein lipase.
infarction, or heart attack occurs. As a result, The liver secretes very high density lipids
part of the heart muscle becomes non- which is formed from endogenous sources.
functional and death may result. The hepatic triacyl glycerol being the main
source. This is contributed by the triacyl
Metabolism of Lipoproteins glycerol reaching the liver as chylomicron
Plasma contains the following lipoproteins: remnants, the fatty acids reaching the liver
1. Chylomicrons derived from intestinal via the plasma from adipose tissue during
absorption of triacyl glycerol. lipolysis and the triacyl glycerol synthesized
2. Very low density lipoproteins (VLDL) in the liver, i.e. extra mitochondrial de novo
derived from the liver. synthesis. The source differs in the fed state
3. Low density lipoproteins (LDL) derived from that in the fasting state. Conditions
from VLDL during its metabolism while associated with increased triacyl glycerol
in circulation. synthesis and secretion of VLDL are feeding
4. Free fatty acids present in combination of high carbohydrate diet in presence of high
with plasma albumin. insulin status (especially high source feeding)
Chylomicrons are formed in the intestine and ethanol ingestion in higher amounts (in
during the digestion of dietary fat. They moderate and severe alcoholics).
contain mostly triacyl glycerol. In the intes- Nascent VLDL that contains triacyl
tinal mucosal cells, they are modified at the glycerol and cholesterol along with Apo-B-
endoplasmic reticulum and secreted into the 100 takes up Apo-C and Apo-E to form VLDL.
lymphatic vessels. Chylomicrons have a half By the action of the enzyme lipoprotein lipase
life of only five minutes. The metabolism of in extra hepatic tissues, Apo-C and some fatty
chylomicrons involves a few steps during acids cleave off, along with glycerol forming
which the nascent chylomicrons with Apo-B- LDL passing through an intermediate stage
48 take up Apo-A followed by combination of VLDL remnants or IDL and fatty acids.
with Apo-C and Apo-E. The lipoprotein lipase The LDL is finally destroyed in the liver or
then acts on the mature chylomicrons and in the extra hepatic tissues, fibroblasts,
degrades to fatty acids and chylomicron lymphocytes etc.
108 Medical Biochemistry for Physiotherapy Students
The liver and intestines also secrete a high ATHEROSCLEROSIS
density lipoprotein HDL. The lecithin
This is a condition associated with hardening
cholesterol acyl transferase enzyme transfers
of the walls of the aorta and arterial walls
the fatty acids from phospholipids lecithin to
due to a variable combination of changes in
free cholesterol thus forming cholesterol
the intima of arteries. This consists of local
esters. Discoid nascent HDL becomes accumulation of lipids, complex carbohy-
spherical HDL, which transfers the choles- drates, blood and blood products, fibrous
teryl ester transfer protein and to the liver tissues and calcium deposits and is associated
(reverse cholesterol transport). HDL helps to with medical changes. Deposition of choles-
dispose of the cholesterol by esterification terol esters needs special mention.
which then enters the liver and gets oxidized Atherosclerosis can occur in diseases with
to bile acids. Thus HDL scavenges the tissue elevated levels of cholesterol and LDL,
cholesterol including that of the smooth lowered levels of HDL and an increase of
muscle cells of the arteries. VLDL. This is because HDL and VLDL have
a reciprocal relationship and a lowered level
OBESITY HDL may cause an increase of VLDL. The
The balance between the rates of deposition various diseases, which could cause athero-
sclerosis, are diabetes mellitus, nephrotic
and mobilization determines the amount of
syndrome, hypothyroidism and familial
fat in the storage depots. An animal becomes
hyperlipoproteinaemias.
fat when the rate of deposition exceeds the
A strong co-relation exists between
rate of mobilization and lean when the
hypertension, hypercholesterolemia, serum
processes are reversed. It has been established
VLDL and LDL and atherosclerosis and
that obesity results purely from the ingestion
coronary heart diseases. Hypercholes-
of more food than is necessary to meet energy
terolemia, increase of LDL, decrease of HDL
requirements; in other words, it is a question
and hypertension are all direct risk factors
of the appetite being improperly balanced
for atherosclerosis.
with energy needs.
LDL carries cholesterol in the plasma and
Alterations in the energy demand of an
deposits it in tissues and thus it is a
animal without simultaneous change in contributing factor in atherosclerosis. So, LDL
appetite may occur for various reasons. This is known as bad cholesterol, whereas HDL
may be true with decreased muscular activity. represents good cholesterol because it
In addition, endocrine disturbances may safeguards against cholesterol deposition and
decrease activity and leads to obesity. prevents its accumulation. HDL efficiently
Certain pituitary deficiencies may lead to removes cholesterol from tissues, esterifies
abnormal distribution of body fat without it and transports it back to the liver where it
changing the total quantity. Some cases of is excreted as bile salts. HDL has the capacity
hyperinsulinism become obese because of the of even stripping the deposited cholesterol
greatly stimulated appetite. It has been from the smooth muscle cell of the arteries.
demonstrated that the hypothalamic region Investigation of total cholesterol, TGs,
of the brain is concerned with regulation of lipoprotein electrophoretic profile, HDL-
appetite. Injuries to this region in experimental cholesterol and HDL/LDL ratio are very
animals may lead to great obesity due to helpful in the diagnostic biochemistry of
appetite stimulation. atherosclerosis.
Chapter 13

Metabolism of Proteins

Amino acids contain N in addition to C, H under ureotelic organisms, the aquatic animals
and O. This nitrogen cannot be stored and are ammoniotelic and reptiles and birds are
amino acids found in excess of the biosynthetic categorized as uricotelic organisms. This
needs of the cell are immediately degraded. category is according to type of ammonia
disposed off the body.
AMINO ACID POOL The formation of urea is divided into 4
processes:
Amino acids released by hydrolysis of dietary 1. Transamination
or tissue protein mix with other amino acids 2. Oxidative deamination
distributed throughout the body and they 3. Ammonia transport
collectively constitute the amino pool. 4. Urea cycle.
In mammalian tissues, the amino groups
of amino acids, derived either from the diet Transamination
or from breakdown of tissue proteins, Transamination involves the transfer of an
ultimately are excreted in the urine as urea. amino group from an amino acid to the keto
Therefore, mammals including man come group of the keto acid forming a new amino

Transamination Reaction
110 Medical Biochemistry for Physiotherapy Students
acid and a keto acid. Enzymes termed trans- (which is a source of nitrogen in urea
aminases or aminotransferases catalyze the reac- synthesis). Liver and kidney contain enzymes
tion. The reaction is reversible and pyridoxal like L-amino acid oxidases, D-amino acid oxidases
phosphate acts as coenzyme. and glutamate dehydrogenase which catalyze the
Transamination mainly takes place in liver, deamination of amino acids.
kidney, heart and brain. There occurs only L-amino acid oxidases catalyze the oxidation
intermolecular transfer of NH2 group without of L-amino acids and have FMN as the
the net loss of the amino group. prosthetic group. There occurs reduction of
Two transaminases, aspartate transaminase FMN to FMNH2. Subsequently amino group
and alanine transaminase are clinically impor- is liberated as ammonia with the help of a
tant. Aspartate transaminase is also known molecule of water. Tissues also contain FAD
as SGOT, i.e. serum glutamate oxaloacetate linked D-amino acid oxidases which catalyze the
transaminase. It catalyzes the following oxidation of D-amino acids.
reaction.

Reaction Catalyzed by SGOT

Alanine transaminase is also known as SGPT


(serum glutamate pyruvate transaminase). It
catalyzes the following reaction.

Oxidative Deamination Reaction

L-glutamic acid which is formed as a end


Reaction Catalyzed by SGPT
product of transamination reactions is not
Oxidative Deamination deaminated by L-amino acid oxidase but by a
It results in the liberation of the amino group specific enzyme called L-glutamate dehydro-
as free ammonia. These reactions occur genase. This enzyme catalyzes the deamination
mainly in the liver and kidney and provide of L-glutamate to form α-iminoglutaric acid,
α-keto acids (which can enter the central which on addition of a molecule of water
pathway of energy metabolism) and ammonia forms NH3 and α-ketoglutarate.

Deamination of L-glutamic acid


Metabolism of Proteins 111
Ammonia Transport Urea Formation (Urea Cycle)
Although ammonia is constantly produced in The formation of urea occurs mainly in the
the tissues, it is present at very low levels in liver. 1 mol. of urea is synthesized from 1
the blood. The ammonia which is formed must mol. of ammonia, 1 mol. of carbon dioxide, 3
be kept low in the blood because even slightly mols of ATP (2 of which are converted to ADP
elevated concentrations (hyperammonaemia) and Pi and 1 to AMP + PP i ), 5 enzymes
are toxic to the central nervous system. This catalyzing the reactions and 6 amino acids
is due to both the rapid removal of ammonia involved in the reaction.
from the blood and the fact that many tissues
release amino acid nitrogen in the form of Reactions of the Urea Cycle
glutamine or alanine rather than as free The first two reactions leading to the synthesis
ammonia. of urea occur in the mitochondria whereas
In addition to ammonia formation by the the remaining cycle enzymes are located in
aminotransferase and glutamate dehydrogenase in the cytosol.
liver, ammonia is also formed in the kidneys 1. Formation of carbamoyl phosphate by
by the action of renal glutaminase. Most of this carbamoyl phosphate synthase I is driven by
ammonia, which is formed by kidneys, is cleavage of two molecules of ATP. Ammo-
excreted into the urine as NH 4+ which is nia incorporated into carbamoyl phosphate
important mechanism for maintaining the is provided primarily by the oxidative
body’s acid-base balance. deamination of glutamate. Carbamoyl
Formation of urea in the liver is the most phosphate synthase I requires N–acetyl-
important disposal route of ammonia. Urea glutamate for activity.
travels in the blood from the liver to the 2. The reaction product citrulline is trans-
kidneys, where it passes into the glomerular ported to the cytosol.
filtrate. 3. Citrulline condenses with aspartate to
Glutamine (amide of glutamic acid) form argininosuccinate. The α– amino
provides a non-toxic storage and transport group of aspartate provides the second
form of ammonia. The formation of glutamine nitrogen that is ultimately incorporated
occurs mainly in the muscle and liver but also into urea. The formation of arginino-
is important in the nervous system where it succinate is driven by the cleavage of ATP
is the major mechanism for the removal of to AMP and PPi.
ammonia in the brain. Circulating glutamine 4. Argininosuccinate is cleaved to yield
is removed by the kidneys and deaminated arginine and fumarate.
by glutaminase. 5. Arginase cleaves arginine to ornithine and
urea. Arginase occurs almost exclusively in
the liver. Thus whereas other tissues can
synthesize arginine, only the liver can
cleave arginine and thereby synthesize
Transport of Ammonia urea.
112 Medical Biochemistry for Physiotherapy Students
6. Urea diffuses from the liver and is Glucogenic and Ketogenic Amino Acids
transported in the blood to the kidneys Amino acids are classified as ketogenic and
where it is filtered and excreted in the glucogenic amino acids according to the
urine. A portion of the urea synthesized nature of their metabolic end products.
in the liver diffuses from the blood into Glucogenic amino acids are those whose
the intestine and is cleaved to CO2 and catabolism yields pyruvate or one of the
NH3 by bacterial urease. The ammonia is intermediates of the TCA cycle. These
partly lost in the faeces and is partly intermediates are substrates for gluconeo-
reabsorbed into the blood. genesis and therefore can give rise to the net
formation of glycogen in liver and muscle.
Ketogenic amino acids are those whose
catabolism yields either acetoacetate or one
of its precursors, acetyl CoA or acetoacetyl
CoA are termed ketogenic amino acids.
Examples are lysine and leucine.

METABOLISM OF IMPORTANT
AMINO ACIDS

Phenylalanine and Tyrosine


Phenylalanine is an essential amino acid but
tyrosine is non-essential amino acid, i.e. it can
be synthesized in the body. Tyrosine is
Reaction of Urea Cycle
formed from phenylalanine by phenylalanine
hydroxylase. The reaction requires molecular
Catabolism of the Carbon oxygen, NADPH and the coenzyme tetra-
Skeletons of Amino Acids hydrobiopterin. The reaction is irreversible
and occurs in liver.
The catabolism of the 20 carbon amino acids
involves the removal of α-amino groups
followed by the breakdown of resulting
carbon skeletons. The catabolism of carbon
skeleton converges to form 7 products which
includes oxaloacetate, α-keto glutarate,
pyruvate, acetyl CoA, etc. These are the
intermediates of TCA cycle, so they result
either in the synthesis of glucose or lipid or
production of energy. Synthesis of Tyrosine
Metabolism of Proteins 113
Fate acid. The reaction requires ascorbic acid
and vitamin B12.
Phenylalanine is catabolised via tyrosine, so
they have same metabolic fate. c. Homogentisic acid oxidase oxidizes
1. Formation of fumarate and acetoacetate. homogentisic acid to maleyl acetoacetate.
2. Formation of melanin pigment. d. Maleyl acetoacetate is isomerised to
3. Formation of thyroid hormones. fumaryl acetoacetate by isomerase.
4. Conversion to epinephrine. e. Fumaryl acetoacetate on hydrolysis by
fumaryl acetoacetate hydrolase forms
Formation of Fumarate and Acetoacetate fumarate and acetoacetate.
a. Tyrosine undergoes transamination by
INHERITED DISORDER
tyrosine-β-ketoglutarate transaminase and
forms p-OH-phenyl pyruvate. Pyridoxal Phenylketonuria (PKU)
phosphate acts as a coenzyme in the
reaction. PKU is caused by a deficiency of phenylalanine
b. P-OH-phenyl pyruvate hydroxylase converts hydroxylase. It is the most common clinically
p-OH-phenyl pyruvate to homogentisic encountered inborn error of amino acid
metabolism. The enzyme catalyzes the
conversion of phenylalanine. Due to the
deficiency of the enzyme phenylalanine
hydroxylase, the main pathway for the
metabolism of phenylalanine via tyrosine is
blocked and the minor alternate pathway
takes place. Phenylalanine accumulates in the
blood; it undergoes transamination to form
phenyl pyruvic acid and its products as phenyl
lactic acid and phenyl acetic acid are pro-
duced.

Alternate Metabolic Products of Phenylalanine

Due to increase in the urinary excretion


of these phenyl ketoacids, the disease is called
Fumarate and Acetoacetate Formation PKU.
114 Medical Biochemistry for Physiotherapy Students
Mental retardation, seizures, tremor, Tyrosinaemia type II is due to the defi-
microcephaly and failure to grow are ciency of the enzyme tyrosine transaminase.
characteristic findings in PKU. The patient There may be raised levels of plasma tyrosine.
with untreated PKU typically shows symp- Characteristic skin and eye lesions and
toms of mental retardation by the age of one moderate mental retardation, self-mutilation
year. Patients with PKU show a deficiency of and disturbance of fine co-ordinations have
pigmentation. been reported.
Patients are treated by giving diet having Neo-natal tyrosinosis is due to a hereditary
very low levels of phenylalanine. In this deficiency of enzyme p-OH phenyl pyruvate
condition, tyrosine becomes dietary essential.
oxidase.
Alkaptonuria
TRYPTOPHAN
Alkaptonuria is an inborn error of metabolism
associated with metabolism of phenylalanine Tryptophan is an essential amino acid. The
and tyrosine. There is deficiency of the main pathway for its catabolism is called
enzyme homogentisic acid oxidase, as a result, kynurenine-anthranilate pathway. This
there is a blockage in metabolic pathway at pathway is also important for niacin biosyn-
homogentisic acid. There is accumulation of thesis. 60 mg of Tryptophan gives rise to 1
homogentisic acid in tissues, blood and mg of nicotinic acid in the human body.
excretion in urine.
There is occurrence of dark urine on Formation of Serotonin
standing in air which slowly turns black from
This is another major pathway for metabolism
top to down. Deposition of homogentisic acid
and its derivatives in cartilages of ears and of tryptophan. Chemically, serotonin is called
other exposed places leads to generalized 5-OH tryptamine. It is stimulant of CNS and
pigmentation of connective tissues and its also vasoconstrictor. It produces contraction
deposition in joints leads to arthritis. This of smooth muscles.
condition is known as ochronosis. Synthesis of tryptophan starts with its
hydroxylation to give 5-OH tryptophan which
Tyrosinaemia is further decarboxylated by 5-OH tryptophan
decarboxylase to form 5-OH tryptamine.
Tyrosinaemia is an inherited metabolic
disorder. It is of three types:
Tyrosinaemia type I is due to the defi-
ciency of fumaryl aceto-acetyl hydrolase. Both
acute and chronic forms are known. In acute
form, infants exhibit diarrhoea, vomiting, a
cabbage-like odour and failure to thrive.
Death within 6-8 months from liver failure is
certain in untreated acute form. In chronic
form, similar but milder symptoms lead to
death by the age of 10 years. Treatment
consists of a diet low in tyrosine and phenyla-
lanine. Reactions for Formation of Serotonin
Metabolism of Proteins 115
Hartnup Disease 3. It is also required for the formation of
creatine with other two amino acids, i.e.
Hartnup disease is an inborn error associated
arginine and methionine.
with metabolism of tryptophan. Hartnup 4. It plays an important role in biosynthesis
disease is named after the family in which it of purine nucleus.
was discovered and is due to a deficiency of 5. It is required for the synthesis of tripep-
enzyme tryptophan dioxygenase in the liver. tide glutathione along with glutamic acid
The clinical symptoms are skin rash, and cysteine.
intermittent cerebellar ataxia and mental 6. It is used for the detoxification of benzoic
retardation. The urine of patients with acid which is converted to a non-toxic form
Hartnup disease contains increased amount as benzoyl glycine or hippuric acid.
of indole acetic acid and tryptophan.

Glycine as Detoxifying Agent


GLYCINE

Glycine is the simplest of amino acids. It does Inherited Disorder


not exhibit optical activity as it does not Glycinuria
possess an asymmetric carbon atom unlike
Glycinuria is a rare disorder of glycine
other L- amino acids.
metabolism. It is characterized by excess
urinary excretion of glycine in association
Metabolic Fate
with a tendency of formation of oxalate renal
1. Conversion to serine: Glycine can be stones; although the amount of oxalate
converted to serine which by non- excreted in the urine is normal. The plasma
oxidative deamination can from pyruvic content of glycine is normal in the glycinuric
acid, thus glycine may be glucogenic. patients. The cause may be due to a defect in
renal tubular transport of glycine.

Primary Hyperoxaluria
Primary hyperoxaluria is a metabolic disease
characterized biochemically by a continuous
high excretion of oxalate in urine, the excess
Conversion of Glycine to Serine oxalate probably coming from glycine. There
is recurrent infection of the urinary tract.
2. It is necessary in the first reaction for the
formation of heme of haemoglobin with GLUTAMIC ACID
other substrate succinyl CoA. Glutamic acid is non-essential amino acid.

Metabolic Fate
1. It forms glutamine which is important in
the detoxification of ammonia in the brain.
2. It is decarboxylated in the brain to GABA
Formation of Heme (γ-amino butyric acid) by glutamate
116 Medical Biochemistry for Physiotherapy Students
decarboxylase. GABA is required for
proper neuronal function.

Formation of GABA

3. It is a constituent of tripeptide glutathione


4. Glutamic acid in the form of N-acetyl
Formation of Creatine Phosphate
glutamate is required in the urea cycle.
5. The oxidative deamination of glutamic acid
Creatine phosphate is the form in which
forms α-keto glutaric acid which is an
high energy phosphate is stored in the muscle.
intermediate of the TCA cycle and can be
During exercise, ATP is used as a source of
converted to glucose. Therefore, it is
energy and is converted to ADP. During
glucogenic.
resting conditions, creatine phosphate is
METHIONINE utilised to phosphorylate ADP. The reaction
Methionine is sulphur containing amino acid. is called Lohmann’s reaction and is catalyzed
Nutritionally, it is essential amino acid. It by the creatine kinase. It is a reversible
serves as a precursor for the synthesis of reaction which can also lead to the synthesis
cysteine and cysteine which are therefore non of creatine phosphate from creatine and ATP.
essential.
Transmethylation
The transfer of the methyl group from
methionine to the acceptors is called trans- Lohmann’s Reaction
methylation. Creatine and choline are the two
Only traces of creatine are normally
important non-protein nitrogenous substances
present in urine. Large quantities may be
synthesized by transmethylation by methio-
excreted in muscular dystrophies, during
nine. Before donating its methyl group,
starvation and pregnancy.
methionine has to be converted to S-adenosyl
Creatinine is an anhydride of creatine and
methionine, also called active methionine in
it is in this form that creatine is excreted in
the presence of ATP.
normal health. It is formed in muscle from
METABOLISM OF CREATINE creatine-(P) by an irreversible, non-enzymatic
AND CREATININE reaction. Creatinine is a waste material and
is one of the major nitrogenous excretory
Creatine is a normal constituent of the body.
products.
It is present in muscle, kidney, liver and brain.
Urine of normal healthy adult contains
It can occur in free form as well as phosphory- creatinine but no creatine and this amount
lated form which is known as creatine excreted is independent of amount of proteins
phosphate. Creatine phosphate is formed from taken in the diet. Excretion is greater in
3 amino acids, i.e. glycine, arginine and muscular persons and hence related to muscle
methionine. mass.
Chapter 14

Minerals

Minerals are inorganic substances which The final group contains elements such as
unlike carbohydrates, proteins or fats, do not lead and mercury that are clearly toxic.
provide energy. Of the large number of
chemical elements found in the human body, CALCIUM
only a few have demonstrable biochemical
Calcium is present in largest quantity of all
or physiologic functions. These elements can
minerals in the human body. It comprises of
be considered in 5 groups.
about 2% of the body weight. About 99% of
The first includes carbon, hydrogen,
calcium of the body is present in bones and
oxygen, nitrogen, and sulphur, the major
teeth. The normal serum level is 9-11 mg%.
components of body molecules. These
Calcium along with phosphorous is essential
elements are obtained through intake of
for the formation and development of bones
water and food stuffs.
and teeth.
The second group includes the nutri-
Calcium is present in three forms:
tionally important minerals - calcium,
1. Ionised form (this is physiologically active
phosphorus, magnesium, sodium, potassium,
form).
and chloride—that are required in the diet in
amounts greater than 100 mg/dl. 2. Protein bound form (this is physiologically
The trace elements which forms the third inactive form).
group contains chromium, cobalt, copper, 3. Combination with citrates (calcium-citrate
iodine, iron, manganese, molybdenum, complex).
selenium, fluorine and zinc—are required in
Physiological Functions
the human diet in much smaller amounts.
The fourth group contains additional 1. Calcification of bone and teeth depend on
elements required for animal nutrition but adequate dietary intake of calcium.
having no known essential functions in 2. Ionised calcium is required in blood
humans: arsenic, cadmium, nickel, silicon, tin, coagulation process. It activates the
and vanadium. conversion of prothrombin into thrombin.
118 Medical Biochemistry for Physiotherapy Students
3. It regulates the excitability of nerve and DEFICIENCY STATE
nerve fibres.
4. It regulates the contraction of muscle Rickets
fibres. This is characterized by faulty calcification of
5. It is important in normal transmission of bones in children.
nerve impulses. This occurs due to:
6. It also regulates the permeability of 1. Vitamin D deficiency.
membranes.
2. Deficiency of calcium in the diet or a
7. It is required for the activation of several
combination of both.
important enzymes like ATPase enzyme,
succinate dehydrogenase and certain 3. Poor absorption of the calcium from
proteolytic enzymes. intestine.
8. It acts as second messenger for some 4. Increased serum alkaline phosphatase
hormones. activity.
5. Parathyroid deficiency.
Sources
Osteoporosis
Richest sources are milk and its products.
Also present in egg yolk, nuts, fish, beans and Fractures of bones occur specially in older
lentils. females, even after minor injury.
It occurs due to following causes:
Daily Requirement 1. Decalcification of bones due to calcium
Adult males and females: 800 mg deficiency in the diet.
In females during pregnancy 2. Vitamin D deficiency.
and lactation: 1,200 mg 3. Hypoparathyroidism.
Infants below 1-year of age: 350-540 mg
Children between 1-8 years: 0.8 -1.2 g PHOSPHORUS
Absorption of Calcium Phosphorus is essential for the formation and
1. Vitamin D is necessary in the diet to development of bones and teeth along with
promote absorption of calcium. calcium.
2. Acidic medium favours the absorption of
calcium. Physiological Functions
3. Higher levels of protein in the diet help
1. It is important for the formation of
to increase absorption of calcium.
phospholipids, phosphoproteins and
4. Organic acids, lactose and basic amino acids
in the diet favour calcium absorption. nucleic acids.
5. Optimum ratio of calcium: phosphorous 2. It is essential in the formation of energy
should be 1:1 for the convenient absorption rich compounds like ATP.
of both. 3. It functions in the buffering system in the
6. After the age of 55-60 years, there is cells, e.g. phosphate buffers.
gradual diminution of intestinal transport 4. It functions in the synthesis of RNA and
of calcium. DNA.
Minerals 119
5. It is required for the formation of coen- 2. It is found in certain enzymes like co-
zymes like NADP, AMP, ADP, etc. carboxylases.
6. It is required in the absorption of glucose 3. It also functions as a cofactor for oxidative
by phosphorylation. phosphorylation.
Sources
Sources
Phosphorous is present in milk and milk
products, egg yolk, nuts, fish and meat. Milk, eggs cabbage, cauliflower and fruits.

Daily Requirements Daily Requirement


Adults: 800 mg Adults: 200-300 mg
Women during pregnancy Infants: 100-150 mg
and lactation: 1,200 mg Children: 150-200 mg
Infants: 240-400 mg
Children: 800-1200 mg Absorption
Absorption Parathyroid hormone increases its absorption.
Absorption of phosphorous is associated with
calcium. High calcium intake diminishes Deficiency State
absorption of phosphorous by forming
insoluble calcium phosphates. Deficiency of magnesium causes depression,
muscular weakness and even convulsions.
Deficiency State Low values of serum magnesium are found
in uraemia, rickets and kwashiorkor.
1. Low phosphate level is associated with
rickets.
SODIUM
2. Serum phosphorous levels are decreased
in hyperparathyroidism and increased in Sodium is the main cation of the extracellular
hypoparathyroidism. fluid.
3. The deficiency of Vitamin D is the cause
of low serum phosphorous and results in
Physiological Functions
decreased calcification of bones.
1. It is largely associated with the regulation
MAGNESIUM of acid base balance.
Magnesium is present largely in bones in the 2. It maintains the osmotic pressure of the
form of salts along with calcium and phos- body fluids and thus protects the body
phorous. from excessive fluid loss.
3. It maintains normal neuromuscular func-
Physiological Functions tion.
1. It acts as activator for many of the phos- 4. It maintains normal permeability of the
phate group transfer enzyme. cells.
120 Medical Biochemistry for Physiotherapy Students
Sources 3. It maintains proper osmotic pressure.
4. It functions in water retention.
Major source is common salt used in kitchen.

Daily Requirement Sources


Adults: 5-15 g. Bananas, meat, chicken, fruits, potatoes, etc.
In tropical countries, intake is more.
Hypertensive patients should not take Daily Requirement
more than 1 g of sodium per day. Adults: 4 g.
Absorption
Absorption
Sodium is completely absorbed from the
gastrointestinal tract in normal conditions, Normally potassium is practically completely
however in diarrhoea large quantities are absorbed from gastrointestinal tract and only
excreted in faeces. about 10% of potassium is eliminated in urine.
In conditions like diarrhoea, a large amount
Disease State of potassium is lost in faeces.
An increased level of sodium in the serum is
called hypernatraemia. Disease State
It occurs in: Increased level of potassium in serum is
1. Hyperactivity of adrenal cortex as in known as hyperkalaemia. It occurs in
Cushing syndrome. following conditions:
2. Prolonged treatment of cortisone, ACTH 1. Renal failure
and sex hormones. 2. Severe dehydration
3. Increased retention of water in the body. 3. Addison’s disease
4. In case of increased blood pressure. 4. Shock
Decreased level of sodium in the serum is Decreased level of potassium in serum is
called hyponatraemia. It occurs in: known as hypokalaemia. It occurs in following
1. Use of diuretics conditions:
2. Sweating 1. Severe diarrhoea and vomiting.
3. In kidney disease 2. Prolonged use of diuretics.
4. In Addison’s disease. 3. Overactivity of adrenal cortex (Cushing
POTASSIUM syndrome) which causes increased excre-
tion of potassium in urine.
Potassium is the main cation of intracellular
4. In familial periodic paralysis.
fluid.
TRACE ELEMENTS
Physiological Functions
Iron
1. It is largely associated with the regulation
of acid-base balance. The total content of iron in a normal adult is
2. It aids in muscular contraction particularly only 4 to 5 g. About 60-70% of total iron is
cardiac muscles. present in haemoglobin.
Minerals 121
Physiological Functions DEFICIENCY STATE
1. Iron mainly functions in the transport of Iron Deficiency Anaemia
oxygen to the tissues by haemoglobin.
2. It is an essential component of haemo- Iron deficiency anaemia is fairly common in
globin, myoglobin, cytochromes, and the children, menstruating females and lactating
respiratory enzymes like cytochrome mothers. In children the appetite is poor.
Growth and development is retarded. In
oxidases, catalases and peroxidases.
females of child bearing age there is pallor of
3. It is also involved in the process of cellular
skin, breathlessness on exertion and giddi-
respiration.
ness.
Sources Treatment: Ferrous sulphate tablets for
anaemic women. A mixture of ferrous
Egg yolk, fish, meat, nuts, beans, spinach,
ammonium citrate sweetened with glycerine
fruits like apple, bananas, etc.
for children below the age of 12 months.
Daily Requirement
Iron Toxicity
Infants: 10-15 mg
The presence of the excess of iron in the body
Children: 1-3 years 15 mg
results in its deposition as haemosiderin in
4 to 10 years 10 mg
various tissues such as in liver, spleen, skin,
Older children and adults (Males) etc. This is called haemosiderosis.
11 to 18 years: 18 mg
After 19 years of age 10 mg COPPER

Female Copper is present in almost all tissues of the


body, liver is the richest source.
11-50 years of age and during pregnancy or
lactation: 18 mg Physiological Functions
After 51 years of age: 10 mg
Iron demand is more during pregnancy 1. It is required in the utilization of iron for
and lactation. Iron loss is more during the biosynthesis of haemoglobin.
menstruation. A monthly loss of 15-30 mg of 2. It is needed for bone formation as well as
for the maintenance of myelin sheath with-
iron occurs.
in the nervous system.
Absorption 3. It is a constituent of many important enzy-
mes like cytochromes a3, catalase, tyrosi-
Absorption of iron occurs mainly in the nase and ascorbic acid dismutase.
stomach and the duodenum. The absorption 4. It is involved in biological oxidation.
of iron depends upon several factors such as
the form of iron and other constituents of the Sources
diet. Vitamin C, HCl and alcohol promotes Milk and its product are poor sources,
iron absorption while consumption of tea however liver, kidney, meat, nuts and leafy
decreases it. vegetables are richest sources.
122 Medical Biochemistry for Physiotherapy Students
Daily Requirement Physiological Functions
Adults: 2.5 mg Iodine is required in the body for the
Infants and children: 0.05 mg/kg body formation of thyroxine and triiodothyronine
weight hormones of the thyroid gland. Functionally,
triiodothronine is more active than thyroxine.
Absorption These thyroid hormones are responsible for
About 10-30% of dietary copper is absorbed growth, reproduction, cellular oxidation and
from duodenum. After absorption, copper is also in the activity of central and autonomic
excreted in the bile by the liver. The major nervous system.
portion of the dietary copper appears in the
faeces. Very little of it is excreted in urine. Sources
Sea foods, marine vegetables and fruits
Deficiency State grown on sea belt. High altitudes are deficient
Severe deficiency of copper causes skeletal in iodine content. Table salts contains good
changes like demineralization of bones, amount of iodine.
demyelization of neural tissue, anaemia,
graying of hair and hypopigmentation of skin. Daily Requirement
Adults: 100-150 μg/day
COPPER TOXICITY Adolescents and pregnant women: 200
Wilson’s Disease: (Hepato-lenticular μg/day.
Degeneration)
Absorption
Wilson’s disease is a rare hereditary disorder
Iodine is quickly absorbed from food and
of copper metabolism. It is characterized by
water as inorganic iodide by the gut. A
the following manifestations:
portion of it is taken up by the thyroid gland
1. Copper is deposited in abnormal amounts
for the formation of thyroxine and tri-
in liver and lenticular nucleus of the brain
iodothyronine.
which causes hepatic cirrhosis and necrosis
of brain. Excretion
2. Copper deposition in kidney causes renal Excretion of iodine occurs mainly through
tubular damage and affects glomerular kidneys. It is also excreted through bile,
filteration. saliva, skin and milk (lactating mothers).
3. Low levels of copper and ceruloplasmin
in plasma and increased excretion of urine. Deficiency State
Treatment: Administration of copper chelating The deficiency of iodine causes goitre.
agent pencillamine. This causes marked (discussed in details under Chapter: Hor-
increase in urinary excretion of copper. mones).

IODINE FLUORINE
About 80% of total body iodine is present in Physiological Functions
thyroid gland. The total body contains about 1. Fluoride, in trace amounts is necessary for
20 mg of iodine. proper development of teeth and bones.
Minerals 123
2. Fluoride prevents development of dental ZINC
caries, by forming a layer of acid resistant
fluoroapatite with hydroxyl apatite of the Zinc is mainly an intracellular element. The
enamel and prevents tooth decay by total amount of zinc in the body is about 2 g.
bacterial acids.
3. It also inhibits activities of certain enzymes Physiological Functions
like fluoroacetate inhibits aconitase of 1. Zinc is an essential constituent of several
citric acid cycle and sodium fluoride enzymes like carbonic anhydrase, carboxy-
inhibits enolase of glycolysis. peptidase, alkaline phosphatase, etc.
2. It plays an important role in the healing
Sources of wounds.
Drinking water is the main source of fluoride. 3. It helps in maintaining normal concentra-
tion of vitamin A in plasma.
Daily Requirement 4. It plays a role in the storage and secretion
of insulin from the β-cells of pancreas.
Intake of less than 2 ppm per day is sufficient
to meet the requirement.
Sources
Absorption Meat, liver, eggs, oysters, fish, beans are good
Soluble fluorides are rapidly absorbed from sources. Milk is a poor source.
small intestines.
Daily Requirement
Deficency Adults: 10-15 mg/day
Deficiency of fluoride promotes the develop- Pregnancy and lactation: 25-30 mg/day.
ment of dental caries in children and osteo-
porosis in adults, particularly in postmeno- Absorption
pausal women.
Zinc present from animal foods are well
Fluoride Toxicity absorbed from duodenum. Zinc present from
Excess amounts of fluoride causes fluorosis. plant sources are poorly absorbed due to
The beneficial effects of fluoride in trace presence of phytic acid which interferes in its
amounts are overshadowed by the harmful absorption.
effects during excess amounts. The enamel of
teeth loses its colouration, chalky white patches Deficiency
with yellow or brown pigmentation are found Zinc deficiency is associated with poor
over the surface of the teeth. Very high
growth, retardation, dwarfism, hypo-gona-
concentration of fluoride causes hypercalcifi-
dism, anaemia, etc.
cation of bones of spine, pelvis and limbs.
Ligaments of spine and collagen of bones are
Zinc Toxicity
calcified. These changes lead to skeletal fluorosis.
In advanced stages, the individuals are Zinc toxicity occurs in welders because of
crippled and unable to do daily routine work. inhalation of zinc oxide fumes and causes
Neurological disturbances are also common. nausea, vomiting, gastric ulceration, etc.
Chapter 15

Fluid and Electrolytes


Balance and Imbalance

Water is the most abundant compound in averages about 15 litres in a 70 kg adult. It is


living cells which contains 65 – 95% of water heterogeneous and can be subdivided into:
by weight in normal individuals. Water 1. Plasma
possesses several unique features due to its 2. Interstitial and lymph fluid
polarity and hydrogen bonding properties. 3. Dense connective tissue, cartilage, bone
1. It is a powerful solvent for many ionic 4. Transcellular fluids.
compounds and neutral molecules.
2. It has strong influence along with dilute 1. Plasma: It is noncellular portion of the
salt solutions on the state of dissociation blood. It is the part of the extracellular
of macromolecules of the cell. fluid and communicates continually with
3. It exerts a major influence on the structural the interstitial fluid through pores in the
and functional components of the cell. capillaries.
4. The high heat of vaporization of water 2. Interstitial and lymph fluid: They may be
helps in cooling by evaporation of moisture considered to represent the actual fluid
in the lungs and from the skin. environment outside the cells. Continuous
mixing and exchange of nutrients and
BODY FLUID DISTRIBUTION metabolic waste products takes place
between plasma and interstitial fluid, since
Total body fluid is distributed between two
they intermingle through pores of blood
main compartments:
capillaries.
A. Extracellular fluid
3. Dense connective tissue, cartilage, bone, does
B. Intracellular fluid.
not exchange fluid or electrolyte readily
with the remainder of the body water due
Extracellular Fluid (ECF)
to difference in the structure and relative
All of the fluid outside the body cells is avascularity.
collectively termed the extracellular fluid. The 4. The transcellular fluids are the fluids found
total amount of extracellular compartment in the salivary glands, pancreas, liver,
Fluid and Electrolytes Balance and Imbalance 125
biliary tract, kidneys, gastrointestinal change by the osmotic forces which direct the
tracts, skin, gonads, thyroid gland, movement of water from one compartment
cerebrospinal fluid and fluids within the to another in the body. These osmotic forces
spaces of the eye and within the lumen of are produced and controlled by the presence
the gastrointestinal tract. of following substances in the body fluids.
1. Electrolytes: These are the most important
Constituents of ECF substances which influence the distribution
ECF both of blood plasma and of interstitial and retention of water. Sodium (chief
fluid contains large quantities of sodium and cation of ECF) and potassium (chief cation
chloride ions, reasonably large quantities of of ICF) are the most important osmotically
bicarbonate ions, but only small quantities of effective electrolytes.
potassium, calcium, magnesium, phosphate, 2. Organic substances of large molecular weight
sulphate and organic acid ions. In addition, and size (serum proteins): An important
plasma contains a large amount of protein, function of the serum proteins is the
whereas interstitial fluid contains much less. maintenance of osmotic balance between
the circulating blood and the tissue spaces.
Intracellular Fluid (ICF) Albumin is of major importance in
maintaining serum osmotic pressure. One
The fluid within the body cells is called the gram of albumin holds 18 ml of fluid in the
intracellular fluid. This is larger of the two blood stream.
major fluid compartments. The concept of a 3. Organic substances of small molecular size
single intracellular fluid is used, since the fluid (glucose, urea, amino acids, etc.): These
within each individual cell is fairly constant substances diffuse relatively freely across
in composition. cell membranes hence they are not impor-
tant in the distribution of water. They influ-
Constituents of ICF
ence total body fluid, if present in large
ICF contains only small quantities of sodium quantities.
and chloride ions and almost no calcium ions;
but it does contain large quantities of Fluid Output
potassium and phosphate and moderate Fluid loss occurs via the following four routes:
quantities of magnesium and sulphate ions, 1. Urine excretion: It normally amounts to
all of which are present in only small 1-1.5 L/24 hrs but this figure may rise to
concentrations in the ECF. In addition, cells clear a heavy fluid load or fall if water
contain very large amounts of protein approx. supplies are limited.
4 times as much as in plasma. 2. Insensible fluid loss: It refers to the continuous
evaporation from the surface of the skin
Factors which Influence the
and airways. Daily losses of this approxi-
Distribution of Body Water
mates to 400 ml, although increased
Water is retained in the body in constant ventilation during heavy exercise may
amount. Its distribution is subjected to double the respiratory component.
126 Medical Biochemistry for Physiotherapy Students
3. Faecal excretion: It accounts for approxi- 2. Another important cause is excessive loss
mately 100 ml of fluid each day. of potassium from the body.
4. Sweating: It leads to additional fluid and Another regulatory mechanism is secretion
electrolyte losses, separate from insensible of ADH (antidiuretic hormone) or vaso-
skin losses about 600-800 ml/24 hrs. With pressin.
strenuous exercise, hot environmental 1. An increase in osmolarity that excites
conditions or during a fever, it can be the osmoreceptors located in the anterior
dominant fluid output. hypothalamus.
2. Excitation of the osmoreceptors in turn
Fluid Intake
causes the posterior pituitary gland to
Water is added to the body fluid compart- release ADH.
ments by two main mechanisms. 3. The ADH increases the permeability of the
1. Intake of water and other fluids: These are
distal tubules, the cortical collecting ducts
taken by mouth as water and beverages.
to water and therefore cause increased
It amounts to about 1200-1500 ml/day.
conservation of water by the kidneys.
2. Metabolic production: It is a byproduct of
4. The conservation of water but loss of
substrate oxidation in body cells e.g.
sodium and other osmolar substances in
carbohydrate metabolism. This normally
the urine causes dilution of sodium and
generates about 300 ml of metabolic water
other substances in the ECF, thus correc-
each day.
ting the initial, excessively, concentrated
3. Water taken in cooking in cooked foods: It
ECF.
averages about 700 ml/day.
Conversely, when the ECF becomes too
REGULATION OF FLUID AND dilute (hypo-osmotic), less ADH is formed
ELECTROLYTE BALANCE and excess water is lost in comparison with
Any mismatch between fluid intake and the ECF solutes, thus concentrating the body
output will lead to fluid accumulation or fluids back towards normal.
depletion within the body. This alters the Another important regulatory mechanism
extracellular fluid volume or may change its is by aldosterone. Aldosterone is a hormone
osmolarity, and it is these effects which are secreted by adrenal cortex. Aldosterone is
detected by receptors, leading to compen- stimulated in response to the following three
satory changes in fluid intake and loss. factors:
Fluid intake is mainly controlled by 1. Decreased ECF sodium ion concentration.
altering drinking behaviour that causes 2. Increased angiotensin II in blood.
drinking. This is regulated through the thirst Whenever either the ECF volume or the
centre in the hypothalamus. Basic stimulus for ECF sodium ion concentration falls below
exciting thirst is intracellular dehydration. normal (normal is 142 mEq/l), aldosterone is
1. The most common cause for intracellular secreted and the renal tubules reabsorb extra
dehydration is increased osmolar concen- quantities of sodium and water, leading to
tration of the ECF, especially increased return of both ECF sodium and ECF volume
sodium concentration. back towards normal.
Fluid and Electrolytes Balance and Imbalance 127
Increased angiotensin II has a direct effect Treatment
on the zona glomerulosa cells to increase the
The treatment involves consuming plenty of
secretion of aldosterone.
plain water or water containing sugar and
The combined ADH-thirst mechanism is
salt which depends upon the cause of
powerful controller than the aldosterone
system so, ADH-thirst mechanism greatly dehydration. If the condition is very severe,
overshadows the aldosterone system for intravenous infusion of fluids (normal saline)
regulation under normal conditions. is required.

DEHYDRATION WATER INTOXICATION


Dehydration is a condition in which there is
loss of water from the body in excess Water intoxication is a condition in which
amounts. It can be due to loss of water alone body water content is increased. It can be
or due to deprivation of water and electro- caused by:
lytes. 1. Renal failure
Dehydration can be caused by: 2. Hypersecretion of ADH following the
1. Severe diarrhoea and vomiting administration of anaesthetics.
2. Excessive sweating 3. Excessive administration of fluids paren-
3. Loss of fluid from skin in case of burns
terally.
4. Diabetes insipidus (due to deficiency of
ADH) 4. Excess of aldosterone may lead to an
5. Non-availability of drinking water as in overhydration of the body and subsequent
the case of desert. water intoxication.
6. Heat stroke.
Features of Water Intoxication
Features of Dehydration
1. Decreased plasma electrolytes
1. ECF volume decreases and there is relative
2. Decreased plasma proteins
rise in electrolyte concentration and
osmotic pressure. 3. Mental confusion
2. Changes in the plasma proteins, urea 4. Incoordination
occurs. 5. Muscular weakness
3. Dryness of skin and tongue occurs. 6. Nausea.
Chapter 16

Biophysics

HYDROGEN ION CONCENTRATION (pH) pOH is defined as negative logarithm of


hydroxyl ion concentration to the base 10.
The hydrogen ion concentration or pH is a Ionization exponent of water (pKw) is the
measure of the acidity or alkalinity of a negative log of Kw to the base 10. It is constant
solution. It is defined as the negative at a given temperature.
logarithm of hydrogen ion concentration to pKw = –log Kw
the base 10. It is expressed as follows: = –log ([H+] X [OH–])
1 = pH + pOH
pH = -log [H+] = log ________
For a dilute neutral aqueous solution at
+
[H ]
room temperature,
+
[H ] is the hydrogen ion concentration of pKw = pH +pOH
the solution in moles per litre. The pH scale = 7 +7
ranges from 0 to 14. This pH scale was = 14.
introduced by Sorensen. An acid solution has
a pH value less than 7.0 and a basic solution Henderson-Hasselbalch Equation
has a pH value greater than 7.0. A neutral
Let us take a weak acid HA. The dissociation
solution has a pH value of 7.0. A change of
of it can be represented as follows:
one pH unit corresponds to a 10-fold change
HA = H+ + A–
of hydrogen ion concentration.
The dissociation constant (K) is written as
If a solution is described as pH 6, its
hydrogen ion concentration is 10–6 and its [H + ] [A - ]
K=
hydroxyl ion concentration is 10–8 at room [HA]
temperature. The value of using pH can be By rearrangement, we get
seen in the case of human blood which has an K [HA] = [H+] [A–]
extremely low hydrogen ion concentration.
K [HA]
Blood [H+] = 0.398 × 10–7 [H + ] =
Blood pH = –log (0.398 × 10–7) [A  ]
= 7.4 Taking logarithm of both sides
Biophysics 129
complementary base pairs in DNA and
[HA]
log [ H + ] = logK + log RNA.
[A  ] 6. The pH dependence of the charged forms
Change sign of both sides of proteins and amino acids is utilized in
separating and isolating them from
[HA] biological materials by chromatography
 log [ H+ ] = - log K - log -
[A ] and electrophoresis.
[A ]-
pH = pK + log Measurement of pH
[HA]
[salt] pH is measured by the following ways:
= pK + log
[acid] 1. By pH papers
2. By indicators
This is known as Henderson-Hasselbalch 3. By pH meters
equation. It follows that when [A-] = [HA], The pH of blood is normally 7.4 (7.36 –
pH equals pK. 7.44). It is maintained mainly by three
mechanisms:
Biological Significance of pH
1. Buffer systems in blood and body fluids
1. pH influences the ionization of polar 2. Respiratory mechanisms
groups of amino acids, proteins, nucleic 3. Renal mechanisms.
acids, phospholipids etc. at a specific pH
called isoelectric pH, each molecule exists Buffer Systems
as a zwitter ion having both anions and Buffers are mixtures of weak acids and their
cations. Below and above this pH, molecule corresponding salts or weak bases and their
exists as anion or cation. corresponding salts. Buffer solution is defined
2. pH affects the ionic and hydrogen bonds as a solution which prevents marked changes
which stabilize the three dimensional in pH when either an acid or base is added
structure of proteins. Each protein has an to it. For example, if only a few drops of conc.
optimum pH at which it can best maintain HCl are added to a beaker of pure water, the
its structure. Drastic changes can result in pH of the water might immediately fall from
its inactivation and denaturation. a neutral 7.0 to as low as 1.0. However, if a
3. There is optimum activity of enzyme at satisfactory buffer is added, the HCl com-
which its activity is best. At above or bines instantaneously with the buffer and the
below this pH, activity of enzyme declines. pH falls only slightly.
4. pH also affects the membrane structure Buffer systems of body are as follows:
by influencing the ionization of membrane In ECF (extracellular fluid), following
proteins and lipids. buffer systems are present:
5. Purine and pyrimidine bases exist as a. Bicarbonate buffer system
tautomers according to pH which are b. Phosphate buffer system
essential for the hydrogen bonding of c. Protein buffer system.
130 Medical Biochemistry for Physiotherapy Students
In ICF (intracellular fluid), following Therefore, the net result is exchange of
buffer systems are present: the strong base for the weak base NaHCO3.
a. Phosphate buffer system This system is more important than other
b. Protein buffer system. buffer systems of the body because the
In RBCs, haemoglobin buffer system is concentration of each of the two elements of
present. the bicarbonate system can be regulated,
carbon dioxide by the respiratory system and
Bicarbonate Buffer System the bicarbonate ion by the kidneys. As a
result, the pH of the blood can be maintained.
The bicarbonate buffer system consists of a
mixture of weak acid carbonic acid (H2CO3) Phosphate buffer system
and its corresponding base sodium bicar- The phosphate buffer system consists of a
bonate (NaHCO3) in the same solution. Salt mixture of weak acid dihydrogen mono-
to acid ratio is 20:1 in blood. sodium phosphate or acid phosphate (NaH2
If any strong acid like HCl enters in the PO4) and its corresponding base disodium
ECF, then it reacts with weak base, i.e. monohydrogen phosphate (Na2HPO4) in the
NaHCO3. The following reactions take place same solution. Salt to acid ratio is 4:1 in
plasma.
If any strong acid like HCl enters in the
ECF, then it reacts with weak base, i.e.
NaHPO4. The following reactions take place

Effect of Bicarbonate Buffer System


on Strong Acid

The net result is conversion of strong acid


hydrochloric acid into weak acid carbonic
acid and salt. It means addition of the HCl Effect of Phosphate Buffer System
lowers the pH of the solution only slightly. on Strong Acid
The carbonic acid is volatile; it is converted
to water and gas CO2 which is exhaled out of The acid phosphate thus produced are
body through lungs. excreted by the kidneys, hence urine becomes
Similarly, if strong base like NaOH enters more acidic.
ECF, it is fixed up by the acid component of Conversely, if a strong base like NaOH
the bicarbonate buffer system, i.e. carbonic enters blood, it is fixed up by acid component
acid, as given below:

Effect of Bicarbonate Buffer Effect of Phosphate Buffer System


System on Strong Base on Strong Base
Biophysics 131
The alkaline phosphate thus produced is When the blood returns to the lungs, these
excreted in urine which cause increased H+ ions are released because of formation of
alkalinity of urine. a stronger acid (oxyhaemoglobin) and the
newly released H+ ion is promptly neutra-
Protein Buffer System lized by HCO3–. This reaction is necessary for
Buffering capacity of plasma proteins is less the liberation of CO2 in the lungs.
than Hb. Buffer system consist of weak acid
H+ Pr– and its corresponding base Na+ Pr–.
Strong acids combine with salt component
and thus produces weak acid.

Effect of Protein Buffer System on Strong Acid

Haemoglobin Buffer System


At the lungs, the formation of oxyhaemo- Haemoglobin as a Buffering Agent
globin from reduced haemoglobin releases
hydrogen ions which react with bicarbonate Respiratory Mechanism
to form carbonic acid. The low CO2 tension The respiratory mechanism involves control
in the lung shifts the equilibrium towards the of CO2 by the lungs by increasing or dec-
production of CO 2 which is continually reasing the rate of respiration.
eliminated in the expired air. The respiration rate is controlled by the
receptors in the respiratory centre which are
sensitive to changes in pH and pCO2 of blood.
Production of CO2 When there is fall in pH of the plasma, the
respiratory centre is stimulated resulting in
In the tissues, the oxygen tension is hyperventilation which eliminates more CO2
reduced and hence oxyhaemoglobin dis- thereby lowering H 2CO 3 concentration in
sociates delivering O2 to the cells and reduced blood. If the blood pH increases, the respi-
haemoglobin is formed. CO2 produced by ratory centre is inhibited so that elimination
metabolism enters blood, where it is hydrated of CO2 is decreased by hypoventilation until
to form H2CO3 which ionizes to form H+ and the blood pH comes to normal.
HCO3–. Reduced haemoglobin acting as an
anion accepts the H + ions forming acid- Renal Mechanism
reduced haemoglobin (HHb). Very little
change in pH occurs because the newly An important function of kidney is to
arrived H+ are buffered by formation of very regulate the function by excreting either an
weak acid. acidic or a basic urine. The pH of urine ranges
132 Medical Biochemistry for Physiotherapy Students
from 4.5 to 9.5 because the renal system plays It occurs in:
a significant role in long-term pH maintenance 1. Damage to the respiratory centre in the
of the blood at 7.4 + 0.05. This is possible by medulla oblongata that causes reduced
its capacity of reabsorption, secretion and breathing.
excretion of the non-volatile acids like lactic 2. Obstruction of the passageways in the
acid, pyruvic acid and inorganic acid, HCl, respiratory tract.
phosphoric acid and H 2 SO 4 which are 3. Pneumonia.
produced in the body cannot be excreted by 4. Decreased pulmonary membrane surface
lungs. area.
Compensatory mechanism is renal mecha-
Acid-base Imbalance
nism. More of HCO3– is reabsorbed from renal
Acid-base imbalance includes conditions tubules which cause ratio to return towards
acidosis and alkalosis. normal as levels of both components is
increased.
Acidosis
Acidosis is increase in H+ concentration or Alkalosis
decrease in pH as pH is inversely proportional Alkalosis is decrease in H+ concentration or
to H + concentration. It is of two types: increase in pH as pH is inversely proportional
metabolic acidosis and respiratory acidosis. to H + concentration. It is of two types:
metabolic alkalosis and respiratory alkalosis.
Metabolic Acidosis
In this condition, there is decrease in the Metabolic Alkalosis
HCO3– fraction with little or no change in the In this condition, there is increase in the
H2CO3 fraction. As a result, the ratio of 20 : 1 HCO3– fraction. As a result, the ratio of 20 : 1
is decreased and pH is decreased. is increased and pH is increased.
It occurs in It occurs in:
1. Uncontrolled diabetes mellitus 1. Vomiting when there is loss of gastric HCl
2. Starvation 2. Ingestion of bicarbonate in the treatment
3. Severe exercise of peptic ulcer
4. Ingestion of acids 3. Excess aldosterone secretions mellitus.
5. Diarrhoea. Compensatory mechanism is respiratory
Compensatory mechanism is respiratory mechanism. There occurs hypoventilation that
mechanism. There occurs hyperventilation slows removal of CO 2 and retention of
that removes CO2 and reduction in H2CO3 . H2CO3. As a result, the ratio returns towards
As a result, the ratio returns towards normal normal as levels of both in blood are
as levels of both in blood are decreased. increased.

Respiratory Acidosis Respiratory Alkalosis


In this condition, there is increase in H2CO3 In this condition, there is decrease in H2CO3
in blood. As a result, the ratio of 20 : 1 is concentration in blood. As a result, the ratio
decreased and pH is decreased. of 20 : 1 is increased and pH is increased.
Biophysics 133
It occurs in: Osmosis Requires
1. Lack of oxygen at high altitude.
a. Semi permeablility of the membrane
2. Fever. separating two solutions (or a solution and
3. In CNS diseases, e.g. meningitis. the pure solvent) so that the membrane is
Compensatory mechanism is renal mecha- permeable to water but not to the solute.
nism. More of HCO3– is lost into the urine b. difference in solute concentration on the
which cause ratio to return towards normal two sides of the membrane.
as levels of both components is decreased. c. In such a case, water molecules diffuse in
both directions across the semi permeable
Osmosis membrane, but a net osmosis of water
Osmosis is defined as a process of net takes place from the dilute to the concen-
diffusion of water from either a dilute trated solution which results from a larger
number of water molecules diffusing in
solution or pure water to a more concentrated
that direction than in the reverse direction.
solution across a semi permeable membrane
Osmosis continues until the hydrostatic
which permits the diffusion of water but not
pressure rises so high on the concentrated
of the solute. For example, if an aqueous side of the membrane as to cause a
solution of sucrose is taken in an inverted diffusion of water in the opposite direction
thistle funnel with its mouth closed by a semi at the same rate as the osmotic inflow.
permeable cellophane or fish bladder
membrane and dipped into water in a beaker, Osmotic Pressure
water flows across the membrane from the
Osmotic pressure is defined as the pressure
beaker to the sucrose solution raising the fluid which has to be exerted on the concentrated
level in the stem of the inverted funnel (Fig. solution, separated from pure water by a semi
16.1). permeable membrane, in order to counteract
and stop the osmotic inflow into the solution.
It equals the difference between the
hydrostatic pressures on the two sides of
membrane.
Osmotic pressure is a colligative property
of a solution. A rise in the number of solute
particles in the solution increases the number
of solvent particles bound by solute particles
in complexes called solvates.
Osmotic pressure of a dilute solution is
directly proportional to other colligative
properties like fall in freezing points, lowering
of vapour pressure, etc. Osmotic pressure is
inversely proportional to the molecular
Osmosis weight of the solute.
134 Medical Biochemistry for Physiotherapy Students
Units 2. The colloid osmotic pressure of plasma
proteins causes water to be retained in
Osmotic pressure is expressed in mm Hg or
plasma.
dynes per square cm. Osmol units (Osm) give
3. Water is absorbed from lumens of intes-
the number of osmotically active particles per
tines and renal tubules by the osmotic
mole of a solute; each mole of a non ionized
solute is equivalent to 1 Osm. effects of absorbed solutes such as sodium
and glucose (obligatory absorption of
Determination of Osmotic Pressure water).

Osmotic pressure is measured by Applications of Osmosis


1. Osmometer
2. Berkeley-Hartley method 1. Pain experienced by the application of salt
3. Freezing point method. on the cut of the skin is due to the osmotic
withdrawl of water from the exposed area
Biological Significance of Osmosis by salty solution.
2. The pain caused by contact of sugar with
1. If red blood cells are kept in isotonic
exposed nerves of teeth is due to the
solution (0.9% NaCl), then neither water
osmotic withdrawl of water from the
flows into them nor water will go out of
exposed area by strong sugar solution.
red blood cells because concentration of
solute is same on both sides. If red blood 3. Water or salts (chiefly NaCl) are excreted
cells are placed into hypotonic solution, by the kidney to keep the blood isotonic
(say 0.3 % NaCl), water passes into cells with the cells. Hence, osmosis is of great
from the hypotonic solution and makes the importance in the process of urine
cells swell. This kind of osmolysis of red secretion.
blood cells is called haemolysis. On the 4. The clinical application of osmotic force is
other hand, if the cells are kept in the injection of hypertonic solution of
hypertonic saline, (say 1.5% NaCl), the cells magnesium sulphate to reduce the volume
get shrunken due to osmotic outflow of of the brain or lower the pressure of
water (exosmosis). This is called crenation. cerebrospinal fluid.
Chapter 17

Metabolism of Specialized Tissues

MUSCLE TISSUE Structure of skeletal muscle: Skeletal muscles are


composed of muscle fibres that are organized
Muscle is the largest single tissue in the into bundles. Each bundle of muscle fibres is
human body and contributes about 25-40% termed a fasciculus. The myofibrils are the
of body weight. Its main function is to convert only contractile elements of the muscle fibre.
chemical energy released by the breakdown The myofibrils are oriented in a longitudinal
of ATP to mechanical energy in the form of direction, parallel to one another and run in
muscular contraction. the entire length of the fibre. Its diameter is
50 to 100 ìm and the length of a muscle fibre
Three different types of muscle tissues varies from a few mm to 60 or 70 cm.
occur: Each muscle fibre is enclosed in a mem-
1. Skeletal muscle brane or covering known as sarcolemma.
2. Smooth muscle Sarcolemma is a thin, selectively permeable
3. Cardiac muscle membrane.
Within each muscle cell is a specialized and
Skeletal muscle or voluntary muscle: Most of this
undifferentiated protoplasm known as sarco-
attached to the skeleton and form ‘somatic’
plasm. As a rule, muscles in constant activity
musculature. These show well developed
(ocular, respiratory) have the greatest amount
cross-striations, therefore also called striated of sarcoplasm whereas muscles that contract
muscle. quickly and fatigue readily contain the least.
Smooth muscle or involuntary muscle: These are The sarcoplasm houses a diversity of granules
present mostly in hollow viscera. These lack of different sizes: mitochondria, fat and
lipoprotein droplets.
cross striations, therefore also called plain
The basic contractile unit of the muscle is
muscle.
a sarcomere which is made up of inter-
Cardiac muscle: These are present in heart digitating filaments—thick filaments made of
only. These have well developed cross- myosin and thin filaments made of actin
striations. (Fig.17.1).
136 Medical Biochemistry for Physiotherapy Students
the I band into the A band on either side up
to the H disc. So there is an overlap of the A
and I filaments in the A band, while only I
filaments are present in the I band.
Sarcomere is the name given to the region
between the centre of one I band to that of
the next I band (includes a complete A band
in the middle, with half of I band on either
side) and is considered as a single functional
segment.

Chemical Composition
75% of skeletal muscle is water, 20% is protein
and the remaining 5% is made up of inorganic
salts and other substances including high-
energy phosphates, urea, lactic acid, the
minerals calcium, magnesium, phosphorous,
various enzymes, ions of sodium, potassium,
chlorides and amino acids, fats and carbo-
hydrates.
The most abundant muscular proteins are
myosin, actin and tropomyosin. They comp-
Fig. 17.1: Structure of skeletal muscle rise about 52%, 23% and 15% respectively of
the muscle’s total protein content. In addition,
about 700 mg of the conjugated protein myo-
Myofibrils globin is incorporated into each 100 g of
muscle tissue.
The fibrils show cross striations - wide bands
called A bands alternating with narrower Proteins of Sarcoplasm
bands called the I bands. The A bands are
The sarcoplasm which bathes all the formed
rich in the fibrous protein myosin and are
elements of the muscle cell contains a complex
strongly birefringent (anisotropic; hence mixture of proteins. They are albumin-like
called ‘A bands’). The I band is isotropic and are called myogens A and B. There is
(hence called I band) and shows two lines — also a globulin-like protein called globulin-X.
the N and Z lines. Each is a mixture of several proteins. It
The A band is said to be made up of contains the glycolytic and other enzymes.
filaments 10 nm diameter. As they enter the
H region in the band, they thicken to 14 nm, Proteins of Myofibrils
thus giving the appearance of a disc to that Myosin, actin and tropomyosin are the main
region. The I band contains only filaments of proteins. The actin and myosin are associated
uniform diameter of 6 nm which enter from as actomyosin.
Metabolism of Specialized Tissues 137
Myosin molecule is shaped as a long rod the muscle. The released calcium binds to the
with a globule at one end. Each myosin TpC subunit of the troponin complex and are
molecule consists of 6 polypeptide chains. transmitted to tropomyosin and then to actin.
Molecular weight of myosin is in the range This allows actin to act with myosin giving
of 520,000 to 600,000. It occurs in thick rise to muscular contraction with the
filament. hydrolysis of ATP which acts as energy
Actin occur in two forms: globular G-actin source. This process continues until calcium
and fibrous F-actin. The molecular weight is ion is removed.
60,000. It is major constituent of thin filaments.
Actomyosin is a contractile system of Energy Source for Muscular Contraction
skeletal muscle and is made up of F-actin and The only immediate source of energy for
myosin. Formation or cleavage of this muscular contraction is ATP. This ATP must
complex is involved in all phases of the muscle be continually renewed. In muscle cells, ATP
cycle i.e. contraction, relaxation, rigor etc. is produced at rest and during light exercise,
Tropomyosin and troponin complex are mainly because of oxidation of stored fat in
proteins present in the thin filaments of the form of free fatty acids. As intensity of
muscle. Tropomyosin is a double stranded exercise increases, fats alone cannot supply
helical rod of molecular weight 70,000 and energy fast enough and so much of the energy
present between the two strands of F-actin. comes from the breakdown of glucose and
Troponin molecules consists of 3 subunits its stored form glycogen (aerobic glycolysis).
which occur at regular intervals along the
With strong muscular contractions, this
length of the thin filament.
process of aerobic glycolysis may be inade-
The three subunits are:
quate. There are three further processes
1. Troponin T (TpT), which binds to tropo-
which can take place to provide energy.
myosin.
2. Troponin I (TpI), which inhibits the F-actin 1. Oxygen supply: Glycogen requires oxygen
myosin interactions. for its oxidation. Muscle contains a red
3. Troponin C (TpC) which binds calcium. pigment, myoglobin which is similar to
haemoglobin. Myoglobin forms complexes
Skeletal Muscle Shows with oxygen and acts as a store. When the
Two Types of Fibres concentration of oxygen in the cells is low,
Type I: Red (slow) and type II: White (fast). myoglobin releases its oxygen.
Type I uses oxidative metabolism for its 2. Anaerobic metabolism: In addition to getting
function (citric acid cycle) and is rich in ATP from the complete oxidation of
mitochondria and myoglobin. glycogen, a muscle cell can breakdown
Type II uses glycolysis (anaerobic meta- further glycogen part in the way, with the
bolism). There is no myoglobin and very few formation of lactic acid. This reaction does
mitochondria. not need oxygen and is called anaerobic
glycolysis.
Role of Calcium 3. Storage of high-energy phosphate: In a single
Calcium ions are released from the sarco- muscle fiber, there is comparatively
lemma by the excitation of a motor nerve to little ATP storage. It can, however, be
138 Medical Biochemistry for Physiotherapy Students
regenerated quickly from high-energy Ground Substance
creatine phosphate, also called phospho-
Ground substance is distributed throughout
creatine, which is stored in large amounts.
the extracellular space. It is a gel like sub-
stance containing water, salt, proteins and
During exercise
polysaccharides in solution. The polysaccha-

ADP + Creatine phosphate   ATP + Creatine
At rest or during
rides are also described as muco-polysaccha-
muscle relaxation rides. Hyaluronic acid, chondroitin sulphate
This reaction takes place when ATP levels and heparin are a few examples. They are
are low. It can continue until all the creatine complex polysaccharides usually containing
phosphate is used up. It is later restored by glucosamine or galactosamine and glucuro-
means of the converse reaction during muscle nate units. The amino group is usually acety-
relaxation; some ATP in the mitochondria lated or sulfated. Due to the ionization of
transfers its phosphate to creatine. these acid groups, they are strongly anionic
and combine with cationic proteins to form
Applied Aspect muco-proteins (glycoproteins). The muco-
When muscle fibres are completely depleted proteins have very large molecules and very
of ATP and phosphocreatine, they develop a high molecular weights.
state of extreme rigor. When this occurs after The enzyme, hyaluronidase, which is
death, the condition is called rigor mortis. In present in some micro-organisms, snake
this, almost all of the myosin heads attach to venom and seminal fluid, hydrolyzes the
actin but in an abnormal, fixed and resistant polysaccharide hyaluronic acid and thus
way. makes the ground substance readily perme-
able to the micro-organisms or their toxins.
CONNECTIVE TISSUE Infection thus spreads in the tissue. Hence, it
is called the ‘spreading factor’. In case of
Connective tissue is distributed throughout
seminal fluid, it helps in clearing the way for
the body. It makes up the capsules and
the spermatozoa to reach the ovum.
framework of organs and the sheaths of
tendon and muscle. The skin is attached to Adrenal cortical hormones and growth
the rest of the body by connective tissue and hormone regulate the synthesis of the
connective tissue forms the supporting mucopolysaccharides. Vitamin C deficiency
framework of blood vessels, nerves and and diabetes mellitus decrease their synthesis.
lymphatics tissue. It anchors the organs in Hence delayed wound healing occurs in these
position. two conditions.

Composition Fibres
It is made up of three parts: Three types of fibres are present:
1. Ground substance a. Collagen
2. Fibres b. Elastin
3. Cells. c. Reticulin.
Metabolism of Specialized Tissues 139
Collagen characteristic feature is its long-range
reversible deformability similar to that of
Collagen is the major fibrous protein of extra-
rubber.
cellular connective tissues. It is a component
It is a scleroprotein. Glycine, alanine,
of most connective tissues including bone,
valine, proline and leucine are the chief amino
and is responsible for the tensile strength of
acids present. Glycine occurs in high concen-
the tissues. About 25-30% of the total body
trations as in collagen.
protein is collagen. It contains high content
of glycine. Also, proline and hyroxyproline
Reticulin
are present in greater amounts.
It is a fibrous, elongated molecule, There are two types of reticulin in connective
2900 Å × 15 Å. The major portion consists of tissue. One type is present in developing
three polypeptide chains, each in a left handed connective tissue and represents thin collagen
helical conformation. The three helixes are fibres. Another type of reticulin is associated
wound about each other to form a three with basement membranes, nerve and muscle
stranded rope-like structure. The three chains fibres and between connective tissue and
are cross-linked by covalent, (ester and special epithelium. It is a complex of collagen, lipid
ß-glutamyl and ß-aspartyl linkages). and polysaccharide.
It is synthesized by fibroblasts and other
mesenchymal cells, but, finally becomes Cells
extracellular. The aged fibres gradually They consist of fibroblasts, osteocytes,
become less soluble, tougher and less elastic. chondrocytes, mast cells and reticuloendo-
Tropocollagen, the precursor of collagen, thelial cells and occur depending on the nature
as synthesized by the cells, contains lysine of the tissue.
and proline. These are hydroxylated to form
hydroxylysine and hydroxyproline by INHERITED DISORDERS OF COLLAGEN
enzymes to form collagen. A disaccharide of
galactose and glucose is added covalently to Ehler-Danlos Disease
some of the hydroxylysine residues. Several types are described. There may be a
Deficiency of vitamin C results in a defect in the normal synthesis of collagen, in
disease called scurvy, a disease mainly the hydroxylation of lysine, in the conversion
involving connective tissue. The hydroxy- of procollagen to collagen etc. The disease is
lation of proline to hydroxyproline requires manifested by fragility of skin, hyperexten-
vitamin C. sibility and hyperflexibility of joints and
skeletal deformities. Most of the cases are due
Elastin to defect in hydroxylation of lysine.
Elastic tissue is yellow and refractive and has
Osteogenesis Imperfecta Congenita
a fibrillar, membranous or fibrillomembra-
nous structure arranged as a three dimen- Increased fragility of bones and pathological
sional meshwork. This tissue is found in aorta, fractures are common in this. The defect is in
skin, lungs and ligaments. Its important the formation of normal procollagen.
140 Medical Biochemistry for Physiotherapy Students
Cutis Laxa by far than any other tissue except adipose
tissue itself. The composition of the different
Skin is lax and joints hypermobile. There is a
parts of the brain may vary within wide
deficiency in the hydroxylation of lysine.
limits. For example, the water content of the
Marfan’s Syndrome gray matter of adult human brain is about
85% while that of the white matter is only
Skeletal deformities, cardiac disease, aortic 70%. The white matter thus contains about
aneurysm, long and thin digits and dislocation twice as much as solids as the gray matter.
of the lens are some of the manifestations of The water content of brain also varies with
this condition. This is an abnormality in age, being highest in young brains.
‘fibrillin’, a glycoprotein component of
microfibrils. Brain Lipids

NERVE TISSUE Phospholipids account for more than half of


the lipids, cholesterol forms about 20%,
Composition of Nerve Tissue cerebrosides 14% and other lipids (sulpho and
amino lipids) about 18% of the lipids. The
This constitutes about 2.4% of the body
phospholipids are mainly cephalins, lecithins
weight of man, the brain representing the
and sphingomyelins. All the cholesterol is in
major portion of this amount. The following
the unesterified form.
distribution of nerve tissue is given for an
The myelin sheath is made up of a series
adult man.
of layers of the external membrane of the
Brain 1400 g
Schwann cell around the axon. The membrane
Spinal nerves 150 g
is double layered (protein-lipid, lipid-
Spinal cord 30 g
protein).
Cranial nerves 10 g
Total 1,590 or 1,600 g
Brain Proteins
Nerve tissue is highly specialized in
function and composition. The passage of Proteins constitute about 40% of the dry
substances to the brain from the blood is weight of nerve tissue. Proteins present
restricted and regulated by the blood- includes albumins, globulins like α1, α2, β1,
cerebrospinal fluid barrier and blood brain β2, γ and fibrinogen. However, the albumin
barrier. These barriers in particular limit the forms a minute fraction (2% compared to 55%
passage of negatively and positively charged in plasma proteins) while the β-globulins form
ions, both organic and inorganic, from the the major fraction (65% compared to 13% in
blood to the brain, as well as the passage of plasma proteins). The other fractions do not
large organic molecule such as lipids, show so much variation from plasma pro-
polypeptides and polysaccharides. teins.
The most striking fact about the com- A protein named cerebrocuprein I contain-
position of the brain and nerves is the large ing two atoms of copper per molecule with a
amount of lipid material relative to protein molecular weight of 35,000 is present in brain.
in the brain solids. Brain contains more lipid This is increased in Wilson’s disease. There
Metabolism of Specialized Tissues 141
is a corresponding decrease of the copper citric acid cycle is maintained converting
containing protein in plasma called cerulo- glutamic acid to α-ketoglutarate, an inter-
plasmin and an increase in the plasma levels mediate in the cycle.
of free copper. Glutamic, aspartic acid and their deriva-
tives occupy a unique position in brain meta-
Amino Acids of Brain bolism and they constitute 70% of the non-
The total free amino acid concentration of nitrogenous nitrogen of brain. Both glutamic
brain is high relative to that of plasma, that is and aspartic acid can be converted to the
about eight times higher. It is particularly rich corresponding amide as
ATP
in aspartic and glutamic acids and their Glutamic acid + NH3———————→Glutamine
derivatives (N-acetyl aspartic acid, glutamine ATP
and γ-amino butyric acid). Taurine and Aspartic acid + NH3———————→Asparagine
glutathione also are present in fair amounts.
The amides so formed are related to the
Metabolism of Brain control of brain metabolism because gluta-
mine is readily permeable through blood-
Brain exhibits a very high rate of oxygen
brain barrier than glutamate. In the above
metabolism which accounts for 25% of total
reaction, ammonia is utilized. This is an
oxygen utilized by the body at rest. The
intracellular mechanism for the detoxification
respiratory quotient of brain is close to unity
of ammonia.
which indicates carbohydrate as the main
Gamma amino butyric acid formed as a
substrate for the brain metabolism.
Brain contains very little glycogen, about result of decarboxylation of glutamic acid is
0.1%. Hence, there must be a continuous said to be a synaptic transmitter for inhibitory
supply of glucose from blood to maintain neurons.
normal function of this vital tissue. Glucose Serotonin (5-hydroxytryptamine) formed
is readily permeable through the blood brain from tryptophan is an excitor. It is produced
barrier. A decrease in O2 supply or glucose within the brain itself and is not permeable
supply will markedly decrease oxidative to blood-brain barrier. Hence, serotonin
metabolism and ATP and creatine phosphate produced in the blood platelets or gastro-
levels of brain. During sleep and anaesthesia, intestinal tract has no cerebral effect.
there is an increase in ATP and creatine Reserpine — an antihypertensive and sedative
phosphate due to lowered activity of the drug, acts by releasing the serotonin from
brain. In convulsions, spontaneous or induced protein combination and facilitating its rapid
by drugs, there is rapid depletion of these destruction by the enzyme monoamine
energy rich phosphates; inorganic phosphate oxidase.
and lactic acid accumulation. Lysergic acid diethylamide (LSD) causes
Glycolysis and citric acid cycle seem to be excitation and hallucinations by inhibiting
the main pathways of metabolism in brain. monoamine oxidase and thus prolonging
In the absence of adequate carbohydrate, the serotonin action.
142 Medical Biochemistry for Physiotherapy Students
BONE Mineral of bone consists chiefly of micro-
crystals of a compound of calcium and
Composition of Bone phosphate with the structure of the mineral
Bone consists of cells (osteoblasts and apatite as hydroxyapatite.
osteoclasts) and an organic matrix in which Calcium ions in bone mineral are replaced
characteristic bone mineral is deposited in by lead ions in cases of chronic poisoning and
orderly arrangement. Inside the bone cavity are mobilized into the blood under conditions
is present the marrow which is rich in lipids that cause bone calcium to be mobilized (diets
(25%), mostly triglycerides. Bone is composed low in calcium and phosphorous, excessive
of about 1/3rd organic matrix and 2/3rd bone doses of vitamin D, parathyroid hormone,
mineral. The greater strength of bone is due etc.)
to the hardness of the mineral component and Formation of Bone
the toughness of the protein matrix in which Osteoblasts are the cells which are primarily
the mineral is embedded. concerned with bone formation and produce
The matrix contains three main groups of the proteins and mucopolysaccharides making
proteins: up the connective tissue fibres and the
i. A collagen-like protein — ossein. cementing gel in which the fibres and mineral
ii. An elastin-like protein — osseoalbumi- components are embedded. This organic
noid. matrix of fibres and cementing substance is
iii. A mucoprotein, rich in chondroitin first formed and then deposition of bone
sulfate—osseomucoid. mineral takes place. This mineral deposition
The minerals present are: calcium, sodium, in orderly arrangement is related to the
potassium, magnesium, phosphate, carbonate, association of chondroitin sulphate with the
citrate, chloride and fluoride. collagen fibres.
Chapter 18

Hormones

Hormones are chemical substances which are The peptide hormones, which may have 3
produced in one part of the body, enters the to over 200 amino acid residues, include all
circulation and are carried to distant organs the hormones of hypothalamus, all the
and tissues to modify their structure and hormones of pituitary and the pancreatic
function. They are similar to enzymes in three hormones.
ways: The amine hormones are derived from the
1. They act as catalysts. amino acid tyrosine and include water soluble
2. They are needed in small amounts. epinephrine and nor-epinephrine of the
3. They are not used up in the reaction. adrenal medulla and thyroid hormones.
The differences between enzymes and Steroid hormones (fat-soluble) include the
hormones is as follows:
adrenocortical hormones, androgens and
Enzymes Hormones estrogens (male and female sex hormones
1. They are utilized at 1. They are produced in one respectively).
the site where they are organ and they perform
produced. their action in other organs
(target organs). MECHANISM OF HORMONE ACTION
2. Structurally they 2. Structurally they all are
all are proteins. not proteins. Only some Activation of Intracellular Receptors
hormones are protein in
nature. Steroid hormones and thyroid hormones
3. Most of the enzymes 3. These are discharged in easily pass through plasma membrane because
are synthesized at the the blood their function
cellular level where prior to use. they are lipid soluble. Upon entering a target
they perform cell, the hormone binds to and activates an
intracellular receptor commonly located
Classes of Hormones within the nucleus. The activated receptor
Three chemically distinct classes of hormones then alters gene expression, that is, it turns
are: specific genes of the nuclear DNA on or off.
1. Peptides As the DNA is transcribed, new mRNA
2. Amines forms, leaves the nucleus, and enters the
3. Steroids. cytosol. There it directs synthesis of new
144 Medical Biochemistry for Physiotherapy Students
proteins usually enzymes on the ribosome. peptides and several sensory transduction
The new gene products cause the physio- mechanisms also act through second messen-
logical responses that are characteristic of the gers.
hormone.
Second Messengers
The best known second messenger is cyclic
AMP (cAMP). It is synthesized from ATP, the
main energy providing chemical in cells. The
enzyme that catalyzes formation of cAMP is
adenylate cyclase and it is attached to the inner
surface of the plasma membrane. When the
first messenger (hormone) binds to a receptor
on the outer surface of the membrane,
adenylate cyclase on the inner surface is
activated. Then, adenylate cyclase converts
ATP into cAMP in the cytosol of the cell. Cyclic
AMP acts as a second messenger to alter cell
function in specific ways e.g. elevation of
cAMP causes adipose cells to break down
Mechanism of Hormone action
triglycerides and release fatty acids more
rapidly but stimulates thyroid cells to secrete
Activation of Plasma more thyroid hormone. After a brief period
Membrane Receptors of time, an enzyme called phosphodiesterase
Catecholamine, peptide and protein hormones inactivates cAMP. Thus, the cells response is
are not lipid soluble and thus cannot diffuse turned off unless new hormone molecules
through the phospholipids bilayer of the continue to bind to their receptors at the
plasma membrane to attach to intracellular surface of the plasma membrane.
receptors. The receptors for these water- Many hormones exert at least some of their
soluble hormones instead are at the external physiological responses through the increased
surface of the plasma membrane. After a synthesis of cAMP. These include antidiuretic
water-soluble hormone is released from an hormone (ADH), oxytocin, follicular stimula-
endocrine gland, it circulates in the blood, ting hormone (FSH), luteinizing hormone
reaches a target cell and brings a specific (LH), thyroid stimulating hormone (TSH),
message to that cell. Since such a hormone can adrenocorticotrophic hormone (ACTH), calci-
deliver its message only to the plasma tonin, parathyroid hormone (PTH), glucagon,
membrane, it is called the first messenger. A epinephrine, nor epinephrine and hypo-
second messenger is needed to relay the thalamic releasing hormones. In other cases
message inside the cell where hormone such as growth hormone inhibiting hormone,
stimulated responses can take place. the level of cAMP decreases in response to
Eicosanoids, some neurotransmitters, neuro- binding of hormones to its receptor. Besides
Hormones 145
cAMP, several other substances are known Different protein kinases exist within
second messengers. These include calcium different target cells and within different
ions, cyclic guanosine monophosphate organelles of the same target cell. Thus, one
(cGMP), inositol triphosphate and diacyl- protein kinase might be involved with
glycerol. A given hormone may use more than glycogen synthesis, another within lipid
one second messenger. breakdown, another with protein synthesis
and so on. Moreover, protein kinases can
Role of G-proteins inhibit as well as activate enzymes, e.g. some
In the cAMP second messenger system, of the kinases unleashed when epinephrine
hormone receptors do not connect directly binds to liver cells inactivate an enzyme
to adenylate cyclase. Rather molecules called needed for glycogen synthesis.
G-proteins link receptors on the outer surface
Hormonal Interactions
of the membrane to the adenylate cyclase
molecules on the inner surface. Binding of a The manner in which hormones interact with
hormone to its receptor activates many G- other hormones is important. One type of
protein molecules which in turn activate interaction is called permissive effect. In this
molecules of adenylate cyclase. Unless further interaction, the effect of one hormone on a
stimulated by binding of more hormone target cell requires a previous exposure to
molecules to receptors, G-proteins slowly another hormone. Such previous exposure
become inactivated, thus helping to stop the enhances the response of a target cell.
hormone response. G-proteins are a common Another type of hormonal interaction is
feature of most second messenger systems. known as synergistic effect. Here, two or
more hormones complement each other’s
Protein Kinases actions and both are needed for full ex-
pression of the hormone effects, e.g. the
Cyclic AMP (cAMP) does not directly pro-
production, secretion and ejection of milk by
duce a particular physiological response.
the mammary glands require the synergistic
Instead, it activates one or more enzymes
effects of estrogens, progesterone, prolactin
collectively known as protein kinases, which and oxytocin.
may be free in the cytosol or bound to the
plasma membrane. Protein kinases are phos- Antagonistic Effect
phorylating enzymes that mean they remove
a phosphate group from ATP and add it to a Here, the effect of one hormone on a target
protein, which is usually another enzyme. cell is opposed by another hormone. An
example is insulin which lowers blood sugar
Phosphorylation activates some enzymes and
level and glucagon which raises it.
inactivates others. It is like an on-off switch.
The result of phosphorylating a particular
PITUITARY GLAND
enzyme could be regulation of other enzymes,
secretion, protein synthesis or a change in The human pituitary gland is a reddish gray
plasma membrane permeability. oval structure. Its diameter is about 10 mm.
146 Medical Biochemistry for Physiotherapy Students
It is located in the brain just behind the optic Clinical Significance
chiasma as an extension from the floor of the
Prolonged hypersecretion of GH usually by a
hypothalamus. Control of hormone secretion
hormone secreting tumours causes gigantism
from the pituitary is in part modulated by
and acromegaly.
regulating hormones from the hypothalamus.
Gigantism occurs when there is excess GH
secretion while epiphyseal cartilages of long
Hormones of the Anterior Pituitary
bones are still growing, i.e. during childhood
Growth Hormone (GH) before ossification of bones is complete. It is
evident mainly in the bones of the limbs and
It consists of a single polypeptide, with a
affected individuals may grow to heights of
molecular weight of about 21,500.
2.1 to 2.4 m, although body proportions
Functions remain normal.
Acromegaly occurs when there is excess
1. Growth hormone stimulates overall GH secretion after ossification is complete.
protein synthesis resulting in pronounced The bones become abnormally thick and there
increase in nitrogen and phosphorus is thickening of the soft tissues. Enlarged
retention. It stimulates protein synthesis. tongue, coarse facial hairs, and excessively
2. GH is mildly lipolytic. large hands and feet are noticeable features.
3. In muscle, GH antagonizes the effect of Deficiency of GH results in dwarfism. The
insulin. Impairment of glycolysis may individual is of small stature but is well
occur at several steps with inhibition of proportioned and mental development is not
transport of glucose. affected. Puberty is delayed and there may
4. GH increases intestinal absorption of be episodes of hypoglycaemia.
calcium also and plays an important role
in the retention of sodium, potassium, Prolactin (Lactogenic hormone)
magnesium, phosphate and chloride ions. It is a protein hormone with a molecular
5. GH stimulates mammary glands and weight of approximately 23,000.
lactogenesis.
Functions
Control of Secretion
It activates the corpus luteum and stimulates
Much of the control of growth hormone continued progesterone production by the
secretion occurs by a specific growth hormone developed corpus luteum. It increases during
releasing factor (GHRF) or (GRH) from pregnancy and may stimulate mammary
hypothalamus. The production of growth development and growth hormone like
hormone in man is about 500 ng/day and in metabolic changes.
plasma, it ranges from 0-5 ng/ml. Plasma
growth hormone concentration is increased Prolactin Pathophysiology
following exercise and stress (apprehension, 1. In women, tumours of prolactin secreting
pain, surgical stress, cold and severe insulin cells cause galactorrhea (breast discharge)
hypoglycaemia). and amenorrhea (cessation of menses).
Hormones 147
2. In men, excessive prolactin is associated Low levels are observed in hyper-
with breast enlargement and impotence. thyroidism and the levels are increased in
hypothyroidal myxedema.
Gonadotrophins (FSH and LH) ACTH is a polypeptide. It regulates
Gonadotrophins are glycoproteins. They adrenal function. It increases the synthesis of
include FSH (follicle stimulating hormone) corticosteroids by the adrenals and stimulates
and LH (luteinizing hormone). They influence their release from the gland. It increases total
protein synthesis. ACTH stimulation results
the function and maturation of the testes and
in an increase in mineralocorticoids, gluco-
ovary.
corticoids and androgens. It is controlled by
FSH: In females, this hormone promotes corticotrophin releasing hormones found in
follicular growth, prepares the graffian follicle the hypothalamus. Excess production of
for the action of LH and enhances the release ACTH produces Cushing’s syndrome.
of estrogen induced by LH.
In males, it stimulates seminal tubule and Hormones of Intermediate
testicular growth. It plays an important role Lobe of Pituitary
in the early stages of spermatogenesis. It secretes MSH (melanocyte stimulating
LH: In the female, it stimulates the final hormone). It is polypeptide and is regulated
maturation of the graffian follicle, ovulation by MSH release inhibitory hormone. MSH is
and the development of the corpora lutea. essential for skin pigmentation and is involved
LH also stimulates progesterone and estro- in the deposition of melanin by the melano-
cytes. It is present in a very active state in
gen levels.
species able to change the skin colour.
In males, it stimulates testosterone
production by the testis.
Hormones of the Posterior Pituitary
Thyrotropic Hormones Hormones of the posterior lobe of pituitary
include oxytocin and vasopressin. Both these
Thyrotropic hormones include TSH (thyroid
hormones are peptides.
stimulating hormone) and ACTH (adreno-
Oxytocin causes contraction of the smooth
corticotropic hormone).
muscles of the uterus and lactating mammary
TSH is a glycoprotein. It increases thyroid glands. It is used in routine obstetrics to
growth and general metabolic activity initiate labour and thus helps in parturition
including and ejection of milk.
- glucose oxidation Vasopressin is also known as ADH
- oxygen consumption (antidiuretic hormone). It is concerned with
- synthesis of phospholipids and RNA the regulation of water balance by reabsorp-
- increases iodine uptake and thyroxine tion of water in the distal tubules of the
metabolism. kidney (facultative reabsorption). In its
Its release is controlled by TRH (thyro- absence, diabetes insipidus with polyuria
tropic releasing hormone) secreted by results, in which large volumes of urine 15-
hypothalamus. 20 litres/day are excreted.
148 Medical Biochemistry for Physiotherapy Students
THYROID GLAND AND ITS HORMONES Transport of Thyroid Hormones

Thyroid gland is a small gland weighing about The thyroid hormones are carried in plasma
25 g in adults located on either side of the in combination with albumin and two specific
trachea below the larynx. It is made up of plasma proteins.
closed vesicles lined by a single layer of One of them is the TBG i.e. thyroxine
epithelial cells and filled with a colloid binding globulin. It is synthesized in liver,
material. this synthesis being decreased by the
The main hormone of the thyroid gland, androgens and glucocorticoid therapy.
thyroxine was Ist isolated by Kendall. In However, estrogens increase its synthesis.
addition to thyroxine, other compounds such Second of the plasma protein is TBPA, i.e.
as mono-iodotyrosine, di-iodotyrosine and thyroxine binding pre-albumin. A small
tri-iodothyronine (T3) are also secreted. amount of thyroxine is always present in the
Physiologically, only thyroxine and tri- free state which is the metabolically active
iodothyronine are active. The activity of the hormone.
tri-iodothyronine is about 5-10 times more
than that of thyroxine, although the thyroxine Actions of Thyroid Hormones
is present in greater quantity in circulating 1. Thyroid hormones accelerate the rate of
blood. glucose oxidation, promote intestinal
absorption of glucose and increases
Biosynthesis and Secretion glycogenolysis in the liver.
As told above that thyroid gland is made up 2. At the physiological levels, thyroid
of closed vesicles and is filled with a colloid hormones increase the incorporation of
material. This colloid material contains an amino acids into proteins and help in
iodine containing glycoprotein which is a protein synthesis. They also stimulate the
precursor for T 3 and T 4 formation. The production of the growth hormone. Above
thyroglobulin molecule contains about 140 the physiological levels, thyroxine has the
tyrosine molecules which can serve as opposite effect on protein metabolism, i.e.
substrates for the formation of thyroid increasing the breakdown of amino acids
hormones. The conversion of iodide to active depressing protein synthesis and bringing
iodine is an essential step for its incorporation about negative nitrogen balance.
into thyroid hormones. This occurs in thyroid 3. Thyroxine increases the deposition of fat
tissue itself. TSH promotes the oxidation of in adipose tissue as well as its release and
iodide to active iodine. oxidation. The cholesterol level in blood
Tyrosine is first iodinated at position 3 to increases in hypothyroidism.
form mono-iodotyrosine (MIT), then at 4. Thyroid hormones increases considerably
position 5 to form di-iodotyrosine (DIT). Two oxygen consumption and oxygen co-
molecules of DIT couples to form thyroxine efficient of almost all metabolically active
(T4). One molecule of MIT couples with DIT tissues. There is increase in heat produc-
to form T3. tion and basal metabolic rate (BMR).
Hormones 149
5. Thyroxine is one of the factors essential for 4. Certain organic compounds are present in
normal growth and skeletal maturation. In vegetables like cabbage and turnip which
hypothyroid children, bone growth is act as natural goitrogens and depress
slowed and epiphyseal closure delayed. thyroid activity. These are present as
goitrins in the raw foods and are however
Metabolic Fate and Excretion are destroyed on cooking.
Thyroxine undergoes the following altera-
Hypothyroidism
tions in the liver.
1. Transamination and deamination: Trans- Hypothyroidism is a deficiency of thyroid
amination with the help of α-keto acids hormones and leads to myxedema in adults
gives the corresponding pyruvic acid and cretinism in children. Such children are
analogues of the hormones. dwarf and mentally retarded. Adults
2. Conjugation with glucuronic acid and complain of undue sensitivity to cold with
sulphate: The phenolic group of the puffiness of face and extremities, thick and
iodothyronines gets conjugated with dry skin and hypercholesterolaemia.
glucuronic acid or sulphates and conjugates
are excreted through bile into the intes- Hyperthyroidism
tines.
3. De-iodination: De-iodination may take Hyperthyroidism is due to the excessive
place in liver in the presence of enzyme secretion of the thyroid hormones and
de-iodinase. The de-iodinated thyronines hyperactivity of the gland resulting in
are metabolized by transamination thyrotoxicosis. It is characterized by
followed by conjugation with glucuronic - nervousness
acid and excreted by the kidney. - irritability
- weight loss
Antithyroid Agents - increased body temperature
- increased heart rate
Substances that inhibit the normal functioning
- fatigue
of thyroid gland are termed as anti-thyroid
- increased appetite
agents. They are:
- fine tremor of the outstretched fingers.
1. Agents which retard the synthesis of
The most common form of the hyperthy-
thyroid hormones, e.g. thiocyanate,
roidism is the Grave’s disease where the
thiocarbamide, sulpha drugs, perchlorate
thyroid gland is diffusely enlarged with
etc.
protrusion of the eyeballs. The BMR is raised
2. Deep X-ray therapy destroys the thyroid
tissue and this depresses the thyroid and T3 and T4 levels are increased in this
activity. condition.
3. Thyroxine itself can inhibit the production
Simple Goitre
of TSH (thyroid stimulating hormone) of
the anterior pituitary gland and check the The enlargement of the thyroid gland without
release of thyroxine by a feed back signs of hyperthyroidism is called simple
mechanism. goitre. Secretion of T3 and T4 is reduced and
150 Medical Biochemistry for Physiotherapy Students
low levels stimulate secretion of TSH resulting ABNORMALITIES OF
in hyperplasia of the thyroid gland. Causes PARATHYROID FUNCTION
are iodine deficiency, some genetic abnor-
mality. Hyperparathyroidism
In this, there is over secretion of PTH. It
Toxic Nodular Goitre
occurs due to a tumour of the gland. The
In this condition, one or two nodules of a symptoms are
gland that is already affected by goitre secrete
- hypercalcaemia
excess hormones causing the effects of
- low serum phosphorus
hyperthyroidism.
- increased serum alkaline phosphatase
PARATHYROID GLAND - decalcification of bones causing pain
Closely associated with the thyroid gland are - renal stones.
two pairs of small glands called the para-
thyroids. These glands produce a parathyroid Hypoparathyroidism
hormone or parathormone (PTH). It is a In this, there is diminished secretion of PTH.
polypeptide containing 84 amino acids. It is
The symptoms are
synthesized as a prepro-hormone. PTH
secreted in the body is degraded very rapidly - hypocalcaemia
in the circulation. - hyperphosphataemia
- muscular weakness
Actions - tetany.
1. It mobilizes calcium and phosphorus from Tetany is the most characteristic feature
bones. seen in hypoparathyroidism. In this, neuro-
2. It increases serum calcium and lowers the muscular hyperexcitability is observed which
serum phosphorus. is due to decreased calcium. There are very
3. It increases urinary excretion of phosphate strong painful spasms of skeletal muscles,
but decreases excretion of calcium.
causing characteristic bending inwards of the
4. It elevates serum alkaline phosphatase
hands, forearms and feet.
activity.
5. It increases the absorption of calcium and
phosphorus from the intestine. Calcitonin
6. In the kidney, it affects renal tubular
Calcitonin is secreted by the C-cells of the
reabsorption of calcium and phosphorus.
thyroid gland. It is a polypeptide consisting
7. It stimulates protein synthesis in the
osteoclasts which affect resorption of of 32 amino acids. Calcitonin release is stimu-
bone. lated by a high calcium levels in serum.

Control of Secretion Actions

PTH secretion is controlled by serum calcium 1. It is directly effective on bone. It inhibits


levels by a negative feedback mechanism bone resorption and mobilization of
which is also regulated by calcitonin. calcium and phosphorus from bone.
Hormones 151
2. It lowers the serum calcium levels. the number of glucose transporters in the
3. In the kidney, it increases calcium cell membrane. The intravenous adminis-
excretion. tration of insulin thus causes an immediate
decrease in the concentration of blood
PANCREATIC HORMONES glucose.
2. Insulin decreases the level of circulating
Insulin fatty acids by inhibiting the activity of
Insulin is a hormone secreted by the β-cells hormone–sensitive lipase in adipose tissue.
of the islets of Langerhans of pancreas. Insulin Insulin acts by promoting the dephos-
plays an important role in metabolism causing phorylation and hence inactivation of the
increased carbohydrate metabolism, glycogen enzyme. Insulin increases the transport and
storage, amino acid uptake, fatty acid metabolism of glucose into adipocytes
synthesis and protein synthesis. It is thus an providing the substrate glycerol–3–
anabolic hormone that acts on a variety of phosphate for triacylglycerol synthesis.
tissues including liver, muscle, and adipose Insulin also increases lipoprotein lipase
tissue. activity of adipose tissue, providing fatty
acids for esterification.
Chemistry 3. Insulin stimulates the entry of amino acids
Insulin is a protein hormone. Insulin molecule into cells by enhancing the rate of synthesis
is composed of two polypeptide chains called of membrane transporters for amino acids.
A chain and B chain. The A chain has 21 amino Insulin increases protein synthesis by
acids while B has 30 amino acids. Both the providing more amino acids in cells.
chains are held together by two interchain 4. Insulin causes decrease in concentration
disulphide linkages. In addition, there is an of K+ ion and inorganic P in blood.
intrachain disulphide link in chain A between
the amino acids 6 and 11. In A chain, N- Metabolism of Insulin
terminal amino acid is glycine and C- terminal
Insulin is synthesized from prepro-insulin.
is asparagine. In B chain, N- terminal amino
This prepro-insulin is converted to pro-
acid is Phenylalanine and C- terminal amino
insulin which is finally converted to insulin.
acid is Threonine.
Insulin is degraded in liver and kidney
Metabolic Effects of Insulin by the enzyme glutathione insulin trans-
hydrogenase which brings about reductive
1. The effects of insulin on glucose meta- cleavage of the S-S bonds that connect A and
bolism are most prominent in three tissues: B chains of the insulin molecule. Reduced
liver, muscle and adipose. In the liver, glutathione acts as a coenzyme. The A and B
insulin decreases the production of
chains are further degraded by proteolysis.
glucose by inhibiting gluconeogenesis and
the breaking down of glycogen. In muscle
Glucagon
and liver, insulin increases glycogen syn-
thesis. In muscle and adipose tissue, insu- Glucagon is secreted by α-cells of Islets of
lin increases glucose uptake by increasing Langerhans of pancreas.
152 Medical Biochemistry for Physiotherapy Students
Chemistry and glucagon secretion. Hypothalamic
somatostatin acts as a regulator of GH
Glucagon is a polypeptide with a molecular
secretion. GIT somatostatin inhibits the
weight of 3485. It contains 29 amino acids in
secretions of gastrin and motilin.
a single chain. It does not contain cysteine,
proline or isoleucine. ADRENAL GLAND AND ITS HORMONES
Actions Adrenal gland is also known as suprarenal
gland. The adrenal glands in the human adult
1. Glucagon increases glycogenesis in liver.
are situated close to the upper pole of the
It also stimulates the conversion of lactic
kidneys and weigh about 10 g each. The
acid and glucogenic amino acids to form
adrenal gland is divided both histologically
glucose. Glucagon also increases the pool
and physiologically into two distinct portions:
of glucogenic amino acids in liver, so that
adrenal cortex and adrenal medulla.
they can be used for gluconeogenesis.
2. Glucagon increases the breakdown of Adrenal Cortex
triglycerides to produce free fatty acids
and glycerol. It increases fatty acid The adrenal cortex is the outer portion of the
synthesis. gland and contains three structurally distinct
layers:
3. Glucagon reduces protein synthesis by
1. Zona glomerulosa
depressing incorporation of amino acids
2. Zona fasciculata
into peptide chains. It also stimulates
3. Zona reticularis.
protein catabolism especially in liver thus
Cells of all the three layers can form the
increasing the hepatic amino acid pool
steroid hormone corticosterone, cells of the
which is utilized for gluconeogenesis. zona fasciculata and zona reticularis can form
4. Glucagon increases heat production and cortisol and sex hormones. Outer zona
causes rise in BMR. glomerulosa can synthesize aldosterone only.
About 50 steroid hormones have been
Somatostatin isolated from adrenal cortex but out of them,
Somatostatin is secreted by δ-cells of Islets of only 7 are important and known to possess
Langerhans of pancreas. physiologic activity. They are all derived from
cholesterol and contain the steroid nucleus
Chemistry called cyclopentanoperhydrophenanthrene
nucleus.
It is a peptide hormone. It consists of 14
Seven important hormones are:
amino acids. There is an intrachain S-S linkage 1. 11-dehydrocorticosterone
joining cysteine 3 and cysteine at position 14. 2. corticosterone
3. cortisone
Actions
4. cortisol
In addition to its secretion from pancreas, it 5. aldosterone
is also secreted from hypothalamus and GIT. 6. androstenedione
Pancreatic somatostatin inhibits both insulin 7. dihydroepiandrosterone.
Hormones 153
Steroids are divided into three types Mineralocorticoids
according to their function: 1. Mineralocorticoids cause retention of
1. Glucocorticoids NaCl and water and so it regulates blood
2. Mineralocorticoids volume and blood pressure.
3. Cortical sex hormones. 2. They cause increased excretion of
1. Glucocorticoids: These primarily affect potassium.
metabolism of carbohydrates, lipids and 3. Intracellular potassium is lowered and
proteins and have relatively minor sodium is raised by the action of mineralo-
effects on electrolyte and water meta- corticoids.
bolism, e.g. cortisone, corticosterone 4. They have little action on metabolism of
and cortisol. carbohydrates and proteins.
2. Mineralocorticoids: These primarily affect 5. They are effective in protection against
the reabsorption of Na+ and excretion stress.
of K + and distribution of water in The blood potassium level regulates the
tissues, e.g. aldosterone. amount of aldosterone produced. When
3. Cortical sex hormones: These primarily blood potassium level rises, more aldosterone
affect secondary sex characters, e.g. is secreted and vice versa. Angiotensin also
androgens and estrogens. stimulates the release of aldosterone.

Functions of Adrenal Cortex Renin-angiotensin-aldosterone System


Glucocorticoids When renal blood is reduced or blood sodium
1. Glucocorticoids stimulate formation of levels fall, the enzyme renin is secreted by
glycogen in the liver and muscles. It kidney cells. Renin converts the plasma
increase gluconeogenesis in liver and protein angiotensinogen, produced by the
depress glucose uptake and oxidation by liver to angiotensin I. Angiotensin converting
tissues. enzyme (ACE) converts angiotensin I to
2. Glucocorticoids increase rate of break- angiotensin II, which stimulates secretion of
down of tissue proteins to amino acids. aldosterone. It also causes vasoconstriction
Body proteins are lost thereby increasing and increases blood pressure.
nitrogen excretion.
Cortical sex hormones: Cortical sex hormones
3. Glucocorticoids stimulate lipid absorption
affect sex organs and secondary or accessory
from the intestine. It mobilizes lipid from
sex characters.
the depot and disintegrate it to form
ketone bodies. Hypofunction of adrenal cortex: Hypofunction
4. Cortisol and cortisone relieve muscular of adrenal cortex causes Addison’s disease.
weakness found in hypofunction of Addison’s disease is usually due to tuber-
adrenal cortex. culosis of the gland involving both cortex and
5. Excess of cortisol causes osteoporosis due medulla. Both the effects are mainly due to
to decalcification and interference in cortical damage and consequently insufficient
formation of protein matrix. secretion of cortisol and aldosterone.
154 Medical Biochemistry for Physiotherapy Students
Clinical features of Addison’s disease are Hyperaldosteronism: Hyperaldosteronism
1. Muscular weakness and easy fatiguability may be primary or secondary. An excessive
2. Vomiting, anorexia and hypochlorhydria production of aldosterone due to a tumour
3. Low blood pressure of the zona glomerulosa tissue of the adrenal
4. Pigmentation of the skin and mucous cortex is termed Conn’s disease or primary
membrane aldosteronism. Hypertension, muscular
5. Low BMR and subnormal temperature weakness, retention of sodium, alkalosis are
6. Disturbed ionic balance found. Prolonged potassium depletion causes
7. Decreased blood volume muscle weakness. Secondary hyperaldoste-
8. Increased capillary permeability. ronism occurs in congestive cardiac failure,
Hyperfunction of adrenal cortex: Hyperfunction cirrhosis of liver, etc.
of adrenal cortex causes Cushing’s syndrome
ADRENAL MEDULLA
and hyperaldosteronism.
Cushing’s syndrome: Cushing’s syndrome is Two hormones are secreted by the medullary
caused when there is adrenal tumour or portion of the adrenal gland. These are
adrenal hyperplasia and there is excessive epinephrine and nor epinephrine. Both these
secretion of cortisol. Pituitary tumours hormones belong to the group of catechola-
causing excess secretion of ACTH may be the mines as they possess the catechol residue (o-
reason for this syndrome. dihydroxy benzene) in their structure.
Epinephrine differ from nor epinephrine by
Clinical Features having a methyl group attached to the
nitrogen atom.
1. Increased deposition of fat on the trunk
These hormones are related to the
2. Characteristic pad of fat at the back of
aromatic amino acid tyrosine and are derived
the neck (a buffalo hump) and abdomen
from it. The naturally occurring hormones are
3. Purple striae are usually found over the
abdomen, thigh, etc. This is due to loss L-isomers.
of protein matrix.
Biosynthesis and Secretion
4. In males, excessive hair growth
5. In females, masculisation with growth The chromaffin granules of the adrenal
of beard, moustache etc. medulla are concerned with the biosynthesis,
6. Wasting of the muscles of the limbs storage and secretion of catecholamines. The
7. Osteoporosis of bones due to decalcifi- hormone is released from these granules by
cation and loss of protein matrix the exocytosis of cells which is a calcium
8. Mental derangement dependent process. Neural stimulation, fright,
9. In males, impotence with atrophy of emotional conditions like anger, etc. are
testis responsible for a quick release of the
10. In females, amenorrhoea, sterility, etc. catecholamine synthesis.
Hormones 155
Physiological and Biochemical Functions function. These metabolites are excreted in
large amounts in patients with pheochromo-
1. Epinephrine causes a rise in blood pressure
cytoma (tumours of the adrenal medulla).
due to arteriolar vasoconstriction parti-
cularly in the skin, mucous membrane.
Hyperactivity
However, the arterioles of the skeletal
muscles undergo vasodialation. The Pheochromocytoma
overall effect is a rise in blood pressure, Tumuors of the chromaffin tissue result in
increase in pulse rate, heart rate and this condition observed in man. It is charac-
cardiac output. Nor epinephrine causes terized by hyperglycaemia, glycosuria,
rise in blood pressure by increasing hypertension and hyperlipidaemia. The
peripheral resistance. It is an overall plasma levels rises several times above normal
vasoconstrictor and has no effect on value and urinary excretion of epinephrine
cardiac output. metabolites increases remarkably.
2. Epinephrine dilates bronchial musculature,
relaxes the musculature of the gastro- SEX HORMONES
intestinal tract and contracts the pyloric
sphincter. These effects are exhibited very The organs producing sex hormones in the
weakly by nor epinephrine. males and females are testis and ovaries
3. Epinephrine raises blood sugar due to respectively. The testis and ovaries, in
breakdown of glycogen in the liver, addition to their function of providing
muscle, and depresses utilization of spermatozoa or ova, manufacture steroid
glucose by the tissues. hormones that control secondary sex charac-
4. Epinephrine and nor epinephrine act as ters, the reproductive cycle and the growth
lipolytic hormones. They stimulate and development of the accessory repro-
lipolysis in adipose tissue and release free ductive organs.
fatty acids into the body, thereby raising The sex hormones are of three types:
the blood free fatty acid level. The blood 1. Androgens or male hormones
cholesterol and phospholipids are also 2. Estrogens or female hormones
increased. 3. Gestogens or progestational hormones

Metabolic Fate of Epinephrine Androgens


and Norepinephrine The male sex hormones or androgens are
Catecholamines once elaborated and circu- produced by the testis and also isolated from
lated in blood are very quickly destroyed and urine. Testosterone is the principal male
excreted in urine. The enzymes that act on hormone synthesized by the interstitial
catecholamines are catechol-o-methyl transferase (Leydig) cells of the testis from cholesterol.
(COMT) and monoamine oxidase (MAO). The It is 10 times more potent than androsterone
metabolic products, metanephrine and (occurs in urine).
vanillyl mandelic acid (VMA) are inactive and The testis also produces another androgen
are useful indices for the adrenal medullary known as androstene-3, 17-dione. The
156 Medical Biochemistry for Physiotherapy Students
metabolic products of testosterone are present Most mammals have a plasma β-globulin
in urine as epiandrosterone, androsterone, that binds testosterone with specificity and
dehydro-epiandrosterone, androsterone 3, high affinity. This is called the sex hormone
17-dione and etiocholane 3-ol-17-one. binding globulin (SHBG) or testosterone–
Epiandrosterone is five times more active than estrogen binding globulin (TEBG) produced
androsterone while etiocholane-ol-one is in the liver.
physiologically inactive.
Biochemical Effects of Androgens
Testosterone is converted to its more
active form dihydro-testosterone in the target 1. Androgens exert a striking anabolic effect
on protein conservation and retention of
tissues. It is then converted to androsterone
nitrogen and thereby muscle growth and
and other 17-ketosteroids by the liver where
maintenance of muscle mass. Androgens
they are further conjugated with sulphate and
help in increasing the storage of creatine
excreted through urine.
in muscles.
2. Androgens stimulate the growth of bones
Biosynthesis of Androgens
before the closure of epiphyseal cartilage.
The androgens are synthesized from choles- 3. Testosterone exhibits a mineralocorticoid
terol by the gonadal tissues, chiefly the testes effect by promoting the reabsorption of
and to a small extent by the adrenal cortex. sodium and chloride ions and water by
the kidney tubules.
Cholesterol 4. Androgens increase the fructose produc-
tion by seminal vesicles and the utilization
of this sugar by the seminal plasma.
Δ5 -pregnenolone 5. The citric acid cycle and fatty acid
synthesis are stimulated by androgens.
The activities of the glycolytic enzymes are
Progesterone also enhanced by this hormone.
The principal secreted metabolites of
testosterone are androsterone and etio
Hydroxyprogesterone cholan-ol-one. Small amounts of dehydro-
epiandrosterone are excreted as the sulphate.
The principal pathway of degradation of
Androstenedione testosterone involves oxidation in the liver
to androstenedione.
ABNORMALITIES RELATED WITH
Testosterone Androsterone MALE SEX HORMONES

Small amounts of androgens are also Hypogonadism


produced in the ovaries and adrenal cortex. It is a disorder characterized by a defect in
The adrenal cortex produces a male hormone testosterone synthesis which may be due to
called androsterone whose production is failure of testes to produce it or impairment
increased in adrenal tumours. in the release of gonadotropins.
Hormones 157
Female Hormones followed by calcification and hyper-
ossification of long bones.
Two main types of female hormones are
secreted by the ovary. Synthetic Estrogens
1. The follicular or estrogenic hormones A number of synthetic estrogens have been
produced by the cells of the developing produced like ethynyl estradiol, diethyl-
graffian follicle and stilbestrol, etc.
2. The progestational hormones derived
from the corpus luteum that is formed in Progestational Hormones
ovary from the ruptured follicle.
Progesterone is the hormone of the corpus
The Estrogenic Hormones luteum. It is also formed by the placenta
during the later part of the pregnancy. It is
These hormones are responsible for the also formed in the adrenal cortex. Proges-
regulation of menstrual cycle as well as the terone is bound in plasma to the corticosteroid
reproductive cycle. The principal estrogenic binding globulin.
hormone in the circulation and the most
important active form of the estrogens is Functions of Progesterone
estradiol. It is bound to a specific plasma 1. Progesterone appears after ovulation and
carrier protein (sex steroid binding protein) causes extensive development of the
which also transports the androgens. Estra- endometrium, prepares the uterus for the
diol is the principal estrogen found in the reception of the embryo and for its
urine of pregnant women and in the placenta. nutrition.
2. Progesterone also suppresses estrus,
Effects of Estrogenic Hormones ovulation, production of pituitary hor-
1. In the lower animals, the estrogenic mones and LH.
hormones induce estrus, a series of 3. Progesterone promotes the growth of
changes in the female reproductive system glandular tissue in uterus and mammary
associated with ovulation. gland.
2. In women, the estrogenic hormones The chief excretory product of proges-
prepare the uterine mucosa for the later terone is pregnanediol which is present as the
action of the progestational hormones. The glucuronide in the latter half of the menstrual
changes in the uterus include proliferative cycle.
growth of the endometrium. Changes in About 75 percent of progesterone (or its
the epithelium of the uterine tubes and of metabolites) are excreted in the intestine by
the vagina also occur. All these changes way of the bile and eliminated in the faeces.
begin immediately after menstrual blee- Large amounts occur in the urine only if the
ding has ceased. biliary route of excretion is blocked.
3. Estrogens bring down the hyperlipaemia
ABNORMALITIES ASSOCIATED WITH
including hypercholesterolaemia and may
FEMALE SEX HORMONES
be related to the prevention of athero-
sclerosis. Coronary heart disease is less Hypogonadism
common in women as compared to men. This occurs due to deficiency in ovarian
4. Estrogen administration causes elevation function causing decreased ovulation and/or
of calcium and phosphate levels in serum insufficient hormone production.
Chapter 19

Diet and Nutrition

Nutrition is the first need of man and his Different foodstuffs when consumed,
general health and well being are much release different amounts of heat. The total
dependent on his nutritional status. The heat produced by the combustion of any
availability of food is not uniform throughout foodstuff in the presence of oxygen can be
the world and due to this unequal distri- measured in a bomb calorimeter. By this
bution, malnutrition and under nutrition method, the energy values (caloric values) of
prevails in certain regions of the world, carbohydrate, protein and fat have been
whereas excess food is available in certain measured. The average values are given
countries where there are diseases due to below:
overeating and over-nutrition.
Energy Value of Foods
CALORIMETRY
Foodstuff Energy value (kcal/g)
Calorimetry is the measurement of energy In bomb In the body
requirements of the body under various calorimeter
physiological conditions and the determi- Carbohydrates 4.1 4.0
nation of energy values of foods. Fats 9.4 9.0
Proteins 5.4 4.0
Energy Value or Caloric Value of Food
When foodstuffs are oxidized in the body,
Energy value of foods and energy require- the oxidation is almost complete as far as
ment for the body are measured usually in carbohydrates and fats are concerned.
calories. A calorie is defined as the heat However, proteins are not completely
required to raise the temperature of 1 g of oxidized in the body because end products
water by l°C (say, from 15°C to 16°C). This of protein catabolism yields urea, ammonia,
unit is small, so a unit 1,000 times of the calorie etc. which still contains oxidizable carbon
is called kilocalorie or simply calorie is used and/or hydrogen. Hence, their caloric value
for this purpose. Calorie in biological science in the body is only 4.0 kcal/g as compared to
always means a kilocalorie (C or kcal). the bomb calorimeter value of 5.4 kcal/g.
Diet and Nutrition 159
Respiratory Quotient (RQ) dioxide are liberated while 163 volumes of
oxygen are required.
It is the ratio of the volume of CO2 produced
by the volume of O2 consumed (i.e., CO2/
O2) during a given time. RQ is simply a ratio.
It gives no idea as to the absolute quantity of
gaseous interchange.

Respiratory Quotient

The RQ of foodstuffs depends upon the


type of components and their varying RQ for Fats

proportions. The RQ of a mixed diet


The oxidation of proteins cannot be readily
containing carbohydrate, fat and protein is
expressed. By indirect methods, the RQ for
around 0.85.
proteins has been calculated to be about 0.8.
The RQ for the oxidation of carbohydrate
For example, oxidation of alanine:
is unity (1). Because in carbohydrate diet, the
volume of CO 2 produced is same as the
volume of oxygen consumed. This is because,
in the carbohydrate molecule, the amount of
O2 present is just sufficient to oxidize the H
present in the same molecule. Hence, external
oxygen is necessary only to convert the C of
the molecule into CO2. Therefore, the volume
of O 2 consumed and the volume of CO 2,
produced will be same. This is represented
in the following equation:
RQ for Proteins

When carbohydrates are converted into


fats in the body, RQ will rise because in this
process, an oxygen rich compound is
RQ for Carbohydrates converted into an oxygen poor compound,
so some amount of O 2 liberated from
Fats have a lower RQ of about 0.7 because carbohydrate will be utilised for purposes of
fat is an oxygen-poor compound. Therefore, oxidation. Consequently, less oxygen will be
fats require more oxygen for their oxidation. needed from outside. Hence, the amount of
For example, for the oxidation of two CO2 produced will be more than the amount
molecules of tristearin, 114 volumes of carbon of O2 consumed. Therefore, the RQ will rise.
160 Medical Biochemistry for Physiotherapy Students
When fats are converted into carbohydrates the surface area, greater would be the
just the opposite effects will be produced and BMR.
RQ will fall. 2. Age: The BMR is inversely proportional
to age. Children have larger BMR than
Importance of Respiratory Quotient adults.
1. RQ acts as a guide as to the type of food 3. Sex: Males have higher BMR than females.
burning or the nature of synthesis taking 4. Climate: The BMR is lower in warm
place in the whole body as well as in a climates.
particular organ. 5. Habit: Persons accustomed to heavy
2. RQ is very helpful in determining meta- exercise or hard physical work have a
bolic rate. higher BMR than those involved in
3. RQ helps in finding out the proportion of sedentary work.
the three foodstuffs that are being utilised 6. State of Nutrition: The BMR is decreased
in the body. in starvation and undernourishment.
7. Disease: The BMR is increased in infectious
Basal Metabolic Rate (BMR) and febrile diseases. The increase is
The metabolic rate determined when the sub- usually proportional to the rise of the
ject is in complete mental and physical resting temperature. The BMR is also increased
state as possible, in a room with comfortable in increased activity of cells and therefore,
temperature and 10- 12 after the last meal is it increases in leukemia, cardiac failure,
called basal metabolic rate and essentially hypertension, polycythemia, dyspnea, and
implies the minimum energy required to keep some types of anaemia.
the body going. 8. Effects of Hormones: The BMR is increased
BMR is proportional to lean body weight in hyperthyroidism and decreased in
and thus to surface area. Its units are kcal/ hypothyroidism. In adrenal insufficiency
m2/hr. The BMR for normal male is 40 kcal/ (Addison’s disease), the BMR is subnormal.
m2/hr and for female is 37 kcal/m2/hr. The
BMR of an individual is expressed as a Specific Dynamic Action
percentage of the normal.
The BMR between –15% and +20% is Specific dynamic action (SDA) is the extra heat
considered normal. The BMR may exceed +50 production when food is used by the body
to +75% in hyperthyroidism. In hypothyroi- over and above the calculated caloric value.
dism, the BMR may be –30 to –60%. This is also known as calorigenic action or
BMR is measured by the apparatus of thermogenic effect of foods.
Benedict and Roth or by Douglas bag method. When 25g of protein undergoes metabo-
lism in the body, the heat production from
Factors Affecting BMR the caloric value is 25 × 4 kcal = 100 kcal.
Many factors affect the BMR which are as However, it is found that the actual heat
follows: production may be 130 kcal. This extra 30 kcal
1. Surface area: The BMR is directly related is due to the specific dynamic action of protein
to the surface area of the subject. Larger in the body. Similarly, it is found that 11.1 g
Diet and Nutrition 161
of fat instead of producing 11.1 × 9 ≈ approx category. Jobs like plumber work, coal
100 kcal of heat, actually produces 113 kcal, mining, stone crushing and rickshaw pulling
whereas 25 g of carbohydrate actually are included in the heavy category. The
produces 105 kcal instead of 100 kcal. SDA average daily calories requirement for
for protein, fat and carbohydrate are 30, 13 different categories is given in the following
and 5% respectively. Thus, proteins have the table.
highest SDA value whereas carbohydrates Category of worker Requirements
have the lowest. in kcal/day
For a mixed diet, the SDA is around 10%. Males
Therefore, it is observed that the SDA due to Sedentary 2,560
food components is greatest when they are Moderate 2,960
Heavy 4,160
fed individually, but the SDA is lowered Females
when the components are mixed. Sedentary 2,040
Moderate 2,340
Energy Requirement of Individuals Heavy 3,140

Dietary energy requirements are determined


Nutritional Importance of Carbohydrates
in large part by the physical activity of the
individual. Some jobs involve moderate Dietary carbohydrates are the chief source
physical activity, others heavy and light. of energy. They contribute to 60 to 70 percent
The following table provides estimates of of total caloric requirement of the body.
the energy output during various activities. 1. Carbohydrates are the major sources of
energy.
Activity Requirement 2. Carbohydrates have a protein sparing
(kcal/hr)
action.
Reading/sitting/standing 35-40 3. Carbohydrates are the main source of
Typing/writing 70-75
energy for the brain and other parts of
Household work 100-110
Painting/carpentry 140-150 central nervous system. Prolonged hypo-
Light exercise/slow 200-250 glycemia may lead to irreversible brain
to moderate walk damage.
Active exercise/walking 350-400 4. Carbohydrate as glycogen in muscle is
downstairs broken down to lactic acid (glycolysis) to
Fast walking/running/ 450-500
swimming provide energy for muscle contraction.
Walking upstairs 1,000-1,100 5. Pentoses, which are one of the constituent
of nucleic acids, are produced in carbo-
Three different categories can be defined hydrate metabolism.
on the type of work like sedentary, moderate 6. Excess consumption of carbohydrates
and heavy category of workers. Sedentary leads to the formation of fat which is
workers include shopkeeper, carpenter or stored in the adipose tissue.
painter, etc. that perform light physical 7. Non-digestible carbohydrates are impor-
activity. Students, farmers doing mechanized tant in the body because they improve
farming are included under moderate bowel motility and prevent constipation.
162 Medical Biochemistry for Physiotherapy Students
Nutritional Importance of Fats 3. It helps to correct hypercholesterolaemia
by binding the bile salts and sequestering
Fats provide about 20 to 35 percent of the
them. As a result, the concentration of bile
total calorie requirement of the body.
salts reaching the liver by entero-hepatic
1. These provide energy about 9 Cal/g.
2. These are the necessary constituents of circulation is decreased. The feed-back
body tissues, contributing to about 14 to inhibition by the bile salts on the catabo-
18 percent body weight. lism of hepatic cholesterol to bile acids is
3. Fats provide the essential fatty acids such decreased. So, there is greater degradation
as linoleic, linolenic and arachidonic acids. of cholesterol to bile acids and disposal
4. Fats can be stored in the adipose tissue of from the body.
the body and is available as a source of 4. Fiber prevents colon cancer. One possible
energy during periods of shortage. factor for colon cancer is through toxins
5. Fats are essential for the absorption of fat- elaborated by some intestinal microbes.
soluble vitamins. These toxins are adsorbed by dietary fiber
6. A lack of fat causes a feeling of hunger and eliminated from the body.
shortly after a meal due to rapid digestion 5. Fiber is also beneficial to diabetics since it
in its absence so; they have a high satiety slows stomach emptying and delays the
value. post-prandial rise in blood glucose.
7. Excess of saturated fat and cholesterol Good sources of fiber are rice bran, wheat
consumption is associated with hyper- bran, legumes, fruits, vegetables and leafy
cholesterolaemia and atherosclerosis. greens.

Nutritional Importance of Dietary Fibers Balanced Diet


Dietary fiber is a collective term that includes A balanced diet is defined as the diet which
all indigestible plant cell wall components: contains different types of foods, possessing
celluloses, hemicelluloses, pectins, lignins and the nutrients, i.e. carbohydrates, fats,
gums etc. proteins, vitamins and minerals in a
Human beings cannot digest, but herbi- proportion to meet the requirements of the
vores such as ruminants can digest cellulose body. A balanced diet should be based on:
and that constitute the major source of energy - locally available foods
for them. - should be easily digestible
1. The indigestible dietary fibers add bulk - should contain enough roughage materials.
to the diet and absorb water in the The basic composition of balanced diet is
intestinal lumen, thus aiding in the highly variable, as it differs from country to
elimination of larger and softer faeces. country, depending on the availability of
2. It helps to avoid hyperglycemia by reduc- foods. Social and cultural habits, economics
ing glucose absorption in the intestines. status, age, sex and physical activity of the
This is because the transit speed of individual largely influence the intake of
digested food is increased in the intestines. diet.
Diet and Nutrition 163
The balanced diet for an adult male (70 kg) OPTIMAL NUTRITION FOR EXERCISE
requiring about 3,000 calories/day is as
follows: The Pregame Meal
Protein 70 g The pregame meal provides the athlete with
Fats 50 g adequate food energy and assure optimal
Carbohydrates 440 g hydration. As a general rule, foods that are
Calcium 500-700 mg high in fat content should be eliminated from
Phosphorus 1.4g the diet on the day of competition because
Iron 30-40 mg these foods are digested slowly and remain
Vitamin A 600-700 μg in the digestive tract for a longer time than
Vitamin B1 1.2-1.5 mg foods containing amounts of similar energy
Vitamin C 60-70 mg in the form of carbohydrates. So, the pregame
meal should be high in carbohydrate to assure
Malnutrition peak storage of liver and muscle glycogen.
Timing of the pregame meal should also
Protein-calorie malnutrition (PCM) results be considered because with the increased
from protein and/or calorie deficiency in stress and tension that usually accompany
varying proportions. It is the most important competition may cause a significant decrease
nutritional disorder in the developing in blood flow to the stomach and small
countries in the world. Malnutrition increases intestines and an accompanying decrease in
the susceptibility of the host to infection. The absorption from the digestive tract. A 3-hr
cause may be atrophy of lymphoid tissue and period should be sufficient to provide for
depression of synthesis of antibodies. adequate absorption of the pregame meal.
Kwashiorkor and marasmus are the two Commercially prepared liquid meals are
forms of PCM. also effective as pregame meal. These foods
Kwashiorkor occurs due to the deficiency are well balanced in nutritive value. These
of proteins. It affects children aged 2-3 years. are high in carbohydrate yet contain enough
It is characterized by oedema, hypopro- fat and protein to contribute a feeling of
teinaemia and some degree of growth satiety. They also contribute’s to the athlete’s
retardation. In general, the appetite is poor. fluid needs as they are in liquid form. The
There is mild to moderate anaemia. Fatty liquid meal is also advantageous because it is
liver, atrophy of the pancreas, salivary gland, digested rapidly and completely. This type
and intestine are some of the other changes. of nutrition on the day of competition is
Marasmus is due to the insufficient intake effective especially during long day meets
such as in swimming and track, or in some
of food (calorie deficiency). It mainly occurs
tennis and basketball tournaments.
in children under one years of age. Growth
retardation and muscle wasting are the most
Metabolism During Exercise
dominant features. There is no oedema as in
kwashiorkor so; this helps in distinguishing During high-intensity exercise, stored muscle
it from kwashiorkor. The appetite is very glycogen and blood-borne glucose are the
good. Plasma chemistry is normal. main sources of energy. About 30 to 40
164 Medical Biochemistry for Physiotherapy Students
percent of the total energy is supplied by is performed to the point where muscle
blood glucose. During initial stages of glycogen becomes severely lowered, fatigue
exercise, the uptake of circulating blood can easily occur, even though sufficient
glucose by the muscle increases sharply and oxygen is available to the muscles and the
continues as exercise progresses. After some potential energy from stored fat remains
time, the glucose uptake rises between 7 and almost unlimited. This is because the glyco-
20 times the uptake at rest depending on the gen stored in the muscles becomes depleted.
intensity of the exercise. Main use of During moderate levels of exercise,
carbohydrate during high intensity exercise sufficient oxygen is available to the cells.
is because it is the only nutrient which Consequently, most of the hydrogens stripped
provides energy when the oxygen supplied from the substrate and carried by NADH are
to the muscles is insufficient relative to the oxidized via ETC. So, in moderate level
oxygen needs. exercise, steady rate exists because hydrogen
During moderate exercise, energy is is oxidized at about the same rate as it is made
supplied mainly from the breakdown of the available. Even at rest or in mild exercise,
body’s stores of fat and carbohydrate. In the some lactic acid is continually formed. Lactic
initial stages, about 40 to 50 percent of the acid does not build up in this situation because
energy requirement is supplied by the its removal rate equals its rate of production
glycogen stored in the liver and exercising whereas in high-intensity exercise, when the
muscles. As exercise continues and the energy demands either the oxygen supply or
glycogen stores become reduced, then greater the rate of utilization, all of the hydrogen
percentage of energy is supplied through the joined to NADH cannot be processed through
metabolism of fat. Fat is mobilized from the respiratory chain. Under anaerobic
storage sites such as the adipose tissue and conditions, hydrogens combine with pyruvic
the liver and is delivered via the circulation acid to form lactic acid. Lactic acid is formed
to the working muscles. However, if exercise in the muscles and fatigue sets in.
Chapter 20

Radioactivity and Radioisotopes

ISOTOPES isotopes is unstable and, therefore, undergoes


decay and give out radioactive emanations
Isotopic atoms are atoms of the same element like α, β or γ rays.
with the same atomic number but different The important unstable isotopes are:
atomic weights. They are subspecies of the H3
same chemical element and occupy the same C14
position in the periodic table, but have P32.
different physical properties.
Isotopes are of two types: RADIOACTIVITY
1. Stable isotopes It is the phenomenon where radioactive
2. Unstable isotopes. substances emit α, β and γ rays on disinte-
gration.
Stable Isotopes Radioactive isotopes possess radioactivity
and give out radioactive emanations like
Stable isotopes are naturally occurring and
alpha, beta or gamma rays.
do not emit radiations. They can be identified
and quantified by mass spectrometry or Beta Rays
nuclear magnetic resonance (NMR). Beta rays because of their light mass and low
The important stable isotopes are: electrical charge penetrate into the matter in
H2 a long course. They are less readily absorbed
C13 by matter than alpha particles. Beta particles
N15 from H3, S35, C14 possess less energy, travel
O18. few mm and cannot cross the glass wall of a
container and pose no radiation damage for
Unstable Isotopes
humans. However, beta particles from P32 can
Unstable isotopes are also known as radio- travel across a room and penetrate deep into
active isotopes. The atomic nucleus of these the body tissues.
166 Medical Biochemistry for Physiotherapy Students
Alpha Rays radioactive material. It is expressed in Bq per
unit mass or volume of the material con-
Alpha rays are streams of high velocity
taining a radio isotope. Specific activity is
particles. Alpha rays because of their high
directly proportional to the decaying constant
charge and large mass follow short courses
and inversely proportional to the half life of
through matter. Alpha rays travel upto only
the radio isotope present in the sample under
a few cm in air, fail to cross the keratinized
assay (half-life of a radioisotope is the time
cutaneous epidermis or even a thick sheet of
at the end of which exactly half of its initial
paper and when emitted from internal
number of nuclei still remain unchanged).
sources can penetrate few μm to mm into soft
inner tissues.
Radiation Detection and Measurement
Gamma Rays Radiations are detected and measured from
Gamma rays are monochromatic electro- their electrical, chemical, thermal and light
producing or thermoluminescent effects.
magnetic radiations resembling X-rays. They
are not deflected in electrical fields. In 1. Gas ionization counters
contrast to X-rays, gamma rays generally a. Ionization chamber counter
possess higher frequencies and shorter b. Proportional counter
wavelengths. c. Geiger Muller counter
Due to their high energies and the absence 2. Scintillation counters
of mass and charge, gamma rays photons 3. Semi conductor detectors
penetrate deep into the matter, but they get 4. Pocket dosimeters
exponentially and progressively reduced in 5. Dosimeter badges.
intensity (attenuated) with the increasing
Artificial Radio Isotopes
thickness of the matter traversed by them.
Many man made radio isotopes are used
UNITS OF RADIOACTIVITY extensively in therapy, research and industry.
1. Curie (Ci) These obey the law of radioactive decay.
2. Becquerel (Bq)
3. Specific activity (SA) Production of Artificial Radioisotopes
Curie was an international unit of radio- a. By high voltage ion accelerators such as
activity based on the radioactivity of 1 g of cyclotrons, betatrons and synchotrons
radium. One Curie equals 3.7 × 1010nuclear b. By nuclear fissions in nuclear reactors
disintegrations per second. c. By neutrons from nuclear fusions in
Becquerel is now used as an international neutron generators
unit of radioactivity. It is the quantity of d. By radioactive decay in nuclide generators.
radioactive material in which one nuclear
disintegration occurs in one second. BIOLOGICAL EFFECTS
1 Bq = 2.7 × 10-11Ci.
Specific activity is the radioactivity Alpha particles, protons and gamma rays
possessed by a unit mass or volume of a emitted from atomic nuclei by the interaction
Radioactivity and Radioisotopes 167
of neutrons with matter, produce ionizing Ejection of other orbital electrons ionizes
effects on biological tissues. The biological the affected molecule, breaking or changing
effects of radioactivity on tissues etc. are its non covalent bonds and altering its
expressed as rad (Roentgen absorbed dose) molecular conformation. In both cases, the
and rem (Roentgen equivalent man). The unit biological activity of the molecule is altered
of radiation actually absorbed is defined in or destroyed.
terms of energy absorbed: 1 rad = 100 ergs/g. Indirect actions follow its direct actions
As an example of the use of this unit, the on mainly water molecules in the organism
maximum occupational exposure to radiation which produce free hydrogen, hydroxyl,
should not exceed 5 rems/year on an average. hydroperoxyl radical and hydrogen peroxide
which in turn result in dissociation of bonds.
Stochastic and Non-stochastic Effects
Effect on Rare Biomolecules
Stochastic or zero threshold effects of
Chromosomal DNA may suffer radiational
radiation include mainly the genetic effects
damage. A DNA molecule may suffer a single
and malignancies. No minimum dose can be
stranded or double stranded break due to
identified for their production and they may
the reaction and binding with free radicals.
result from the exposure to even a single
Single stranded breaks may be repaired
gamma or X-ray photon.
in a few hours but a double stranded break
Stochastic effects seem to result from
resulting from the binding of two free radicals
changes in some essential biomolecules of
side by side to the two strands of DNA, may
very low concentrations like nucleic acids and
frequently escape repair and leads to
enzymes due to direct ionizing actions of
mutations, chromosomal aberrations, cellular
radiations on them.
death or malignancy.
Non-stochastic or threshold effects of
radiation include most of the other biological Salient Acute Effects
effects. Production of each non stochastic
Acute effects appear in a short interval after
effect requires a minimum or threshold dose
a single exposure to a high dose of radiation
of radiation. Non stochastic effects seem to over a short period of time.
result from the indirect actions of radiation
Acute effects of whole body irradiation
which produce free hydrogen, hydroxyl,
vary with the magnitude of the dose because
hydroperoxyl radical and hydrogen peroxide
all tissues are not equally radio sensitive.
which in turn affect other biomolecules like
Nausea, vomiting, hyperthermia, fatigue,
enzymes and nucleic acids.
cutaneous erythrema and dermatitis occur at
all levels of radiation. Other acute effects
Direct and Indirect Actions
consist of haemopoietic, gastrointestinal and
Direct actions of radiations on biomolecules central nervous system syndromes.
mostly result in the ejection of orbital
electrons from their atoms. Ejection of valence Delayed Effects
electrons from their orbits dissociates cova- Delayed effects follow either long after a
lent bonds to cause cleavage of the bio- single high dose of radiation or after a
molecule. prolonged low level radiation.
168 Medical Biochemistry for Physiotherapy Students
Prolonged irradiation may result from course of radioactivity changes is studied
either an external source of radioactivity or in different parts of the body. Instead of
an ingested or inhaled radioisotope of long a steady and uniform distribution in the
half life deposited in the tissues. In the long entire circulation, radioactivity rises
range, slow depopulation of endothelial cells progressively in an organ suffering from
and consequent capillary occlusions may internal hemorrhages because the Cr51—
impede the repair and cellular repopulation erythrocytes accumulate extravascularly
of tissues having high mitotic rates, may also there.
slowly deplete the populations of long living 2. Thyroid function investigation: Thyroid
cells having low mitotic rates and may even function and the metabolic pathway of its
cause a secondary loss of non dividing cells hormones are investigated after adminis-
like neurons, cardiac cells and striated muscle tration of I131 which is concentrated by the
gland, used in thyroid hormone synthesis
cells. Such secondary delayed effects include
and secreted as thyroid hormones in the
1. cutaneous lesions, ulcers, scars and
blood.
atrophy of skin
3. Radiopyelography: Renal functions may be
2. pericarditis, pericardial fibrosis
investigated by injecting I 131 labelled
3. pneumonitis, pulmonary fibrosis and
diodrast intravenously and measuring the
progressive dyspnoea radioactivity subsequently in each kidney
4. late secondary damage of intestinal by a Gieger–Muller counter placed over it
muscle and serous coats leading to for a short given interval. A normal kidney
fibrotic changes on the intestinal wall as eliminates diodrast much more rapidly
well as intestinal absorption than one suffering a failure.
5. loss of microvilli, flattening of renal 4. Cerebral circulation: A gas mixture containing
tubular epithelium, glomerular damage, Xe133 is inhaled for a few minutes. Geiger-
nephritis, loss of nephrons and even Muller counter or scintillation counters are
renal hypertension then placed over specific sites of the skull
6. demyelination and loss of neurons, focal to determine the times of disappearance
necrosis in the white matter of the central of the radioisotope at the underlying sites
nervous system. of the brain. The mean blood flow in
Delayed effects not dependent on different cerebral areas may be estimated
capillary occlusions include from these findings.
1. cancers 5. Radiocardiography: Following the injection
2. cataract of Ba137 into the subclavian vein, Geiger-
3. genetic effects Muller counters or scintillation counters
are applied over the body to study the
4. shortening of life span.
blood flow between heart chambers, from
DIAGNOSTIC USES OF RADIOISOTOPES left ventricle to aorta and between the
heart and lungs. This method rapidly
1. Detection of internal hemorrhages: The patient diagnoses congenital or pathological
is intravenously infused with a suspension defects and malfunctions of cardiovascular
of Cr 51 labelled erythrocytes and the and pulmonary systems.
Radioactivity and Radioisotopes 169
6. ECF volume: Sodium ion (Na+) diffuses A radioisotope may be implanted in a
freely from plasma to tissue fluids, but is malignant tissue, so controlling the dose of
largely retained by kidneys and diffuse radioactivity and period of implantation that
little into cells. So the total ECF volume is its ionizing radiations destroy the malignant
given by the dilution of Na24 in plasma cells with minimum damage to healthy cells
following an intravenous injection of a or patient can be injected with a radioisotope
known amount of NaCl24 into the patient. which is selectively concentrated by the tissue
7. Cardiac output: A measured amount of a to be irradiated.
non diffusible radioisotope substance is 1. Co 60 is the most important isotope in
injected into the forearm vein. The clinical medicine. It emits β and γ rays
concentrations of the radio isotope are which can penetrate deep into the tissues
determined in several successive blood and treat internal cancers without causing
samples on the basis of their radioacti- severe skin reactions.
vities. The data is used in calculating the 2. P 32 is administered to polycythemia
cardiac minute volume.
patients and gets rapidly concentrated by
8. Circulating blood volume: The patient is
bone marrow cells, leading to their
intravenously infused with a measured
selective destruction by the emitted
volume of an isotonic fluid emitting a
β-rays.
known amount of radioactivity due to the
3. I131 is selectively concentrated by thyroid
presence of either I131 -albumin or Fe59 –
cells and helps to destroy thyroid tumors
labelled erythrocytes in it. The radio-
activity of a blood sample collected is by its β and γ rays.
subsequently measured. The dilution of 4. Au48 is used in used in the treatment of
the infused isotope is given by the ratio malignant pleural and peritoneal effu-
of radioactivities of the infused fluid and sions.
the blood sample. The circulating blood 5. Sr90 is used for lesions of cornea, conjuctiva
volume is calculated from this ratio and and sclera.
the volume of the infused fluid. 6. Heavy particle proton irradiation of the
pituitary gland has been used in diabetic
THERAPEUTIC USES OF RADIOISOTOPES retinopathy in an attempt to improve
Radioisotopes have a useful role in the treat- vision or slow down the rate of deterio-
ment of diseases, particularly malignancies. ration of vision.
Chapter 21

Normal and Abnormal


Constituents of Urine

Urine is an excretory product of the body, clean, dry vial is adequate. Container should
formed in the kidneys. Examination of urine be free of disinfectant and properly labelled.
for changes in physical characteristics and the Investigation should be done immediately,
detection of abnormal constituents is an in case of ketone body estimation or within 2
important test for suspected renal damage as hours of its collection.
well as systemic diseases. The patient should be instructed to void
directly into the container. During the collec-
COMPOSITION OF NORMAL URINE tion, the initial portion of the urine stream is
allowed to escape while the midstream
Urine is a fluid containing water soluble waste portion is collected.
products excreted from the blood via the For qualitative tests, the morning urine is
kidneys. Urine is mainly composed of 90 to useful; however, quantitative tests are
95 percent water and the rest being made up performed only on urine specimen collected
of solid material which may be organic or for 24 hours.
inorganic like urea, uric acid, creatinine, Urine collection requires preservatives to
sodium, potassium, chloride, calcium, prevent some changes which urine can
phosphates etc. The composition of urine undergo on standing (keeping):
varies and is affected by three factors: 1. conversion of urea to ammonia due to
1. Nutritional status of the individual. bacterial action
2. State of metabolic processes. 2. action of micro-organisms, both bacteria
3. Ability of the kidney to selectively handle and yeast on glucose
3. precipitation of phosphates
the material presented to them.
4. deposition of uric acid on standing.
Specimen Collection and Preservation Preservatives
For most of the routine investigations, fresh 1. Hydrochloric acid: 100 ml of M-hydrochloric
midstream specimen of 10- 20 ml urine in a acid is a satisfactory preservative for urine
Normal and Abnormal Constituents of Urine 171
collections for calcium, phosphate, - Diabetes insipidus
nitrogen, ammonia and vanillyl mandelic - Later stages of chronic glomerulonephritis
acid (VMA). - Compulsive polydypsia.
2. Toluene: 10 ml of toluene is commonly used
for 24-hr collections for sodium, potassium, Oliguria
uric acid, creatinine and protein analyses.
The term refers to the excretion of an
Toluene collections are not suitable for the
abnormally small volume of urine (upto
determination of calcium, phosphate,
VMA, ammonia etc. 500 ml) in 24 hours. Causes are:
3. Hibitane: Hibitane (chlorhexidine diace- - diarrhoea
tate) is a satisfactory preservative when - severe oedema
glucose is to be estimated in urine, since it - fever
inhibits the action of bacteria but does not - acute nephritis
interfere with the analysis either by - early stages of glomerulonephritis.
enzymatic or reduction methods. 5 ml of
a 5 percent aqueous solution are added to Anuria
the container. This is the complete absence of urine
4. Acetic acid: 100 ml of 25 percent acetic acid formation which is unusual. Causes are:
is used as a preservative for 24-hour - acute tubular nephrosis
collections for 5-hydroxyindoleacetic acid - blood transfusion reaction
estimation. 20 ml of glacial acetic acid is
- surgical shock
used for the daily 24-hour collection
- bilateral renal stones.
required for the ascorbic acid saturation
test.
Nocturia
PHYSICAL EXAMINATION OF URINE The excretion, by an adult, of urine more than
500 ml at night is called nocturia. Cause is
Physical examination of urine is also referred chronic glomerulonephritis.
to as macroscopic examination and it involves
the assessment of physical properties such as Colour
volume, colour, appearance, odour, specific
gravity and pH. The colour of normal urine varies from
colourless to pale or straw yellow. This colour
Volume of normal urine is due to the presence of three
For an adult, the normal daily volume of urine pigments: urochrome (a yellow pigment
is about 1,200-1,500 ml. More urine formation present in large concentrations), uroerythrin
takes place during the day than at night. (a by-product of red blood cell degradation)
and urobilin (another pigment resulting from
Polyuria red blood cell degradation).
This term refers to consistent elimination of There may be variation in colour which
an abnormally large volume of urine may be physiological or pathological. Physio-
(> 2,500 ml) in 24 hours. Causes are: logical – Colour may be dark yellow due to
- Diabetes mellitus vitamin B complex therapy. It may be orange
172 Medical Biochemistry for Physiotherapy Students
due to certain drugs. Pathological- Colour Causes of high specific gravity are:
may be reddish or reddish brown due to the - excessive sweating
presence of blood. This condition is called - glycosuria
haematuria. If colour is milky, it is due to fat - acute nephritis
globules and condition is known as chyluria. - albuminuria
If urine turns black on standing, it is due to - all causes of oliguria.
metabolic disorder known as alkaptonuria.
Yellow colour is due to the presence of bile Causes of low specific gravity are:
pigments mainly bilirubin and biliverdin. - excessive fluid intake
- chronic nephritis
Odour - diabetes insipidus
- all causes of polyuria except diabetes
Normally urine gives aromatic or ammoniacal mellitus.
odour due to presence of volatile organic sub-
stances. Foods like onion and garlic impart pH
their smell to urine. In diabetic ketoacidosis,
there is acetone like smell. Bacterial infections Freshly voided normal urine has a pH range
give foul smell. of 5.0- 6.8. On standing at room temperature,
urea is converted to ammonia by the action
Appearance of bacteria in the urine, thus raising the pH.
A diet rich in meat proteins causes
Normal urine is usually clear. It may appear
acidification of urine while consumption of
cloudy if amorphous phosphates are present
vegetables and citrus fruits makes the urine
in alkaline urine or amorphous urates are
alkaline.
present in acidic urine. Urine may appear
cloudy or turbid from the presence of Acidic urine may be seen in different types
leucocytes and epithelial cells. Presence of red of metabolic disorders, especially diabetic
blood cells may give urine turbid and smoky acidosis resulting from an accumulation of
appearance. ketone bodies in the blood. Alkaline urine
may be seen in some infections, metabolic
Specific Gravity disorders and with administration of some
drugs.
Specific gravity of urine is the measure of the
amount of dissolved substances in the urine. NORMAL CONSTITUENTS OF URINE
It is the weight of the urine compared to the
weight of an equal volume of distilled water Organic Constituents
at a constant temperature. The normal range Urea
is 1.003-1.030.
Specific gravity is measured in several Urea is formed in the liver as the end product
ways; of protein metabolism and so its excretion
- urinometer depends on protein intake. Among the
- refractometer organic constituents, urea is the main. About
- reagent strips 80- 90 percent of nitrogen in the urine is in
- osmometric method. the form of urea.
Normal and Abnormal Constituents of Urine 173
Urea excretion is increased on a high occurs in muscle in the form of creatine
protein diet and in protein catabolic states phosphate. This leads to the formation of
such as during fever. A decrease in urea creatinine in blood which eventually gets
excretion is seen in a diet low in protein, excreted through urine.
diseases associated with impaired liver
functions (such as cirrhosis) and in kidney Test for Creatinine
diseases (such as nephritis).
The presence of creatinine in urine is routinely
checked by using alkaline picric acid solution.
Test for Urea
This is known as Jaffe’s test (Jaffe’s reaction).
When urea is treated with sodium hypo- Take 5 ml of urine in a test tube and add 1 ml
bromite, it decomposes to give nitrogen. Take saturated solution of picric acid to it. Make
5 ml urine in a test tube and add 3- 4 drops of the mixture alkaline with 1 ml of sodium
hypobromite regent. There will be marked hydroxide solution. A reddish-orange colour
effervescence as the gaseous nitrogen is appears due to the formation of creatinine
evolved. picrate.
Uric Acid
INORGANIC CONSTITUENTS
Uric acid is derived from nucleic acids by the
oxidation of purine bases. It is found in urine Chloride
normally to the extent of 0.5 to 1.0 g/24 hrs
Chloride is the chief inorganic constituent of
but its excretion varies widely depending
urine. Excessive sweating, diarrhoea,
upon the diet and in certain other conditions.
vomiting, oedema are causes for low urinary
On a purine free diet, uric acid output may
be reduced upto 0.1 g/24 hrs while on a high chlorides. Excessive water drinking,
purine diet, uric acid excretion may be Addison’s disease are causes for increase of
increased upto 2.0 g/24 hrs. Pathologically, urinary chlorides.
uric acid excretion is increased in gout and
leukaemias. Test for Chloride
Take 2 ml of urine in a test tube; add 0.5 ml
Test for Uric Acid of concentrated nitric acid and 1 ml of silver
Uric acid is a reducing agent in alkaline nitrate. White precipitates of silver chloride
condition. It reduces phosphotungstic acid to indicate the presence of chloride.
tungsten blue. Take 2 ml of urine in a test
tube; add a few drops of phosphotungstic acid Phosphates
reagent followed by a few drops of 20 percent Urinary phosphates are present as inorganic
sodium carbonate. The development of blue as well as organic form. Increase of urinary
colour shows the presence of uric acid. phosphate excretion occurs in rickets,
osteomalacia, hyperparathyroidism etc.
Creatinine
Decreased excretion occurs in diarrhoea,
Urinary creatinine is derived from muscle nephritis, acute infections, and hypopara-
creatine. Considerable amount of creatine thyroidism.
174 Medical Biochemistry for Physiotherapy Students
Test for Phosphates ABNORMAL CONSTITUENTS OF URINE
To 5 ml of urine in a test tube, add few drops Abnormal urine may contain protein, reducing
of concentrated nitric acid and pinch of sugar, bile pigments, ketone bodies, uro-
ammonium molybdate and warm it. Yellow bilinogen, bile salts, blood, etc.
colouration shows the presence of phos-
phates. Proteins
Normally adults excrete small amount upto
Sulphates
160 mg/day of proteins in urine which consist
Sulphates excreted in the urine arise mainly of albumin, plasma proteins and glyco-
from the oxidation of the sulphur of proteins proteins which cannot be detected by routine
(from sulphur containing amino acids) within tests.
the body. Its excretion may be increased on Increased amount of proteins in urine i.e.
high protein diets, excessive tissue break- proteinuria can be caused by increased
down while decreased in renal functional glomerular permeability, reduced tubular
impairment conditions. reabsorption, increased secretion of protein
in renal tract. Most common type of pro-
Test for Sulphates teinuria is due to albumin, i.e. albuminuria.
Proteinuria may be physiological and patho-
Take 5 ml of urine in a test tube and add 3-4
logical.
drops of concentrated hydrochloric acid.
1. Physiological proteinuria: Causes are
Then, add few drops of barium chloride
- postural, i.e. present on standing and
solution to it. A white precipitate of barium
absent on lying down
sulphate is formed due to the presence of - severe stress
sulphates in urine. - lasts weeks of pregnancy
2. Pathological proteinuria: Causes are
Calcium - pre renal
The greater proportion of calcium is excreted - renal
- post renal
in the faeces. The urinary output is between
Pre renal causes: Kidneys are affected in such
0.1 to 0.3 g. Increased urinary excretion occurs
a way that they secrete albumin as such in
in hyperparathyroidism, hyperthyroidism,
urine. It can be due to cardiac disorder in
multiple myeloma and renal stones.
which the circulation of blood to kidneys is
affected like toxaemia of pregnancy, heart
Test for Calcium
disease, abdominal tumour, severe anaemia,
Take 2 ml of urine in a test tube; add 5 drops fever, etc.
of 1 percent acetic acid and 5 ml of potassium Renal causes: In all forms of kidney diseases
oxalate. White precipitates of calcium oxalate like nephrotic syndrome, pyelonephritis, and
are formed. glomerulonephritis.
Normal and Abnormal Constituents of Urine 175
Post renal causes: Proteins are added to of pathological glucosuria. Positive reaction
urine as it passes along the urinary tract in of urine can also be seen in the cases like
the conditions like severe urinary tract alimentary glycosuria, hyperthyroidism, in
infection, inflammation. emotional disturbances, in severe infections.
Certain protein cells called Bence Jones
protein are present in the urine of patient Test for Sugars
suffering from multiple myeloma. Bence Jones Benedict’s Test
proteins are light chains of immunoglobulins
and have specific characteristics that appear Take 5 ml of Benedict’s solution and add to
in urine when heated for 50-60ºC and it 8 drops of urine and set in a boiling water
disappear when urine is further heated. bath for 5 minutes or else boil it over a flame
for 2 minutes. A light green, yellow or brick
Test for Proteins red colour is produced depending on whether
urine contains 0.5, 1 or more than 2 percent
Heat Coagulation Method
glucose.
Take 5 ml of urine, heat the upper half edge
of the tube. Add 2-3 drops of 3 percent glacial Ketone Bodies
acetic acid and heat it. Formation of white Normally about 1 mg/ dl is the blood level
precipitates indicates the presence of proteins. of ketone bodies. Small amounts are also
excreted in urine, i.e. less than 1 mg/ 24 hrs.
Sulphosalicylic Acid
Ketone bodies are acetone (2%), acetoacetic
Take 2 ml of centrifuged urine in a test tube acid (20%) and β-hydroxy butyric acid (78%).
and equal amount of 5% sulphosalicylic acid. Synthesis of ketone bodies occur in liver and
Stand it for 10 minutes at room temperature. they are utilized in extrahepatic tissues like
If protein present, then white precipitates will brain, kidney, muscle, etc. Causes of ketonuria
be seen. are uncontrolled diabetes mellitus, starvation,
and intake of high fat, low carbohydrate diet.
Heller’s Test
Test for Ketone Bodies
Take 1 ml of nitric acid in a test tube and add
urine by touching the sides of the test tube. Rothera’s Method
If white ring is observed at the junction, it
Take 5 ml of urine in a test tube, saturate it
indicates proteins presence.
with ammonium sulphate, and then add 1
crystal of sodium nitroprusside and mix. Add
Reducing Sugars
liquor ammonia by touching the side of the
Normal urine contain small amount of test tube. A purple ring at the junction
reducing sugar, i.e. 1-1.5 g/24 hrs. Out of this, indicates the presence of ketone bodies.
glucose is present in concentration of 50- 300
mg/24 hrs. These are not detected in urine Gerhardt’s Test for Acetoacetic Acid
by routine test. Glucose in urine is called Take 2 ml of urine in a test tube; add 5-6 drops
glucosuria. Diabetes mellitus is an example of ferric chloride. Red colour development
176 Medical Biochemistry for Physiotherapy Students
indicates the presence of acetoacetic acid. Test for Urobilinogen
Ehrlich’s Aldehyde Test
Bile Salts
Take 10 ml of urine in a test tube, add 2.5 ml
These are sodium and potassium salts of
of barium sulphate and filter. Take 2.3 ml of
glycocholic and taurocholic acid. They are
filtrate in a test tube and add 0.5 ml of Ehrlich’s
synthesized in liver from cholesterol. These aldehyde reagent. Allow to stand for 3 min.
help in the digestion of fats. Cause of After 3 min., observe for the pink colour
appearance in urine is obstructive jaundice. which indicates its presence.

Test for Bile Salts Blood


Presence of blood in urine is called haema-
Hay’s Sulphur Test turia. Causes are injury of kidney, violent
To 2 ml of urine in a test tube, add a pinch of exercise, tumour of kidney and various drugs
sulphur powder. If it sinks towards the like salicylates and barbiturates.
bottom, it indicates presence of bile salts. Test for Blood
Benzidine Test
Urobilinogen
To a small amount of benzidine, add 0.5 ml
It is formed from bilirubin in the intestine by of glacial acetic acid and then add hydrogen
the action of bacteria. In patients suffering peroxide. Mix well and add 1 ml of urine and
from haemolytic jaundice, it appears in urine. again mix it. Deep blue colour indicates the
presence of blood.
Index

A factors controlling active optical activity 31


transport of glucose 77 zwitter ion and isoelectric
Abnormal constituents of urine Absorption of fats 78 pH 32
174 Absorption of proteins 80 neutral amino acids 28
bile salts 176 Adrenal gland and its hormones acidic amino acids 29
blood 176 152 neutral amino acids 28
ketone bodies 175 adrenal cortex 152 nutritional classification 30
proteins 174 functions 153 essential amino acids 30
reducing sugars 175 glucocorticoids 153 non-essential amino
test for proteins 175 mineralocorticoids 153 acids 30
heat coagulation method Adrenal medulla 154 semi essential amino
175 biosynthesis and secretion 154 acids 30
Heller’s test 175 hyperactivity 155 physical properties 32
sulphosalicylic acid 175 pheochromocytoma 155 structure 34
test for sugars 175 metabolic fate of epinephrine primary structure 34
Benedict’s test 175 and norepinephrine 155 quaternary structure 35
urobilinogen 176 physiological and secondary structure 34
test for urobilinogen 176 biochemical functions 155 tertiary structure 35
Abnormalities associated with Amino acid pool 109 Atherosclerosis 108
female sex hormones formation of urea 109
157 ammonia transport 111 B
hypogonadism 157 oxidative deamination 110
Abnormalities of parathyroid reactions of the urea Balanced diet 162
function 150 cycle 111 Basal metabolic rate (BMR) 160
actions 150 transamination 109 factors affecting BMR 160
calcitonin 150 urea formation (urea age 160
hypoparathyroidism 150 cycle) 111 climate 160
Abnormalities related with male Amino acids 28 disease 160
sex hormones 156 biological importance 32 effects of hormones 160
estrogenic hormones 157 classification 32 habit 160
effects 157 conjugated proteins 33 sex 160
female hormones 157 derived proteins 33 state of nutrition 160
hypogonadism 156 simple proteins 33 surface area 160
synthetic estrogens 157 classification of proteins specific dynamic action 160
Absorption 75 based on the shape Biochemistry 1
general principles 75 of the protein animal biochemistry 1
adsorption 76 molecule 34 medical biochemistry 1
diffusion 75 fibrous or fibrillar plant biochemistry 1
hydrostatic pressure 75 proteins 34 Bioenergetics 69
hydrotropy 76 globular or corpuscular ATP- energy rich compound
osmotic pressure 76 proteins 34 69
passive and active denaturation 36 concept of energy 69
transport 76 functions 29 free energy 69
Absorption of carbohydrates 76 general characteristics 31 Biological oxidations 70
active transport of glucose 76 isoelectric pH 32 aerobic dehydrogenases 71
178 Medical Biochemistry for Physiotherapy Students
anaerobic dehydrogenases 71 Cellular functions of DNA 42 chyluria 79
hydroperoxidases 71 Cerebrosides 24 lipid malabsorption 79
oxidases 70 clinical aspect 24 Defect in digestion and
oxygenases 71 Gaucher’s disease 24 absorption proteins 81
Biosynthesis of fatty acids 104 lipoproteins 25 Dehydration 127
de novo synthesis of fatty functions 25 features 127
acids 104 sulpholipids 25 treatment 127
microsomal chain elongation types 24 Derived lipids 25
105 cerebron 24 fatty acids 25
mitochondrial chain kerasin 24 saturated fatty acids 25
elongation 105 nervon 24 properties 26
Biotin 65 oxynervon 24 Diabetes mellitus 97
chemistry 65 Chargaff’s rule 40 insulin-dependent diabetes
daily requirement 66 Cholesterol metabolism 105 mellitus 97
deficiency 66 blood cholesterol and non-insulin-dependent
functions 66 cardiovascular diabetes mellitus 97
sources 66 disease 107 Diagnostic importance of
Body fluid distribution 124 fate of cholesterol 106 enzymes 51
constituents of ICF 125 metabolism of lipoproteins 107 Diagnostic uses of radioisotopes
constituents of ECF 125 synthesis 106 168
extracellular fluid (ECF) 124 Composition of normal urine 170 cardiac output 169
factors which influence the Connective tissue 138 cerebral circulation 168
distribution of composition 138 circulating blood volume 169
body water 125 ground substance 138 detection of internal
fluid intake 126 fibres 138 hemorrhages 168
fluid output 125 collagen 139 ECF volume 169
intracellular fluid (ICF) 125 elastin 139 radiocardiography 168
plasma 124 reticulin 139 radiopyelography 168
Bone 142 Copper 121 thyroid function investigation
composition 142 absorption 122 168
formation of bone 142 daily requirements 122 Digestion and absorption of
deficiency state 122 carbohydrates 74
C physiological functions 121 Digestion and absorption
sources 121 of lipids 77
Calcium 117 Copper toxicity 122 dietary sources of lipids 77
absorption of calcium 118 Wilson’s disease 122 animal sources 77
daily requirement 118 Cushing syndrome 154 vegetable sources 78
deficiency state 118 Cyanocobalamin 67 Digestion and absorption of
osteoporosis 118 chemistry 67 proteins 79
rickets 118 daily requirement 67 Digestive system 74
physiological functions 117 deficiency 67 functions 74
sources 118 functions 67
Calorimetry 158 sources 67 E
energy value of foods 158
energy value or caloric value Ehrlich’s aldehyde test 176
D
of food 158 Electrolytes 8
respiratory quotient (RQ) 159 Defect in digestion and acids 9
Carbohydrates 12 absorption of inorganic acids 9
classification 12 carbohydrates 77 organic acid 9
functions 12 Defect in digestion and precautions while hand-
Cardiolipin 24 absorption of lipids 79 ling acids and bases 9
Index 179
properties of acids 9 deficiency 67 metabolic fate 115
safe use 10 functions 67 primary hyperoxaluria 115
storage 10 sources 66 Glycogen metabolism 89
bases 10 Fructose metabolism 94 Glycogenesis 90
importance 10 Functions of essential fatty acids steps 90
properties 10 26 Glycogenolysis 90
salts 11 alcohols 27 Glycogen storage diseases 81
importance 11 steroids and sterols 27 Glycolipids 24
medical salts 11 Functions of nucleotides 39 types 24
theory of arrehenius Functions of phospholipids 24 cerebrosides 24
ionization 11 gangliosides 24
Electron transport chain 72 G Glycolysis 83
inhibitors of the ETC 73 importance 83
location 72 Galactose metabolism 94 regulation 83
reactions 72 Genetic code 42 steps 83
Enzyme kinetics 48 characteristics 42
nonoverlappin and H
enzyme inhibition 49
feedback inhibition 50 commaless 42 Hay’s sulphur test 176
irreversible inhibition 50 redundancy 42 HMP shunt 87
Michaelis-Menten equation 48 specificity 42 metabolic significance 89
revesible inhibition 49 universality 42 importance of NADPH 89
competitive inhibition 49 Gluconeogenesis 81 importance of pentoses 89
non-competitive inhibition gluconeogenesis from amino steps 87
49 acid 92 Hormones 143
Enzymes 43 gluconeogenesis from mechanism of hormone action
chemical nature 43 glycerol 94 143
classification of enzymes 44 gluconeogenesis from activation of intracellular
propionate 94 receptors 143
hydroases 45
glucose-6-phosphate to activation of plasma 144
isomerases 45
glucose conversion 92 antagonistic effect 145
ligases 45
glyconeogenesis from hormonal interactions 145
lyases 45
pyruvate 92 membrane receptors 144
oxidoreductases 44
importance 92 protein kinases 145
transferases 45
pyruvate to phospho- role of G-proteins 145
coenzymes 44
enolpyruvate second messengers 144
functions 44
conversion 92 Hydrogen ion concentration 128
properties 43
reactions 92 acid-base imbalance 132
ribozymes 44
Glucose tolerance test (GTT) 98 acidosis 132
Enzymes in clinical diagnosis 50
increased glucose tolerance 99 metabolic acidosis 132
indications 98 respiratory acidosis 132
F interpretation 98 alkalosis 132
Fluorine 12 normal glucose tolerance metabolic alkalosis 132
absorption 123 curve (GTC) 98 respiratory alkalosis 132
daily requirements 123 intravenous glucose tolerance biological significance 129
deficiency state 123 99 Henderson-Hasselbalch
fluoride toxicity 123 method 98 equation 128
physiological functions 122 Glutamic acid 115 measurement 129
sources 123 metabolic fate 115 mechanisms 139
Folic acid 66 Glycine 115 buffer systems 129
chemistry 66 inherited disorder 115 renal mechanism 131
daily requirement 66 glycinuria 115 respiratory mechanism 131
180 Medical Biochemistry for Physiotherapy Students
I M Muscle tissue 135
chemical composition 136
Inherited disorder 113 Magnesium 119 myofibrils 136
alkaptonuria 114 absorption 119 protein of myofibrils 136
phenylketonuria (PKU) 113 daily requirements 119 proteins of sarcoplasm 136
tyrosinaemia 114 deficiency state 119
Inherited disorders of collagen physiological functions 119
139
N
sources 119
cutis laxa 140 Malnutrition 163 Nerve tissue 140
Ehlers-Danlos disease 139 Mechanism of enzymes action 47 amino acids of brain 141
Marfan’s syndrome 140 antithyroid agents 149 brain lipids 140
osteogenesis imperfecta hyperthyroidism 149 brain proteins 140
congenita 139 hypothyroidism 149 composition 140
Inorganic constituents 173 metabolic fate and excretion metabolism of brain 141
calcium 174 149 Niacin 62
test for calcium 174 simple goitre 149 chemistry 63
chloride 173 specificity 47 clinical feature 63
test for chloride 173 toxic nodular goitre 150 daily requirement 63
phosphates 173 Metabolism of carbohydrates 82 deficiency 63
test for phosphates 174 anabolism 82 functions 63
sulphates 174 catabolism 82 sources 63
test for sulphates 174 fate of carbohydrates 82 Normal constituents of urine 172
Iodine 122 amino acid synthesis 83 creatinine 173
absorption 122 ATP production 82 organic constituents 172
daily requirements 122 uric acid 173
glycogenesis 83
deficiency state 122 Nucleoside 39
lipogenesis 83
excretion 122 Nucleoties 37
Metabolism of creatine and
physiological functions 122 nitrogenous bases 37
creatinine 116
sources 122 purine bases 38
Metabolism of important amino
Islets of Langerhans 151 adenine (A) 38
acids 112
Isoenzymes 51 guanine (G) 38
fate 113
Isotopes 165 pyrimidine bases 37
formation fumarate and
stable isotopes 165 Nutritional importance of
acetoacetate 113
unstable isotopes 165 carbohydrates 161
phenylalanine and tyrosine
Nutritional importance of dietary
112
K fibers 162
Methionine 116
Nutritional importance of fats
Ketone bodies 102 transmethylation 116
162
excessive production of Monosaccharides 12
ketone bodies in biological importance 14
O
diabetes mellitus 103 D-series and L-series 13
synthesis 103 epimers 14 Obesity 108
utilization 103 general properties 13 Optimal nutrition for exercise
asymmetric carbon atom 163
13 metabolism during exercise
L
isomerism 13 163
Lipids 21 Optical activity of sugars 13 pregame meal 163
importance 21 properties 14 Osmosis 133
classification 21 physical properties 14 applications of osmosis 134
simple lipids 21 reactions due to the alcoholic biological significance of
compound lipids 23 groups 16 osmosis 134
Index 181
determination of osmotic Pituitary gland 145 Regulation of blood glucose 96
pressure 134 clinical significance 146 by hormones 96
osmosis requires 133 control of secretion 146 by metabolic processes 96
osmotic pressure 133 hormones of the anterior epinephrine 96
units 134 pituitary 146 glucagons 96
Oxidative phosphorylation 73 functions 146 glucocorticoids 96
P:O ratio 73 growth hormone (GH) 146 growth hormone 97
Sites of oxidative Plasma membrane 3 thyroxine 97
phosphorylation 73 centrioles 5 Regulation of fluid and
endoplasmic reticulum 4 electrolyte balance 126
P Golgi apparatus 4 Renin-angiotensin-aldosterone
lysosomes 5 sytem 153
Pancreatic hormones 151 microfilaments and clinical features 154
actions 152 microtubules 5 Riboflavin 62
chemistry 151 mitochondria 3 chemistry 62
chemistry 152 nucleus 3 daily requirement 62
glucagons 151 ribosomes 4 deficiency symptoms 62
insulin 151 Potassium 120 functions 62
metabolic effects of insulin 151 absorption 120 sources 62
metabolism of insulin 151 daily requirements 120 Role of metal ions in enzymes 45
Pantothenic acid 64 disease state 120 factors affecting enzyme
chemistry 64 physiological functions 120 action 45
daily requirement 65 sources 120 effect of activators 46
deficiency 65 Progestational hormones 157 effect of enzyme
functions 65 functions 157 concentration and
sources 65 Prolactin (lactogenic hormone) substrate concentration
Parathyroid gland 150 146 46
actions 150 functions 146 effect of ionic strength 47
control of secretion 150 gonadotrophins (FSH and effect of oxidation 46
Phosphatidyl ethanolamine LH) 147 effect of pH 45
(cephalin) 23 hormones of intermediate effect of product
Phosphatidylinositol 24 147 concentration 46
Phospholipids 23 hormones of the posterior effect of radiation 47
Phosphoric acid 38 pituitary 147 effect of temperature 46
Phosphorus 118 lobe of pituitary 147
absorption 119 S
prolactin pathophysiology
daily requirements 119 146 Serum enzymes in muscle
deficiency state 119 thyrotropic hormones 147 diseases 51
physiological functions 118 Pyridoxine 63 CPK 51
sources 119 chemistry 63 SGOT 52
Physical examination of urine 171 daily requirement 64 SGPT 52
appearance 172 deficiency 64 Sex hormones 155
color 171 functions 64 androgens 155
odour 172 sources 64 biochemical effects 156
pH 172 biosynthesis 156
specific gravity 172 Sodium 119
R
volume 171 absorption 120
anuria 171 Radioactivity 165 daily requirements 120
nocturia 171 alpha rays 166 disease state 120
oliguria 171 beta rays 165 physiological functions 119
polyuria 171 gamma rays 166 sources 120
182 Medical Biochemistry for Physiotherapy Students
Somatostatin 152 physiological functions 121 deficiency 57
actions 152 sources 121 rickets 57
chemistry 152 Tricarboxylic acid cycle 85 functions 57
Sphingomyelins 24 biological significance of citric hypervitaminosis D 57
β-oxidation of fatty acids 100 acid cycle 87 osteomalacia 57
energy yield from β-oxidation energy production during sources 56
101 citric acid cycle 87 Vitamin E 57
oxidation of fatty acids with steps of citric acid cycle 86 absorption 58
odd number of yeast fermentation 87 chemistry 57
carbons 102 Tryptophan 114 daily requirement 58
β-oxidation proper 101 formation of serotonin 114 deficiency disease 58
transport of fatty acids into Hartnup disease 115 functions 58
the mitochondria 100 Vitamin K 58
Structure and function of cell U absorption and storage 59
constituents 1 chemistry 58
Units of radioactivity 166 daily requirement 59
Structure of DNA 40
artificial radio isotopes 166 deficiency diseases 59
circular DNA 42
production of artificial functions 59
denaturation of DNA 41
radioisotopes 166 hypervitaminosis K 59
different structural forms 41
radiation detection and sources 58
Structure of nucleic acid 40
measurement 166 Vitamins 53
primary structure 40
secondary structure 40 classification 53
tertiary structure 40 V
Sugar derivatives of biological Vitamin A 54 W
importance 16 absorption and storage 54 Water 6
chemistry 54 chemical properties 6
T daily requirement 54 classifications 7
deficiency signs 55 hard water 7
Thiamine 61 on bones 56
chemistry 61 soft water 7
on epithelial tissues 56 functions/physiologic
daily requirement 61 on eye 55 importance of water 8
deficiency 61 on general growth 55 impurities of water 7
functions 61 functions 54 methods of purification 8
sources 61 bones and teeth 55 necessity of water 6
Thyroid gland and its hormones epithelial tissue 55 properties 6
148 growth 55 purification of water 7
actions of thyroid hormones reproduction 55 sources 6
148 sources 54 Water intoxication 127
biosynthesis and secretion Vitamin C 59 causes 127
148 biosynthesis and metabolism features 127
transport of thyroid 60
hormones 148 chemistry 59
Trace elements 120 Z
daily requirement 60
absorption 121 deficiency diseases 60 Zinc 123
daily requirements 121 functions 60 absorption 123
deficiency state 121 sources 60 daily requirement 123
iron deficiency anaemia Vitamin D 56 deficiency 123
121 absorption and storage 57 physiological functions 123
iron toxicity 121 chemistry 56 sources 123
iron 120 daily requirement 56 zinc toxicity 123