Physiology Biochemistry and Biotechnology

PHYSIOLOGY, BIOCHEMISTRY
AND BIOTECHNOLOGY
"This page is Intentionally Left Blank"
Physiology,
Biochemistry
and
Biotechnology
Dr. Madan Lal Bagdi
MANGLAM PUBLISHERS & DISTRIBUTORS
DELHI - 110 053 (INDIA)
Published by
MANGLAM PUBLISHERS & DISTRIBUTORS
L-2111, Street No. 5, Shivaji Marg, Near Kali Mandir,
l.P. Nagar, Kartar Nagar, West Ghonda, Delhi -53
Phone: 22945678, Cell.: 9868572512
Physiology, Biochemistry and Biotechnology
Reserved
First Published 2007
ISBN <)78-81-89972-0 I -I
[No Part of this book may be reproduced in any form by
photocopying or by any electronic or mechanical means, including
information storage or retrieval systems, without permission in
writting from publisher of this book.]
PRINTED IN INDIA
Published by D.P. Yadav for Manglam Publishers & Distributors, Delhi
and Printed at Sachin Printers. Delhi-
Preface
The text book entitle, Physiology, Biochemistry and l!iotechnology as a
distinct subject developed in the middle ofthe 20th century as researchers
started to understand the molecular approach of the various systems of
the body. It is designed to introduce the principles of physiology and
biochemistry and their application to the students aiming for a career as
a nurse, medical assistant, physician's assistance, medical laboratory
technologists, radiologists, pharmaceutical and molecular biologist. It
encompasses all the chemical reactions carried out by the cells, organ
and systems. In clear and compelJing language this text book describes
the ideas and techniques that are creating a new domain of science and
technology. It also provides a real service to the layman interested in this
exploding technology.
Contents
1. Alimentary System and Nutrition ............................ 1-32
1. 1 Gastrointestinal System ....................................... 1
1. 1. 1 Gastric Activity ..................................... 8
1.1.2 Gastric secretions .................................. 9
1. 1. 3 Protein Digestion. . . . . . . . . . . . . . . . . .. .. . . . . .. .. .... 11
1.1.4 Control of Gastric Secretion.................... 11
1.1.5
1.1.6
1.1.7
1.1.8
1.1.9
1.1.10
1.1.11
1.1.12
1.1.13
1.l.l4
1.1.15
1.l.l6
1.1.17
1.l.l8
Histamine Receptors ............................. 14
Gastric Emptying. .............. .. . .... .. . . . . . . . .. 14
tntestinal Absorption ............................. 15
oarbohydrates .................................... 15
Proteins... ... ...... . .... ... . ....... .... ... . . .... ... 17
Fats ................................................. 18
The Structure and Catalytic Mechanisms
of Pancreatic Enzymes .......................... 19
Control of Pancreatic Secretion ................ 21
Control of Bile Secretion ........................ 22
Motility of Small Intestine ., .................. " 24
Absorption of Water and Electrolytes ......... 24
Vitamins ........................................... 25
The Large Intestine .............................. 25
Defaecation ....................................... 26
1.2 Metabolic Rate ............................................... 27
2. Respiratory System ............................................. 33-63
2.1 Respiration .................................................... 33
2.1.1 Diffusion of Gases in Respiration .............. 33
(ii)
2.1.2
2.1.3
2.1.3
2.1.4
2.15
2.1.6
2.1.7
CONTENTS
Ventilation of the Lungs ..... ., ................. 34
Pulmonary Circulation .......................... 36
Mechanics of Respiration ....................... 38
Lung Volumes .................................... 45
Gas Mixtures and Pressures .................... 46
Respiratory Dead Space and
Alveolar Ventilation ............................. 48
The Transfer of Gases Between
Alveoli and Blood ................................ 50
2.1.8 Gravity Effects on Blood
and Gas Flow to the Lungs ..................... 51
2.1.9 Carriage of 02 and CO
2
by the Blood ......... 52
2.1.10 Oxygen Dissociation Curves ................... 53
2.1.11 The Carriage of CO
2
and
the CO
2
Dissociation Curve .................... 54
2.2 Regulation of Respiration ................................... 57
2.2.1 Control System ................................... 57
2.2.2 Non-chemical Influences on Respiration ...... 58
2.2.3 Chemical Control of Respiration .............. 59
2.2.4 Ventilatory Response to Pure
Changes in CO
2
, 02 or [H+] .................... 61
2.2.5 Ventilatory Response to Combined
2
, CO] and [H+] Changes ..................... 62
2.2.6 Non-respiratory Features of
Pulmonary Circulation .......................... 62
2.2.7 Nutritional Blood Supply to the Lungs ........ 63
3. Blood and Body Fluids ......................................... 64-88
3.1 Cells of the Blood ............................................ 64
3.1 . I Erythrocytes ...................................... 65
3.1 .2 Erythrocyte Production and Destruction ...... 65
3.1.3 Haemolysis of Erythrocytes .................... 66
3.1.4 The blood pigments .............................. 66
3.1.5 Abnormal Haemoglobins ........................ 67
3.1.6 Synthesis and Destruction of
Haemoglobin .......... " ............... " ....... " 69
3.1 .7 Substances Essential to Erythropoieses ....... 69
3. 1. 8 Vitamin B72 and Folic Acid .................... 70
CONTENTS (iii)
3. 1.9 Death of erythrocytes, degradation of
haemoglobin and metabolism of
bile pigments ...................................... 72
3. 1.10 Blood Groups ............... ,..................... 72
3. 1.11 The Leucocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 75
3.2 The Lymphoreticular Organs ..... " ........................ 77
3.2.1 Lymphocytes and Immunity .................... 78
3.2.2 Two Populations of Lymphocytes:
T - and B-cells ..................................... 79
3.2.3 The Platelets ...................................... 81
3.3 Plasma ......................................................... 81
3.3.1 Salts ................................................ 82
3.3.2 Plasma Proteins .................................. 82
3.3.3 Haemostasis ....................................... 83
3.3.4 Anti-clotting Mechanisms ....................... 85
3.3.5 Lymph ............................................. 87
4. The Cardiovascular System ................................. 89-140
4.1 Circulation of the Blood ..................................... 89
4.2 The Heart ..................................................... 89
4.2.1 Outline of Function .............................. 89
4.2.2 Physiological Properties of the Heart ......... 90
4.2.3 The Cardiac Action Potential.. ................. 92
4.2.4 Conduction of the Action
Potential Over The Heart ....................... 96
4.2.5 Contraction of Heart Muscle Cells ............ 97
4.2.6 Events of the Cardiac Cycle .................... 98
4.3 The Vascular System ....................................... 103
4.3.1 Arteries ........................................... 103
4.3.2 Veins .............................................. 110
4.3.3 Arterioles and Capillaries ...................... 113
4.3.4 Peripheral Circulation as a Whole ............ 118
4.3.5 Methods of Measuring Blood Flow ........... 120
4.4 The Overall Control of the Circulation .................. 122
4.4.1 Control of Blood Pressure ..................... 123
4.4.2 Sensory Receptors in the Heart and Aorta .. 125
4.4.3 Regulation of Cardiac Output ................. 126
(iv)
4.4.4
4.4.5
4.4.6
4.4.7
4.4.8
CONTENTS
Control of Heart Rate ........... :: ............. 127
Control of Stroke Volume ..................... 127
Regulation of the Peripheral Resistance ..... 129
Coronary Circulation ......... , ................. 133
Cerebral Circulation ............................ 133
4.4.9 Effects of Posture on the Circulation ......... 135
4.5 Placental and Foetal Circulation .......................... 137
4.5.1 Uterine Circulation ............................. 137
4.5.2 Placenta ........................................... 137.
4.5.3 Foetal Circulation ............................... 137
4.5.4 Changes at Birth ................................. 139
5 The Kidney and Micturition .............. 141-161
5. 1 Gross Structure of the Kidney ............... " ............ 141
5.1.1 The Structure of the Nephron ................. 142
5.2 The Control of Urine Formation .......................... 144
5.2.1 Glomerular Filtration ........................... 144
5.2.2
5.2.3
5.2.4
5.2.5
5.2.6
5.2.7
5.2.8
5.2.9
5.2.10
5.2.11
5.2.12
5.2.13
5.2.14
The Concept of Clearance ..................... 146
The Urea Clearance ............................ 148
Control of Blood Osmolality .................. 149
Iso-osmotic Reabsorption in the Proximal
Tubule ............................................ 149
Trans-tubular Potentials ........................ 150
Reabsorption in the Distal Parts
of the Nephron .................................. 151
Hormonal Control of Sodium
Reabsorption ..................................... 154
Urea Excretion .................................. 154
Reabsorption of Sugars and
Amino Acids ..................................... 155
Tubular Secretion ............................... 157
Potassium Regulation ........................... 157
Renal Regulation of Hydrogen
Ion Concentration ............................... 158
Mechanism of Micturition ..................... 160
6 Reproductive Physiology .................................... 162-189
6.1 Sex Differentiation. . . . . .. . .. .. . .. .. . .. . .. .. .. .. .. .. .. .. .. ... 162
CONTENTS (v)
6.2 Female Reproductive System .............................. 164
6.2.1 Ovarian and Uterine Cycles ................... 166
6.2.2 Ovarian Cycle ................................... 166
6.2.3. Uterine Cycle .................................... 169
6.).4 Ovarian Hormones .............................. 169
6.2.5. Anterior Pituitary and Hypothalamus ........ 171
6.2.6 Control of the Ovarian Cycle .................. 171
6.2.7. Oral Contraceptives ............................. 174
6.2.8 Pregnancy ........................................ 174
6.2.9 Hormones in Pregnancy ........................ 175
6.2.10 Pregnancy Diagnosis ............................ 177
6.2.11. Parturition ........................................ 179
6.2.12 Lactation .......................................... 180
6.3 Male Reproductive System ................................ 183
6.3.1 Spermatogenesis ................................. 183
6.3.2 Control of Testicular Function ................ 184
6.3.3 Fertilisation ...................................... 187
6.4 Puberty ....................................................... 188
190
7. Endocrinology and Metabolism ....... .................... 190-246
7.1 Hormones in General ....................................... 190
7.1.1 Factors which Determine the Blood,
Concentration of Hormones ................... 191
7.1.2 Blood Transport ................................. 192
7.1.3 Hormone Action on Target Cells ............. 193
7.1.4 The Way in which the Primary
Event Alters Cellular Functions ............... 195
7.1.5 Levels of Complexity of
. "
Hormone As;tlon ................................. 197
7.1. 6 Endocrine Investigations ....................... 198
7.2 The Hypothalamus and Pituitary .......................... 199
7.3 Growth Hormone ........................................... 202
7.4 Prolactin and the
Pituitary Gonadotrophins ................................... 206
7.5 Hormones of the Posterior
Pituitary Gland .............................................. 206
\
(vi) CONTENTS
7.6 The Thyroid Gland ......................................... 209
7.7 The Control of Growth ..................................... 213
7.8 Adrenal Gland ............................................... 2 I 6
7.9 Adrenal Cortex .............................................. 217
7.9. 1 Glucocorticoids .................................. 219
7.9.2 Mineralocorticoids .............................. 223
7.10 Organic Metabolism and
the Glucose Regulating Hormones ........................ 227
7.10.1 Outline of Absorptive and
Post -absorptive Events ......................... 229
7.10.2 The Absorptive State ........................... 230
7.10.3 Post-absorptive or Fasting State .............. 231
7.10.4 Insulin ............................................. 234
7.11 Bone and the Calcium
Regulating Hormones ....................................... 239
7.11.1 Calcium ........................................... 239
7.11.2 Calcium Regulation ............................. 242
7.11.3 Other Modulators of Ca ........................ 246
8. Introduction to Metabolism and
Energy Metabolism ............................. 247-257
8.1 Characterization of Metabolism ........................... 247
8.2 Energy Cycles in Animate Nature ........................ 249
8.3 Energetics of Biochemical Reactions ..................... 252
8.4 Hight-Enegry and Low-Energy Phosphates: General
Considerations ............................................... 254
8.5 Energy Transfer in
Biochemical Processes ..................................... 256
9. Carbohydrate Metabolism .............................. 258-272
9.1 Carbohydrage Catabolism in Tissues ..................... 258
9.1. 1 Pentose Phosphate Cycle ....................... 259
9. 1.2 Interrelation of the Pentose
Phosphate Cycle and Glycolysis .............. 262
9.1.3 The Biological Function of
the Pentose Phosphate Cycle .................. 263
9.2 Biosynthesis of Carbohydrates in Tissues ............... 265
9.2.1 Gluconeogenesis ................................. 265
CONTENTS (vii)
9.2.2 Biosynthesis of Glycogen
(Glycogenogenesis) ............................. 268
9.2.3 Biosynthesis of Other
Oligosaccharides and Polysaccharides ....... 269
9.3 Carbohydrate Metabolism
Control in the Organism ................................... 270
10. Lipid Metabolism ............................................ 273-293
10. I Degradation of Lipids in Tissues ............. " ........... 273
10.1.1 Intracellular Hydrolysis of Lipids ............. 273
10.1.2 Oxidation of Glycerol .......................... 274
10.1.3 Oxidation of Fatty Acids ....................... 274
10.2 Biosynthesis of Lipids in Tissues .......... '" ............ 279
10.2.1 Biosynthesis of Fatty Acids .................... 279
10.2.2 Biosynthesis of Triglycerides .................. 282
10.2.3 Phospholipid Biosynthesis ...................... 283
10.2.4 Biosynthesis of Ketone Bodies ................ 285
10.2.5 Biosynthesis of Cholesterol .................... 287
10.3 Regulation of Lipid
Metabolism in the Organism .............................. 288
10.4 Pathology of Lipid Metabolism ........................... 290
10.5 Applications of Lipids and Their
Components in Pharmacotherapy .... '" .................. 292
,
f
1
Alimentary System
and Nutrition
1.1 GASTROINTESTINAL SYSTEM
The tract is, in effect, a long muscular tube of variable diameter
lined with mucous membrane, running from the mouth to the anus.
Into ~ h lumen of this tube various organs and glands add their
secretions and these, together with those of the mucous membrane
itself, are responsible for the digestive process whereby the complex
foodstuffs we ingest are broken down into much simpler molecules.
This is necessary as only simple molecules can be absorbed from the
intestinal lumen and carried in the blood to supply the nutritional needs
of cells.
Structure. Although the structure of the alimentary canal varies
somewhat along its It!ngth, it does have a basic form which can be
recognised in most parts. From within outwards, the gut wall consists
of four layers:
1. Mucosa.
2. Submuoosa.
3. Muscular layer.
4. Serosal or adventitous coat.
The mucosal lining has an epithelial layer with supporting
connective tissue beneath it in which the blood capillaries run, and
this in turn is in contact with a thin layer of smooth muscle called
the muscularis mucosae. This muscle is innervated by sympathetic
nerve fibres and contraction of the muscle results in folding of the
mucosa.
The submucosal layer is composed of more dense connective tissue
in which run larger blood vessels and a plexus of nerves (Meissner's
2 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
plexus) to supply the mucosal glands. The main muscle layer consists
of an outer longitudinal layer and an inner circular one of smooth
muscle with a nerve network between them (Auerbach's plexus) .

.. __ . ____ Parotid
gland
___ Pharynx
1+-_______ Oesophagus
Gall
bladder
Liver
Stomach

__ Splenic
flexure
Duodenum --t----:;>..co<-+ WQ:::::;;?"-:;)'"
Hepatic
flexure
Ascending
colon __ -+
Jejunum
Terminal
ileum
Caecum /
Appendix
Figure 1.1 Gross anatomy of alimentary tract.
-H.<;;j--- Transverse
colon
1---- Descending
colon
'r .... '-+-___ Taenia
colon
The whole tube in most parts covered with several layers of
squamous epithelium and connective tissue.
Outline of digestion. The digestion of food to smaller molecules
occurs at different points in the intestinal tract, although mainly in
the small intestine. Enzymes, the substances involved in the breakdown
of food molecules, are to be found in the secretions of the salivary
ALIMENTARY SYSTEM AND NUTRITION 3
glands of the mouth, in gastric juice, bile from the liver, in pancreatic
juice and in secretions from the mucous membrane of the small
intestine.
Layers
Mucosal_
Longitudinal
muscle
Circular
muscle
Muscularis
mucosae

layer
Muscosal
blood vessels
Submucosal
nerve plexus
Myenteric
nerve plexus
Figure 1.2 Cross-sectton showing basic layers of the gastrointestinal tract.
The start of carbohydrate digestion occurs in the mouth because
of the presence of salivary gland secretion of the enzyme ptyalin.
The main carbohydrate in our diet is starch, a polymer of glucose.
either in the unbranched chains of amylose or in the branched form,
amylopectin. We also ingest the disaccharides sucrose and lactose.
Only very small amounts of monosaccharides are normally ingested.
The digestive enzymes found in the human intestinal tract can
break the 1,4-a type of linkage found in the straight chains and also
the 1,6-a linkage found at the branches of amylopectin. We do not,
however, secrete enzymes capable of breaking the linkages between
the glucose subunits of cellulose (1, 4-f3 linkage) and so we are unable
to digest this and it passes through the gut adding bulk to the faeces.
Saliva. The active enzyme in saliva is ptyalin, an amylase, which
hydrolyses l,4-a linkages. The optimal pH for amylase activity is 6-
7 and so its action is to some extent limited because food does not
stay long in the mouth and, once it has passed into the stomach, the
food bolus is subjected to a high acidity which inhibits amylase activity.
4
PHYSIOLOGY. BIOCHEMISTRY AND BIOTECHNOLOGY
The enzyme activity will of course continue in the middle of a bolus
until penetration by the gastric acid.
H OH
lal
(bl
Figure 1.3. (a) Maltose .. u 1.4 linkage of glucose subumts. (b) Celiobiose-j3 J.4 linkage
o. glucose subunits. .
Saliva is also important in that it moistens the food, enhancing the
p .,..:ess of chewing and making food more easily swallowed; in addition
it has a bacteriostatic action. Saliva comes partly from the many mucous
glands in the mucosa of the mouth but mainly from discrete glands
which lie outside the mouth and pass their secretions into the mouth
by ducts. These acinar glands are the parotids. the sub-mandibular and
sub-lingual glands and they secrete both mucous and serous fluid. The
secretion is continuously fonned at a slow rate. even when the mouth
is empty, at an average of 0 2 ml/minute in man; this may increase 20
times when eating. As with gastric secretion, the increase in flow of
saliva is partly caused by a conditioned reflex. The thought of food
acts as a stimulus to secretion. but the presence of food in the mouth
also acts as a stimulus. Impulses from receptors in the oral mucosa
pass in the lingual and nerves to the medulla from
whence efferent impulses pass to the salivary glands. The presence of
anything in the mouth. for example a piece of gnt, acts as a stimulus
in this unconditioned reflex.
ALIMENTARY SYSTEM AND NUTRrnON 5
The salivary glands receive both sympathetic and parasympathetic
nerve supplies. Stimulation of the latter causes an increased blood
flow through the glands and a copious watery secretion is formed.
This does not mean that the secretion from the gland is merely a
filtrate of blood. The increased oxygen consumption of the gland which
accompanies this increased activity shows that saliva is being formed
by a metabolically active process in the gland. When the sympathetic
supply is stimulated these glands produ.ce a viscid saliva with a high
concentration of mucus.
Chewing. When food is taken into the mouth it is broken into
smaller particles which can be more easily swallowed by chewing.
This is obviously a voluntary process but also occurs as an involuntary
reflex. The contact of food on the palate and tongue causing a reflex
opening of the mouth, but this results in an anti-gravity reflex closing
it, thus setting up a rhythmic movement. Chewing continues until
ingested food is soft, small enough and sufficiently well mixed with
saliva to be swallowed; but this is greatly influenced by habit and
external forces such as conversation. The amount of chewing probably
does not influence the digestion of food very much.
Swallowing. Once food has been chewed and mixed thoroughly
with saliva, it is formed into a small ball or bolus which can be
swallowed. Swallowing is a complex series of contraction and
inhibition of many voluntary and involuntary muscles. The tongue
initiates the process by pushing the bolus into the back of tbe mouth,
resulting in the stimulation of receptors there. You can convince
yourself that swallowing is not a purely voluntary process but requires
stimulation of receptors by trying to swallow your saliva three times
in succession; after swallowing twice, no saliva remains in the mouth
and you cannot initiate a third swallowing movement. Impulses from
these swallowing receptors in the mouth pass to a 'swallowing centre'
in the medulla which coordinates the train of events. Once swallowing
has been initiated it cannot be voluntarily stopped.
As the tongue pushes the bolus backwards in the midline the soft
palate is raised to shut off the naso-pharynx, respiration is inhibited,
the larynx raised and the glottis closed to prevent food entering the
trachea.
This movement of the bolus backwards and downwards folds the
epiglottis back over the glottis although this is not sufficient by itself
to protect the trachea from food; closure of the glottis is essential.
Occasionally a small particle of food or fluid may reach the respiratory
tract before closure of the glottis and cause coughing by stimulation
of respiratory tract receptors.
Oesophagus. The oesophagus is the tube which carries food from
the pharynx to the stomach. It has skeletal muscle around its upper
third and smooth muscle in its lower two-thirds. Before and after
,
swallowing both the upper and lower ends of the oesophagus are
closed, but swallowing results in relaxation of both sphincters. Once
food has passed through the upper sphincter it closes, the glottis opens
and respiration starts again.
Perisfollic
wove
Figure 1.4. Peristalsis in oesophagus. Muscle fibres below food bolus relax, those
behind contract.
Once a bolus has entered the oesophagus a wave of contraction
passes along the smooth muscle of the wall from the oral to the
stomach end. This wave of contraction is immediately preceded by a
wave of relaxation and results in the food bolus being pushed onward
through the relaxed gastro-oesophageal or cardiac sphincter into the
stomach. This process is called peristalsis.
Peristalsis is a feature of many different parts of the gastrointestinal
tract and its coordination is the result of local nerve-plexus activity in
the gut wall. In the oesophagus, however, the nerve network in the
wall is coordinated by the swallowing centre and occurs only if the
vagal efferent nerves from the centre to the oesophagus are intact.
Normally a peristaltic wave passes along the oesophagus in about 10
seconds. (You can time this for yourself by listening for liquid reaching
your empty stomach.) The passage of a bolus is normally assisted by
gravity, but it is possible to swallow while standing on one's head
when movement of food along the oesophagus depends entirely on
peristalsis. If a bolus sticks in the oesophagus this distending stimulus
results in a secondary wave of peristalsis, and this again requires an
intact extrinsic nerve supply.
High - pressure

+ ve pressure
+ ve pressure
Figure 1.5. Part of the oesophagus lies below the diaphragm am so is exposed to the
positive intra-abdominal pressure which helps to squeeze it am prevent reflux.
The lower end of the oesophagus normally remains closed and so
prevents reflux of gastric contents. Although the muscle fibres of the
lower end of the oesophagus do not form an anatomical sphincter, the
lowest 4 cm is normally in a state of contraction and acts as a
physiological sphincter. This region has been called the high-pressure
zone (HPZ). Intra-abdominal pressure is always higher than the normally
negative intrathoracic pressure, and at times becomes very much higher,
so it is surprising that stomach contents do not get pushed back into
the oesophagus. This is prevented not only by this high-pressure-zone
sphincter but also by the fact that part of the oesophagus lies below
the diaphragm and any increase in intra-abdominal pressure which
would squeeze the stomach also 'squeezes this part of the oesophagus
and keeps it closed. The oesophagus runs diagonally through the
diaphraj!m to enter the abdomen; it has been suggested that a part of
8 PHYSlOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
the muscle of the diaphragm may pinch the oesophagus shut, particularly
during a deep inspiration. The oesophagus enters the stomach at an
acute angle; perhaps a fold of mucosa acts as a flap-valve to prevent
ret1ux of contents into the oesophagus.
1.1.1 Gastric Activity
One function of the stomach is to act as a hopper, collecting
swallowed food so that it can be held there until it is passed on to
the duodenum at a controlled rate for digestion and absorption. Gastric
muscle shows 'receptive relaxation'; this means that as the quantity
of food in its lumen increases during a meal the muscle in the wall
relaxes to enlarge the cavity, so preventing an increase in intraluminal
pressure.
Figure 1.6. Gastric peristalSIS. As the move of contraction o e ~ not close the lumen
of stomach completely, some contents escape back into the fundus and cause mixing.
Another function of the stomach is to help mix the swallowed
food with gastric secretions forming a semi-liquid substance called
chyme. This mixing process is accomplished by rhythmic contractions
of the gastric smooth muscle. The longitudinal layer of smooth muscle
has a basic electrical rhythm of 3 beats/minute; the propagation of
this electrical activity produces a wave of contraction spreading across
the stomach to the pyloric area. This peristaltic wave pushes the more
liquid contents through the pylorus into the duodenum, but as the
pyloric muscle contracts its lumen closes and this forces the antral
contents back into the body of the stomach resulting in further mixing
of food and gastric juices.
1.1.2 Gastric secretions
Gastric secretions consist of mucus, pepsinogens, hydrochloric acid
and intrinsic factor, all of which arise from cells in the gastric mucosa.
Their total volume is about 3 litres/day. The mucous membrane is
Surface
Neck
mucous
cell
Parietal
cell
Lumen __
Chief
cell " ___ -...
epithelial
cell
Figure 1.7 Diagram of cells to be found in the body of the stomach.
not the same throughout the stomach, the epithelium of the body of
the stomach being arranged in gastric pits and glands. The different
components of gastric secretion arise from different types of cells in
these pits and glands. Seventy-five per cent of the gastric mucosa is
of this type. The antral area has larger pits and branching coiled glands
with no parietal or chief cells but instead rounded cells lying between
the mucus- secreting cells. These rounded cells secrete gastrin.
The main enzyme in gastric juice is secreted as inactive
pepsinogen from serous or chief cells and in the presence of acid or
pepsin the inactive pepsinogen becomes active pepsin. Hydrochloric
acid is secreted by the parietal or oxyntic cells which have small
channels or canaliculi in them into which Hel is secreted, carried to
the lumen of the gastric glands and so to the stomach. The process
of acid production by these cells is not fully understood but as the
hydrogen ion concentration in gastric juice can be as high as 150
mmol compared with 000004 mmol in blood, hydrogen ions have to
be secreted by the parietal cells against a very large gradient. Separate
pumps are thought to deal with the hydrogen and chloride ions.
Blood Parietal
Lumen
vessel
cell
of
Cl- Cl-
stomach
Cl-
COz
CO
2
+ HzO
1 eo. boo;<
anhydrase
~ C 3
!
HCOi+ H+ H+
Figure 1.8 Secretion of He) by parietal cell.
The mucous neck cells of the gastric glands secrete both mucus
and intrinsic facto. Mucus, by adhering to the surface of the mucosa,
plays a role in the protection. of gastric mucosa from digestion by
gastric juice. It has lubricating properties which allow the chyme to
move over the mucosal surface and the proteins and bicarbonate in it
act as local buffers protecting the mucosa from the hydrochloric acid
without altering the pH of the bulk of the contents.
1.1.3 Protein Digestion
The pepsins secreted by the chief cells of the stomach start the
digestion of protein. Enzymes break down proteins by attacking the
bonds between the amino-acid units of the molecule with certain
enzymes attacking specific bonds. There are several different active
pepsins-pepsins I, II and III-derived frum the inactive pepsinogens
secreted by the gastric cells. These hydrolyse proteins into polypeptides
of various sizes by attacking the bonds within the protein molecule
between aromatic and other amino acids. The optimal pH for this
reaction is between I and 3, so their activity is stopped on entry of
stomach contents into the alkaline milieu of the duodenum. Further
digestion of the polypeptides depends on enzymes in the small intestine
w h i c h
are active in an alkaline environment.
1.1.4 Control of Gastric Secretion
The experimental techniques of 'sham feeding' and 'gastric
pouches' have proved very useful in the investigation of the control
of gastric secretion. If the eosophagus of an animal is divided and
brought to the body surface, then that animal may chew and swallow
food but it comes out from the exteriorised oesophagus and does not
reach the sto . Gastric pouches are basically of two types: the
Figure 1.9 Oesophagus is exteriorised to allow 'sham' feeding. When food is chewed
and swallowed it does nOI reach the stomach. Gastric juice can be collected easily and
is uncontaminated by food. This preparation also allows food to be placed directly in
the stomach without chewing having taken place.
Pavlov pouch and the Heidenhain one. In the Pavlov pouch part of
the stomach is made into a pouch so that its lumen is not in continuity
12 PHYSIOLOGY. BIOCHEMISTRY AND BIOTECHNOLOGY
with the main lumen but the blood and nerve supply to both is the
same. In the Heidenhain one the nerve supply to the pouch has been
cut but it still has an intact blood supply. Pouch secretions can thus
be collected easily, uncontaminated by food and the factors controlling
these secretions can be studied.
, .
'.
CNS
B
ostrin _+ .......... Secretion
'.
+
Distension

Polypeptides
SecretoCJQQues
VOQus
Figure 1.16 Interrelationship of factors causing gastric secretion. Increased vagal activity
causes'release not only of Hel but also of gastrin which in turn stimulates He!.
Distension stimulates HeI secretion directly and through gastrin released from G cells.
Gastrin is also released by polypeptides and secretogogues in the gastric lumen.
There is a small, continuous gastric secretion, which increases
markedly during a meal; in adult man this amounts to 2-3 litres/day.
The first phase of increased secretion has been called the cephalic
phase because it is mediated from the brain via the vagus nerve. This
can be shown by sham' feeding when secretions appear in the
innervated Pavlov pouch but not in the denervated Heidenhain one.
This reflex is partly a conditioned one as the thought of food is
sufficient stimulus to produce an increase in gastric secretion, but the
smell and taste of food also act as stimuli.
The presence of food in the stomach also causes an increase in
secretion, the gastric phase; this secretion occurs in both tbe Pavlov
and Heidenhain pouches. Since an increased secretion is found in the
denervated pouch, the stimulus to secretion must be blood borne. The
presence of food in the stomach causes the release of a local hormone,
gastrin, from the antral mucosa into the blood by which it is carried
to the fundal mucosa to stimulate acid secretion. The main stimulus
for gastrin release is the presence of polypeptides in contact with antral
mucosa, but distension of the stomach and the presence of substances
like alcohol and other secretogogues also act as stimuli. These two
controlling mechanisms for gastric secretion are interrelated and indeed
synergistic. Vagus
Acid
Hormonal
inhibition
Acid
Gastrin
Duodenum
Inhibition of
vagal activlty
Figure 1.11 Inhibition of gastric secretion. Acid inhibits gastrin release; its presence
in the duodenum causes the reflex inhibition ~ He! release from the parietal cells.
There is evidence that the presence of food in the duodenum may
also cause an increase in gastric secretion; gastrin has been isolated
from the proximal duodenal mucosa but there is no agreement on the
importance of this intestinal phase of gastric secretion.
We have seen how gastric secretion is stimulated, but is there
any mechanism for turning it off? It has been observed that when the
pH in the antrum falls below 3 inhibition of acid secretion occurs.
This results from the inhibition of gastrin release which in turn leads
to less stimulation of the parietal cells. An additional factor, however,
is that the passage of acid, fat and to a lesser extent hyperosmolal
solutions into the duodenum also inhibit gastric secretion, and although
the vagal nerve plays a role in this mechanism the evidence points to
the main inhibition being due to the release of an inhibitory hormone.
The hormone responsible is not known and although secretin and
cholecystokinin-pancreozymin, hormones involved in the control of
pancreatic and bile secretion, have inhibitory effects on gastric
secretion there is additional evidence that a separate inhibitory hormone
may exist.
1.1.5 Histamine Receptors
We have seen that both acetylcholine, released by vagal nerves,
and gastrin cause an increase in gastric acid secretion. For many years
it has also been known that histamine will cause an increase in acid
secretion by the stomach; the current view is that it has this action
by virtue of histamine receptors on the parietal cells of the stomach.
Experiments with drugs which block the action of histamine suggest
that there are two classes of these receptors: H2 in most areas of the
body and on the parietal cells. Drugs are currently available which
block these receptors-and thereby heal peptic ulcers-but the
physiological role for histamine in normal gastric secretion is not yet
clear.
1.1.6 Gastric Emptying
The rate at which the stomach empties depends, to some extent,
on the volume of its contents, which suggests that stretching of the
gastric wall stimulates stretch receptors which then increase motility.
The main control over gastric emptying, however, is exerted by the
duodenum. The presence of acid, fat or hyperosmolal solutions in
contact with duodenal mucosa results in decreased activity of gastric
muscle and, in consequence, a decrease in theamount of gastric
contents entering the duodenum. Passage of water into the lumen of
the duodenum through the intestinal wall soon renders the contents
iso-osmolal, bicarbonate in the pancreatic juice neutralises the acid
and the inflow of bile and lipase results in the digestion and absorption
of the fat. This mechanism has the logical effect of controlling the
rate of gastric emptying so that the duodenum is not at any time
overloaded with chyme.
How is this inhibitory int1uence of the duodenum on gastric muscle
activity mediated? It seems to be due to both nervous and humoral
mechanisms. Vagal denervation of the stomach markedly reduces the
inhibitory effect of acid or fat in the duodenum on gastric motility;
this has been called the enterogastric reflex. The fact that the inhibitory
effect, although reduced, is still seen after denervation indicates that
the effect must also be produced by humoral means; the name
enterogastrone has been suggested for this agent. There is still doubt
about the n ~ t u r of this hormone: both secretin and cholecystokinin-
pancreozymin have been shown to decrease gastric motility as well
as decreasing secretion; polypeptides such as motilin and gastro-
inhibitory peptide (GIP) , released from the mucosa of the small
intestine, also alter gastric emptying rates, so at present it is probably
best to use the term enterogastrone to describe the active agents in
this humoral feedback mechanism.
1.1. 7 Intestinal Absorption
The main site of digestion and absorption is in the small intestine.
This part of the alimentary tract consists of about 30 cm of duodenum
and 240 cm of jejunum and ileum. The small intestine is adapted for
absorption in that the surface area of its mucous membrane is increased
by infolding and by a large number of finger-like projections, the villi,
which project into the lumen. These villi are up to 1 mm in length and
covered with epithelium, with capillaries, lymph vessels and smooth
muscle fibres in their core. The epithelial cells have a brush border of
hundreds of microvilli which increase their surface area and absorptive
surface even more. Hydrolytic enzymes are located in this brush border
and it is here that the [mal stages of carbohydrate and protein digestion
occur. All epithelial cells are formed in the depressions between villi,
the crypts of Lieberkiihn, and migrate up the villus to be extruded
from its tip over a period of 2 days. These extruded cells add to the
enzymic content of the lumenal contents.
The transfer of substances across the intestinal epithelium from
the lumen to blood and lymph occurs in different ways. Molecules
can diffuse passively down concentration or electrochemical gradients
but, for most, an active transport mechanism is required to move
them against a gradient. This movement of substances also depends
on their physical characteristics; lipid, soluble substances, for example
ethanol, move readily through the lining epithelium of all parts of
the gut. Watersoluble molecules, unless they have a molecular weight
less than 100, do not pass through readily and require carrier
mechanisms.
1.1.8 Carbohydrates
The second stage of carbohydrate digestion occurs in the duodenum.
Smooth
muscle __
Micra-
vi \Ii --I--"'I!1'111.
Ly:nphotic
vessel
.'IDIt=-1l>
Crypt of
lieberkUhn
Artery
Terminal
web
To Iymphatlcs

junction
__ .. _t-Na+
transport
Microvillus
Figure 1.12 Intestinal villus supplied with a large lymph vessel in addition to a capillary
network. Note the smooth muscle fibres from the muscularis mucosa layer which cause
shortening vf villi and so aid movement within the lacteals. Inserts show details of
cells and a single microvillus; each microvillus has a central cone of microfilarnents
which intermingle with the mesh work of microtubuies and ftIaments called the 'terminal
wel>'. In some species actm has been idenllfied in the microfilaments and the addition
of ATP causes them to contract.
Here the bicarbonate in the pancreatic secretion ensures an optimal pH
for amylase activity in the subsequent breakc'own of carbohydrate to
dextrins and maltose. The maltose is hydrolysed to monosaccharides
by maltase and the 1-6 linkages are hydrolys':d by oligo 1-6 glucosidase.
The final breakdown occurs mainly in thf. microvilli of the epithelial
cells-which contain the enzymes concemed, i.e. maltase, sucrase,
lactase, etc. -and not to any great extent within the lumen of the
duodenum.
AUMENTARY SYSTEM AND NUTRITION 17
Carbohydrates ingested
__________ I
Lactase Starch Glucose Sucrose
Maltolriose Maltose
I Maltase I
Galactose Glucose Glucose Glucose Glucose Glucose Fructose
[ ENZYMES
Figure 1.13 Digestion of carbohydrates by various enzymes. The amylase activity takes
place in the lumen of the gut: that of the uther enzymes in the brush border.
Absorption. An active carrier mechanism is responsible for the
absorption of the monosaccharide end-products of carbohydrate
absorption. There is competition between different sugars and they
show different absorp tion rates, glucose and galactose having the
highest rate. A fll.:tor which affects the rate of absorption is the
concentrati0n of sodium ions in the lumen; the fact that ollabain inhibits
absorption of S\l2,:!I's points to the probable importance of sodium
pumps in the process.
1.1.9 irotebns
Secretions from the pancreas entering the duodenum contain trypsin
and chymotrypsin, enzymes which break down protein into dipeptides
and amino acids; these enzymes, unlike gastric pepsin, are active in
alkaline conditions. AllY remaining polypeptides are broken down to
their final amino acids by carboxy peptidase and amino peptidase from
the intestinal mucosa. Some of this final digestion occurs within the
lumen of the gut but much of it is carried out in the brush border of
the mucous membrane epithelial cells.
Absorption. Only amino acids are absorbed into the bloodstream
from the lumen of the gut; once again this is by an active process.
There appears to be three- carrier mechanisms involved in carrying
the amino a,cids according to whether their side chains are neutral,
basic or acidic. These carriers also show a preference for laevo amino
acids. As with the carriage of sugars, the carriage of amino acids is .
linked with the active transport of sodium.
1.1.10 Fats
The main fat in our diet is the ester of long-chain fatty acids
and glycerol; this is digested and absorbed in the small intestine. Under
normal conditions almost all the fat we ingest is absorbed and only
in abnormal conditions do we find fat in the faeces .
. Fat leaves the stomach in large droplets within the aqueous
solution of chyme. If it remained in this form the water-soluble lipase,
which digests it, would have great difficulty in getting into contact
with most of it; however, bile salts emulsify these large droplets,
breaking them down into much smaller ones, so enabling lipase to
come into contact over a much larger area. Agitation within the
duodenum helps break up the large droplets, the lipid part of the bile
salt molecule dissolves in the fat and the electric charge on the polar
part of its molecule faces outward towards the aqueous phase of the
mixture, preventing these droplets from coalescing. The action of
pancreatic lipase on the triglycerides in these small droplets results
in the formation of di- and monoglycerides, free fatty acids and a
little glycerol. Free fatty acids and glycerol help the bile salts in the
formation of even smaller droplets called micelles. These are small
water-soluble droplets of up to 10 nm diameter. These micelles come
into contact with the intestinal mucosa enabling the lipid-soluble free
fatty acids and mono glycerides to pass through the epithelial
membranes into the interior of the epithelial cells. Within these cells
triglycerides are synthesised again from fatty acids and glycerol by
the activity of the endoplasmic reticulum. These triglycerides form
chylomicrons which are small droplets with over 90% triglycerides
with a surface film of lecithin, cholesterol and protein. They are
released by the epithelial cells into the interstitial spaces and from
there they pass not into the blood capillaries but into the lymphatic
channels called lacteals.
Short-chain fatty acids are directly absorbed into blood capillaries;
the bile salts which have been left in the lumen of the intestine during
this process pass to the terminal ileum where they are actively absorbed
and returned to the liver in the blood.
ALIMENTARY SYSTEM AND NUTRITION
la)
Ester
linkage
"Fatty+
acid
Bile
(b) salts
Lipase
Glycende

Monoglycerides Free
fatty acid

Fatty
acid

Free
fatty acid
Monoglycendes
19
Figure 1.14 (a) Pancreatic lipases show specificity for the I-ester linkages on the
rriglycerides. The end products are: (i) free fatty acids; (ii) 2-monoglvcerides; and
(iii) a small amount of glycerol. (b) Formation of a micelle. The water-insoluble fatty
acids and mllnoglyceTldes are solubilised in the hydrophobic centre of the micelle.
1.1.11 The Structure and Catalytic Mechanisms
of Pancreatic Enzymes
The enzymes secreted by the pancreas need (a) to be activated
from precursors, (b) to recognise the peptide bond which is to be
attacked, (c) attack this bond and (d) be inactivated when the process
is over. Recent evidence using the techniques of molecular biology
has given much new information on this process; this will now be
outlined.
+ Bile salts
o Free folly acids
Glycerides
Ta ileum
++
Micelles
Protein
@) @ +- Chylomicron'3
Lymphatic
f<'igure 1.15 Intracellular synthesis of triglycerides in epithelial cells of the. villus.
There are three main proteolytic enzymes secreted by the
pancreas: chymotrypsin, trypsin and elastase; these split peptide bonds
next to auuno aCIQS with aromatic, basic and short side chains,
respectively. The system works as follows: the substrate is recognised
by a pocket which lies next to the active site in the enzyme; this
pocket is so tailored that it attracts the side chain of the amino acid
preceding the peptide bond to be split. In the inactive precursors of
the enzymes this side pocket is closed; trypsin activates these
precursors by opening this pocket. The pancreatic trypsin inhibitor
mimics a substrate by presenting to the enzyme a side chain which
the pocket 'recognises', but which has the peptide bond following
that side chain shielded from water so that the enzyme cannot split
it. The inhibitor thus remains firmly attached to the enzyme in the
position fleetingly occupied by the normal substrate during catalysis.
ALIMENTARY SYSTEM AND NUfRITION 21
This mt:chanism of activation, recognition, catalysis and inhibition
has been described in some detail because it appears to be exploitt:d
over and over again for different functions; examples are: the c1ottinr,
of blood; the dissolution of blood clots; the disposal of complexes of
antigen and antibody by complement; and in some functions of kinins.
1.1.12 Control of Pancreatic Secretion
The exocrine secretions of the pancreas are of two types: a watery
solution with a high bicarbonate content which is secreted by the cells
lining the ducts of the glands; and a secretion rich in enzymes which
comes from the acinar cells at the base of the glands.
Recognition
pocket closed
RecOgnition
! Trypsin
pocket open I f o ; ~ ~
Recognition
pocket occupied
by side chain
of protein
~ + protein
Protected bond
Inactive enzyme
I
Active enzyme
1
Enzyme blocked
by molecule
Figure 1.16 Diagram showing activation. n:cognition. catalysis and inhlbll'<ln <It
proteolytic enzymes.
22 PHYSIOLOGY. BfOCHEMISTRY AND BIOTECHNOLOGY
As with most glands the pancreas has a resting secretion level
which increases when food is eaten. The initial increase is due to
increased vagal activity as in the cephalic phase of gastric secretion;
the presence of food in the stomach causes a further increase in secretion
due to the release of gastrin which stimulates the release of a
bicarbonate-rich secretion. The main secretion from the gland occurs,
however. when gastric contents enter the duodenum. TIle characteristics
of the secretion depends on the type of chyme entering the duodenum;
if the chyme is acid then the pancreatic secretion is rich in bicarbonate
which neutralises the acid; this response is mediated by the release of
the hormone secretin from the duodenal mucosa. (Incidentally secretin
was the first gastro-intestinal hormone to be discovered.) The presence
of amino acids and fatty acids in the duodenal contents results in a
pancreatic juice rich in enzymes being released, the mediator in this
case is another duodenal hormone cholecystokinin-pancreozymin (CCK-
PZ), which as its name implies was described originally as two separate
hormones.
1.1.13 Control of Bile Secretion
Bile, which is required for the digestion and the absorption of
fat, is secreted by liver cells into small ducts which eventually
converge into the common bile duct by which bile is conveyed to the
duodenum. As the bile duct leaves the liver it is joined by another
duct coming from the gall-bladder, a small sac lying under cover of
th: liver. Just before it enters the duodenum the bile duct is usually
joined by another duct coming from the pancreas, and the resultant
('1let carries both bile and pancreatic secretion into the gut. At a point
of entry the duct is surrounded by smooth muscle, the sphincter of
Oddi.
Bile is secreted continuously by liver cells but does not enter the
duodenum because the sphincter of Oddi is normally closed. This
results in bile being diverted into the gall-bladder where sodium is
actively absorbed from bile into blood, with water following by
osmosis. The bile is therefore concentrated in the gall bladder. Bile
consists of (a) bile salts, (b) cholesterol, (c) lecithin-a phospholipid-
and (d) bile pigment. The bile pigment, which gives bile its yellow
colour, is bilirubin, a breakdown product of haemoglobin. This is
changed in the intestine to a brown pigment which gives faeces their
normal colour. If bile pigment is prevented from reaching the intestine
faeces become greyish-white.
The secretion of bile by the liver is a continuous process; up to
1000 mllday being formed. This rate of secretion is increased by
substances called cholertics, one such substance being the bile salts
themselves. After bile salts have assisted in the absorption of fat they
are absorbed in the ternlinal ileum and carried in the venous blood
from the intestine which ends in the portal vein. This vein enters the
Gall bladder
Na+ and H20
to bloodstream
Sphincter
ofOddi __ ./
__ ___ Duodenum
Figure 1.17 Storage and concentration of bile in the gall bladder and the route to the
duodenum.
liver and breaks up into a second type of capillary bed in the liver
before returning from the liver to the right atrium in the hepatic vein.
This portal circulation ensures that substances absorbed from the
intestine are carried first to the liver. The bile salts are thus returned
to the liver where they act as cholertics and are released again into
the intestine in bile. This has been called the entero-hepatic circulation.
As the total amount of bile salts in the body is about 3 g, and up to
8 g may be required for the digestion and absorption of a fatty meal,
it is very necessary that this recirculation exists.
Although bile is secreted continuously, in man it is stored in the
gall-bladder until required for the digestion of a meal. Contraction of
a gall-bladder and relaxation of the sphincter of Oddi results in bile
gaining entry to the duodenum. The mechanism controlling this
discharge of bile is twofold, first the increased activity of the vagus
which accompanies eating causes contraction of the gall-bladder and
later the cholecystokinin-pancreozymin released from the duodenum
also has this effect.
1.1.14 Motility of Small Intestine
Various types of movement of the small intestine have been
described; their purpose is to mix the contents with digestive
secretions, allowing the digestive end-products to come into contact
with the mucosa so they may be absorbed, and to move the residue
onwards into the large intestine. The main mixing movement is
segmentation in which rings of intestinal wall contract, dividing the
contents into segments. These rings of wall then relax and adjacent
rings contract, breaking up the first segments. One of the reasons for
chyme moving along the intestine is that the frequency of segmentation
Figure 1.18. Segmentation; as the rings of contraction occur at different points in the
intestine the contents are moved in opposite directions and mixed.
is greater at the duodenal end than the ileal end of the intestine. In
addition, there is peristaltic activity in the small intestine which always
proceeds towards the large intestine, although these waves of
contraction only travel a few centimetres and then cease. Peristaltic
waves do not normally travel along the whole length of the intestine
as in the oesophagus. Distension by the intestinal contents causes an
increase in muscle activity. The motility is coordinated by the internal
nerve network; parasympathetic nerve activity increases and
sympathetic activity decreases the general level of activity.
1.1.15 Absorption of Water and Electrolytes
Water moves freely in either direction through the intestinal wall,
depending on the osmotic gradient. Although the average intake of
water only amounts to about 1200 mI/day, a large amount of water is
found in the secretions poured into the intestine each day, and as the
average water content of faeces is only about 100 mI/day a large
quantity must oe absorbed. This takes place mainly in the small intestine.
In most parts of the gut the contents are nearly isotonic as water
quickly follows the osmotic gradient (mainly due to sodium); as in
other areas of the body where sodium ions are absorbed, this process
is affected by blood aldosterone levels. Chloride and bicarbonate ions
may be actively absorbed or may follow passively. Sulphate and
magnesium ions are not well absorbed and their presence in lumenal
fluid will tend to hold water there, so producing a large volume of
intestinal contents and thus acting as a purgative.
Water
reabsarbed =::::::;E:---
into blood
Mouth
Food (8009)
+
Fluid (1300 mll

__ Saliva
(1200ml)
Gastric juice
(3000ml)
Bile (500mll
- Pancreatic juice 11000 mll
Intestinal secretions (2000 ml)
Faeces [Water (lOOml) + Solids (
60
91]
Figure 6.19 Although large volumes of water are secreted into the gastrointestinal
tract only 100 rnl remain in faeces.
1.1.16 Vitamins
Water-soluble vitamins are freely diffusable and so rapidly
absorbed in the small intestine. One of them does depend for
absorption on the presence of a mucopolysaccharide from the stomach,
the intrinsic factor. This absorption takes place in the terminal ileum.
Fat-soluble vitamins are absorbed along with fats in the jejunum.
1.1.17 The Large Intestine
The colon or large intestine forms the final part of the gastroint-
estinal tract and, as its name implies, it is wider than the small intestine
and not so coiled. Its function is to absorb water and electrolytes from
chyme, and to store the resultant faeces until the time is convenient
for their excretion. About 500 ml of chyme enters through the ileocaecal
valve daily and normally 400 ml of water would be absorbed, although
this does depend on the length of time the contents stay in the large
intestine. Active absorption of sodium is again the _ mechanism
responsible for the removal of water. The potassium content of the
intestinal contents is greater than the plasma level, potassium being
exchanged for the sodium absorbed.
Hepatic
flexure_
Ascending
colon __
Taenia
Ileo-caecal
valve
Caecum
Transverse
colon
Figure 1.20 The large intestine.
Splenic
flexure
Descending
colon
Sigmoid
colon
t--_______ Rectum
Peristaltic movements travel for greater distances in the large
intestine than in the small intestine, propelling the contents into the
sigmoid colon. A non-propulsive movement like segmentation also
occurs and is responsible for mixing.
1.1.18 Defaecation
When the contents of the sigmoid colon are carried by peristalsis
into the rectum, the distension of the rectal acts as a stimulus for
defaecation. If conditions are appropriate, further peristalsis occurs in
the sigmoid colon, the rectal walls contract and both the internal and
external anal sphincters relax resulting in the elimination of faeces
through the anus. This is usually assisted by inhalation of a deep
breath, closure of the glottis and an increase in intra-abdominal pressure
due to the forced contraction of abdominal muscles. If circumstances
are not suitable for defaecation, the external anal sphincter-which is
under voluntary control-remains closed, the intestinal muscle activity
is depressed, the rectal wall relaxes and so the urge to defaecate
disappears.
1.2 METABOLIC RATE
Catabolism, or the slow controlled combustion of molecules of
carbohydrates, fat and protein which occurs in the body, results in
the release of energy from the breaking of carbon and hydrogen
linkages. The energy thus liberated is either given off immediately as
heat or stored as energy in the form of A TP. This energy store is
used to perform both external work (movements through the
contraction of skeletal muscle), or internal work (functional activities
of cells). The units used for the measurement of energy are kilocalories
(240 kcal=l06 J=lMJ).
Total energy = energy stored + energy used + heat
When foodstuffs are burned outside the body the heat produced,
and so its potential energy, can be measured. The amounts of energy
produced in the body in the catabolism of food is not exactly the
same but are approximately equal to 4 kcal/g for carbohydrates and
protein, and 9 kcal for fat. The amount of energy liberated by the
body in unit time is called the metabolic rate.
The heat output from man can be measured directly in a human
calorinleter, but is more often done by indirect calorimetry. This
depends on the fact that the amount of heat produced when 1 mole
of oxygen is consumed in the oxidation of different foodstuffs is
known. To do this calculation the relative amount of fat, carbohydrate
and protein needs to to be known. This additional information can be
obtained from the respiratory quotient (RQ). This is the ratio of the
volume of CO
2
produced to the volume of 02 consumed when a
substance is oxidised,
e.g. C
6
H
1
P6 + 602 = 6C0
2
+ 6Hp
so the RQ for carbohydrate is 1. The RQ for fat is about 0-7 and for
protein approximately 0 82.
The RQ of the whole body can be calculated from analysis of
expired air, but factors other than metabolism, for example an increase
in ventilation, will vary the ratio of CO
2
exhaled to 02 inhaled.
Measurement of the CO
2
exhaled to 02 inhaled ratio is now usually
referred to as the respiratory exchange ratio (R).
As the body consumes a mixture of carbohydrate, fat and protein,
the energy liberated when I litre of oxygen is consumed is approximately
482 kcal. If we therefore measure the oxygen consumed by a subject
- 28 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
we can calculate the energy production or metabolic rate by multiplying
the 02 consumed per minute (STPD) by 482.
The metabolic activity of an individual will vary with activity,
so that in order to make comparisons of the metabolic rates it must
be determined under standardised resting conditions. It is then known
as the basal or resting metabolic rate; in an average adult male this
amounts to 2000 kcal (84 MJ) per day.
There must be a balance between the energy output of a person
and the potential energy or calorific value of food ingested. If the
calorific value of ingested food is greater than his energy output, the
energy is stored and a person gains weight. Food intake and energy
expenditure vary widely from day to day and, although body weight
varies over time, it is constant enough to suggest there must be a
mechanism which regulates it.
Many mechanisms are advanced to account for the close matching
of caloric intake to energy expenditure in normal animals; it has,
however, proved difficult to decide which are the most important.
From a consideration of other control systems it is thought that there
must be sensing devices which monitor food intake and the current
amount of foodstuffs in the body; also neural mechanisms in the
hypothalamus to integrate these signals, and effector mechanisms which
cause the animal to hunt and eat food.
Possible sensory inputs involved in these mechanisms are:
I. A system in the mouth or eosophagus which monitors the
food intake; for 'sham feeding' leads to cessation of feeding
even though the food is not absorbed into the body.
2. A system which responds to distension or partial filling of
the stomach; for distension of the stomach leads to cessation
of feeding, and if the stomach is partly filled (through a
stomach tube) the animal, or man, only eats enough to make
up its normal intake. 3 A system of receptors (perhaps in
the hypothalamus) which respond to the circulating levels of
glucose and fatty acids in the blood.
The hypothalamus is thought to contain a 'feeding' or 'hunger'
area and a separate 'satiation' area; destruction of the former produces
animals which will not eat, destruction of the latter produces animals
which overeat and become obese.
The control of food intake is not always successful in keeping
the body weight constant; when errors occur they are in the direction
of too much weight gain rather than too little. Perhaps, as with the
control of fluid intake, our mechanisms are more successful in
guarding against deficits rather than excesses. Some animals (rat and
possibly man) gain much more weight if fed a constant amount of
food in a few big meals than in many small ones, but there is no
resulting alteration in food intake-a puzzling finding. It has been
suggested that appetite control is more successful at high rather than
low exercise levels; this may explain Western man's difficulty in
controlling his weight. In man it has been suggested that eating habits
together with some voluntary control (clothes getting tighter, etc.) is
enough to regulate body weight.
Our diet is composed of carbohydrate, protein, fat, vitamins,
minerals and water. In the diet of most people carbohydrate forms
the biggest portion; the poorer the individual or community the greater
the percentage of carbohydrates. Most of it is eaten in the form of
starch in grain or root vegetables, although an increasing amount in
the diet of the more 'advanced' societies is in the form of sugars.
Most of the sugars are the disaccharides, sucrose, lactose and maltose
with little as the monosaccharides, glucose and fructose.
Proteins not only act as a source of energy, like carbohydrates
and fat. but supply the nitrogen and sulphur required for the synthesis
of proteins to replace the body proteins which are continuously being
broken down. This is a constant need for normal maintenance, but
the body's protein requirement is obviously increased in children for
growth and in mothers during pregnancy and lactation. A simple guide
to our protein requirements is that for each kg of body weight we
require I g in our diet. This is a generous allowance and, as protein
is expensive and its supply is inadequate in many areas of the world,
many people exist on much less, but it is a good level to aim for in
order to provide a safety margin. Some amino acids are not synthesised
in the body and so must be present in our diet; these are called
essential amino acids. Because all proteins do not contain all possible
amino acids, we find that some are more suitable as a human dietary
protein than others and for this reason it is wise to provide a mixture
of proteins in our intake.
Fats, because of their high calorific value, make up a large
proportion of the energy we derive from our diet. This is not
necessarily taken as obvious fa( but forms part of much of the food
we eat; indeed it is necessary to improve the palatability of our food,
as can be vouched for by anyone who has had to take a low-fat diet
for any length of time.
Oesophageal
monitoring
of food intoke
F"""'''ol ~
h . ~ ' h /
Receptors for
measuring the
blood
concentration
of glucose
and fatty
acids
? Voluntary
control (man)
1
Figure 1.21 Suggested mechanisms for appetite control.
_____ Appetite
'&
Behavioural
challQes
Our daily calorific requirement obviously varies with our age,
size and activity but for an adult male of sedentary disposition 2800
kcal (11 7 MJ) would suffice. Remembering that the calorific value
of carbohydrate and protein is 4 calories and of fat 9 callg, a
reasonable diet might be as follows:
cal MJ
375 g of carbohydrate 1500 6-27
100 g of protein 400 1-67
100 g of fat 900 376
2800 11-70
Vitamins. These are organic substances in our diet which are
essential, as the body cannot usually synthesise them. They are
required in many enzymatic processes and their absence leads to
deficiency diseases. Since they have been isolated and synthesised they
have been given their correct chemical names but are still often
referred to by letters. They can be classified into two groups, the
fat-soluble Vitamins A, D, E and K, and the water-soluble Vitamins
Band C.
Mineral and water. The normal minerals in our diet, sodium,
potassium, calcium, phosphorus and iron, are dealt with in other
sections of the book.
Table 1.1 Fat-soluble vitamins.
Vitamin Chemical Source Deficiency disease
name
A Retinol Dairy products Night blindness
Liver
Some can be formed Keratinisation of
in gut from carotene epithelium
D Cholecalciferol Dairy products Rickets in
Eggs children
Liver oil
Some can be formed Osteomalacia in
in skin from action adults of UV light on
7-dehydrocholesterol
E Tocopherol Wheatgerm Unknown
F Naphthoquinone Green vegetables Not primary
derivatives Synthesised by gut failure to absorb
bacteria it in disease of G-I
tract or sterilisation
of gut with antibiotics
leads to failure of
prothrombin
Table 1.2 Water-soluble vitamins.
Vitamins Chemical Source Deficiency disease
name
C Ascorbic acid Citrus fruit Scurvy
Blackcurrants Fragility of blood
Vegetables vessels
B complex B, thiamine Wheatgerm, pulses, Beriberi, polyneuritis,
yeast heart failure
B2 riboflavin Meat, milk, Degeneration of
wholemeal flour, skin-mucous
membrane
beer junctions
Nicotinic acid Milk, liver, yeast Pellagra, dermatitis,
dementia, diarrhoea
Pantothenic acid Widely distributed None known
Pyridoxine Widely distributed None known
B,z cyano- Foods of animal Pernicious anaemia,
cohalamin origin lack of intrinsil:
factor, thus failure
of absorption
Trace elements. Certain elements are needed in our diet in very
small amounts and are present in tissues in as small a concentration as
1 part in 20,000. These are called trace elements; the most important
are iodine, copper, cobalt, manganese and zinc.
2
Respiratory
System
2.1 RESPIRATION
Respiration is the transfer of gases between body cells and the
environment. It goes hand in hand with metabolism, the process by
which food produces energy for the needs of the body. In metabolism
oxygen is taken from the environment and carbon dioxide added to
it; in our bodies these processes proceed at 37'C due to the presence
of catalysts-the enzymes. Lavoisier thought that similar processes
occurred in a flre and in animal metabolism as in both q is absorbed,
CO
2
given off and energy produced. We now know that, though the
end results of these processes are similar, the detailed reactions are
very different; in metabolism hydrogen is removed from substrates
whereas in a flre oxygen is added to them. The reason for these
differences is probably that animal metabolism evolved before the
atmosphere contained oxygen to act as an electron acceptor; so
metabolism predates combustion.
2.1.1 Diffusion of Gases in Respiration
Most living animals ultimately gain their 02 and lose their CO
2
by diffusion. '1 hIS IS the process whereby a net movement of molecules
occurs from an area of higher concentration to one of lower
concentration. The rate at which substances diffuse increases with the
driving force and the area available for diffusion and decreases with
the distance. Diffusion in gases is much faster than in liquids because
the molecules in a gas are less tightly packed. Such a transfer of
substances is fast over short distances but very slow over large
distances. As animals increase in size there are two reasons why the
33
supply of gases by diffusion becomes inadequate. The first is that the
distance for diffusion becomes too great and the second IS that the
surface area of the animals becomes relatively smaller than the mass
of the metabolising tissue. With spherical animals this point is reached
at sizes of about 1 mm. All animals with sizes greater than this require
a specialised respiratory system together with an associated circulatory
system. In man this respiratory system consists of lungs in which gas
movement occurs by a bellows action of the chest wall. which creates
a negative pressure in the chest and so draws air in. As we require a
large gaseous exchange with the environment the surface area of our
lungs is very large due to their division into hundreds of millions of
small air spaces, the alveoli.
2.1.2 Ventilation of the Lungs
Distribution of gas and blood to the lungs. Air is filtered by hairs
in [he nose-which remove the larger dust particles-moistened and
warmed by the mucosa of the nose and pharynx so that by the time
F'1gure 2.1. Schematic dlagrdm of lung function.
Bulk flow
(external "breathing")
respiration
Diffusion
Bulk flow
(transport in
blood)
Diffusion
Internal respirotion
it reaches the trachea it is saturated with water vapour and is at body
temperature. The hairs in the nose only tilter out gross particles from
the inspired air, and particles smaller than I pm stay in suspension
and are breathed out again. Dust of size, that is from
10 down to 1 pm, is removed from the ai during its passage along
the respiratory tract, the larger particles in the nasopharynx and upper
respiratory tract and the smaller in tP..! bronchial tree itself. An
important factor in this filtration is thl' presence of mucous on the
surface of the respiratory tract epithelium. Turbulence occurs in the
air flow as it changes direction, particles impinge on the walls and
RESPIRATORY SYSTEM
Table 7.1 Symbols used in respiratory physiology.
Primary symbols
P pressure or partial pressure
V volume of gas
V volume of gas per unit time
F fractional concentration in dry gas
Q volume of blood
Q volume of blood per unit time
C concentration of gas in blood
S percentage saturation of haemoglobin with 02
Secondary symbols
1 Gas phase
I inspired air
E expired air
A alveolar air
T tidal air
D dead space air
B barometric
2 Blood phase
a arterial blood
v venous blood
c capillary blood
v mixed venous blood
c' end capillary blood
Examples
Partial pressure of CO
2
in mixed venous
blood Pv, co
2
Fractional concentration of N2
in inspired gas PI' N2 02 concentration in
arterial blood Ca, 02
35
are trapped in the mucous. This particle-laden mucous must then be
disposed of and this is the task of the ciliated epithelium. The
epithelium of the respiratory tract is ciliated. that is it has minute
hair-like projections from its surface on which the sheet of mucous
is carried, These cilia move in a coordinated fashion with a fast stroke
in one direction and a slow one in the other. This has the eHect of
carrying the sheet of mucous, with its entrapped particles, ill onc
dire('tioll, downwards in the nasal mucosa towards thl' pharynx and
upwards in the bronchial tree to the pharynx. Once in the pharynx
the mucous is unconsciously swallowed.
The ciliated epithelium ends at the terminal bronchioles, so what
happens to particles which are deposited in the respiratory bronchioles
and the alveoli? In the main they are mopped up by wandering alveolar
macrophages which get to the mucociliary 'escalator' and so to the
pharynx, or are carried away in lymph vessels or are sequestered in
lung tissue by a tissue reaction.
The trachea divides into right and left branches and then continues
dividing into smaller and smaller bronchi, fmally reaching the terminal
bronchioles which lead to the alveoli. The incoming air thus eventually
reaches the alveoli where the main exchange of CO
2
and 02 occurs.
These alveoli are some 100-300 pm in diameter and consist essentially
of a bundle of capillary vessels suspended in air. Most of the alveolar
surface is covered by these blood vessels, which are some 1500 miles
in length spread out over the 40-60 m
2
of diffusing surface. At rest
the capillary blood is renewed every second and during exercise every
third of a second or less. To do so the total blood flow is about 5
titres/minute at rest and during exercise about 20-30 litres/minute.
The air flow to the alveoli is about 4 litres/minute at rest and up to
100 litres/minute during exercise.
The details of the blood and air flow to the lungs will be
considered in more detail, separately.
2.1.3 PuImonary Circulation
The pulmonary circulation, being in series with the systemic one,
receives the total cardiac output. The right ventricle pumps this blood-
mixed venous blood-through the pulmonary artery to the lungs, and
its branches accompany the bronchioles to the terminal bronchioles
where they break up into the capillary bed.
02 removal from lungs = arterial-venous oxygen concentration
x blood flow
or
blood flow =
02 uptake
arterial - venous oxygen cOllcentratio
all per unit of time.
If 02 usage is 250 mllminute and arterial and venous contents are
200 and 150 mlllitre the cardiac output = 5 litres/minute. This neglects
the nutritional blood flow.
The main difference hetween the pulmonary circulation and the
RESPIRATORY SYSTEM
37
systemic one is that it is a low-pressure system with a systolic/diastolic
pressure of 25110 mmHg (33/13 kPa) and a mean pressure of IS
mmHg (2 kPa). The left atrialpressure is about 6 mmHg (08 kPa), so
the driving force for blood flow through the pulmonary circulation is
only some 10 mmHg (13 kPa) compared to about 80 mmHg (106
kPa) for the systemic one. As the same volume of blood flows through
both circulations this means that the resistance to flow in the pulmonary
circulation is much less than iri the systemic one; this is because the
walls of the pulmonary arterioles are much thinner and contain less
smooth muscle than those elsewhere.
__ Oxygen consumption V
02
Blood flow 0
Figure 2.2 Measuring cardiac output by the Fick prinCIple. Blood leaving th.
contains more oxygen than blood entering the lungs. This difference in amount IS equal
to the oxygen added by the lungs. So
When cardiac output rises, as in exercise, the pulmonary arten
pressure does not rise and this must mean that resistance to blood 0(1\\
through the lungs decreases. There is both an increase in the diamelel
of pulmonary vessels and an opening up of some which have heen
closed under resting conditions; this results in a decreased resistance
to flow, so enabling the pressure to remain low.
The pulmonary circulation is also different from the systemic onc
in other respects:
1. The capillary network is more dense in lung than elsewhere
in order to give a large interface for gas exchange.
2. Because the hydrostatic pressure in lung capillaries is always
much lower than the oncotic pressure of the plasma protein
there is less tiltration of fluid out of the capillaries into the
lung interstitial space.
3. The pulmonary arterioles constrict to a fall in P0
2
or a rise
in the opposite of what happens in the systemic
circulation. This has the 'useful' effect of diverting blood
away from poorly ventilated segments where less exchange
of gases could occur.
Pulmonary
vein ___
\
Lymphatics
Vasomotor
nerves
Pulmonary
(
artery
;
_ Vasometer
I nerves
/ Lymphatics
Figure 2.3 Composite diagram (Jf thf lung s!ructul'el>. The diagT"dm on the left is taken
from a ca,t of the of a human lung. showing airways from rhe trachea to the
nght terminal bronchioles (the alveoli are removed). The diagram on the right continues
the airways down to the alveoli. Note the difference in scales.
4. Because mean pressure is so low the difference due to gravity
above and below heart level is significant, particularly in
the upright posture. Blood is therefore normally distributed
preferentially to dt:pendem parts.
2.1.3 of Respiration
Air moves into and (Jut of the lungs as a result of changes in
... res<;ure within the chest cavity, the lungs themselves expanding and
RESPIRATORY SYSTEM
39
shrinking passively. The energy for these changes comes from
contraction of the diaphragm at the bottom of the chest cavity and
the muscles of the chest wall. The lungs do, however, contribute an
elasticity to the system by virtue of their tendency to collapse.
Nonnally they are prevented from doing so by adhering to the inside
surface of the barrel-like thoracic cage.
Blood flow
~ ? ~ m l L
Interstitial
space
Figure 2.4 Forces and fluid movement in lung capillaries. Bulk flow of plasma filtrate
occurs into the interstitial space, as elsewhere; it is then either reabsorbed or forms
lymph. Little fluid passes into the alveoli mainly because of the presence of tight
junctions between the alveolar epithelial cells. Gas exchange occurs at the 'thin regions'
shown.
About 500 rn1 of air are taken into the chest during each breath
as a result of descent of the diaphragm and the elevation of the rib
cage. The descent of the diaphragm occurs because of nerve impulses
in the phrenic nerve supplying it; the rib cage is altered mainly by
the contraction of external intercostal muscles. During quiet respiration
movements of the diaphragm account for 75% of the air movement.
Because of the way the ribs are articulated, the breadth as well as
the depth of the chest is increased during a breath.
At the end of a normal quiet inspiration the nervous activity
driving the muscles of the diaphragm and chest wall falls off, and
the chest size decreases due to the elastic recoil of the lungs. So at
the end of a normal expiration the elasticity of the lungs tending to
cause them to collapse is exactly balanced by the elastic tendency of
the chest wall to expand. During vigorous respiration this passive
collapse of the chest and diaphragm is too slow, and active expiration
occurs. This involves various thoracic and abdominal muscles which
pull the chest wall-down and push the diaphragm up, thus increasing
the rate of expiration.
Figure 2.5 Diagram showing the tendency of the lungs to collapse and the tendency
of the chest wall to expand. At the end of a normal expiration these forces are balanced.
Quantitative information on the mechanical factors modifying
breathing has been obtained by measuring pressures in various parts of
the system, together with the volume of gas breathed into and out of
the chest and the rate of gas flow. The important pressures are:
1. The intrapleural or intrathoracic pressure, which is measured
in the 'space' between the pleural membrane lining the inside
of the chest wall and that lining the outside of the lung.
(These layers are normally only separated by a thin film of
pleural fluid.)
2. The intrapulmonary pressure, which is the pressure inside an
airway. The place where it is measured must be specified as
there is a pressure gradient between the mouth and the alveoli,
with the alveolar pressure most negative during inspiration.
It is this pressure gradient which 'pulls' the gas along the
airway.
3. Atmospheric pressure is the reference level to which the
others are compared.
RESPIRATORY SYSTEM
Inspiration: Expiration
o I
-6
..
-7
- 8 1--'---"'''1''''''-------4
+0.5
'-----+0
-0.5
+1
'------.0
-I
41
Figure 2.6. Alveolar pressure, intrapleural pressure and volume of air breathed. plotted
during one respiratory cycle.
The effect of the inspiratory effort is to reduce the intrathoracic
pressure, which causes a reduction in the alveolar pressure and thus
causes a gas flow to occur.
The intrapleural pressure may be measured by inserting a hollow
needle, connected to a manometer, through the chest wall so that the
needle tip lies between the two pleural layers. Normally it is negative
(or subatmospheric) throughout the cycle, with a value of about -5
cmH20 (-05 kPa) at the end of a quiet expiration. At the end of a
deep inspiration it may become -20 cmH20 (-20 kPa), and during
forced expiration against a resistance, for example, blowing bagpipes,
it may reach + 70 cmH20 (+ 69 kPa) or so. The relation of the
change in intrapleural pressure to the volume of air inhaled is about
02 litres/cmH20 (i.e. if the pressure drops by I cmHp, 02 litres of
air are inhaled).
Air flows through the airways because there is a pressure
difference between the alveoli and the atmosphere. This circumstance
may be used to calculate a resistance to flow, using Ohm's law. In a
normal subject this is about 2 cm/litre/second, i.e. if air is breathed
in at a rate of I litre/second, there is a 2 cm Hp drop of pressure
between the alveoli and the atmosphere.
During one respiratory cycle the events are quite complex. Figure
7.6 shows these various pressures, volume changes and rate of gas
flow during one breath. Air flow into and out of the lungs depends
on the pressure difference between atmospheric air and that in the
alveoli. The alveolar pressure depends on the algebraic sum of the
pressure created by the lung elasticity, and the superimposed pressure
due to respiratory muscles. Pressure swings are much greater during
heavy breathing leading to more rapid flows of air. To create a
pressure difference requires work and larger pressure differences
require more work than small ones. During vigorous breathing the
work of breathing is therefore greater than at rest.
Lung elasticity
+ sur face tension
of alveolar
fluid
~ __________________________
+
Pressure
Figure 2.7 The behaviour of isolated lungs to changes in pressure. The lung elasticity
was obtained by inflation with saline whereas the lung+ surface tension was obtained
by air inflation.
The elastic behaviour of lung is made up of two different
components; that due to the elasticity of the lung tissue and that due
to the surface tension of the thin layer of fluid lining the alveoli.
These may be shown by inflating an isolated lung with air-which
measures both componentsand then with saline, which removes the
surface tension effect and so measures that due to the tissue elasticity.
The elastic property of lung tissue is fairly constant with distension,
whereas that due to lung and surface tension is complex and differs
depending on whether the lung is being inflated or deflated, i.e. shows
hysteresis. This complexity is due to the effects of the surface tension
of the fluid lining the alveoli and small bronchioles.
RESPIRATORY SYSTEM 43
We all know that it is more difficult to start inflating a balloon
than to continue inflating it once it is big. This is partly because the
wall becomes thinner as the size increases, but largely because of
the physical properties of spheres. This can be studied most easily in
gas bubbles which have a constant wall thickness; small bubbles require
a greater force to stop them collapsing than larger bubbles. 'Fhis can
be shown by connecting a large and small bubble together, the small
one collapses and empties into the larger one. It would clearly be
inconvenient if small alveoli in the lungs collapsed and emptied into
larger ones. This effect is counteracted in normal lungs by the presence
of surfactant, which not only diminishes the surface tension but also
diminishes it more in the smaller alveoli than in the larger ones.
Surfactant therefore adjusts the surface tension of the wall of an
alveolus to correspond with its size. Reducing surface tension also
decreases the force causing the exudation of fluid from the capillaries.
The alveoli can be considered as spheres of various sizes. The
relationship between the internal pressure (P) applied to them, the
wall tension (T) and radius (rJ is described by Laplace's law:
P ~
The work of breathing is of two kinds: non-elastic required to
move air through the airways; and elastic required to overcome the
elasticity of the lungs and chest wall. The non-elastic work depends
on the rate of breathing, for as the rate increases the velocity of air
flow in the airways increases and so there is an increase in frictional
work; whereas the elastic work depends 011 the depth of breathing
for as the depth increases the lungs need to be stretched more. The
amount of air breathed depends both on the rate and depth, so that
any particular ventilation can be obtained by various combinations of
these. If the total work of breathing is measured over a range of
different combinations of rate and depth at constant ventilation then a
minimum value is found. In natural breathing rate and depth are set
to this minimum value. It is an interesting but so far unresolved
question as to how the body works this out; perhaps it is related to
volume information via the vagus and length-tension infomIation from
respiratory muscles. It is likely that the total work of breathing rarely
exceeds 3 % of the total energy expenditure of the body.
The non-elastic frictional work is small during quiet respiration,
for the frictional resistance to air moving through the airways is then
not very great. The friction is enough, however, to mean that the air

,
A
!

interface
Some
surfactant
B present
0---0---0
c
!
0-------0
Figure 2.8 Surface tension is due to intermolecular attraction forces (A). Surfactant
(the black circles) reduces this attractive force between molecules (B); the smaller the
surface area, the less is surface tension (C). The higher pressure in the alveolus (A)
would cause air to pass into the other two, with collapse of A.
\
III
,
,
\
\
'-
'-
,

Increasing volume
decreasing rate
[slow deep breaths]
.....
/
./
,./
,/ Non-elastic
or frictional
Decreasing volume
increasing rate
[fast shallow
FIgUre 2.9 The work of breathing at constant total ventilation but differing combinations
of rate and volume.
flow, and so the change in volume, lags slightly ,pn the pressure so
that if a pressure-volume curve is plotted it has the form. The shaded
area is the frictional or non-elastic work required to do this. With
increases in air flow this increases.
o
Intrapleural pressure
Figure 2.10 Pressure-flow relationship in an intact chest during quiet respiration. If
air flow followed pressure change instantly then this diagram would be reduced to the
centre sloping line; but air flow into and out of the lungs l a g ~ behind the pressure
change producing it so giving the curves shown.
2.1.4 Lung Volumes
The total lung capacity of a 30-year-old 70-kg man is about 6
litres. This is divided into the various components.
/,
Residual
volume
~
Expiratory
reserve
volume
lidal
volume
Inspirotory
reser.e
volume
5.0
4.0 ~
l
3.0
2.0
1.0
'"
---Rest'"O--
: tidal
~ ---yolume- --- --
{!
Functional
1-----residual--...1----'-L..:....L-rIL-....L-
capacity
Residual
volume
Figure 2.11 The lung volumes and capacities. Left: illustrates the four primary lung
vulumes and their magnitude; right: typIcal record obtained by spirometry. TV, Tidal
volume, or the depth of breathing. IRV, Inspiratory reserve volume. ERV, Expiratory
reserve volume. RV. Residual volume.
2.1.5 Gas Mixtures and Pressures
Certain physical properties of gases must be considered in relation
to the movenlent of gases' in the body. These physical properties
determine the molecular concentration of oxygen and carbon dioxide
and so control their diffusion between various parts of the body and
their reaction with chemicals in the blood.
The concentration of a gas in a mixture of gases may be indicated
in a number of different ways, for example moles/litre or volumes
percent. When a gas has access to both gas and liquid phases it is
necessary to use the concepts gas tension and partial pressure. The
relationship between pressure and volumes of gases is given by Boyle's
law and between volume and temperature by Charles's law. Thus the
pressure of a gas at a given temperature defines its concentration.
PV = nRT
where n=number moles of gas, V=volume, T=absolute temperature
and R = gas constant
Concentration (nIV) =-
R
With mixtures of gases the partial pressure of an individual
component is defmed as the contribution of a gas to the total pressure.
The composition of dry air at standard temperature (2 73 K) and
pressure (760 mmHglOI 3 kPa) is
%
02 2093
N2 7904
CO
2
0-03
The partial pressures of the gases, indicated by the symbols P0
2
PN
2
and Pc0
2
are derived from these proportions. Thus the partiai
pressure for oxygen will be given by
20.93
--x 760 = 159 mmHg (21.2 kPa
100
Similarly, the partial pressure for nitrogen will be 600 7 mmHg
(8009 kPa) and Pc0
2
, 023 mmHg (003 kPa). When the air is moist
then water vapour contributes its partial pressure to the whole. At 3
7C the partial pressure of water vapour in saturated air is 47 mmHg
(62 7 kPa). So air entering the alveoli of the lung has a lower partial
pressure of 02 than dry air. For example,
RESPIRATORY SYSTEM
20.93
--x (760 - 47) = 149 mmHg (19.9 kPa
100
47
However, the actual partial pressure of 02 in alveolar air is less
than inspired air even making allowances for water vapour pressure
as 02 is removed by the blood and a steady state is reached with a
partial pressure of around 100 mmHg (133 kPa).
Inspired oir
Alveoli
Veins
Tissues
Eltpired gos
Dead
space
Arteries
Figure 2.12. Partial prt'Ssures of gases in mmHg in respiration. The total pressure III
the l s ~ u e s and veins is subatmospheric due to less CO
2
being produced than 02 used.
The tIssue 02 is often less, and the CO
2
greater, than shown here but IS not easy to
measure. The normal percentages of 02' CO
2
and N2 m air are 20-9, 003 and 7903
respectively
It will be noticed that alveolar gas differs from inspired air in
hav!ng less 0, and more CO
2
; this is due to 02 diffusing from the
alveoli into the blood and cq diffusing from the blood into the alveoli.
The normal average values of P0
2
of 100 mm (133 kPa) and Pc0
2
of 40 mm (5 3 kPa) are remarkably constant in resting man, due to
the fact that the respiratory control system seems to be 'set' to a
Pea.' value near to 40 mm (53 kPa).
When gases are dissolved in a liquid the molar or molecular
concentration depends on the partial pressure of the gas in the liquid
(which at equilibrium is the same as that of a gas in contact with it)
and the solubility of the gas in the liquid. Commonly we use the
partial pressure of the gas (tension) as a measure of the molar
concentration; this is convenient and correct for a single gas but some
care is necessary when comparing different gases. CO
2
is 23 times
more soluble than 02 in water so that equal molecular concentrations
occur with a partial pressure of CO
2
23 times less than that of 02. In
arterial blood the concentrations of 02 and C02 are 94 mmoillitre
and 21-7 mmoi/litre, in saline equilibrated with blood they are 2 and
1 2 mmolllitre respectively.
Oxygen lack at high altitudes can be explained by these
considerations, for with air breathing at the reduced atmospheric
pressure the alveolar P0
2
falls to levels too low to adequately oxygenate
the haemoglobin in blood.
2.1.6 Respiratory Dead Space and Alveolar Ventilation
Because the alveoli are situated deeply within the body at the end
of the conducting airways not all the air breathed in reaches them.
These conducting passages constitute the 'anatomical dead space' of
the respiratory system and, by their presence, reduce the efficiency of
respiration. The anatomical dead space may be measured in a cadaver
by filling the air passages with water, and in life by continuously
monitoring the N2 concentration of expired air after a single breath of
pure 02 and measuring the volume exhaled before the N2 concentration
rises due to alveolar gas. Normally, the anatomical dead space is
about 150 m1 and so represents about one-third of the volume of an
average breath at rest.
The total, or physiological, dead space includes both gas in the
anatomical dead space and that ventilating alveoli with relatively poor
or no blood flow, so it is larger than the anatomical dead space and
more relevant to a consideration of the efficiency of breathing. If an
alveolus receives no blood supply then the gas in it will be unchanged
and so, when exhaled, will be like atmospheric air. If an alveolus
receives an adequate blood supply the gas within it will equilibrate
with the blood and so have a similar gas composition to blood. The
composition of expired air could thus vary between that of the inspired
air (=no blood supply to the lung) and that of the arterial blood leaving
the lungs (=all the alveoli receive an adequate blood supply and there
is no dead space). Bohr's formula for calculating the physiological
dead space is based on this idea:
physiological dead space volume arterial Pco - expired gas P
e
= '
tidal ,volume arterial P
eo
,
The result is expressed as a ratio with a normal value of 03.
The volume of air which enters the alveoli per minute and so
causes effective ventilation is therefore the product of the breathing
rate x (the tidal volume-physiological dead space). This is the alveolar
ventilation, V A, and varies from some 4 litres/minute at rest up to
100 litres/minute during severe exercise.
_ ~ ~ ... ___ Pulmonory
capillary
.... /.'---_ Alveolar
epithelium
............ , ~ , . . . . . - - - - _ Basement
membrane
Figure 2.13 Pulmonary capillary in the alveolar wall. The arrow Indicates the diffusion
path from alveolar gas to the interior of the erythrocyte which includes the layer of
surfactant (not shown in the preparation), alveolar epithelium. interstitium, capillary
endothelium and plasma. Note the extremely thin blood-gas hamer of less than 0 5
/Lm.
Gas exchange between the alveoli and the blood occurs continu-
ously, whereas that between the alveoli and the air is intermittent.
Therefore the partial pressures of gases in the alveoli vary around a
so PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
mean value during respiration. This variation is kept to a value of
4% of the mean because the total alveolar volume is much larger
thaIl each single breath. The overall effect of the respiratory control
systems discussed later is to minimise changes in this mean level during
varying metabolic activity of the body.
2.1. 7 The Transfer of Gases Between Alveoli and Blood
Although it was at one time thought that 02 was 'secreted' by
the lungs, it is now clear that the movements of 02 and CO
2
between
the alveoli and blood are determined only by the pressure differences
between them and the nature of the intervening membranes. The
diffusion path between alveolar air and blood is short, and consists
of two layers of cells with very little interstitial fluid in the thinnest
parts. The net transfer of a gas from the alveolus to capillary blood
continues so long as there is a pressure gradient. For 02 under normal
resting conditions Ihis only occurs in the first third of the capillary
network which it reaches; thereafter no net transfer occurs as there
is no pressure gradient. An individual erythrocyte (and the plasma
aroWld it) spends about I second in an alveolar capillary under resting
conditions and perhaps a third of this time during vigorous exercise.
Capillary
IOOmmHg
I lory
Figure 2.14 Net 0, transfer between an idealist:<. alveolus and lung capillary under
resting conditions. Incomplete oxygenatioll may' ccur if the 0, tensioll in the alveolus
is reduced (tor ex.ample, high altitude) or the membrane is thickened (by disease),
particularly it the tim\: available for equilibration is increased (in ex.ercise). The
horizontal arrows indicate the relative size and direction of 02 tran&fer.
RESPIRATORY SYSTEM SI
At normal oxygen tensions, even this shortened time is sufficient
for full ox:ygenation to occur, but at reduceq oxygen tensions or with
thickening of the membrane (for example, at altitude or in disease)
this may not be sufficient and so less than equilibrium oxygenation
occurs.
2.1.8 Gravity Effects on Blood
and Gas Flow to the Lungs
For most efficient transfer of gases between alveoli and blood
all alveoli should receive an adequate air and blood supply and these
should be matched properly. In fact both the air and blood supply to
alveoli varies greatly from one part of the lung to another. The main
reason for this variation is the effect of gravity, which, in man standing
upright, causes a greater air and blood flow to the base than to the
top of the lungs.
r:cD

Cii:\
e;----
Alveolar and capillory
sizes at the end of
normal inspiration
-+1
~ "-"
0.1
Blood and air flow in lungs
[L/min % lung VOlume]
Figure 2.15 Both blood flow and ventilation increase from top to bottom of human
lung. The inserts show the relative sizes of the alveoli and capillaries at apex and
base, which explains this distribution.
The partial pressure of oxygen and carbon dioxide in the alveoli
of any zone of the lung and of the blood leaving that zone is
determined by the ratio of ventilation to perfusion. Theoretically, if
the ratio decreased to zero one would get the situation where the
alveolar gas partial pressures of oxygen and carbon dioxide are the
same as that of mixed venous blood. With an increasing VIQ ratio,
the extreme situation would be of an alveolus with no blood supply;
the partial pressures of oxygen and carbon dioxide in the lung unit
would be the same as atmospheric air. Obviously, mismatching of
ventilation and perfusion could result in any ratio between these two
extremes.
O
2
" '50 mmHQ
COZ"O

CO
2
" 45
00 ... ---- A
Decreasing ';VQ Inreasino VA";
Figure 2.16 Effect of alterations of the ventilation/perfusion ratio on the P0
2
and Pco}
in a lung unit (after West).
How do variations in the ratio affect blood gas levels? Blood
leaving an area with a low V IQ ratio will have a lower P0
2
than
normal and as more blood comes from the bases in the upright lung
this will tend to decrease the arterial blood Po
2
This is not
compensated for by the areas which have a high VIQ ratio.
As the CO
2
dissociation curve is more linear over its physiological
range, the high VIQ areas do tend to compensate for the low VIQ
areas. Further, any remaining increase in arterial Pc02 will stimulate
the overall ventilation and return the Pco
z
to normal levels, without
adding much oxygen to blood. The final effect will be to lower the
blood oxygen level slightly in normal lungs but VIQ ratios can be
grossly disturbed in lung disease. The difference between top and
bottom is much greater for blood than for air so that the ratio of
ventilation to perfusion changes from 3 at the apex to less than 0,5 at
the base with a mean overall value of 08 for the whole lung,
2.1.9 Carriage of 02 and CO
2
bl the Blood
In man at rest, about 250 ml of 02 and 200 ml of CO
2
is carried
per minute between the lungs and the tissues. The main substance
responsible for 02 carriage is haemoglobin, which, for osmotic
1
O
2
content 146 16'0
19-5
Figure 2.17 Because of the shape of the oxygen dissociation curve, areas with a high
V IQ ratio add very little more oxygen and not enough to compensate for areas with a
low V/Q ratio (after West).
reasons, is carried within' the erythrocytes. Haemoglobin combines
reversibly with oxygen in the reaction
4Hb + 40
2
;=: (Hb0
2
)4
to carry' over 98% of the oxygen present in the blood.
A smaller proportion (25 %) of the CO
2
is carried in direct
combination with haemoglobin as a carbamino compound, but
haemoglobin is also important in buffering C02 carriage as bicarbonate.
2.1.10 Oxygen Dissociation Curves
One gram of haemoglobin can combine with 134 ml of oxygen.
So 100 ml of blood, which normally contains 15 g of haemoglobin,
can carry about 20 ml of oxygen when saturated. This degree of
saturation occurs at the oxygen tension of 100 mmHg normally found
in the alveoli.
The relationship between the partial pressure of oxygen and the
quantity of it combined with Hb is sigmoid shaped because of the
interaction between the four oxygen binding sites on the haemoglobin
molecule. Once a single molecule of 02 has been bound, the affmhy
of the other three sites is increased and so on. Thif sigmoid shape is
of clear physiological usefulness , in that: (l) the amount of 02 carried
is not very sensitive to changes in P0
2
in the lungs over the range of
100 down to 60 nunHg; (2) at tissue P0
2
levels the curve is very steep
so that small changes in P0
2
below 60 nunHg lead to a large release
of oxygen to the tissues. In myoglobin-a molecule found in muscle
containing a single haem and a polypeptide chain-the relationship is
hyperbolic, as expected when no molecular interactions can occur.
100
20
------
~
90
/ /" Myoglobin
80
/
~
c
I
!i
~
.0
70
I
0
I
60
I
E
g
E
E
I
0
.,
c
50
I
10
Q
~
'0
....
-
40
I
E
0
~
c
I
~
~
30
e
:> :>
D
20 <;
III
I
III
Dissolved Oz
20 30 40 50 60 70 80 90 100
po. lmm) po. (mm)
Figure 2.18 Relationship between partial pressure of oxygen and the oxygen carried
by haemoglobin, myoglobin and a simple solution in the plasma. The inset shows the
effect of CO
2
and temperature variations on the steep part of the Hb curve. Myoglobin
is found in muscle; it has a simple hyperbolic type of uptake because it only binds
one O
2
molecule per molecule.
The carriage of oxygen is related to that of CO
2
in that with
increasing concentrations of C02 more 02 is released at a given tissue
P0
2
level (the .Bohr effect). Increases in temperature have a similar
effect. Both effects are clearly 'useful' adaptations in that in active
tissues producing more CO
2
and having a higher temperature more
oxygen will be given off. In active muscle this effect. may be very
large.
2.1.11 The Carriage of COl and
the COl Dissociation Curve
Considerably more CO
2
is held in blood than the small amounts
which are exchanged between the tissues and the lungs on each _
circulation. Our primary concern here is only to consider the carriage
of this 'extra' CO
2
from tissues to lung consequent on tissue
respiration. Although most of this CO
2
is carried as bicarbonate, with
a lesser amount combined with protein, we shall see that this carriage
is greatly influenced by the presence of haemoglobin.
Carbon dioxide diffuses from the tissue cells into the blood and
'8
90
0
1i
E

!.
8'
'0

Simple
:::J solution
:g
---
---
0
40
100
Portio I pressure fA CO
2
Figure 2.19 CO
2
dissociation curve for whole blood. The arterial and venous points
indicate the total CO
2
content of normal blood. The inset shows the fate of the extra
CO
2
which enters venous blood.
Table 2.2 The amounts of CO
2
carried in 100 ml of venous and
arterial blood.
Mixed A-V difference
Arterial venous
blood blood Units %tage
Peo
2
(rrunHg) 41 47'5 65
Dissolved (ml/100 ml) 2-7 31 01 8
As H
2
C0
3
(ml) 439 47-0 31 62
As carbamino compound (ml) 2-4 3-9 1-5
30
thence mainly into the erythrocytes, driven by its concentration
gradient. The most important reactions which occur are that COr in
the erythrocytes forms bicarbonate and also combines with the local
proteins (mainly haemoglobin) as a carbamino compound. The result
of these reactions is that much CO
2
can be taken up by the erythrocyte
without a large rise in the CO
2
tension. Lesser reactions also occur.
The formation of bicarbonate in the erythrocyte is more complex
than might appear at fIrst sight and will now be CO
2
forms bicarbonate as follows:
CO
2
+ H
2
0 H
2
C0
3
W +
The equilibrium point for this reaction is such that little
bicarbonate can be formed unless the reaction products-H+ and HC0
3
-are removed. In the erythrocyte most of the H+ is removed by
56 PHYSIOLOGY, BIOCHEMISTRY AND BIillECHNOLOGY
combination with ionised haemoglobin to give unionised haemoglobin,
i.e.
H++ Hb- ~ HHb
This reaction can proceed because reduced haemoglobin is a weaker
acid than oxygenated haemoglobin (due to a molecular rearrangement
consequent on 02 removal) and so is less ionised. It is clearly very
'useful' that the change in haemoglobin which occurs when 02 is given
up 'mops up' the H+ produced by an uptake of CO;.
Tissues
~
Blood
Tissues
Figure 2.20 Formation of bicarbonate from CO
2
in erythrocytes. For each I mmol/l
Ol '02 released 07 mmolll of COCand hence W-can be taken up by the Hb. A
small amount of bicarbonate is also produced in the plasma.
The bicarbonate produced as a result of CO
2
entering the
erythrocyte diffuses into the plasma in exchange for Cl- (the chloride
shift), so removing this reaction product from the cell. The whole
reaction by which CO
2
is taken up by the erythrocyte is speeded up
because of: (a) the enzyme carbonic anhydrase, present in erythrocytes
which speeds up C02 'hydration; and (b) the presence of a chloride/
bicarbonate carrier system in the erythrocyte membrane which allows
the chloride shift to occur rapidly. These factors ensure that the
reaction goes to completion in the short time the erythrocyte spends
in the tissue and lung capillary.
Carbon dioxide reacts with the terminal amine groups of
haemoglobin and other proteins to produce a carbamino compound.
Reduced haemoglobin again binds more CO
2
than oxyhaemoglobin.
The total amount of CO
2
bound as carbamino is not large, but 30%
of the carriage of CO
2
from the tissues to the lungs is by this method.
If the tissue respiration increases then there is a greater
concentration difference between tissues and blood and so more CO
2
RESPIRATORY SYSTEM
57
diffuses into, and 02 out of, the blood. This automatic adjustment of
the rates at which substances are transferred is an important
consequence of the laws of diffusion. It continues to operate so long
as the capillary blood flow is sufficiently great to keep the blood
CO
2
tension lower than the tissue CO
2
tension.
2.2 REGULATION OF RESPIRATION
2.2.1 Control System
Respiration occurs due to rhythmic discharge of the motor
neurones in the cervical and thoracic spinal cord which supply the
respiratory muscles. This rhythm of discharge is dependent on impulses
which come down from higher centres; neither the spinal cord, nor
the respiratory muscles in the chest, abdominal walls or in the
diaphragm have any inherent rhythmic activity.
Respiration is controlled by two separate but interacting neural
mechanisms: a voluntary system, located in the cerebral cortex, and
an automatic system, located in the pons and medulla. The voluntary
system is used in behavioural activities such as speaking, whereas
the automatic or involuntary system matches respiration to the
metabolic needs of the body. The voluntary system sends nerve
impulses to the spinal motor neurones via the corticospinal tracts,
whereas the automatic system sends impulses to the same neurones
via the reticulo-spinal tracts.
As with other activities which depend on opposing sets of muscles,
there is a reciprocal innervation which ensures that the inspiratory
muscles are inhibited when the expiratory ones are working, and vice
versa. These probably work via collaterals from excitatory pathways
which synapse on inhibitory interneurones, and then send impulses to
motor neurones supplying antagonistic muscles.
Brainstem celUres . . At one time it was thought that a discrete
respiratory 'centre' was located in the brains tern with separate
inspiratory and expiratory components. The current view, based on
further experiments, is that neurones with activity related to the
respiratory cycle are more diffusely located throughout the reticular
formation. We think it convenient to retain the term respiratory centre,
but with the new meaning of a diffuse region in the brainstem with a
respiratory rhythm. This rhythm, producing inspiratory and expiratory
is usually modified by various influences from the rest of
the body. The respiratory control system is thus rather like the
pacemaker region of the heart, it has an inherent rhythm which is
normally modified by external influences.
Lung
muscle
Higher centres
vascular -----...
joint
temperature
Intermittent
inspiration
Chemical
1<l4J----- influences
Figure 2.21 Factors altering the rhythmic activity of the brainstem respiratory centre.
Regulation of the respiratory centre-introductory outline. The
activity of the respiratory centre is altered by two kinds of stimuli,
chemical and non-chemical. The chemicals with this effect are the
respiratory gases, 02 and CO
2
, and H+ ion. A rise in its Pco
2
or H+
concentration or a drop in its P0
2
increases respiratory centre activity,
while changes in the opposite direction cause a decrease in activity.
These changes are mediated by two sets of chemoreceptors; situated
either in the medulla or peripherally. These chemical substances act as
mediators between the metabolic activity of the body and the respiratory
activity necessary to maintain the body fluids in a constant state. The
non-chemical influences are mainly afferent nerve inputs from various
other systems. These two kinds of stimuli will now be discussed in
more detail.
2.2.2 Non-chemical Influences on Respiration
Higher centres. The normal automatic pattern of breathing can
be interrupted by pathways from higher centres. This voluntary control
of respiration occurs, for example during speech and playing certain
musical instruments; in both of which lungs are used as air reservoirs
to be emptied at appropriate rates. In these activities there is a
surprising amount of modulation of the respiratory muscles during
stressing of words and musical notes. This kind of control is integrated
with the normal automatic control to keep alveolar Pc0
2
normal. Pain
and emotional events also affect respiration.
Proprioceptors. The main proprioceptor input is from the lungs.
In many animals a maintained inflation of the lungs delays the
appearance of the next inspiration, whereas a maintained deflation
increases the rate and depth of inspiratory efforts (the inflation and
deflation reflexes respectively). These two reflexes-the Hering-Breuer
reflexes-are mediated by vagal afferent fibres which run from stretch
receptors in the smooth muscle of the conducting airways to the brain
stem. In adult man there is some doubt as to its normal role in
ventilation, though it is known that afferent impulse patterns change
continually during the breathing cycle. In certain animals, in young
babies and in diseases which increase the stiffness of the lungs the
Hering-Breuer reflex is probably important.
Joint movements affect respiration. This occurs via the proprioc-
eptor input from joints and tendons and helps to control ventilation
during exercise.
Irritation of air passages. Irritation of these passages leads to
coughs and sneezes. Coughs are due to a large build up of air pressure
in the chest against a closed glottis, followed by sudden opening of
the glottis. The resultant blast of air (about 600 m.p.h.) helps to
dislodge the irritant.
Gut reflexes. During swallowing and vomiting material passing
along the oesophagus is prevented from entering the lungs by closure
and elevation of the glottis.
Spinal reflexes. Respiratory muscles, like other striated muscle,
contain muscle spindles which monitor the length of the muscle. If
an increased load, i.e. an increased resistance to air flow, is imposed
on respiratory muscles, the decreased shortening, i.e. smaller tidal
volume for the same effort, which occurs will be sensed by receptors
on the spindles, resulting in an increased excitation of the alpha motor
neurones and increased muscle contraction. Some control of ventilation
can thus occur at spinal level.
2.2.3 Chemical Control of Respiration
The chemical regulatory mechanisms adjust breathing so that the
Pc02 of the arterial blood is kept constant, the effects of excess H+
ions in the circulation are overcome, and the arterial P0
2
is increased
if it should fall markedly. The total ventilation is proportional to the
metabolic rate, but the link is CO
2
, not 02" A decrease in P0
2
causes
respiratory stimulation via the peripheral receptors, but acts centrally
to depress the respiratory centres, as it does on most neurones. An
increase in Pco
2
stimulates respiration both via the peripheral
chemoreceptors and centrally via the medullary receptors.
Chemoreceptor areas. The peripheral chemoreceptors are located
in the aortic and carotid bodies. They consist of glomus cells, arranged
in islands, surrounded by sinusoidal vessels. The blood supply per
unit of tissue is very large (2000 mli 100 g) compared to all other
tissues. This means that the oxygen needs of the cells can be met
largely by dissolved oxygen alone, without needing to draw on the
oxygen stored in the haemoglobin. It used to be thought that these
receptors were not much affected by decreases in 'chemically combined
oxygen', such as occurs in anaemia and carbon monoxide poisoning,
and only responded to dissolved oxygen, which is proportional to the
partial pressure of oxygen in blood. However, recent experiments have
cast doubt on this idea.
Glosso-
pharyngeal
nerve
Aorta
Aortic
body-__ -/
Figure 2.22 The chemoreceptor areas in the right side of the human neck with insets
showing the structure of the carotid body and their response to anoxia. These
chemoreceptor cells have motor nerve fibres running to them; at present the function
of these fibres is unclear.
The central chemoreceptors, situated on the surface of the medulla,
respond to changes in the [H+] in their immediate environment-the
extracellular fluid. This is readily influenced by changes in blood Pc02,
for CO
2
equilibrates rapidly with cerebral extracellular space and
cerebrospinal fluid. These fluids are less well buffered than blood,
so changes in Pco
z
produce larger changes in rH' J in brain fluids
than in blood. Changes in Peo
z
produce only a small direct effect on
these receptors in the absence of [H+] change.
The present view is that the initial response to a rise in Peo
z
is
through the peripheral chemoreceptors but that the major response is
via the hydrogen ion receptors of the medulla-both seem to depend
on CO
2
producing changes in the extracellular hydrogen ion
concentration at the receptors.
2.2.4 Ventilatory Response to Pure
Changes in COl' 01 or [H+]
If the Pco
2
of venous blood is raised (for example, by transfusing
blood containing a raised concentration of CO
2
), the alveolar Pco
z
rises and the respiratory 'centre' is stimulated, causing an increase in
ventilation which eliminates more CO
2
, This brings the alveolar and
arterial blood Pco
2
back towards normal. It does not quite return' to
the normal value but reaches a new equilibrium level near it. In this
way, a rise in venous Peo
2
is reflected in only small changes in
alveolar Peo
r
A decrease in Peo
2
works the other way in that it
decreases or inhibits respiration.
Alveolar PCOI
\ \
\ \
\ "
\ \.
\ '-
\ ...... - - - - - PA, CO
2
45mL
\
"-
- - - - - - - PA, CO
2
34ml
I I
I 40
60 80 100
Alveolar P
OI
Figure 2.23 Ventilatory responses to 02 and CO
2
changes from normal.
. When the alveolar P0
2
falls there is stimulation of ventilation but
this effect is very small until levels of about 60 mmHg are reached,
when it becomes marked. This non-linearity of response to changes
in P0
2
is in marked contrast to the linear response to changes in Peo
r
The reasons for the small response to changes in P0
2
down to 60
mmHg are not clear; probably the chemoreceptors show a similar
non-linear response. An increase in P0
2
has little effect on respiration.
If the blood pH falls for metabolic reasons then ventilation
increases. This is mediated by the peripheral receptors only because
hydrogen ions do not enter the CSF readily. This mechanism is
effective around the normal body pH of 74.
2.2.5 Ventilatory Response to Combined
2
, CO
2
and [H+] Changes
If the alveolar concentrations of both 02 and CO
2
change, the
ventilatory changes which result are complex. The CO
2
response curve
becomes more steep as POI decreases and minor degrees of hypoxia
now stimulate respiration in the presence of increased Peo
r
Therefore,
when both gases change the resultant change in ventilation is greater
than the sum of each separate change. This complex response
presumably matches ventilation:to metabolism more precisely during
periods of very vigorous metabolism, for example exercise, than would
otherwise be the case.
Changes in the H+ ion concentration of blood due to metabolic
causes, on the other hand, produce effects which are simply additive
with Pc0
2
changes, i.e. the slope o! the Pc0
2
curve does not alter.
2.2.6 Non-respiratory Features of
Pulmonary Circulation
Apart from its main function of gas exchange, the pulmonary
circulatory system serves several other important functions within the
cardiovascular system.
1. At any instant, the pulmonary vessels contain about I litre
of blood, of which only 100 ml are within the capillaries.
This volume of blood within the vessels can vary under
different conditions, increasing, for example, in changing
from the upright to the horizontal position. It is thought to
be one of the important reservoirs for blood in the circulatory
system, from which blood is immediately available to the
left ventricle for increasing the cardiac output.
2. The pulmonary circulation acts as a fIlter for small thrombi,
preventing them from reaching the systemic circulation and
so obstructing the blood supply to other vital tissues.
3. Various constituents of blood are modified in their passage
through the lung. As the total cardiac output passes through
the capillary bed every minute, and as the blood volume is
approximately equal to the cardiac output at rest, all of the
blood is exposed to this large endothelial surface in this time.
Vaso-active substances such as bradykinin, 5-hydroxytryp-
tamine, prostaglandins, histamine and noradrenaline may be
taken up or released from the lung. One example of this type
of activity is the conversion of the polypeptide angiotensin I
to angiotension II by a converting enzyme from lung
epithelium.
4. The large number of mast cells in lung containing heparin
are now thought to play an important part in the clotting
mechanisms of blood.
2.2.7 Nutritional Blood Supply to the Lungs
The lungs, like any other organ, require a nutritional blood supply.
This is carried by the bronchial artery which arises from the aorta,
supplies the needs of the tissue in the bronchial tree, and then largely
drains back into the left atrium via pulmonary veins. This flow is
about 1 % of the total blood flow to the lungs; when this venous blood
is mixed with the oxygenated blood from the alveolar capillaries it
causes a small drop in the P0
2
of the arterial blood. These blood
vessels are systemic ones and so respond to hormones and circulating
metabolites like systemic vessels, rather than like pulmonary vessels.
3
Blood and
Body Fluids
Each cell in the body carries out, within itself, all the chemical
processes necessary for its own maintenance. Therefore all the
substances necessary for this must be carried to, and all the waste
products taken away from, each cell. The system which does this-the
cardiovascular system-transports materials in a closed system of tubes
around the body. Exchanges between the cells and this system occur
from the capillariesthe smallest blood vessels-which lie within a few
thousandths of a millimetre of all body cells.
The main materials which cells require are fats, amino acids,
sugar, vitamins, salts, water, oxygen and hormones. These substances
are carried to the cells by the blood. The waste products of cells are
carbon dioxide and simple soluble compounds of nitrogen such as
urea-which are transported to the lungs or kidneys for excretion.
In the higher animals the blood consists of a fluid, the plasma.
in which cells of various kinds are suspended, each adapted for special
purposes. The blood volume is about 8% of the body volume (or 52
1
litres in a 70 kg man); of this, about 55% is plasma.
3.1 CELLS OF THE BLOOD
The cells of the blood do not originate id the blood; they are
made in organs outside the blood and m e r t ~ y use it as an avenue
while performing their functions, or while ir transit from one area to
another. There are three kinds of such cell.: erythrocytes (red cells),
leu(;ocytes (white cells) and thrombocyte', (platelets-these are really
parts of cells). '
The plesent view is that these three cell types are derived from
64
BLOOD AND BODY FLUIDS 65
a common precursor (stem cell) which originates in the bone marrow;
an organ scattered (in adults) in the flat bones of the skull, the ribs,
the breast bone (sternum) and parts of the hip, and with a total size
similar to the liver. The red cells, platelets and most of the white
cells are made in the bone marrow; the lymphocytes originate both
in the bone marrow and the thymus and multiply in the lymph nodes.
- The monocytes, made in the bone marrow subsequently invade the
lymphoreticular system (reticuloendothelial system).
3.1.1 Erythrocytes
The primary function of the erythrocytes is to carry oxygen; a
function done by the haemoglobin within them. They are thus container
cells continuously circulating in the vascular system, their other
properties being adapted to this rigidly defined role. Each cell in man
is a small non-nucleated biconcave disc (75 pm x 2 pm); there is a
total of about 3 x 1013 cells in the body containing 1 kg of
haemoglobin. The average life span of a cell is 120 days. It has been
calculated that the biconcave shape is the best for allowing diffusion
of oxygen into the cell, but in the small capillaries in which exchange
occurs the cells are often very distorted by the narrowness of the
vessels.
3.1.2 Erythrocyte Production and Destruction
Erythrocyte production (erythropoiesis) occurs in bone marrow,
starting with the stem cell and progressing through various intennediates
to the reticulocyte, which then matures into the erythrocyte in blood.
Normal development takes some 7-10 days. At the end of their lives
the erythrocytes are destroyed by the scavenger cells of the liver
(macrophages or Kiipffer cells) and many of the constituent parts
recycled.

(1 '\
Erythrocyte Granulocytes Lymphocytes Monocyte Platelets
i- --l---- -- - --- ----
Figure 3.1 Originn of the blood cells.
The rate of prodm.:tion of erythrocytes by the bone marrow is
controlled by a feedback system which seems to depend on the level
of the oxygen supply to lhe kidney. If the oxygt"Il supply is decreased-
by anaemia or hypoxia-then more erythrocytes are produced; if it is
increased-by an increase in erythrocytes or high oxygen tension-then
less erythrocytes are produced. The receptors for this effect are
unknown.
The current view of the process is that the kidney releases
erythrogenin, which acts on a plasma globulin, converting it to
erythropoietin; this then controls the number of erythrocytes which
starts to maturate. This view of the control system is over-simplified
and not yet well established.
3.1.3 Haemolysis of Erythrocytes
The size of erythrocytes, like other animal cells, depends on the
osmotic pressure of the solution in which they are bathed. If put into
hypertonic solutions then water leaves them, they shrink and take a
'crenated' appearance. If put into hypotonic solution, water enters
them causing swelling. They thus act, within certain limits, as
osmometers. When erythrocytes are exposed to a range of hypotonic
salt solutions it is found that at each osmotic pressure a certain
proportion of the cells burst, with consequent release of haemoglobin.
Erythrocytes Stem
7
Erythropoietin
""1-
Globulin
Figure 3.2 Control of the production of erythrocytes by the bone marrow. A decrease
in the 02 supply to the kidney (for example due to a low number of erythrocytes)
causes an increase in the production of erythrogenin, etc. and ultimately more
erythrocytes.
A haemolysis curve may thus be obtained for normal cells which
provides an index of the 'fragility' of the cells. In some disease
conditions this fragility is increased.
3.1.4 The blood pigments
Haemoglobin is the pigment found in erythrocytes and is
responsible for their large capacity to carry oxygen. It has a molecular
weight of some 65,000 and is made up of four subunits, each
consisting of a pigment part (haem) joined to a polypeptide part
(globin). The haem part of the molecule is an iron-containing porphyrin
derivative, which binds reversibly to Or The haem molecule is
suspended between two residues of histidine in the polypeptide chain
by links to the central Fe
2
+ atom; one of these links is direct, the
other can be broken so that 02 can be inserted when oxyhaemoglobin
is tormed.
Various different haemoglobins exist. Ultimately these differences
are due to differences in the amino-acid sequences of their constituent
polypeptide chains; but these differences are often restricted to two of
the four polypeptide chains in each haemoglobin molecule. Thus it is
convenient to think of the four polypeptides in each molecule of
haemoglobin as consisting of two pairs. One pair of polypeptides-the
a chains containing 141 amino acids-is constant for nearly all
haemoglobins; variations in the other pair give rise to different
haemoglobins. In normal adult haemoglobin (haemoglobin A) the second
pair of polypeptides are called the (3 chains. When the four parts of
haemoglobin A are put together to fonn the complete molecule there
is a large area of contact between an ex and (3 chain with little contact
between each ex or each (3, so that the molecule exists functionally as
two subunits, each of an ex and (3 chain. On oxygenation these two
halves come together. Thus oxygenation leads to changes in the shape
of the whole molecule; this 'explains' the sigmoid shape of the oxygen
dissociation curve and the buffering power of haemoglobin.
Foetal haemoglobin (haemoglobin F) has Y chains instead of 13
chains, the changeover to haemoglobin A occurring in the first three
months of extra-uterine life; the y chain contains the same number of
amino acids as the 13 chain but 37 are different. The substance 2,3-
diphosphoglycerate. (2,3-DPG), normally present in erythrocytes,
competes with 02 for binding to deoxygenated haemoglobin. Foetal
haemoglobin binds 2,3-DPG less strongly than adult haemoglobin and
so binds more 02 at any given partial pressure of oxygen. Therefore
foetal haemoglobin can receive 02 from the maternal haemoglobin at
the same partial pressure of oxygen.
3.1.4 Abnormal Haemoglobins
The amino-acid sequences of the polypeptide chains of haemoglohin
are, of course, genetically determined. Amino-acid sequence analysis
shows that many haemoglobins exist; these are identified by letter as,
for exan1ple, haemoglobin A (normal), F (foetal), S, J, C, etc. The
differences may be very slight but the consequences very great; the
a chain
(al
Figure 3.3 (a) Outline structure of the a subunit of the haemoglobin molecule showing
the 'pocket' into which the oxygen atom must pass in order to bind with the ferrous
iron. Oxygenation can only occur between the Fee+ and one of the histidines of the
backbone. (b) Three-dimensional structure of two subunits showing the f1at-<lisc shape
of the haem part of the molecule. The shape of the protein moiety varies with the
state of oxygenation
change of one amino acid for another at a critical site leads to a
change in the threedimensional organisation of the haemoglobin with
marked consequences on affinity for 02' physical properties, etc. In
haemoglobin S, for example, substitution of valine for glutamic acid
gives a haemoglobin which has a low solubility at low oxygen tensions,
a circumstance which causes the erythrocytes to become sickle shaped
and makes them more liable to haemolysis. Haemoglobin S persists in
some 40% of the Negro population of Africa because it confers
resistance to one type of malaria, although it gives a severe anaemia.
Most other abnormal haemoglobins are harmless, presumably because
the altered amino acids are present in non-critical parts of the
polypeptide chain.
Carboxyhaemoglobin is a combination of haemoglobin with carbon
monoxide instead of 02" As the affinity of haemoglobin for CO is
much greater than that for O
2
, CO is taken up preferentially so
reducing the 02 carrying capacity of the blood.
Methaemoglobin is the pigment formed when the ferrous iron (Fe
2
+)
is converted to the ferric state (Fe3+). This occurs in the presence of
various drugs and oxidising agents (for example, nicotine), but also
normally at a slow rate. The normal concentration of methaemoglobin
is kept low by an enzyme system (NADH-methaemoglobin reductase)
which converts it back to haemoglobin; for the reaction is not reversible
otherwise.
3.1.5 Synthesis and Destruction of Haemoglobin
Haemoglobin is manufactured by the developing erythrocytes
within the bone marrow. Before losing its nucleus the erythrocyte is
much like any other cell in that it can synthesise RNA and lipid in
addition to haem and proteins. After maturing to the non-nucleated
form metabolism becomes very low, consistent with its container ,
cell function.
3.1.6 Substances Essential to Erytbropoieses
Protein. Amino acids are required for the globin part of the
haemoglobin. When the supply of amino acids is limited, as in
starvation, haemoglobin manufacture seems to have priority so that
severe anaemia (Hg < 8 g/dl) rarely occurs in starvation.
Iron. Iron is required for erythrocyte formation because of its
essential place in the haemoglobin molecule, if lacking an iron-
deficiency anaemia (Hb < normal) occurs. Normally very little iron is
lost from the body, so that only about 1 mg/day is required for
70 PHYSIOLOGY, BIOCHEMIS1RY AND BIOTECHNOLOGY
replacement purposes. A normal diet provides much more than this,
thus more may be absorbed if required. It is known that uptake from
the gut can occur against a concentration gradient and is controlled
by the needs of the body. Iron metabolism therefore works as 3 closed
system in that none is excreted, and so the body must have an efficient
sytem of control of intake for once absorbed it cannot be excreted.
The mechanism for control of uptake is, at present. not understood.
12-I'5mg
daily
, '
I",
I,'
'un.
"
,11'
Img/day
Stored as ferritin
in mocrophoges In
these tissues (20"10)

'j)-- ,
-; -< Liver
.,
Spleen
C::::=::::=J
Bone morrow
,/)'
,-------,
Blood
transferrin
<0'1%

.-------.,
MYOQlobin
5%
1t
Haemoglobin
75%
Figure 3.4 a. Iron metabolism; iron is taken up by an active process in the small
intestine and normally only lost when cells are desquamated from the skin (of
menstruation in the female). The uptake procC'ss is 'acti\le' and controlled b:v bOlHy
needs. Only ferrous iron absorbed; the current role of hydrochloric acid in
iron uPtdke is uncertain. There is a total of 34 g iron in the body.
3.1. 7 Vitamin 872 and Folic Acid
These substances are essential for the normal maturation of
erythrocytes; in their absence the maturation of the nucleus of the
erythrocyte occurs more slowly than the cytoplasm. leading to cells
which are iarger in size and fewer in number than norma!.
Like other vitamins neither substance is made in the body,
although B2 is stored in large amounts in our liver. In the absorption
of B\2 an intrinsic factor produced by the stomach is essential.
BLOOD AND BODY FLUIDS
Green
leafy I
vegetables
(folate)
Ipg/day
Micro -organisms
to
Animal
proteins
(B
I2
)
1
___ + Folate----..... Blood
\
Bone morrow
50pg/doy
7l
Figure 3.4 b. B'2 and folate metabolism; BI2 requires an 'intrinsic factor' for ansorption
to occur; once absorbed it is stored in the liver (1 year's supply). Both are concerned
with the metabolism of one-carbon radicals concerned in transmethylation.
DIET
Amino aci 5
Some iron loss
Figure 3.4 c. Erythrocyte formation and distribution (after Ganong).
3.1.8 Death of erythrocytes, degradation of
haemoglobin and metabolism of bile pigments
The precise mechanism of erythrocyte death is not yet clear-ideas
ranging from sudden haemolysis to gradual fragmentation have been
suggested-but eventually they are cleared from the circulation by the
mononuclear phagocytic system. These phagocytes split off globin,
the haem molecule is split open and the iron removed for recycling.
The residual pigment is biliverdin which, in man, is converted to
bilirubin and then transported (bound to albumin) to the liver where
it is made water soluble and excreted via the bile. In the gut the bile
pigments are further converted to stercobilinogen which colours the
faeces.
3.1.9 Blood Groups
The erythrocytes of all races have an identical appearance. Their
membranes, however, contain a variety of antigens-sometimes called
agglutinogens-which vary in type in different individuals and to some
extent in different races. When these antigens combine with the
appropriate antibodies-called agglutinins-the cells become sticky and
clump together like bunches of grapes (agglutination). The different
antigens give rise to the different blood groups. The common blood
groups, A, B, AB and 0 are due to the presence, in various
combinations, of two antigenic determinates A and B. Thus group A
has the antigen A present on its surface, group B the antigen B on
its surface, group AB has both and 0 has neither present. These
antigenic specificities are due to the carbohydrate chains of glycolipids
and result from the action of a series of glycosyltransferase enzymes
determined by genes at a number of loci. Type A has the sugar residue
acetylgalactosamine as part of its skeleton, whereas Type B has
galactose.
It is normally found that the absence of A or B antigen in a
person's erythrocytes is always associated with the presence of the
appropriate antibody in their serum. Thus the absence of A in the
erythrocyte means that anti-A or a-antibody is present in the plasma.
The reasons for this seem obscure but it can be remembered that for
the ABO system there are always antibodies to A and B present unless
their appearance is inhibited by the presence of A and B antigens in
the cell membrane. The practical importance of this is that cells from
one person transfused into another must be 'matched', so that they
are not agglutinated by the antibodies present in the recipient's plasma.
If agglutination does occur, then haemoglobin is released causing
jaundice in severe cases. The donor's serum has little effect on the
recipient's cells because of the large dilution which takes place. The
effects of transfusion are tested for directly, by testing the donor's
cells on the recipient's serum before transfusion (,cross-matching').
Blood group Red cells
(antigens I
A
0
8
o
Plasma
(ant ibodyl
=-=
anti -8

anti-A
Frequency
(%1
(UK and USA)
40
10
5
45
Figure 3.S Diagram of the antigens and antibodies found in the erythrocytes and plasma
in the ABO blood-group system. There are several subgroups of A and B. The most
important are the Al and A2; about 20% of group A people belong to A2 which has
a much weaker antigen than AI.
These antigens are also found on the surface of other body cells,
for example liver, lungs, salivary glands, sperm, and in the amniotic
fluid. There are also other antigenic molecules present on the surface
of erythrocytes and other cells, so that probably each of us could be
identified by our cellular surfaces. The most important of these
antigens is the Rh system; discovered when it was shown that a serum
prepared against rhesus (Rh) monkey erythrocytes also agglutinated
the erythrocytes of most humans.
The Rh system of antigens is complex as it is really a group of
many antigens. The most important quantitatively is called the D
antigen and this is the one usually tested for. Thus Rh-negative
individuals have no D antigen and normally no D antibody in their
plasma. Different races have slightly different proportions of Rh
___ Antibody
Figure 3.6 a Agglutination occurs by cross-linking of many red cells with serum
antibodies. In transfusion the important reaction to avoid is the clumping of a donor's
erythrocytes by a recipient'S antibodies.
positive, for example, Caucasians 85%, Orientals 99%. The Rh factor
is important in transfusion, but it is also of great importance in
pregnancy when an Rh-negative mother has an Rh-positive foetus.
Occasionally foetal cells enter the maternal circulation during
childbirth. The Rh-positive foetal cells induce antibodies in the mother
so that subsequent Rh-positive children will have their red cells
haemolysed by antibodies from the mother, leadmg to haemolytic
disease of the newborn. Usually first children are not affected and
(for some unknown reason) only a small percentage of those who
mip,ht be affected are, in fact, so affected. Current practice is to kill
the foetal cells which enter the maternal circulation before they can
induce antibodies. This is done by injecting the mother with Rh-positive
antibody within the first 36 hours after childbirth.
The blood groups are inherited according to Mendelian laws. In
a given person the blood group is a complex (phenotype) of the
mother's and father's blood-grouping genes. For example, a person
Attachment Engulfment Endocytosis
Figure 3.6 b Phagocytosis hy engulfment and membrane flow-this occurs in
granulocytes and monocytes.
with type B blood may have inherited a B antigen from each parent
or a B antigen from one parent and an 0 from the other; an individual
of group B may therefore be B x B or B x 0, i.e. may be
homozygous or heterozygous. Studies of blood-group inheritance are
important in tracing the migration of populations and in paternity cases,
3.1.10 The Leucocytes
The white (i.e. colourless) cells of the body are one of the main
agents in the body's defence against infection. They do this in two
ways: either by scavenging (phagocytosis), which is the process of
ingestion and retention of foreign matter, or by the production and
distribution of antibodies, molecules which combine with foreign
molecules to inactivate them. White cells can move between body
tissues and the bloodstream; at anyone time only a small fraction of
the total (say 5%) are present in the blood, so that very large increases
in the blood concentration of granulocytes can occur by mobilisation
of cells from the bone marrow. They can be thought of as policemen
most of whom are stationed at various parts of the body, but some
of whom patrol the bloodstream ready to deal with blood-borne
infections and ready to rush to sites where required. The main classes
of white cells are the granulocyte series (neutrophils, eosinophils and
basophils: distinguished by their granules), the monocytes and the
lymphocytes. The blood contains about 10,000 white cells per yl so
that there is only one white cell per 500 erythrocytes.
The granulocyte series of leucocytes are produced in the bone
marrow from stem cells and remain there in store. They then enter
the circulation for a few hours and finally reach the tissues, from
which they do not return. They are expended performing body-defence
functions; eventually being destroyed by the tissue macrophages or
by passing into the lungs, saliva or gut. There are not usually many
granulocytes in the tissues, except in acute infections.
The neutrophils in the blood-which are active phagocytes-are the
body's first line of defence against bacterial invasion. Their numbers
in the blood increase rapidly in such infections due to mobilisation
from the marrow pool; this is used diagnostically. The eosinophils are
rather more common in the tissues than in the blood but are not normally
very numerous; they are thought to be concerned with parasitic infection
and the degradation and removal of protein. Their numbers in the
blood incre'ase greatly in allergic conditions when the amount of antigen-
antibody complex increases. The functions of the basophils are unclear;
they contain the substances 5HT, histamine and heparin.
76
Cell type
10fL
m
[
Origin and
life cycle
Life
Function
PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
Granulocyte series Monocytes Lymphocytes

i'

.,,;,,"'''
Small
(7f'ml
Neutrophil
Monocyte
(20f'm)

-..
Basophil
Large
Polymorphs
(14fLml

Lymphocytes
Stem cell Stem cell
Stem cell

Bone marrow
Circulation Circulotion
pool
! !
Circulation
Fixed in Lymph nodes

sinusoids of
Tissues
liver, spleen
Short lived, say'
I week of which 6 hours ---Long lived ----.
spent in blood ? 100 days
_ Antibody production_
...---- .. ...--- and distribution
Neutrophils - bacteria
Eosinophils - proteins
Antigen - antibody
complexes
Figure 3.7 Appearance, life style and function of the white cells.
The monocytes are produced in the bone marrow from stem cells
and migrate via the blood to the tissues where they become the tissue
macrophages. In the tissues they may divide to increase their numbers.
They act as scavengers, retaining some of the breakdown products
and help in the immune response. Their lifespan and eventual fate
are unknown.
The lymphocytes originate from stem cells in the bone marrow.
They migrate via the blood to the lymph nodes where they perform
their immunological role. In blood about 85 % are small (5-10 pm) in
77
diameter) and the rest medium (15 pm). Small lymphocytes are the
final stage in the series and they circulate via the lymphatic system.
Large lymphocytes do not recirculate but can proliferate rapidly to
form the small lymphocytes.
3.2 THE L YMPHORETICULAR ORGANS
The body, like a city, needs a disposal system which will cope
with waste particles. This waste matter may arise from within the
body itself-as worn-out cells (mainly erythrocytes) and macromolecular
complexesor by invasion from outside-as dust or bacteria. Disposal
works by a system of scavenger cells which eat up these particles
and then either concentrates them or breaks them down (phagocytosis).
Figure 3.8 Sttucture of the wall of a sinusoid showing the fenestrated layer of flattened
endothelial cells, with interspersed scavenger cells (shaded). These cell types are difficult
to identify histologically; but the endothelial cells have a basement membrane visible
under electron microscopy.
If the waste materials consist of foreign proteins then the body produces
antibodies specific for each protein; these antibodies react with them
to produce a complex, with subsequent precipitation and phagocytosis.
The scavenger cells are either free in the blood or tissues and can
'hunt' the particles, or are fixed in various organs and 'seize' the
particles as they pass. The fixed scavenger cells are found in a system
of lymphoreticular organs which consist of thin-walled supporting tubes
with the phagocyte cells in their walls. Blood and lymph are passed
along these tubes, allowing the phagocytes to act on the suspended
particles. The lymphoreticular organs contain lymph tissue as well as
phagocytes. These organs include lymph nodes, spleen, thymus, bone
marrow and the alimentary tract. The proportion of lymphoid tissue to
that of structural and phagocyte cells varies in these different tissues.
In the liver a similar system of phagocytic cells exists with no lymphoid
tissue.
Both fixed and wandering phagocytes in tissues are part of the
same system as the monocytes and so originate in the bone marrow.
Their tissue functions are:
1. Macrophages of the spleen and bone marrow remove wom-
out cells, bacteria and macromolecules such as fibrin and
antigen-antibody complexes.
2. Kupffer cells of the liver handle important breakdown
products of the above, such as iron for recycling and ingest
chylornicrons from the portal blood.
3. To co-operate with the lymphoid system to produce immunity
to foreign proteins of bacteria, etc.
3.2.1 Lymphocytes and Immunity
Protection against infection is produced by both immediate and
longer term measures. The immediate response is phagocytosis, the
longer term response (2-3 weeks) is the production of antibodies to
the infecting agents. A common way for this to happen is that foreign
material is captured and processed by the macrophages and then
reactive changes made in the nearby lymphocytes. The close
association of both phagocytosis and lymphocyte production in the
same organ greatly facilitates this process.
Lymphocytes recirculate from the blood through the lymph nodes,
the lymph and thoracic duct, back to the blood. In the spleen they
circulate through the white pulp back into the sinusoids. The
lymphocyte population of the spleen and lymph nodes are thus changed
every few hours. The process of recirculation is important, for it
distributes immunologically competent cells round the body and ensures
their adequate exposure to antigen trapped by the scavenger cells.
The immune response is the protective reaction to a specific
protein (called an antigen). It is so specitic that it can be used as the
basis of the assay of hormones and other substances in biology. The
phagocyte response, on the other hand, is non-specific and can be
evoked by many antigens, although the body has various ways of
Superior vena cova
Ii- - 1
Thoracic .
Postcapillary

Cortex
Lymph node
_-;-_Lungs
heart
+-___ Aorta
Fip", 3.9 Circulation of lymphocytes. Both B- and T-Iymphocytes travel to the
Iyrnpho .. ytes of th" cortex (B areas) paracortex (T areas).
makmg antigens 'tasty' to the phagocyte. On first exposure to an
antigen it takes 2-3 weeks for a specific immunity to arise (the primary
response); on subsequent exposure to the same antigen, the
inununological memory laid down previously can be called upon to
give a large, quick, second response.
3.2.2 Two Populations of Lymphocytes: T - and B-cells
Two types of specific immunological responses are set up by the
body, either cell-mediated or humorally mediated; t.hese are caused
by two small lymphocyte popUlations, the T- and B-cells, respectively.
On appropriate stimulation by antigen both populations proliferate and
undergo morphological changes as follows.
The T-lymphocytes regress to lymphoblasts; cells which have litle
rough endoplasmic reticulum but many free ribosomes. They have
'receptorlike' molecules on their surface for binding antigen. They
do not secrete antibodies. They act as 'killer' cells which attack
foreign grafts, etc.
Bone marrow
~ stem cells -Al\
~ ~ ~ ~ ~ . ..
Thymus
processing
T -lymphocyte
Cell- mediated
reactions
"Bursa"
pracessin9
B -lymphocyte
Humoral antibody
synthesis
Figure 3.10 Processing of bone-marrow cells by thymus and gut-associated lymphoid
tissue to become immunocompetent T- and B-lyrnphocytes, respectively. Proliferation
and transformation to cells of the lymphoblast and plasmacell series occurs on antigenic
stimulation.
The B-lymphocytes develop into plasma cells which have a well-
marked rough endoplasmic reticulum capable of producing and
'exporting' antibodies into blood and tissue fluids. The B-Iymphocytes
often require the presence of T-Iymphocytes in order to be triggered
by antigens; it seems that the receptor properties of the T-Iymphocytes
are required for antigen recognition.
3.2.3 The Platelets
81
The platelets are produced from stem cells via megakaryocytes;
these are large (200 p,m) cells lying outside the sinusoids of the bone
marrow. The megakaryocytes push into the blood through
GI
Exterior

Unit
membrone
Microfilaments
(condensed in the
sub membranous
region)
Golgi zone
_-"" .... A .j\"li ........... "'Wi''--- Dense
Dense body
in granule
body

tubular
system
Figure 3.11 Outline and more detailed sttucture of platelets showing biconcave form
and equatorial arrangement of microtubules.
the walls of the sinusoids; these processes break off to form the tiny
platelets. The number of circulating platelets (about 0-3 x 1()6/p,1) is
probably controlled by a hormone, thrombopoietin. The platelets are
rich in enzymes and contain large amounts of ATP, 5HT and
phospholipids. Their life span is about 10 days. Their main function
is in the clotting mechanism. Normally it is thought that minute clots
are continuously being formed and destroyed. This is probably the
main way platelets die, but 'old age' may also occur.
3.3 PLASMA
The plasma is a clear, straw-coloured fluid, except after a fatty
meal when it is milky in appearance due to the presence of fatty
globules. It is rather like raw egg-white diluted with a salt solution;
its main constituents are protein (10%) and sodium chloride (09%).
The high water content (90%) it contains dissolves the other substances
and allows the blood to pass easily through the capillaries. Plasma
lefl in a test tube clots, unless substances are added to prevent it.
When whole blood is allowed to clot the fluid remaining is serum,
which has a similar composition to plasma without the clotting agents
(fibrinogen and various factors).
3.3.1 Salts
The main salt in plasma is sodium chloride (NaCI), with smaller
amounts of NaHC0
3
, KCI, CaCI
2
, MgCI, etc. In addition to their
specific functions mentioned elsewhere, such as action potential
mechanisms, sodium pumps, etc . these salts act as a protein solvent.
allowing the plasma proteins to be carried in solution. The concentration
of sodium chloride and the other salts in plasma is regulated within
narrow limits by intake and excretory mechanisms.
Bluod is very well buffered at a normal pH of 7-4. This is
ach:eved mainly by the interaction of CO
2
with the sodium bicarbonate
in the plasma and erythrocytes.
3.3.2 Plasma Proteins
Traditionally the plasma proteins were fractionated into three
groups: albumin. globulin and fibrinogen; the first two by salt
precipitation and the last because it could be readily identified in the
clutting mechanism. Modern methods of separation have demonstrated
the presence of a multitude of different proteins. Quantitatively the
most important is albumin. with the various globulin fractions (0.1, 0.2,
13 and immuno-) contributing much smaller amounts. Most are made
iT} the hepatocytes of the liver; but the antibodies-inununoglobulins-are
produced by the cells of the lymphOId system. About 10% of the
plasma albumin is replaced per day. The p l ~ . m a proteins have various
functions, summarised here and considered elsewhere: (1) they provide
an osmotic pressure of about 25 mmHg '.cross the capillary wall. for
they do not readily cross into the inte .stitial fluid. and so-with the
blood pressuredetermine fluid distribul ton between blood and the rest
of the extracellular compartment; (2) they provide about 15% of the
total buffering capacity of the blood, due to the weak ionisation of
their constituent COOH and NH2 groups; (3) their specific functions
include the clotting mechanism of the fibrinogen fraction. the carrier
mechanism of the binding proteins which provides a reservoir of
hormones (e.g. thyroid) in the blood and prevents their rapid loss
through the kidneys, the carriage of antibodies to the tissues as the
globulins and the carriage of metals, ions, fatty acids, amino acids,
etc. by albumin.
(0)
,10nm,

No", CI-, glucose
Albumin
69,000
-
Haemoglobin
68,000

y-Globulin
900,000
Fibrinogen
350,000
Figure 3.12 (a) The relative dimensions and molecular weights of haemoglobin and
some of the plasma proteins.
3.3.3 lIaennostasis
If a small blood vessel is cut a repair mechanism is activated
which seals off the cut; tllis process involves the conversion of
fibrinogen to fibrin in the blood which forms a clot at the site of the
injury. The clot then forms a scaffold on which new tissue is built.
The series of events-haemostasis-which leads to this final result
(bl
Albumin
Globulin fractions
Cathode Anode
Point of application of sample
Figure 3.12 (b) Densitometer tracing of an electrophoretic separation carried out on
cellulose acetate with a specimen of normal serum and stained to show the presence
of protein.
involves several stages: (1) constriction of the blood vessel to narrow
the lumen; (2) formation of the plug of platelets; and (3) the conversion
of this plug into a clot of fibrin.
The trigger for haemostasis is an injury to the endothelium of
the blood vessel, exposing the underlying layer of collagen. Platelets
in the blood stick to this exposed collagen and secrete granules
containing 5-HT, adrenaline, ADP and several clotting factors. The
5-HT and adrenaline cause the blood vessel to constrict, reducing the
blood escape, particularly if the cut is 'clean' rather than irregular.
The ADP released by the first few platelets make others sticky so
that they form a loose plug of aggregated platelets in the vessel. This
loose plug is bound together and converted into a clot by fibrin. Fibrin
is a long chain protein molecule composed .of many subunits; it is
rather like a long necklace made up of beads. In its manufacture
each 'bead' is derived from tibrinogen molecule by splitting off a
few polypeptides; the residues are then joined together to form fibrin.
The mixture of platelets an fibrin molecules is initially loose network
of strands, but is soon stabilised by covalent cross-linkages to form a
tight, a strong clot.
Damaged
vessel
_____ ..... Release
thromboplastin
+
Expases
collagen
1
Platelets
stick to
collagen,
release
factors
1
(R
d
5tHT}-+contraction
eucase.
ADP -+ More -+ Loose
platelets plug
1
+-__ Fibrin laid down
forms true clot
Figure 3.13 Steps in the formation of a clot in a damaged vessel.
Extr:nsic
path
Thrombin
The conversion of tibrinogen to fibrin is catalysed by thrombin
which in turn is formed in the plasma from its circulating precursor.
prothrombin. This latter conversion depends on the presence of the
activated form of factors X and V, lipids and Ca", The formation of
the active form of factor X can occur either by an intrinsic pathway,
which probably starts with activation of factor XII by collagen and
proceeds by a pathway involving lipids released by platelets, or an
extrinsic pathway which depends on the release of a substance-
thromboplastin-from the damaged walls of the blood vessel.
3.3.4 Anti-clotting Mechanisms
Blood does not normally clot in undamaged blood vessels of the
body. This is partly due to mechanisms which tend to stop clotting
and partly to mechanisms which break up any clots which do form.
The former mechanisms include: (I) the removal of activated factors
by the liver; (2) the presence in circulating blood of heparin, an anti-
Collagen
+ platelets Tissue damage
Fibrinogen
:lZ:1 Ca+
2
------..... Thrombin ---.... 1
Prothrombin
Fibrin (loose)
lxm
Fibrin (tight)
Figure 3.14 Proposed pathways and reactions involved in the activation of fibrinogen
t" fibrin.
c0agulant which stops the activation of one of the factors (IX) in the
intrinsic pathway and which, with a plasma cofactor, inhibits the action
of and (3) factors which reduce the adhesiveness of platelets,
1>0 that their 'stickiness' at any time is detemlined by a balance
between those which increase and those which decrease it. Once a
clot is formed spread is limited because it releases anti-clotting factors.
Clots are removed by a fibrinolytic system, which degrades fibrin
and fibrinogen to products which themselves inhibit thrombin. The
active component of this system is fibrinolysin (plasmin) which is
formed from profibrinolysin (plasminogen) by the action of thrombin
and other substances. Fibrin is therefore made and broken down by
systems which depend on the activation of thrombin; normally a balance
exists with little clot formation. During injury the reactions seem to be
biased to favour clot formation. The coagulation mechanism therefore
represents a delicate balance between many factors, the specific
functions of which are still poorly understood. The current view is
that these sequences of events, platelet aggregation, coagulation and
then fibrinolysis. occur continuously at a multitude of sites throughout
the body where small damage has occurred due to daily traumas.
87
~
Fibrinogen
~ ~ - - - ~
.to
Fibrinolysin (plasmin)
Figure 3.15 Fibrin is formed from fibrinogen subunits by the actions of thrombin;
fibrin is broken down into its degradation products by fibrinolysin (plasmin). The amount
of fibrin present at any onetime depends on the balance between these processes.
In medical practice clotting in blood vessels does sometimes
happen, for reasons which are at present poorly understood. Such
thrombosis tends to occur in blood vessels damaged by arteriosclerotic
plaques (e.g. coronary vessels) and in vessels with a sluggish blood
flow (perhaps due to a build up of activated clotting factors). Parts
of these thrombi may break off (emboli) and lodge further down the
vascular tree. Abnormalities of clotting also occur, due to a variety
of causes often genetic in origin. For example, congenital absence of
factor VIII is classical haemophilia, a sex-linked disorder, and of factor
IX is Christmas disease.
Blood may be prevented from clotting in test tubes, etc. by
removal of Ca2+l, thus preventing the formation of thrombin from
prothrombin (this explains non-clotting of menstrual blood). Common
methods are to add oxalates, which forms an insoluble salt with Ca
2
+,
or to add citrate or other chelating agents-which bind Ca
2
~
3.3.5 Lymph
The lymphatic system acts both as an overflow mechanism which
drains excess tissue fluid and protein back into the veins, and as a
system for circulating 'competent' lymphocytes for immunological
purposes. Most lymph returns via the thoracic duct (Figure 4.13).
To perform these functions the lymph vessels must have a high
permeability to protein and must remain patent when the pressure
around them increases. The current view is that the high permeability
is due to the absence of basement membrane and the presence of
numerous 'flaps' between the cells of its wall: the patency is due to
the anchoring of the lymph vessel walls by numerous filaments to
the surrounding structures. The amount of protein in lymph is variable,
from about 80% of that in plasma in the liver to 10% in the legs;
the concentration is sufficient to cause lymph to clot when left standing.
This surprisingly large amount of protein reflects the relatively high
permeability of capillary walls to proteins and other large particles.
: ~ f t e r a fatty meal lymph is milky due to the presence of large numbers
of fatty globules absorbed from the intestine.
4
The Cardiovascular
System
4.1 CIRCULATION OF THE BLOOD
The function of the vascular system is to convey nutrient and
other materials to and from the various parts of the organism; the
blood serving as the intermediary between the environment and the
cells. To fulfil this function the blood must be kept in a state of
continuous circulation. The heart provides the energy for this
circulation and the blood vessels are the channels through which it
takes place. The force of cardiac contraction drives the blood to the
tissues through thick-walled vessels, the arteries, and back to the heart
by a system of thinner-walled vessels, the veins. In the tissues the
blood is driven through a fme meshwork of vessels, the capillaries,
whose walls consist of a single layer of cells which allows interchange
of materials between the blood and the tissue fluid.
4.2 THE HEART
4.2.1 Outline of Function
The heart becomes filled with venous blood during its relaxation,
or diastole, and forces the blood into the arteries during its contraction,
or systole. The energy for moving the blood comes from the muscular
walls of the chambers of the heart; the direction of flow of the blood
is largely determined by the presence of valves at the entry to and
exit from the ventricles. Each ventricle is precededd by another
chamber, the atrium, which 'receives' the blood entering the heart.
Under resting conditions the atrium probably does not take much part
in moving the blood onwards.
The valves of the heart are fibrous flaps of tissue covered with
89
90
1
PHYSIOLOGY. BIOCHEMISTRY AND BIOTECHNOLOGY
Pulmonary
circulation
.-..
Systemic
circulation
---.
Figure 4.1 Diagram of the mammalian circulation. The capillary beds can be considered
as two 'trees' connected together by their fine branches (=capillaries).
endotheliwn. Those guarding the atrioventricular opening have strands
of fibrous tissue connecting them to small muscles within the
ventricles; these papillary muscles strengthen the valves during
contraction and help the ventricle to empty later in the cycle. The
valves on the exit from the ventricles (aortic and pulmonary) are rather
more fIlamentous than the atrioventricular valves and float open during
ejection of blood from the ventricle.
4.2.2 Physiological Properties of the Heart
Heart muscle cells have properties somewhere in between those
of skeletal and smooth muscle. They are small, striated and branching
cells with a single nucleus. Each cell is connected to its neighbour
by intercalated discs. within which are 'gap-junctions'. These junctions
allow the passage of electrical currents and small molecules between
adjacent cells. A piece of heart muscle therefore behaves as a single
unit (syncytium) to electrical stimulation, rather than a group of
THE CARDIOVASCULAR SYSTEM
(a 1
\-.- -----..J.
-_/ , ...... _-
..

-
\------------.J
c ;;r.
Inlet t .. --------... Outlet
( b)
Inlet
valve
I
Muscular
walls
(c )
Fillino
valve \ valve
Outlet
valve
Emptyino
91
Figure 4.2 Principle by which the ventricles impart motion to the blood. Compressing
a rubber bulb (a) causes the contained fluid to move out of it along both tubes. If
valves are positioned at each end of the chamber (b) then fluid is moved in one way
only. In the ventricle (c) the walls are muscular and the chamber is bent back on
itself so that the inlet and outlet valves are quite close to each other.
Tricuspid
(Rt AV) (l AV)
"
"---- Left coronary
artery
Right coronary /
artery_J
Aorta

Figure 4.3 View of the fibrous framework of the human hean from above showing
the valves. These valves are surrounded by fibrous tissue rings which form the fibrous
skeleton of the hean. To this skeleton are attached the muscles of the two ventricles
and the two atria, the aorta and pulmonary artery.
isolated units as does skeletal muscle. Certain heart cells are specialised
to conduct action potentials faster than normal; they also contract with
less force than normal heart cells; these cells make up the conducting
tissues of the heart. As in other muscle cells the visible contraction
of the cell is preceded by an electrical event-the action potential-which
depolarises the cell and triggers the mechanical events.
The properties of heart muscle will be discussed in terms of the
current views of the heart, first by considering the cardiac action
potential an d then the contraction.
4.2.3 The Cardiac Action Potential
The cardiac action potential is basically rather similar to that in
nerve and muscle but lasts for a much longer time; its precise shape
and duration also depends on which part of the heart it comes from.
It is usually divided into four components: depolarization, plateau
phase, repolarization and pacemaker potential. In general, the
properties of the cardiac action potential are similar to that in nerve
in particular there are permeability differences between heart and nerve
which 'explain' the differences in shape of the action potential.
Depolarization in heart, as elsewhere, is due to a large increase
in sodium permeability, which causes the membrane potential to be
reversed producing the overshoot.
THE CARDIOVASCULAR SYSl EM
Atrioventricular
node
1
1
1
1
1
____ I
1- - - - - - --
1
1
1
50
fl
m
Branches of
A-V bundle
93
Figure 4.4 Diagram of cardiac muscle cells in various parts of the heart. Each cell is
short and has several branching processes. The processes of adjacent cells are joined
end to end at structures known as intercalated discs. The size of these discs and the
desmosomes are slightly exaggerated.
The plateau phase is much longer than that in nerve because of
three main reasons: (1) the sodium permeability does not return quickly
to normal but stays above the resting value; (2) the potassium
permeability drops at the onset of the plateau and rises slowly towards
its resting value during the plateau; (3) there is a rise in the calcium
permeability at the start of the plateau with some decline during the
plateau; (4) the chloride permeability of heart is low, so that there is
little short circuiting of the potential during the plateau. The combined
effect of these changes is to produce a long plateau with a small gain
of Na+ and Ca
2
+ and a small loss of K+; this means that the cardiac
action potential is not too 'wasteful' in ion changes. The gain in Ca
2
+
is available for coupling to the actomyosin mechanism. The cell remains
refractory during the action potential and so may only be stimulated
(by a large stimulus) after its end; the refractory period is thus some
hundreds of milliseconds long rather than the few milliseconds of nerve
or skeletal muscle.
i
-@

90mV---.....
i
o
I
.,/ ......... .. .. .
... \ .
100 2 0
Time (miUlseconds)
..
----K+
I \
,
Figure 4.5 Diagram of twitch, ion movements, action potential and permeability changes
in a ventricular muscle cell.
TIlE CARDIOV ASC'ULAR SYSTEM 95
Repo/arisation is associated with a rise in potassium penneability
and a return of the sodium and calcium perrneabilities to normal.
PaceflUlker activity. Atrial and ventricular muscle cells have high
resting potentials (-SO mV or more) and show no spontaneous activity;
they are normally excited by impulses spreading from adjacent cells.
Cells in the sino-atrial (SA) node, the atrioventricular (A V) node and
the Purkinje fibres do not have a steady resting potential but show
pacemaker activity. This means that the potential is at a maximum
negative value just after an action potential, but then drifts slowly
towards the threshold potential; an action potential being set up when
it is reached. Usually SA and AV node cells have potentials which
uever reach more than 60 rn V, whereas Purkinje fibres have potentials
of - 70 to -90 mV. The current explanation for this behaviour is that
(a) there is a decline in the potassium permeability during the
pacemaker potential and (b) that the sodium permeability of pacemaker
Prepotential
or
pacemaker
potential
o ----f-t------Hi-----Hr--
Threshold - - - -
-bOmV
maximum
negative
potential
Pacemaker
cell
Driven
cell
Figure 4.6 Diagram showing electrical recordings from adjacent cells in the SA node.
cells is higher than in other parts of the heart; the decreasing potassium
permeability in the presence of the high sodium permeability leads to
the decreasing potential across the cell membrane. Normally the slope
of the pacemaker potential in the SA node is steeper than that of the
A V node which is steeper than that of the Purkinje fibres; therefore
if these parts could be separated the SA node would beat at the fastest
rate and the Purkinje fibres at the slowest rate. In, intact heart the
SA node, therefore, usually determines the heart rate, and drives all
the others at its rate.
These modem observations on pacemaker potentials 'explain' the
old observation that isolated heart is spontaneously active whereas
skeletal muscle is not.
The transmitter chemical ACh, released by the vagus, increases
the potassium permeability of heart cells and so decreases the slope
of the pacemaker potential; the threshold for firing an action potential
is reached later and so the heart slows down. Noradrenaline, released
by the sympathetic nerve endings, or adrenaline, circulating in the
blood, decreases the potassium permeability and increases the calcium
permeability of pacemaker cells, it therefore steepens the prepotential
and so speeds up the heart rate.
4.2.4 Conduction of the Action
Potential Over The Heart
The SA node makes contact with adjacent atrial cells and so causes
them to be depolarised by conduction through the gap-junctions of
the intercalated discs. These atrial cells in tum depolarise their
neighbours so starting action potentials. In this way a wave of electrical
activity spreads throughout the right atrium and, as the atria are
connected together, also spreads over the left atrium. This wave has
been likened to the ripples of a pool following a stone being thrown
into it. Many experiments have confirmed that the atria have no
specialised conducting tissue but simply spread electrical activity from
one cell to the next.
The atrioventricular (A V) node lies at the base of the right atrium
near the wall between the ventricles. It COllilects with the Purkinje
fibres which lie in the bundle of His in the interventricular septum;
this connection is the only electrical link between the atria and the
ventricles. Purkinje fibres conduct action potentials at higher velocities
than normal ventricular cells and have only very feeble contractile
properties.
The wave of electrical activity reaches the A V node, passes slowly
SA node
AV node
Bundle of His - - , f - - - - = - - - l ~
Right Left bundle
branChes
(a) Atrial spread (b) Ventricular spread
97
Figure 4.7 Spread of electrical activity throughout the human heart. The conduction
velocity in atrial and ventricular muscle is about 1 mlsecond in the A V node, 11 ml
second and, in the bundle of His. 5 mlsecond, i.e. in the ratio of I, and 5. Ventricles
from small animals (for example, a mouse) do not have Purkinje fibres, presumably
because conduction time is fast enough without it.
across it and then rapidly down the conducting bundle to reach the
ventricular muscle. At the ventricular muscle the wave of activity
again passes slowly from cell to cell. This arrangement ensures that:
(a) there is a delay between atrial and ventricular excitation (say 01
second), allowing time for blood to leave the atria; and (b) excitation
reaches most ventricular cells simultaneously, ensuring a single
coordinated contraction.
4.2.5 Contraction or Heart Muscle Cells
In heart muscle the coupling between electrical and mechanical
activity is triggered by the Call' which enters the cells during the
depolarisation phase of the action potential; the prolonged length of
the action potential causes the prolonged contraction. This is clearly
a 'good thing' in that blood has a high inertia and so a prolonged
force must be applied to it to cause it to be ejected from the heart;
short sharp contractions, like those seen in skeletal muscle, would be
ineffective. Although Call, which enters heart cells during the action
potential, is involved the detailed way in which electrical events are
coupled to mechanical events in the heart is still not clear. It has
recently been suggested that variation in the concentration of the cyclic
nucleotides may explain the variable coupling which occurs between
action potentials and their associated mechanical twitches.
Skeletal muscle has a short refractory period because the
associated action potential only lasts a few milliseconds. Skeletal
muscle can therefore be tetanised. whereas heart muscle cannot.
Clearly this is a valuable property of heart muscle; to have a
continuous circulation the heart must contract and relax alternatively,
a tetanised heart muscle would stop the circulation and kill the animal.
4.2.5 Events of the Cardiac Cycle
Our heart beats in a more or less regular fashion some 3.000
million times throughout our lives. It is convenient to discuss the
electrical and mechanical events which occur during a single cardiac
cycle as representative of all of them.
4.2.5.1 Electrical events
Each cell in the heart maintains a resting potential of some -60
to -90 mV at rest which changes to +20 to +40 mV during activity.
This change in potential. occurring across the millions of individual
cells in the heart, causes currents to flow throughout the body which
appear at the body surface as small potentials. The electrical activity
of the heart may thus be examined either by recording directly from
heart cells with intracellular electrodes, or by recording from the rest
of the body or the body surface with extracellular electrodes. The
electrocardiograph (ECG) is an instrument for recording these electrical
potentials from the skin surface. The letters P. Q, R, S and T (first
used by Einthoven) refer to the electrical waves set up by atrial
depolarisation (P), ventricular depolarization (QRS) and ventricular
repolarisation (T). The atrial repolarisation wave is normally hidden
by the QRS complex. The physiological points shown by the ECG
are:
,
1. The PR interval, of about 0 1 ,econd, measures the
conduction time of the impulse from the atria to the
ventricles. It thus measures th! time required for the
following events: (a) passage of t'.Ie atrial depolarisation wave
to the vicinity of the AV nod.:; (b) the delay imposed by
the AV node; and (c) the rapid spread of activity in the AV
bundle and its branches. The main delay is actually that in
the AV node.
2. The duration of the QRS complex, about 012 second,
measures the time of spread of the impulse over the
THE CARDIOVASCULAR SYSTEM 99
ventricles. In high-fidelity recordings a notch can often be
seen in this wave due to excitation reaching one ventricle
before it reaches the other.
3. The RT interval, of about 03 second, measures the mean
duration of the action potential in the ventricles.
4. Inspection of a series of R waves gives information about
the regularity of the heart beat. Two kinds of variation in
heart rate are described in nonnal-particularly young-subjects.
(a) Sinus arrhythmia is a variation of the heart rate with the
phases of respiration; during inspiration the heart rate speeds
up, during expiration it slows down. Two explanations are
advanced for this phenomenon; either that the cardiovascular _
centre is affected by irradiation of impulses from the
respiratory centre or that it is affected by impulses from
stretch receptors originating in the lungs. (b) Vagus-halt
phenomenon after exercise the heart rate frequently slows
in a series of oscillatory changes rather than smoothly. This
is due to sudden bursts of vagal activity-which can be
detected by their effects on the P waves of the BCG. In
s!1me subjects this phenomenon may continue for many
minutes before the heart settles to its resting rate. There is
still some controversy as to whether this is a manifestation
of sinus arrhythmia or a separate phenomenon.
Clinically the BCG is used to detect alterations in impulse
propagation, cell damage, etc., which occurs ill various heart defects.
4.2.5.2 Mechanical events
The events occurring during a single heart cycle can be
demonstrated by measuring the pressures in the various chambers of
the heart, together with volume measurement and measurement of
the BCG and heart sounds.
Students should examine and accompanying text and work their
way through a single cardiac cycle, understanding the various changes
which occur. Remember that blood, like all substances, flows from a
region of higher pressure to one of lower pressure. (In this account
the same numbers are used on the main figure, the inset diagrams
and in the text.) This account describes the left heart, events of the
right heart are similar but the pressures are lower.
Diastole. (1) In early diastole the ventricle is at its lowest volume
(65 rnl). The pressure in the pulmonary vein is slightly greater than
that in the left atrium which in turn is slightly greater than that in
100
Ventricular
ejection
PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY
Diastole of atria
and ventricle
and aortic
pressure 40
(mmHQ)
ElectrocardioQram
First and second
heart sounds
t------\Iot--'\,Y (phonocardiaoram)
Ventricular I ~
o 1----,[
I so metric ventricular
contraction
Isometric ventricular
relaxation
Figure 4.8 Diagram of electrical and mechanical events in the heart (left side) during
a single cycle at a heart rate of 75 contractions/minute. The insets show the changes
diagrammatically. The small effect of atrial contraction has heen omitted.
the left ventricle; blood therefore flows into the ventricle down this
pressure gradient and distends the ventricle. By mid-diastole the
ventricle is almost ftlled although no atrial contraction has yet occurred.
In late diastole, electrical activity, followed by contraction of the atria,
occurs; this drives a small, extra volume of blood into the ventricle
(particularly at fast heart rates) and also causes a back wave in the
venous system.
Isometric ventricular contraction. (2) Electrical activity spreads
to the ventricle, followed by ventricular contraction. The pressure in
the ventricles starts to rise quickly, exceeds that in the atria and so
the AV valves close (=the first heart sound). The ventricles are now
closed cavities and so the pressure rises rapidly. When it exceeds
that in the aorta then:
Ventricular ejection (3) of blood from the left ventricle occurs.
The ventricular pressure remains just greater than that in the aorta
(the pressure difference is not large as the filamentous aortic valves
offer little resistance to blood flow) causing the ventricles to eject
blood rapidly. The aorta receives blood so fast that: (a) its pressure
goes on increasing; and (b) it expands, thus 'storing up' some blood
in it. Gradually the rate of ejection of blood falls until the ventricle
is repolarised and stops contracting. The ventricular pressure now falls
below that in the aorta, the pulmonary and aorta valves snap shut
(=the second heart sound) and so the phase of:
Isometric ventricular relaxation (4) occurs; in this the ventricle
is once more a closed cavity as both inlet and outlet valves are again
closed, so the pressure drops from about 100 mmHg towards zero.
When it is less than the atrial pressure the A V valve opens and the
cycle restarts.
4.2.5.3 Additional points
1. The ventricle does not empty completely on contraction; the
difference between the end-diastolic and the
volumes is the stroke volume.
Diastole Systole
75/min
200/min
Figure 4.9 Relative durations of ventricular diastole and systole at nor;nal and fast
heart rates. With increasing heart rate diastole and systole both shorten-to minimum
values of 26 and 60%, respectively. The durations of the ventricular action potential
is almost the same as that of systole; the refractory period is slightly shorter Times
are in seconds.
2. The rate of filling of the ventricles is exponential not linear
and so fiiling is largely complete by mid-diastole. This means
that the marked shortening of diastole which occurs with
increase in heart rate does not greatly dt:.crease the amount
of blood entering the heart per beat. The rate of emptying
of the ventricle also has an exponential time course and so
is not much affected by the moderate shortening of systole
at high heart rates; in addition the rate of rise of tension is
greater at fast heart rates.
3. The arterial pulse. The ventricular contraction not only pushes
blood into the aorta but also sets up a pressure wave which
travels down the arterial walls. This wave travels much faster
(about 5 mlsecond) than the blood which is ejected into the
aorta; so it reaches the wrist in about 01 second. The
strength of this 'pulse' is related to the pulse pressure rather
than to the actual pressure within the artery and so usually
varies with the stroke volume of the heart. If the stroke
volume i! low, for example, following blood loss or
weakening of the heart, then the pulse feels weak; after
exercise it is large and can be felt by the individual as a
'thumping' in the body.
Figure 4.10 Observation of pulsations in the neck. Either examine someone else Qr
yourself in a mirror (to increase venous pressure hold your breath and blow). Venous
pulsations can be seen but not felt. carotid artery pulsation can be seen and felt.
4. Venous pulsation'). There are no valves on the inlet side to
the atria (although folds of muscle partly act as such) so
that pressure changes in the atria are transmitted back up
the veins, and may be observed in the neck. Classically three
waves, a, c and v, are described due to atrial systole, the
bulging of the A V valve during ventricular contraction and
the rise in atrial pressure before the A V valve opens during
diastole. The venous pressure waves can readily be seen in
the neck of someone lying flat and can :en be noticed in
'close-ups' of someone singing. particularly when sustaining
a long note. (The reason is that in this procedure the
103
intrathoracic pressure rises and so 'dams back' the blood in
the neck.)
5. Heart sounds. Heart sounds were originally detected by the
physician putting his ear to the patient's chest (perhaps using
a silk handkerchief), later by listening through a solid wooden
stick or trumpet and now via a stethoscope. They are the
first and second heart sounds, traditionally called 'lub' and
'dup'; they are caused by the closure of the AV valve and
of the aortic and pulmonary valves respectively.
4.3 THE VASCULAR SYSTEM
The left heart creates a pressure in the aorta which drives blood
round the various parts of the circulation back to the heart: the pressure
in the aorta is therefore greatest and that in the right atrium least.
The right heart drives blood through the pulmonary circulation; as
the resistance to flow is thus much lower than that of the systemic
., circulation it requires a much smaller pressure to do so.
Functionally, the systemic circulation may be divided into:
1. A distributory system consisting of the aorta and arteries
which contain a small amount of blood, held at high pressure.
2. A variable resistance system consisting of the arterioles,
where most of the arterial pressure is dissipated.
3. A capillary system with a vast surface area, where the
interchange of substances with the extracellular fluid of the
tissues occurs.
4. A collecting and reservoir system of veins, venae cavae and
right atrium which contains most of the blood, at low
pressure.
4.3.1 Arteries
The arteries have two main functions; first they act as low-resistance
elastic pipes taking blood from the heart to the arterioles in the various
tissues; secondly they store blood during diastole. Blood is ejected in
spurts from the heart at each contraction, is partly stored in the aorta
and large vessels and runs away through the arterioles at a continuous
rate. This 'smoothing out' function of the arterial system is due to the
combination of the capacity effect of the elastic vessels and of the
resistance to flow offered by the arteriolar resistance. You may examine
this as follows: take a very hot bath, thus dilating the skin arterioles,
you will find that a pulsation appears in your nail-beds; this is because
the arteriolar resistance of the skin has been decreased thus transmitting
pulsatile pressure changes to the capillaries.
4.3.1.1 Structure
Arteries, arterioles and veins have three coats, an inner smooth
layer, a middle layer which contains a variable amount of elastic and
smooth muscle cells and an outer layer of connective tissue. The
proportions of each component varies with the function of the vessel.
The smooth layer reduces friction to blood flow; the elastic tissue
gives the vessel a storage function, the muscle regulates the vessel
size; and the connective tissue outer coat prevents over distension as
well as anchoring the vessel to surrounding tissue.
4.3.1.2 Blood flow in arteries
The blood flow through arteries and the large veins near the heart
is pulsatile; the fact that this occurs in vessels whose walls have elastic
and viscous properties leads to a complex relationship between pressure
and flow, with some odd phenomena. In this section we shall fIrst
consider the much simpler relationships which hold if pressure and
flow are considered as non-pulsatile, and later mention some of the
more complex fIndings in pulsatile systems.
Figure 4.11 Cartoon of the circulation. The upper pipe represents the aorta and arteries
(high-pressure reservoir), the taps represent the arterioles and the tubs the organs
supplied. By varying the size of the arterioles the blood flow to an organ is controlled;
opening all the taps fully causes circulatory collapse.
Various physical relationships need to be considered before dealing
with this:
Pressure, wall properties and radius. Laplace showed that the
pressure within a vessel required to keep it open depended both on
the properties and on the radius of the vessel walls. Four consequences
which are applicable to the vascular system follow from this:
1. Critical closing pressure. If the properties of the walls of a
tube remain constant as the tube is made smaller then the
pressure required to keep it open increases. This is the reason
why small vessels close at a relatively high internal pressure,
called the critical closing pressure. A somewhat similar
situation occurs in trying to blow up a balloon; at small sizes
large pressures are required, at larger sizes little pressure is
required.
2. Very small vessels lequire much thinner walls to balance
internal- pressure than larger vessels do; this is why capillaries
can be just one cell thick, while veins (at a lower pressure)
have to be relatively thick.
I
I
Artery and
vena comitans
Endothelium___ .1
intima
Internal elastic
lamina
Smooth muscle
External _ _ _
elastic _.
lomina--,,= .- .. ,
.. -;": ... }
1/ ..
v ... ,;':,:-
Tunica "Endothelium

adventitia } Smooth muscle
and
collagen
Tunica media
Tunica
adventitia
Figure 4.12 The structure of a small muscular artery and its accompanying veins.
The tunica adventitia of the veins and artery are continuous with each other so that
pulsation within the artery compresses the veins and so forces the venous blood towards
the heart.
3. Efficiency of cardiac pumping. If the heart expands, then
any particular degree of contraction produced by the muscular
walls (= T) will produce a smaller rise in internal pressure
(for T=IRP). Dilated hearts are therefore less efficient than
normal sized ones.
4. Curved vessels. Any curvature reduces the effect of internal
pressure trying to distort a vessel wall; so when a vessel is
Shape of surface
Components of force
01161110.
(al (bl
Figure 4.13 (a) Relationship between the pressure (P) required to distend an elastic
sphere or tube and the tension in the walls (T). T is a measure of the elastic tension
in the wall (shown as a spring) and so must be overcome to keep the vessel open; the
force required to do so depends on the radius of curvature (R) and the pressure in the
vessel (strictly it should be the pressure difference between inside and outside).
For a sphere p= n For a tube P= T
2T
For a sphere P = R
T
ForatubeP=/i
(b) An approximate reason for this behaviour is as follows: a flat spring only exerts a
force in its own plane, a curved spring also exerts a force at right angles to the
surface; as the curvature increases the inwards component increases and must be
overcome to keep the tube open.
curved (for example arch of the aorta) the internal pressure
has a greater effect on the convex side rather than the
concave side. To oppose this pressure the wall is thicker on
the inside curve rather than the outside one.
-
-
QU
~

SG

--+

--

---
- -
(01 Turbulent flow (b) Laminor flow (cl Plug flow
Figure 4.14 The nature of fluid flow in a tube. Frictional resistance is produced by
molecules jostling each other and the walls of the tube.
The likelihood of turbulence occurring increases as the velocity increases, but it occurs
at much fower velocities with steady (b) than pulsatile (c) flows and so hardly ever
happens in a normal circulation.
b b
'I'ty f tu bit (radius of tube x velocity of flOW)
pro a I I 0 r u ence as - . ~
vISCOSity
/
Laminar and turbulent flow. Fluid can either flow along a tube
in a laminar or turbulent way. Laminar flow can be considered as a
series of concentric layers sliding past each other, the outer ones nearly
stationary, the inner ones moving faster; the energy required to
overcome the friction to move one layer past the next is at a minimum
and no noise is produced. In turbulent flow the molecules hit each
other and the vessel walls randomly; the energy required rises
disproportionately to any increase in flow and the lateral pressure on
the vessel wall increases; vibrations are set up which may be audible.
Laminar and turbulent flows can be examined with any domestic tap;
with the tap turned on slightly laminar flow occurs, with it turned on
further a noisy, turbulent flow develops.
Turbulent flow hardly ever occurs in a normal circulation largely
because in the situations where the velocity is high flow is also
pulsatile.
The relationship between pressure, resistance and flow. The
vascular system consists of many narrow pipes with fluid flowing along
them; so you need to know something about the relationships involved.
The factors which determine the resistance to flow are:
1. The diameter of the lumen.
2. The length of the tube.
3. The stickiness, i.e. viscosity, of the fluid.
Common sense would suggest that it is more difficult to drive a
sticky fluid, such as treacle, along a long narrow tube than it is to
drive water along a short wide tube. 'Difficulty' is measured as a
resistance to flow and is obtained from:
. ( length x ViscoSity)
resIStance oc 4
radius
The force required to drive fluid through a tube is measured as
the pressure gradient between the ends of the tube; the refutionship
linking all these factors is similar to Ohm's law, i.e.
therefore
driving force = flow x resistance to flow
or flow =
driving force
resistance to flow
pressure difference x radius
4
blood flow oc ""------------
viscosity x Length
This is known as Poiseuille's equation.
Normally the length of the blood vessels and viscosity of blood
are fairly constant and so need not concern us. The radius of blood
vessels does vary, and as flow is proportional to the fourth power of
the radius quite small changes in radius lead to large changes in flow.
i
o
LL
(0 I Rigid lube
Pressure--+
( b I Blood vessel
1
Pressure--+
Figure 4.15 Pressure-flow relationships in a rigid tube (a) and in a blood vessel (b).
A blood vessel collapses below a certain 'critical' pressure and has no flow, with
increasing pressure the vessel enlarges (exaggerated in the diagram) giving a larger
flow. In blood vessels the resistance to flow is therefore not constant but depends on
the pressure.
Thus a 16% increase in the size of a vessel doubles the blood flow;
doubling the radius increases the flow 16 times (or reduces the
resistance to 6%). As we shall see, the main way in which the
distribution of blood within the body is controlled is by variation in
the size of blood vessels, particularly the arterioles.
In rigid tubes the flow increases linearily with pressure so that a
'resistance' can be calculated which is constant; in elastic tubessuch
as blood vessels-increasing pressure distends the tube and so a non-
linear pressure-flow relationship is obtained, this means that resistance
to flow varies with the applied pressure.
Pressure-volume relationships; the stiffness of the vascular system.
The 'stiffness' of different parts of the vascular system varies, arteries
are very stiff, veins less so and the atria least. This means that arteries
are difficult to distend whereas veins and the atria are easily distensible.
4.3.1.3 Effects of pulsatile rather than steady flow
Blood flow is pulsatile not steady. This means that each 'particle'
of blood near the heart is accelerated during systole and then slowed
down during diastole; it also of course rubs against its fellows. Force
needs to be applied to blood to {)vercome these two resistive components,
inertial and frictional. With pulsatile flow the 'resistance' is replaced
by 'impedance'; this contains two components, reactance and resistance.
One consequence of this has already been mentioned, i.e. the flattening
of the velocity profile; others will be mentioned here.
150

0.
J:
100
E
E
!!
::J
'"
'"
!!
50
Cl.
o
Volume (ml)
Figure 4.16 Effects of filling short equal lengths of human aorta and vena cava with
fluid. The vena cava can hold a large volume at low pressure, the aorta develops a
considerable pressure if the same volume is forced into it.
Anyone corpuscle in the aorta moves forward in a jerk with each
heart beat, followed by standstill or even some backwards movement
during diastole; progressively along the aorta, this pulsatile pattern of
flow becomes damped by the storage function of the aorta until, in the
smaller arteries, it is more nearly continuous. Measurement of the
pulse wave at various parts of the arterial system shows that the
maximum size of pulsation increases as the wave travels away from
the heart, though of course the mean pressure decreases. The explanation
for this phenomenon is that when the wave reaches a branch in an
artery some reflection occurs from the junction, this adds to the original
wave so producing the increase.
Arterial pressure pulse. The pressure in the large arteries varies
in a roughly triangular or saw-toothed way. The descending part of
the wave has a notch-the dicrotic notch-corresponding to the closure
of the aortic valve. The peak pressure is the systolic pressure, the
lowest the diastolic pressure and the difference the pulse pressure;
the mean pressure may be found by averaging over the cycle but is
roughly the diastolic + one-third of the pulse pressure.
c;.
J:
E
E
Cl>

, \

60
et
140
"t:I
100

:11
60
......
E

3: 20
0
L;:
-20
, \
"
AORTA
Ascending Abdominal Saphenous
Thoracic Femorol
I"igure 4.17 Pressure and flow pulses in the dog arteries at increasing from
the heart. Pulse pressure increases while flow oscillation decreases markedly. It will
be noticed that the peak of flow slightly precedes the peak of pressure, this is because
of the reactance of the system.
4.3.1.4 Measurement of arterial pressure
Arterial pressure may be measured directly by inserting a hollow
needle into an artery and connecting this to a manometer'. This procedure
is frequently used in experimental animals but infrequently in man.
Normally, in clinical practice, arterial pressure is measured by an
external means. The usual method used is to apply a pressure to the
outside of an artery by means of an inflatable rubber bag held in a
cloth cuff. This procedure interferes with the blood flow through t!te
artery, causing the production of characteristic sounds (Korotkow).
The technique is: the bag pressure is inflated above that of the arterial
pressure so that all blood flow stops; the pressure is then released
slowly; the systolic pressure is taken as that pressure at which sounds
first appear and the diastolic pressure as that pressure at which they
become muffled. The sounds are due to partial occlusion of the vessels
with the production of a noisy, turbulent flow. The method depends
for accuracy on the bag pressure being transmitted to the artery wall;
if the limb is very fat then this may not happen giving consistently
high readings.
4.3.2 Veins
The systemic veins, like the arteries, have two main functions:
THE CARDIOVASCULAR SYSTEM III
first they act as a low-resistance collecting system to carry blood from
the body back to the right side of the heart; secondly they act as a
low-pressure storage system (i.e. are capacity vessels), holding over
50% of all the blood at anyone time at a pressure below say 18
rrunHg. They often lie alongside the arteries going to an organ,
sometimes sharing a connective tissue coat.
Although their structure also consists of three coats they have
much thinner and more distensible walls than arteries, with much less
elastic and muscle tissue in them. They are therefore less stiff (i.e.
more compliant) than arteries. At intervals on the long veins,
particularly in the limbs, the endothelial lining protrudes into the lumen
to form cusps or valves similar in structure to the semilunar valves
in the heart. This arrangement is functionally very useful in that veins,
having thin walls, are easily compressed by surrounding muscle; during
muscular activity the contraction and relaxation of the tissues around
the veins acts as a muscle pump helping blood to return to the heart.
The cluse proximity of arteries to veins also produces a similar effect
at each pulse wave.
Pressure
.. n _ ~ I . __ .
(0)
ArterlCl
pressure
wove
State 01
artery
Figure 4.18 Sphygmomanometer method of measuring blood pressure. A cuff is applied
to the upper arm, this cuff contains a rubber bag winch is connected to a mercury
manometer and can be inflated with air (inset). The bag lies over the radial artery so
that when inflated it compresses it.
The presence of ~ l v e s in the veins and the way in whcih they
work was used by Harvey in 1628 as part of his evidence for a
circulatory system. The experiments he performed are easily repeated
by students and you should do so.
Veins are supplied with autonomic sympathetic nerves which inner
vate the smooth muscles in their walls. Increased activity in the
sympathetic system leads to an increased stiffening of the walls, so
that the vessels become narrower.
2
3
4
Figure 4.19 Venous valves. Harvey noticed during dissections that a probe could easily
be pushed up a vein past the venous valves towards the heart but could not be pushed
backwards down a vein; particularly into side branches. He illustrated their function
by (I) which shows the presence of the valves (as 'knots'). (2) how the blood could
easily be emptied towards the heart but not away from it (3) and how (4) the vein can
fill from 'below' but not from 'above'. He used the rapidity with which veins filled
from below as part of his evidence that the circulation of the blood was very fast and
so must move in a circle. The inset shows a cut-away, enlarged view of a venous
valve.
mE CARDIOVASCULAR SYSTEM 113
4.3.2.1 Venous blood flow and pressure
Flow in veins is less phasic and more continuous than in the
corresponding arteries. It does, however, vary with the heart beat
and with pressure changes in the thorax. The average velocity of blood
increases as it passes from the venules back to the heart, due to the
decrease in the crosssectional area of the vessels.
The pressure in the venous system must be lower than that in
the capillaries and yet still high enough to fill the heart and so maintain
the cardiac output. The pressure at mid atrium is remarkably constant
at a few mmHg 5) in health.
The transmural, or distending, pressure is the pressure difference
between the inside and outside of a vein. Because veins are thin-
walled floppy structures, their shape is very dependent on this pressure;
if high the veins are circular in cross-section, if low they collapse
and are then difficult to feel or see below the skin though they go on
carrying blood normally. This different appearance with change in
transmural pressure is the basis of estimating the 'central venous
pressure' by observation of neck veins.
Changes in pressure in the right atrium and in the thorax are
transmitted back to the veins in the neck and may be examined as
already described. The normal atrial waves are readily seen, and
increases in intrathoracic pressure (for example, blowing up a balloon,
blowing a trumpet, etc.) soon distend the neck veins. Normal
inspiration produces a reduced intrapleural and increased intra-
abdominal pressure; this increases the venous return to the heart giving
an increase in the cardiac output and blood pressure above the
expiratory values.
The mechanism for maintaining the central venous pressure
constant depends mainly on altering the size of the venous reservoir;
this is adjusted by variation in the degree of tension in the smooth
muscle of the walls of the veins. Such changes in tension alter the
compliance of the vein walls and so alter the relationship between
volume and pressure in the system. Changes in cardiac function caused
either by direct action or reflexly also contribute to the constancy of
the venous pressure.
4.3.3 Arterioles and Capillaries
The capillary network and associated vessels are the most complex
and least understood part of the circulation and that part where the
primary function of the circulation, i.e. exchanges of gases, nutrients,
metabolites and heat, takes place. To perform this function the blood
in the capillary bed is slowed and spread out in a vast thin layer
such that no tissue cell is much more than 10 urn away from a
capillary (except in lens and cartilage).
4.3.3.1 Structure and function
Arterioles are small muscular tubes with their walls and lumen
about 20 pm in size. The capillary bed consists of true capillaries
which are long thin vessels (wall 1 urn, lumen 8 /Lm, i.e. just sufficient
to pass an erythrocyte). There are two types of bypass vessels which
connect arterioles directly to venules: (1) the commonest of these is
the thoroughfare vessel or metarteriole which occurs in all tissues.
These are vessels whose walls are discontinuously coated with muscle
cells. The capillaries start as side branches from the thoroughfare
vessels; they have muscle cells around their origins which act as pre-
capillary sphincters to control the blood flow through them. (2) Some
epithelia also contain short A-V shunts. The small venules have no
muscle cells, the larger ones do.
The muscle of the arterioles, metarterioles and precapillary ,
sphincters shows histological differences related to their function and
innervation. The arteriolar muscle is richly innervated, each cell is
insulated from the others and the cells are not much influenced by
locaJ metabolites. Metarteriolar and precapillary muscle is poorly
innervated, has many connections between the cells and is largely
controlled by local metabolites.
Most arterioles are supplied by sympathetic nerves which release
noradrenaline, but there are (alas!) two exceptions to this general rule:
(l) some sympathetic nerves to skeletal muscle release acetylcholine,
which causes an active vasodilation of the arterioles, perhaps at the
start of exercise; (2) the blood vessels of certain areas of the genital
tract are supplied with nerves which release
acetylcholine and cause a dilation. Certain glands which show an
increased blood supply following parasympathetic nerve
probably do so via an increased metaoc,:ism rather than by direct
arteriolar innervation.
The direct rome for blood throuth a .me is from arteriole, to
bypass vessel, to venule; the capillaries are rna side branch controlled
by the precapillary sphincter. The metarter"ole, capillaries and venules
form the microcirculation. All tissues (e,cept the lens and cartilage)
have such a microcirculation; but the (apillaries in spleen and liver
form sinusoids rather than true capillaries. Most organs have so many
capillaries that they may be regarded as sponges filled with blood;
THE CARDIOV ASeULAR SYSTEM 115
when a muscle or organ contracts it expresses some of this blood
into the general circulation and this action materially influences the
tirculation.
The detailed arrangement of the capillaries varies with the tissue
concerned. In skin they appear as loops coming up towards the surface
and then retreating back into the skin (these can readily be seen
in the skin beside your nail-beds under a low-powered microscope).
In muscle they are arranged longitudinally among the muscle fibres.
In membranes such as the pleura or peritoneum (tissues derived from
the mesothelium) they appear as flat networks. If any of these capillary
networks is observed for a length of time it will be noticed to be
'unstable' in that one capillary can be seen to transmit blood at one
time and then close, while another opens and so on (under the
microscope you actually see the erythrocytes in the capillaries rather
than the vessels themselves). This intermittent arrangement of blood
flow means that much of the exchange and equilibrium between tissue
fluids and blood may take place when the blood flow is actually
stopped.
4.3.3.2 Exchanges between tissue fluids and capiUaries
Exchanges of gas and other constituents occur very rapidly
between the capillaries and tissue fluids; this is largely because of
the very large surface area of blood exposed to the tissues rather
than any high permeability possessed by the capillary wall. The route
by which such exchanges take place appears uncertain, perhaps it is
via vesicles in the lining cells rather than through 'pores' between
the cells. The mean pressures in the capillaries at heart level almost
balance the oncotic pressure of the plasma proteins. As the oncotic
pressure is about 2 5 mroHg and the interstitial pressure a few mmHg
negative to the atmosphere, the mean capillary pressure is therefore
about 20 mmHg. As already discussed this capillary pressure is correct
for the centre of the capillary, but proximal (i.e. upstream) to this
point it is higher and distal (i.e. downstream) it is lower giving a
'circulation' of fluid in the tissue space.
4.3.3.3 Control of the capillary circulation
The blood flow through the microcirculation needs to fulfil two
partially distinct functions: (1) tissue exchanges and (2) control of
the peripheral resistance.
1. The primary function is to exchange substances with tissue
cells to meet their current metabolic needs; for this exchange
to he effective the critical factor is the total surface area
available for exchange rather than the volume of blood in
the capillaries. As we have seen this is achieved by having
the capillaries as small as is consistent with allowing
erythrocytes to pass through them.
2. The total cardiac output flows through the microcirculation
so that the size of its vessels (mainly arterioles) determines
the total peripheral resistance of the vascular system and,
ultimately, the blood pressure.
Arteriole ___ 11
Capillary
Bypass vessel -++-1-
1
Arteriolar resistance
controlled by nerves
resistance
controlled by
local metabolites
Capillary
Figure 4.20 Diagram of control of arteriolar and microcirculation. The overall blood
flow is controlled by the nerve supply to the arteriole. the distribution of blood between
the capillary and thoroughfare vessels is determined by metabolites acting on the
precapillary sphincters and muscle of the thoroughfare vessel.
For example, if all the arterioles in the body were fully opened
up then the peripheral resistance would decrease markedly and the
blood pressure drop catastrophically. These conflicting functions require
some degree of independent control over the arteriolar resistance and
the microcirculation. The present view is:
I. The nervous system controls the peripheral resistance. py
altering the tone of smooth muscle in the walls of the
arterioles. Over the whole body this controls the overall
arteriolar resistance; in anyone vascular bed this controls
the blood flow to it.
2. Local metabolites in the tissues control the precapillary
sphincters and hence the blood flow through the large capillary
exchange area; increase in metabolites diverts more blood to
the capillaries thus washing away these metabolites. There is
still some argument about the nature of these metabolites;
they do include a raised Pc0
2
, a decreased pH, an increase in
K+ ions, adenosine and perhaps lactic acid; decreased P02 also
dilates these sphincters. A temperature rise may also be
involved. Students sometimes wonder how metabolites released
in the capillary bed can act on sphincters 'upsteam' of the
point of release; the explanation is that metabolites released
by cells do not only act on their 'own' sphincters, but also
increase the local extracellular concentration of metabolites
and these then dilate adjacent sphincters. Because capillaries
criss-cross a tissue this means that increased activity leads to
a general increase in blood flow in the area.
These two systems interact to some extent; a metabolic rh.c first
produces a local increase in capillary flow and then opens up the
local arterioles. If a severe arteriolar constriction is imposed on an
area by central control then the resulting accumulation of metabolites
may modify and so relieve the ischaemia caused.
4.3.3.4 Effects of various stimuli on perivheral circulation
1. Heat applied locally causes an increased blood flow, and cold
a decreased blood flow. Generalised heating, sufficient to
raise the core temperature, causes a generalised rise in skin
blood flow by a reduction in sympathetic 'tone'. These effects
act on the arterioles.
2. Exercise leads to a 20- to 40-fold increase in blood flow to
the muscles concerned. The increase in blood S U ~ i occurs
because of a direct effect of 'metabolites' on the local
sphincters in the muscle (both arteriolar and precapillary);
this is called active hyperaemia. This is clearly an efficient
arrangement; increased work produces increased concentration
of metabolites, these adjust the local blood flow to a rate
sufficient to remove the metabolites from the muscle. Muscle
work and blood now are related to eae h other. the link is
the local metabolite concentration acting on the arterioles and
precapillary sphincters in the tissue,
3. If the blood supply to a tissue is cut off for a time, release
is followed by a large rise in the hlood flow over the next
few minutes. This increase may 30 times the resting
value; it is probably caused by accumulation of the
metabolites mentio:1ed above. This is called reactive
hyperaemia
4.3.3.5 The response of skin vessels to trauma
The white reaction. When a pointed instrument is drawn lightly
over the skin, a white mark appears on the part touched by the
instrument: this response is said to be due to contraction of the cells
lining the vessels which give the colour to the skin-i.e. the capillaries.
It still xcurs if the circulation is stopped or the capillary pressure is
raised to 70 mmHg. It develops in about 15 seconds and lasts a few
minute", it is not to be confused with the immediate effect of touching
the skin, which is merely a passive emptying of the vessels.
Triple response. When the skin is stroked more firmly, a series
of events ensue called the triple response. (a)_ A red reaction soon
occurs (10 seconds), limited to tlle area of contact, followed by (b)
an irregular, diffuse flare which affects the surrounding area and fmally
by (c) a swelling, or wheal, which is due to local accumulation of
nuid. This sequence of events is probably due to the local release of
a (H suhstance, which probably includes histamine); it is
th.e normal response of tissues to injury, the skin is sUnply a convenient
to observe it. The red reaction is probably due to opening up
of the precapillary sphincters with an increase in capillary blood flow
:so it is very localised); the nare is due to an axon reflex which
causes adjacent arkrioles to dilate, tltusbecause they supply a bigger
area-leading to a diffuse irregular blotch; the oedema is due to
increased vessel permeability allowing protein and white cells to leave
the circulation. The whole response can be thOUght of as a mechanism
for bringing maximum amounts of antibodies and white blood cells
to a site of injury.
4.3.4 Peripheral Circulation as a Whole
In the preceding sections we have discussed the various parts of
the vascular system separately; here we wish to discuss the way in
which these parts work together. To do so we shall consider the blood
supply to a single organ or part of the body (for example, the
adn:na!s), a situation in which all the blood entering through the artery
Spinal Sensory
cord neurone
Skin
endings
Kinins
released
Arteriole
Figure 4.21 Antidromic axon reflex of the flare reaction. Stimuli from skin receptors
travel up the sensory axon to its junction with a sensory neurone from the arteriole.
they then travel backwards to the arteriole and dilate it.
leaves through the vein, without any cross-circulation. Several features
of such a circulation:
1. The pressure in the aorta and large arteries (in mmHg or
kPa), created by the contraction of the heart, drives the blood
through the capillary system back to the veins (flow occurring
from an area of high to one of low pressure). The main
pressure drop occurs in the arteriolesdue to their high
resistance-little drop occurring in the conducting vessels.
2. The volume flow through the system, usually called blood
flow, (litre/minute or m
3
/second) is constant, as fluid is
neither added nor subtracted by the tissue. In the kidney, of
course, the venous outflow is less than the arterial inflow
by the amount of urine produced.
3. The surface/volume ratio of the vascular tree is lowest in
the supply and drainage vessels to reduce friction, and highest
in the capillaries where exchanges with the tissue cells take
place; structure is thus well adapted to function. In man the
surface/volume ratio of the capillary bed is about 500 times
greater than that of the aorta or the vena cava.
4. The velocity of blood flow is the reciprocal of the cross-
sectional area, so that it is highest in the aorta and lowest in
the capillaries. In man the peak aortic velocity may reach
100 em/second (about 4 kmIhour) while that in the capillaries
120 PHYSIOLO(jY, BIOCHEMISTRY AND BIOTECHNOLOGY
is 007 em/second. (This means that an erythrocyte moves its
own length in 10 milliseconds.) Most children know about
the relationship between cross-sectional area of a vessel and
the velocity of fluid flow; if a domestic tap is turned on fully
and the end partially closed with a finger a most spectacular
jet of water is produced.
5. Resistance, as we saw on by analogy with Ohm's law, is
the ratio of the driving force applied to a fluid divided by
the flow (i.e. (PI-P2)1F). As the flow in the system is
constant, then the resistance is higher where the pressure
drop is greatest, i.e. in the arterioles. The energy imparted
to the blood by the heart is dissipated as heat to overcome
the resistance of the vascular system. This frictional loss
occurs due to the jostling of the molecules of blood against
the vessel walls and with each other.
4.3.5 Methods of Measuring Blood Flow
These are two methods of measuring cardiac output in man; these
are the direct Fick method and the indicator dilution method.
A
Time from dye addition
Figure 4.22 Diagram illustrating the principle of the dye dilution technique.
The indicator dilution technique can be readily understood as
follows: imagine you and a friend are standing by a stream separated
by a few metres; your friend, upsteam of you, throws a bucket of
red dye into the stream. You start a stop-watch as he does so and
THE CARUlOVASCULAR SYSTEM
(0)
Pressure
gouge
(e)
12i
Figure 4.23 Interrelationships between pressure, flow and resistance. The high-pressure
reservoir (i.e. the arteries) is represented by a ball with a thick elastic wall. The heart
is represented by a bicycle pump and the peripheral resistance of the arterioles by a
variable leak in the balI.
then time the appearance of the dye when it reaches you; you also
collect samples of the dyed stream at intervals as it passes. You would
expect the dye to take some time to reach you and to be diluted
when it did so. If the stream starts to run faster, then the dye will
reach you sooner and be more dilute (for more water will pass your
friend as he empties the dye into the stream). With an experiment
such as this you could work out the volume flow of the stream. This
example makes it clear that for this method to work adequate mixing
of the dye in the stream is necessary, partly mixed dye would give
false results.
In Clinical practice dye is injected into a vein and blood collected
or monitored in an artery; it is supposed that adequate mixing has
occurred in between. In the body the dye recirculates' so that the curves
do not return to the baseline. A good adaptation of this technique is
to inject cold solution instead of dye and measure change in
temperature. This may be done repeatedly, for the amount of cold
solution does not permanently change the blood temperature.
A modification of this principle is commonly used to measure
blood flow in organs or tissue (especially muscle). In this technique
an injection of a radioactive substance (for example, Na+) is made
into the muscle and then the rate of removal of the substance
measured; the faster the blood flow the faster the removal. This may
be understood by considering the effect of throwing dye into a fast
or slowly moving stream, the fast stream soon carries away all the
dye, the slow stream takes longer.
4.4 THE OVERALL CONTROL
OF THE CIRCULATION
We have already discussed the control of the microcirculation;
we now need to consider the control of the circulation as a whole.
The circulatory system works on the following two basic principles:
1. Maintaining a relatively constant arterial blood pressure.
2. Allowing wide variations in blood flow to the different organs
and tissues when the demand for this arises.
The function of central control is to maintain a high pressure in
the arteries with a minimum cardiac output, this high pressure then
allows blood to flow to the various tissues when the arteriolar taps'
are turned on by local metabolic controls. Such problems and their
solutions are by no means confmed to biology, every electricity and
water supply company face similar ones: the electricity company has
to maintain the supply voltage at a fixed high level (say 5%). This
then allows the users to draw off current for their various appliances
simply by turning a switch. If the mains voltage swings greatly then
our radios, televisions and lamps would also vary in their performance;
if we wanted a constant performance from these devices we would
then need complex local regulatory devices.
Four main systems are involved in the regulation of the vascular
system: (I) control of the blood pressure; (2) regulation of cardiac
output; (3) control of the local circulation; and (4) control of the
blood and extracellular fluid volume.
4.4.1 Control of Blood Pressure
The arterial blood pressure exists because the heart forces blood
into the elastic aorta, from which it has some difficulty in escaping
through the peripheral resistance. The pressure which is produced in
any given circumstance is therefore dependent on both the cardiac
output and on the peripheral resistance; control over the blood pressure
can thus be exercised by actions on either of these. If you have
difficulty with this concept then think about the following.
Imagine a situation in which the heart continued to pump but all
the arterioles were either closed or fully open. If closed the blood
could not leave the large arteries, the blood pressure would rise
markedly and the heart fail; if open the blood would rush out of the
large arteries and the p r s ~ u r would fall towards zero. Figure 5.31
shows this concept in another way.
This argument can be quantified by using the derivation of Ohm's
law-Poiseuille's law-which we used before. You will remember that:
driving force = flow x resistance.
The driving force here is the pressure difference between the
arterial and venous sides of the heart, which is approximately equal
to the arterial pressure (for the venous pressure is very low); the
flow is the cardiac output and the resistance is the total arteriolar
resistance (the peripheral resistance). So
arteriolar pressure = cardiac output x peripheral resistance.
From this it is clear that the arterial blood pressure can be varied
by changing the peripheral resistance-as argued above or by changing
the cardiac output. This relationship is sometimes used to calculate
the total peripheral resistance of the circulatory system; with an arterial
blood pressure of 100 mmHg and a cardiac output of 5 litres/minute
this gives a resistance of
100 nunHg 20 H rl '-1
5 IImin = mm g mm
Of the three interdependent variables of blood pressure. cardiac
output and peripheral resistance the blood pressure is the least variable
in any given individual over short periods of time, and also shows a
long-term stability. As with many other physiological variables it shows
a change throughout the 24 hours. with a minimum during the night.
The 'normal' values for the population show a fairly wide frequency
distribution with (in Western man at least) an increase with age.
Typical daytime values for middle-aged man are a systolic pressure
of 130 mmHg and a diastolic pressure of 80 mmHg (i.e. a blood
pressure of 130/80): 80% of the normal population are within a range
of 10 mmHg of these mean values.
200

.! 150
...

Q.
"0
o
,g
ID
C

'"
ii
...
t!
l!

I
:a
0
120
100
60
11
0
I I
2
I'
1
2
I I I I I
4 6 8
3 5 7
I I I I I
4 6 8
3 5 7
Syslolic
.;
DioSIOlic
I I I 1 1 1 1 1 1 1 I 1 1 I 1 I
10 12 14 16 18 20 22 24
9 11 13 15 17 19 21 23
Tirfle of day (hours)
I 1 I I I 1 I 1 1 I 1 1 1 1 1
I
10 12 14 16 18 20 22 24
9 11 13 15 17 19 21 23
Time of day (hours)
Figure 4.24 Circulation variations in the systolic and diastolic pressures and heart
rates in five normal subjects (mean age 29 years). The records were taken by an
indwelling catheter over 2 days. while the subjects worked normally. Typical standard
error shown on 24-hour reading.
4.4.2 Sensory Receptors in the Heart and Aorta
If we wish to have an automatic heating or cooling system in a
house then we need to arrange to measure the house temperature and
then use this, via some kind of switch, to control the heating or cooling
unit so as to keep the house temperature constant. The vascular system
is controlled in the same way; there are sensory receptors to measure
the performance of the heart and blood vessels, a cardiovascular
centre' which makes decisions about the result and a motor system
which controls the heart and blood vessels.
The sensory input system from the heart and blood vessels appears
to rely on measuring the mechanical events in the heart and blood
,vessels, rather than detecting the electrical events in the heart directly;
thus the mechanical consequences of the heart's action are measured.
All the receptors which do this measurement are 'stretch receptors in
the walls of the blood vessels (tunica media and adventitia) and in
the heart itself. The quantity which they measure depends on the
properties of the vessel or chamber they are situated in; if they are
in a very stiff structure (i.e. low compliance), such as the aortic arch,
then they effectively measure pressure, if they are in a very floppy
structure (i.e. highly compliant), such as the vena cava or atrium,
they effectively measure volume.
High-pressure receptors. The baroreceptors, or high-pressure
receptors, in mammals are in the aortic arch, pulmonary arch, carotid
arteries and other main arteries near the heart (Figure 5.33). In man
the carotid sinus is the main receptor site; this is situated at the
bifurcation of the external and internal carotid arteries in the neck.
These receptors are thus situated between the head and the heart on
the direct route to the brain; they are thus well sited to monitor the
pressure of the blood supply to the head, of obvious importance in
animals which can adopt both an upright or horizontal position. These
receptors are stimulated by stretch of the arterial walls, which in turn
is caused by a rise in intra-arterial pressure; as the pressure rises the
rate of firing of anyone receptor increases, as it falls the rate of
firing decreases. A single receptor will only respond to pressures
within a rather narrow range, so that many receptors are required to
cover the whole pressure range which normally exists. In addition to
responding to the absolute pressure, these receptors are also affected
by the rate of change of pressure and so respond differently to a
pulsatile pressure with.the same mean value as a steady pressure.
The impulses travel to the 'centres' in the upper medulla concerned
with the nervous control of the blood vessels and the heart. The motor
impulses from this centre travel via the parasympathetic nerves to the
heart and the sympathetic nerves to the heart and blood vessels.
Sympathetic control is differential as the brain vessels take little part
in these adjustments. Stimulation of the carotid sinus by raised blood
pressure leads to a slowing of the heart and a vasodilation of the blood
vessels; the former action decreases the cardiac output, the latter
decreases the peripheral resistance, both actions which decrease the
blood pressure.
Low-pressure receptors. It is now clear that low-ptessure receptors
in the artria and great vessels feed impulses into the cardiac centre
in the medulla to alter its activity. An increase in the volume of the
right atrium and great veins causes increased firing in these receptor
nerves and a reflex increase in heart rate and contractility and
peripheral vasodilation (Bainbridge reflex). Thus both the input and
the output from the heart are monitored.
The vasomotor nerves form part of the sympathetic autonoinic
nervous system; they therefore run from the medulla down the spinal
cord (lateral columns) to the thoracic and upper lumbar regions, where
they synapse with the lateral hom cells. The preganglionic fibres leave
in roots Tl-L2 and enter the mixed peripheral nerves to supply the
smooth muscles of the vascular system. (There is no parasympathetic
supply to these muscles.) The cardiac nerves arise in TI-T4, synapse
in the three cervical ganglia and reach the heart via the cardiac branches
of the sympathetic chain; the parasympathetic nerves reach it via the
vagus. Normally there is always some activity in the vasomotor nerves
to the blood vessels-leading to their partial restriction-and in the
parasympathetic supply to the heart-leading to slowing of the heart
rate below its denervated value.
Other inputs. Inputs from the respiratory system and higher centres
also influence the medullary centres; thus rage, fear, excitement, etc.,
can cause large rises in blood pressure. In clinical ,practice this often
means that casual readings of blood pressure are unreliable: the act
of making the observation itself influences the result.
4.4.3 Regulation of Cardiac Output
We have seen that arterial blood pressure depends on the product
of the cardiac output and the peripheral resistance. Normally blood
pressure is kept constant despite changes in peripheral resistance,
because the cardiac output is changed in an inverse way to the peripheral
resistance, i.e. as peripheral resistance decreases (for example during
exercise) the cardiac output increases to keep their product constant.
This section is concerned with the regulation of the cardiac output.
The cardiac output, which is the volume of blood ejected by one
side of the heart per minute, is:
cardiac output Htres/minute = heart rate beats/minute x
stroke volume Iitres/beat
At rest an average man's heart beats about 72 times per minute
and each ventricle ejects 70 ml of blood per beat. The cardiac output
is thus 72 x 70/1000 litres/minute. Each side of the heart has the
same cardiac output when averaged over many beats, but over a few
beats the outputs may be different, for example ,,,,hen bulk movement
of blood from skin to lung vessels is occurring.
Clearly the cardiac output can be varied by changing either the
heart rate or the stroke volume; as we shall see the IlIain control is
exercised by changing the heart rate, lesser control is by changing
stroke volume. The rate can change by more than three times, the
stroke volume by less than twice. These two parameters will be
discussed separately.
4.4.4 Control of Heart Rate
The heart rate is determined by the rhythmic discharge of the
node, which depends on the slope of the pacemaker potentials;
this in turn is influenced by the sympathetic and parasympathetic supply
to rhe heart and by circulatory adrenaline. These influences set the
hCaIi rale to a value within the range of 50-200 beats/minute. At rest
the heart ratc is set by the level of parasympathetic 'tone', a fact
which can be shown by blocking with atropine (this causes a large rise
in rate). Factors such as blood temperature, pH, ionic composition
and other hormone concentrations have a smaller influence on heart
rate.
4.4.5 Control of Stroke Volume
Harvey thought that the heart emptied completely each time it
contracted; we now realise t..'1at only about half of the blood is expelled
at each contraction. The stroke volume is therefore the difference
between the ventricular volumes at the start and end of each
contraction, i.e. the difference between the end-diastolic and the end-
systolic volumes. Control of the stroke volume is obtained by changing
either of these volumes
The Frank-Starling law of the hean. The relationship between
contractility and length of muscle fibres.
The strength of contraction of a heart muscle fibre varies with
its length; ultimately depending on the overlap of the actin and myosin
filaments and therefore on the number of interacting sites between
them. The relationship between the energy of contraction of heart
and the resting length of the muscle fibres was first measured by
Frank and later by Starling; where the stroke volume is used as a
measure of energy of contraction and end-diastolic volume as a
measure of length. As stated above, increasing length of the muscle
fibres produced by increasing distension of the heart leads (within
limits) to increased output.
Stroke
volume
End-diastolic volume
Filling J
pressure l
Stroke
volume
F"JgUre 4.25 Relationship between stroke volume and end-diastolic volume of hean.
The inset shows the heart preparation used to demonstrate this (Starling's heart-lung
preparation). Raising the reservoir increases the filling pressure (and hence the end-
diastolic volume) and eventually leads to a greater stroke volume.
The change does not occur instantaneously; usually for several
beats the diastolic filling exceeds the systolic emptying so that
progressive swelling of the ventricle occurs until the output equals
the input.
This m ~ h n i s m can be regarded as a form of autoregulation,
the more the muscle is stretched the greater the subsequent contraction;
because the length changes it is sometimes known as heterometric
regulation. We argued that a distended heart was less efficient than
one of normal size (because of the Laplace's Law) so that a heart
which increases its stroke volume by distending is less efficient than
one which does so by emptying more. This latter type of response is
sometimes known as homometric regulation (because the muscle fibres
at end-diastole are the same length). It can be most readily studied
by looking at the family of Frank-Starling curves in a heart perfused
with different concentrations of catecholamine; at anyone drug
concentration the heart works along one of these curves. This means
that in the body the stroke volume of the heart can be changed at
constant diastolic volume by changing the concentration of
catecholamine present. It now appears that other variables such as
blood pressure, blood pH, plasma [Ca
2
+], etc. also produce families
of curves in a similar way.

I

1
Decreased contractility
produced in disease
End-diastolic volume (of ventricle)
Figure 4.26 Catecholamine effects on the Starling curves of the ventricle. In intact
animals the level of circulating catecholamines moves the length-tension curve of the
ventricle upwards and to the left. The dotted vertical line shows that stroke volume
can be increased for the same end-diastolic volume by increasing concentrations of
catecholamines; this may come from sympathetic nerves or from the adrenal medulla.
Clinically the right atria] pressure is used as a measure of end-diastolic volume.
How then is the stroke volume normally controlled in the body?
The general rule is that the heart remains the same size during diastole
but empties more during systole, i.e. a homometric regulation occurs.
It is thought, however, that this effect is rather slow in developing
so that sudden changes of work are dealt with by some cardiac
swelling (i.e. heterometric regulation) until the heart can adjust itself
to increased contractility. Thus, a sudden increase in the load on the
heart (or heart rate) leads to swelling and increased output, but after
10-20 beats the heart shrinks again but continues with a higher output.
This system optimises the efficiency of the heart. Cardiac failure is
often associated with a loss of this homometric response, the
heterometric response remaining as the only mechanism.
4.4.6 Regulation of the Peripheral Resistance
Most of the details of this have been dealt with already, here a
sununary only is given. Different organs show both a difference in
their resting rates of blood flow per gram of tissue and in their ability
to change this basic flow; the kidney and brain have a high, fairly
constant blood flow in health, whereas muscle at rest has a low blood
flow which can increase some 40 times during exercise. The actual
flows are, in all cases, dependent on the degree of contraction or
relaxation of the vascular smooth muscle for the blood pressure is
fairly constant.
Autoregulation. Vascular smooth muscle, even if denervated, has
a resting 'tone'. Normally this tendency to contract is opposed by
the production of local metabolites by the tissue, so that the balance
between the two determines the local blood flow; this constitutes the
basis of the autoregulation shown by many tissues. The nervous supply
to the arterioles and microcirculation can be regarded as a central
control system superimposed on this local one.
4.4.6.1 Summary of cardiovascular control
The following features are apparent:
1. There is a well-developed system of local control of blood
flow in the tissues; in general this can override central control
systems.
2. The two pumps which provide the energy for the system,
the right and left hearts, are very sensitive to the venous
pressures on their inputs; normally these input venous
pressures are kept constant and the outputs of these pumps
are accurately matched.
3. The cardiovascular system is controlled by the medullary
'centres' so that a fairly constant blood pressure is maintained
despite large variations in demand by the tissues.
4.4.6.1 Integration of the cardiovascular system
Students usually learn the function of separate parts of the
cardiovascular system but often have difficulty in comprehending how
it works as a whole. The venous reservoir, venous return, cardiac
output and arteriolar resistance are all so intimately related that none
can be altered without affecting the other$. Here we shall concentrate
on the relationship between venous return aT'd cardiac ou:put which
must, of course, be equal over any length c l time.
The central venous pressure is the prf SSUrt: in the intrathoracic
portions of the superior and inferior v',nae cavae: it is of great
importance because the difference betv een it and the surrounding
illtrathroacic pressureille transmural pre',sure-represents the distending
pressure of the heart. The transmural pressure is maintained within
quite narrow limits in health regardless of the position of the body in
space, the redistribution of blood in dilated capillary beds or the
accumulation of blood in distended dependent veins. Its lower limit
is zero otherwise filling of the heart would be deficient during the
diastolic intervals. Its upper limit cannot be too high otherwise it would
raise the gradient of pressure in both the venous and the lymphatic
systems, which would cause accumulation of fluid in the tissues. The
true distending pressures of the heart are the transmural pressures in
the right and left ventricles during diastole; these are slightly lower
than the central venous pressure and difficult to measure.
Venous
return
(5 litres I'mm)
Cardiac
output
(5 litres/mm)
Central venous pressure (mmHg)
Figure 4.27 The relationship between venous return, cardiac output and central venous
pressure in normal man. Transient changes in venous return or cardiac Qutput cause
transient changes in central venous pressure. The effective filling pressure of the heart
is greater than the central venous pressure because the usual intrathoracic pressure is
about -4 mmHg.
Although the venous return must equal the cardiac output over
any extended period of time, for a few beats at a time the two may
be different. Consider the effects of a sudden reduction in venous
return; the cardiac output will continue unchanged for a beat or two,
thus decreasing the central venous pressure and so decreasing the stroke
volume until venous return and cardiac output are equal again.
Conversely, if the venous return increases suddenly the cardiac output
will continue unchanged for a time, leading to a build-up of blood and
an increase in pressure in the venae cavae and heart until the cardiac
output increases to match the new venous return. In this way the
changes in venous return which occur in life are continually matched
to the cardiac output by changes in the central venous pressure.
Quantitatively it can be shown that the venous return is proportional
to the difference between the mean systemic pressure and the central
venous pressure; whereas the cardiac output varies with central venous
pressure according to Starling's law. These relationships can be
illustrated as in Figure 5.40. The steady-state central venous pressure
present in the intact circulation is given by the point where the venous
40
Cardiac output
(exercise)
30
Venous return
(exercise)
c:
B
'e
"
.....
"
'"

"-
20
"-
"-
"-

"
Cardiac output
"
iD
"
(rest)
10
Venous return
"
(rest)
"
----
"
--
"
-
I-::::J:...
"
0
I I I
-4 4 8 12 16
Central venous pressure (mmH91
Fbwre 4.28 Theoretical curves showing the way cardiac output and venous return vary
'''ltIl central venous pressure in an athlete. The actual central venous pressures are
given by points A and B; both just positive. In non-athletes point B is usually several
mmHg positive. The mean systemic pressure at rest is about 7 mmHg, that during
exercise is 16 mmHg. Note the flattening of the venOUR return curves at negative
central venous pressures; this is due to collapse of the veins where they enter the chest.
return and cardiac output curves cross. This figure can be used to
illustrate the effects of exercise on the cardiovascular system. There
are two main factors which act: first, there is a large increase in the
cardiac output, due to the marked sympathetic activity present; and,
second, there is an equally large increase in venous return, due partly
to the increase b the mean systemic pressure consequent on sympathetic
activity and partly to the reduced resistance of the venous system and
partly to the action of the muscle pump. These two effects shift the
cardiac output and venous return curves to higher levels, with a large
rise in cardiac output. Students should consider other examples such
as blood loss, decrease in cardiac contractility, fluid retention, etc.
TIlE CARDIOVASCULAR SYSTEM 133
When doing so try to work out the effects of these conditions on blood
pressure, heart rate, cardiac output, right atrial pressure and capillary
pressures.
4.4.6.2 Circulation through special areas
The blood flow through the various parts of the systemic
circulation is arranged as a number of resistances in parallel; each of
which have special features, some of which will now be described.
The pulmonary circulation differs from others in that it is in series
with the rest of the circulation.
4.4.7 Coronary Circulation
Three small arteries supply blood to the heart. They all arise
from the aorta just past the aortic valve; one from the front and the
other two as a single artery from the rear which then divides into
two. Each artery divides into numerous branches to supply the muscle,
but little communication occurs between branches so that narrowing
of one leads to a lack of blood supply to part of the heart muscle.
Such narrowing or blockage often leads to death of heart muscle (a
'heart attack'). Venous drainage from the ventricles occurs mainly
via the coronary sinus back to the right atrium but partly by direct
drainage into the ventricular cavity.
Increased muscular work requires an increased cardiac output and
so an increase in the work of the heart; to perform this extra work
the heart requires an increase in its supply of free fatty acids, lactate,
glucose and oxygen to its cells. However, that the blood supply to
the heart occurs mainly during diastole and least during systole; so
that during increased cardiac work (Le. increased contraction and
shorter diastolic pauses) the conditions for an increase in coronary
blood flow are less favourable than at rest.
The oxygen extracted from coronary blood is very high-usually
15 ml/lOO ml compared to a mean elsewhere of say 4 ml/lOO ml-so
that increases in 02 supply to the heart can only occur by an increased
coronary blood flow. This occurs in exercise and hypoxia but the
mechanisms for doing so are not well understood; probably nerves
play little part and local control via metabolites and hypoxia is the
main - factor.
4.4.8 Cerebral Circulation
The brain is an area of high and fairly constant metabolism which
is particularly sensitive to hypoxia; its blood supply is correspondingly
large (075 litre/minute) and fairly constant. though the blood supply

to individual parts of the brain seems to vary with activity. The
cerebral circulation is unique in that it occurs in an area which is
incompressible, so that changes in the blood volume in the brain can
only occur if something else (usually cerebrospinal fluid) is displaced.
Cranium _--J"7
Intracranial
pressure -tt:--.....
Local constriction
and dilatalion of
cerebral arlerioles
pressure at
brain level ___ -+_
Mean venous
pressure 01
brain level
Figure 4.29 Factors affecting blood flow.
Brain, spinal cord
and spinal fluid
f+-__ verlebral
column
The factors affecting the total cerebral blood flow are: (1) those
common to all regions, i.e. the perfusion pressure (the arterial-venous
pressure difference at the brain level), the degree of vasoconstriction
or vasodilation of the cerebral blood vessels and the blood viscosity;
and (2) the intracranial pressure. The interactions of the perfusion
pressure with intracranial pressure helps to maintain a constant cerebral
blood flow under various conditions, for example if standing on your
head both arterial and venous pressures increase (due to gravity) but
the intracranial pressure also increases so that blood flow does not
change much.
At present it is thought that the blood flow to the different parts
of the brain is regulated by local factors: thus a rise in Peo
2
or fall in
P0
2
produces a vasodilation and a fall in Peo
2
or perhaps a rise in P0
2
a vasoconstriction; there is thus a considerable degree of autoregulation.
The cerebral vessels are innervated by both sympathetic vasoconstrictor
fibres and parasympathetic dilator fibres but these do not seem to be
involved in the regulation of blood flow. Considerable rises in intracr-
Mean arterial
pressure
(mmHg)
0---
la
20
30
40
50
60
70
Venous 80
pressure
----0
90
. 10
100
20 110
30 120
40 130
50 140
60 150
70 160
80 170
Figure 4.30 Hydrostatic pressure effects in erect man. The arterial and venous pressures
are both increased by some 85 mmHg at the ankle. With the arm held above the head
the arterial pressure at the wrist is' about 40 mmHg and the effective venous pressure
is zero down to a level just above the heart. Blood flows through the collapsed veins
but the internal and eltternal pressures are exactly equal.
anial pressure reduce blood flow due to compression of the vessels; a
reflex causing a rise in arterial pressure has been described in these
circumstances (Cushing's reflex) which helps to maintain normal blood
flow.
4.4.9 Effects or Posture on the Circulation
So far we have considered the cardiovascular system as operating
in 'gravity-free' conditions, a condition approached by man or animals
lying horizontally. About two-thirds of our lives are, however, spent
in the upright position and so this should be considered as the 'nonna!'
for man. Gravity has a considerable effect on the pressures in the
various parts of the vascular system in upright man (Figure 5.43),
increasing the pressure in vessels below the heart and decreasing the
pressure in those above the heart.
4.4.9.1 The main effects of gravity are indirect:
I. There is increased exudation of fluid from the capillaries,
causing an oedema (i.e. swelling) of the tissue spaces with
an increase in the tissue pressure. Our feet swell during the
day, our faces swell at night in bed (due to redistribution of
fluid); men should shave after breakfast not before, allowing
time for the swelling to decrease.
2. On standing upright some 500 ml of blood is transferred
from the chest (i.e. lungs and heart) to the legs. This
(equivalent to a brisk haemorrhage) causes a drop in the
filling pressure to the heart, with a consequent drop in stroke
volume and a drop in cardiac output. In order to minimise
this pooling of blood in the feet a reflex vasoconstriction of
arterioles and venules occurs (this affects vessels in the lower
rather than the upper part of the body); the heart rate also
increases and its volume decreases a little. This reflex is
relatively slow (seconds) in action; this slowness is responsible
for the feelings of faintness and darkening of the visual fields
which you may have sometimes experienced on rising suddenly
from a horizontal position (explanation: the sudden drop in
blood pressure reduces the brain blood flow, producing these
effects). This postural reflex disappears quite quickly if not
used; in space flights it is gone in a few hours and staying
in bed for several days reduces it. It appears that its continual
use during life is necessary for its retention.
This reflex does not, however, completely correct the effects of
posture, as the cardiac output remains some 30% lower when upright
when lying horizontally. As metabolism stays the same, more O
2
is extracted from the venous blood (6 rather than 4 ml per 100 ml)
to keep the supply to the tissues constant. The 'muscle pump' (together
with the venous valves) helps to reduce pooling of blood in the feet
by aiding venous return; this of course only occurs during muscular
activity.
Students are often puzzled as to how blood can 'climb' back up
137
from the feet to the heart. The vascular supply to the legs can be
compared to a u-tube with arterial blood entering one of its sides and
venous blood returning from the other. Common exPerience tells us
that fluid which enters one side returns from the other side. The same
happens in the vascular system except that there is superimposed upon
the gravity gradient a vascular gradient produced by the arterioles,
capillaries, etc. The venous pressure in the feet is always greater
than that in the right atrium if adjusted to the same level.
4.51 PLACENTAL AND FOETAL CIRCULATION
405tt Uterine Circulation
The blood flow to the uterus shows cyclical changes during the
menstrual cycle; during pregnancy it increases with the ihcrease in
size of the uterus and contained foetus. The increase in the mass of
the foetus and placenta is much greater (1 cell-S kg) than the 20-fold
increase in the maternal uterine blood flow which occurs, so that the
01 extracted from the uterine blood is greater at the end of pregnancy
than it is at the start leading to a decrease in the 01 saturation of the
uterine venous blood. Just before parturition the uterine blood flow
decreases sharply.
4.5.2 Placenta
The placenta is the site where 01 and COl exchange occurs,
through which all nutritional materials enter the foetus and where all
waste products are discharged; it therefore acts as the foetal lung,
gastrointestinal tract and kidneys. The maternal portion of the placenta
acl$ as a 'lake' into which the villi of the foetal arteries and veins
prd)ect. Exchanges take place across the four layers which separate
thej maternal pool of blood from the foetal capillaries; as pregnancy
prqceoos the overall thickness of these layers decreases, perhaps to
facjilitate the increased diffusion of materials required as the foetus
beComes larger. The foetal and maternal circulations do not, however,
normally mix.
4.5.3 Foetal Circulation
Blood returning from the placenta is split into a part which goes
directly to the right heart (via the venous duct) and a part which
gere via the liver; this is a sensible arrangement because the
pI ental blood contains both 01 and nutrients (glucose, fats and amino
ac ds). At the right atrium the larger part of this placental blood is
sh through an oval ~ o l into the left heart and is then sent to
the head via the upper aorta; this again is sensible, for the head is
Foetal capillary endothelium
UmblDcal artery and
vein (foetal blood)
Uterine Septum
artery
: : : [ l Z : ~ 3 and vein
'" (maternal
blood)
Foetal surface
(chorionic plate)
Direction of
blood flow In
Intervillous
space
Maternal surface
(decidual base
plate)
Figure 4.31 Diagram of placenta showing two villous units (foetal cotyledons). Each
villous is like a 'tree' whose branches consist of an artery and a vein running side by
side; at its tip the artery runs into a capillary which drains back to the vein. The villi
are closely crowded together and not freely floating in space as shown. Inset shows
the four layers between maternal and foetal blood.
thus supplied with the most bighly oxygenated blood. The smaller
stream of blood remaining in the right atrium is augmented by that
returning from the upper body and enters the right ventricle. It then
largely bypasses the lungs (via the arterial duct) to the descending
aorta and hence to the lower body and placenta again. This latter
arrangement means (a) that the less oxygenated blood goes to the
lower body. and (b) that the lungs. which are of course non-functional.
receive only a small blood supply.
Foetal Hb receives its 02 from the maternal Hb, through the rather
thick exchange membranes of the placenta. To facilitate this exchange
of 02 the foetal form of Hb has a bigher affinity for 02 than the adult
form.
4.5.4 Changes at Birth
At birth there is an immediate need to change from placental to
pulmonary respiration. As soon as the infant ceases to be 'weightless'
Table 4.1. Circulatory values in the human foetus and newborn.
Systemic arterial pressure
(npnHg)
I
Pulmonary arterial pressure
(mmHg)
Heart rate/minute
Blood flow
in pulmonary artery
through ductus arteriosus
Foetus Newborn Adult
70/45
(umbilicall
arterial)
140
small
large to
70/45
35/15
140
large
small from
aorta aorta
12O/SO
30/10
70
nil
the respiratory centre receives a flow of new sensory impulses, notably
from the skin and muscles. The partial anoxia and mild acidosis during
labour may also play a part. These cause the first few respiratory
movements (mainly due to the diaphragm) which give very negative
intrathoracic pressures -40 to - 60 cmH20 (4-6 kPa) during inspiratory
effort and large positive pressures (say 20-30 during expiratory
effort. These overcome the initial resistance to entry of air to the
lungs. When the lungs expand the Pot rises to normal values and the
arteriolar resistance falls to about half. Much more blood
:E:
flOWS through the lungs and so the left atrial pressure rises,
s pping the left to right shunt through the foramen ovale. The ductus
osus closes as the PO
I
rises; a mechanism apparently triggered
by increased oxidative metabolism in the muscle cells.
The placental blood flow decreases after birth due to contraction
of the umbilical vessels; this causes a 'transfusion' of about 100 ml
of blood into the circulation but also causes the right atrial pressure
to drop, thus helping to close the foramen ovale. The ductus venosis
stFts to close soon afterwards, reducing the bypass of blood past the
liver.
Although all these changes start at birth it takes some weeks for
complete changeover to be completed. The unstriped muscle in the
pulmonary arterioles gradually atrophies and so their walls become
thinner. After blood flow through the foramen ovale, ductus arteriosus
and venosis stops. all those channels gradually close. The table given
in this chapter shows normal values in man for various parameters.
5
TJte Kidney
mild Micturition
The composition of blood (and hence of the body) is kept constant
mainly through the selective elimination of water and solutes by the
kidneys rather than by a close control over the intake of these materials
in food. This control involves balancing the body's input of ions and
water with the amounts excreted. As Na+ and Cl- are the most abundant
osmotically active solutes in plasma, control of plasma volume and
tonicity can be largely achieved by controlling the amounts of these
ions and water excreted.
Since the requirements of salt balance to maintain plasma osmolality
may conflict with those of water balance to control plasma volume,
two separate control mechanisms are necessary; one each for water
and sodium excretion. Other simple inorganic substances whose conc-
entration is regulated by the kidney include, K+, Ca2+, Mgl+, H+,
S O ~ , POt. The kidney also has a limited control over the concen-
tra ons of amino acids and sugars in the blpod, although the main
site for the homeostasis of these substances resides in the liver.
CaUlbolism of body protein produces about 30 g1day of urea, which is
excreted in the urine along with other breakdown products of metabo-
lism. These include uric acid derived from the metabolism of nucleic
acids, creatinine from muscle creatine, and the breakdown products of
steroids, haemoglobin, etc. The kidney also functions in the elimination
of foreign chemicals such as pollutants, drugs and food additives.
5 . ~ GROSS STRUCTURE OF THE KIDNEY
I In man the kidneys are paired organs situated on the back wall
of ~ e abdomen on either side of the vertebral column. The basic
I 141
J'.
"
"
.,
"
"
"
.:
IJ
Cortex __ _
Renal
column ____
It----Ureter
-r----_Bladder
'--___ Urethra
Figure 5.1 Gross features of the kidney and associated urinary system.
unit of kidney function is the nephron of which there are about
1,000,000 nephrons in each kidney. Urine formed in the tubular part
of the nephron in the renal pelvis and then flows through the
ureter to the bladder for subsequent elimination via the urethra.
5.1.1 The Structure of the Nephron
Each nephron consists of a vascular and a tubular component.
The tubular component consists of a single layer of epithelial cells
whose basement membrane forms the outer walls of a tube. Filtration
mE KIDNEY AND MICfURITION 143
Loop of
CoIIec:Iing duct---H--If----;/r-
Collecting duct cell
Figure 5.2 Essential features of the structure of a nephron.
takes place in the renal corpuscle at the proximal end of the tubular
S}'SteIlL The remainder of the tubular system consists of a proximal
s e t i ~ t h e loop of Henle. the distal tubule and collecting duct. Cells
linin the proximal tubule have a characteristic brush border of
micrq . . formed by protrusions of their surface membranes, the
junctibns between the cells are formed by tight junctions.
The structure of the loop of Henle differs according to its location
in the kidney. Those nephrons located in outer regions of the kidney
have short loops of Henle with a thin descending limb and a thick
ascending limb. The cells lining the thick segment of the loop of
Henle form a larger lumen and lack prominent brush borders. In
contrast. nephrons situated deeper in the kidney-the cortical!
juxtamedullary nephrons-have somewhat longer loops of Henle and
thin stgments on both sides of the loop.
The distal tube has a similar structure to that of the thick segment
of the loop of Henle. Distal tubules from a large number of different
nephrons drain into a common collecting duct and then via a papillary
duct into a large central cavity called the renal pelvis. The epithelial
lining of the collecting duct is composed of a layer of cuboidal cells
which gradually become taller as the collecting tubule merges into
the papillary duct. In man the nephrons are arranged into some lO-
IS groups called Malpighian pyramids. The nephrons are all orientated
so that the Bowman's capsules with related proximal and distal tubules
are situated in the outer layers of the kidney cortex while the loops
of Henle and collecting ducts converge towards the apices of the
Malpighian pyramids to form the medullary rays.
The vascular component of the nephron is composed of the
capillary system of the glomerulus. Blood enters the glomerular
capillary network via an afferent arteriole and leaves it through an
efferent arteriole rather than a vein. The proximal tubule expands in
the region of the renal corpuscle to form the Bowman's capsule which
allows the capillaries of the glomerulus to come into close contact
with the epithelial lining of the tubular system. It is here that filtration
of the blood takes place. In the majority of nephrons (those situated
in the cortex) the afferent arteriole from the glomerulus divides up
into capillaries which cover the surfaces of the convoluted tubules
and finally empties into the venous system. In the deeper juxtame-
dul1ary nephrons the efferent arteriole from the glomerulus divides to
form loops which lie parallel to the loops of Henle and so run down
into the cortex. These vessels are called the vasa recta; and are
concerned with a system for concentrating the urine.
5.1 THE CONTROL OF URINE FORMATION
The initial stage in urine formation is the filtration of an almost
protein- free plasma through the capillaries into the
Bowman's capsule. During subsequent pr through the component
parts of the tubular system its composition is altered by the selective
reabsorption of certain substances and the sel.:ctive secretion of others.
The rates and extents to which tubular secreion and reabsorption occur
are subject to physiological regulation md form the basis of the
homeostatic mechanisms whereby the hdney is able to control the
composition of the blood.
5.2.1 Glomerular Filtration
Average kidneys filter about 180 Jitres of plasma each day (120
mlImin). Since the total plasma volume is only about 3 litres, this
THE KIDNEY AND MICTURITION 145
means that the total blood volume of a person is filtered about 60
times each day. This allows a very precise control of the internal
environment of the body. The glomerular capillary pressure favouring
ftltration is around SO mmHg. The smaller diameter of the efferent
compared to afferent arteriole (especially in the outer nephrons) helps
to maintain this relatively high capillary pressure in the glomerulus.
Opposing this is the fluid pressure in the Bowman's capsule (-10 mmHg)
and the osmotic pressure produced by the plasma proteins (-25 mmHg)
(oncotic pressure). Therefore the net pressure producing filtration is
allqut IS nunHg. The energy for glomerular filtration comes entirely
froPl the energy transmitted to the blood when the heart contracts
I
I
Descending
IImb_
Ascending
11mb
ThIn &egment--+J--.. ... ,
Straight
collecting
rubure ____
Outer
medulla
Inner
medulla

5.3 Diagram illustrating the two types of nephron. Those in the outer regions
of idney have shorter loops of Henle than those in the inner medulla. Each collecting
due system draws fluid from some 3000 to 6000 nephrons.
,
Figure 5.4 Diagramatic representation of the blood supply to cortical and juxtamedll1l3ry
nepbrons.
(hydrostatic pressure). Thus glomerular filtration is a passive process
with the pores in the glomerular membranes simply acting to sieve out
blood corpuscles, white cells and most of the plasma protein.
It has been found that glomerular capillary resistance varies with
systemic arterial pressure to maintain glomerular filtration rate
relatively constant over a wide range. This 'autoregu1ation' of blood
flow also occurs in denervated kidneys and is thought to arise from a
direct adaptation of the vascular smooth muscle to altered pressures.
Current views of the filtration process involve the concept of the
pressure and osmotic gradients along the glomerular capillaries; under
different conditions these profiles change with the net result that
filtration remains constant under most nonnal conditions. More
advanced texts should be consulted for an account of this.
5.2.2 The Concept of Clearance
In order to understand the way in which th,cr .. kidney works, both
THE KIDNEY AND MlCfURlTION 147
in health and in various disease states, it is useful to be able to estimate
various quantities such as the renal plasma flow, the glomerular
filtration rate, etc. The method used to do so is based on the concept
of 'clearance' which depends on Fick's principle. This idea will be
explained by considering various examples.
Artery
l-----..
Tubular
system
Urinary excretion
Efferent
arteriole
S.S A diagram ilIusuating the factors which control the formation of the urine.
Mean renal arterial pressure
(mmHg)
Figure 5.6 Autoregulation in the dog kidney. This process probably occurs by changes
in ! the resistance of the afferent arteriole.
Renal plasma flow. If a substance, x, can be found which is
completely removed from the plasma and excreted into the urine during
one passage of blood through the kidney, then this can be used to
measure the renal plasma flow and hence (by knowing the
haematocrit.) to measure the blood flow. Clearly. in this circumstance.
the amount of this substance recovered from the urine over a certain
time must be equal to the amo.1}1lt delivered to the kidney in this same
time. Therefore the volume of urine multiplied by the concentration
of x in the urine must equal the volume of plasma multiplied by the
concentration of x in the plasma; by rearrangement of this equation
and division by time the plasma flow per unit of time can be
calculated.
P-aminohippuric acid (PAH) is both filtered by the glomerulus
and excreted by the tubules and is completely extracted by a single
passage through the kidneys. Therefore
the effective renal plasma flow (RPF)=PAH clearance= UpAHV
PPAH
The glomerular filtration rate (GFR). In a similar way if a
substance. y, can be found which is filtered without hindrance at the
glomerulus and not thereafter affected by its passage down the rest
of the nephron. then the glomerular filtratioJl rate could be calculated.
To do so the volume of urine collected in' a certain time multiplied
by the concentration of y is equal to the volume of plasma filtered
multiplied by the concentration in the plasma. The result of this
calculation gives the volume of plasma filtered per unit of time.
Another way of considering this result is to suppose that a certain
volume of plasma has been ~ m p l t l y 'cleared' of y rather than that
a larger volume of plasma hils been partially cleared. Thus the figure
calculated as this 'clearance' is the glomerular filtration rate.
Inulin is filtered by the glomeruli but neither secreted nor absorbed
by the tubules, therefore ..
GFR = inulin clearance = U
1
",
~
Clearance is expressed as millilitres of plasma cleared per minute.
U and V refer to urinary concentrations and volume of the substance
respectively and P is the plasma concentration.
5.2.3 The Urea Clearanl"e
Other possibilities exist; thus if some of the substance, Z, is
reabsorbed by the tubules then the result of this calculation will give
a 'clearance' less than the glomerular filtration rate; if an extra amoWlt
of substance is secreted by the tubules (in addition to being filtered)
then the clearance will be greater than the glomerular filtration rate.
Thus by measuring the clearances of substances from blood information
about their handling by the tubules may be obtained.
i.e.
The urea clearance can be estimated as above:
UV
clearance =
urea Pu
As urea is reabsorbed in the tubules the urea clearance is less
than the inulin clearance; the ratio of these is often used in description
of the processes occurring in the kidney
the clearance ratio = U
a
v" : U
I
~
P
u
~
and as the urine volume (V) is common to both this simplifies to
U
u
.U
I
P
u
~
5.2.4 Control of Blood Osmolality
Control of blood osmolality is ultimately achieved by regulating
the concentration of Nat Sodium from the diet is actively absorbed
from the intestine and the excess is eliminated by the kidney.
Homeostasis of salt and water is achieved by selective reabsorption
of these substances from the glomerular filtrate. Therefore in healthy
animals the osmolality of the urine will accurately reflect the salt
and ~ t e r balance of the individual. Since the final response of the
kidney is a compromise between the potentially conflicting demands
of maintaining the salt and water contents of the plasma constant there
is a dual-control mechanism operating on both the reabsorption of
sodium -and the reabsorption of water.
5.2.5 Iso-osmotic Reabsorption in the Proximal Tubule -
As the glomerular filtrate passes down the proximal tubule Na +
is actively transported out into the adjacent extracellular space.
The rise in osmolality outside the tubule results in a net movement
of water across the epithelium due to osmosis. Some 80% of the
Na+ filtered at the glomerulus is removed during passage down the
proximal tubule. This 80% of Na+ takes 80% of the filtered water
Active transport
[ => Passive diffusion
;,
..
Tubule
lumen
Tight junction
membrane
Capillary
Figure 5.7 Movement of sodium and water from the tubular lumen to capillary in
epithelial cells of all parts of the nephron (except the loop of Henle). In proximal
tubules the tight jllDCtions are low resistance and in distaI tubules they are high
resislmlce. The tubular SIIrface also has microvilli.
with it (iso-osmotic reabsorption), so that fluid leaving the proximal
has the same Na+ concentration as plasma. Since the volume
of filtrate has decreased. the concentration of all other substances is
increased, causing them to move back into the blood by passive
movement down their concentration gradients. The remaining 20%
of the glomerular filtrate is the fraction which is controlled by the
kidney in order to maintain plasma osmolality.
S.1.6 Trans-tubular Potentials
Normally there is a difference of potential between the lumen of
the nephron and the surrounding tissue; the size and direction
depending on the part of the nephron examined. Thus in the
mammalian nephron the potentials are: proximal tubule, 38 mV inside
negative; thick ascending limb of Henle's loop, a few mV inside
positive; distal convoluted tubule and collecting duct a high potential,
inside negative.
mE KIDNEY AND MICfURlTION 151
These potentials are associated with the presence of active
transport processes in the walls of the tubules. They may be explained
thus: if there is a primary transport of Na+ then for each Na+ removed
the tubule becomes more negative, this causes a passive transport of
Cl-, which can be considered as being 'dragged' behind the Na+. If
there is a primary transport of Cl (thick loop of Henle) then the
tubule becomes positive which gives a passive movement of Na + .
The size of the potential depends both on the rate of transport and
on of the wall of the tubule. Thus if transport is stopped
(for e le by ouabain) then the potential disappears. In the proximal
tubul , which has low-resistance tight junctions, the potential is less
than the collecting duct system, which has high-resistance tight
junctions.
5.2.7 Reabsorption in the Distal Parts of the Nephron
The changes in the remaining 20% of filtrate made by the distai
part of the nephron depend on whether antidiuretic hormone (ADH)
is present or not. In the presence of ADH (i.e. dehydration) the kidney
Table 5.1. Changes in water metabolism produced by ADH at a
constant osmotic load of 700 mosmollday. Specific gravity (SG) and
osmolality are not always related to each other because SG depends
on the nature as well as the number of molecules present.
Percentage
of filtered Urine
water 24-hour concent-
GFR reabso- urine ration Specific
(mlImin) rbed vol. (mosmol/t) gravity
Urinl!:toniC
to pi 125 98-7 24 290 1010
ADH I present 125 997 0-5 1400 1035
NoADH
(diabetes insip- 125 88 233 30 1002
. idus)
produces a small volume (1-2 litres/day) of concentrated urine; in its
it produces a large volume of a dilute urine. The process by
whichl the kidney produces a concentrated urine depends ultimately
on the setting up of a large osmotic gradient between the cortex and
the inper part of the medulla. This osmotic gradient is then used to
reabsorb water from the distal tubule back into the renal tissues. In
conditions when the body is adequately hydrated (i.e. no ADH
circulating), this osmotic gradient is reduced or absent so that no
concentration of urine can occur.
In the presence of ADH, CI-, followed by Nat is pumped out of
the thick part of the loop of Henle and water follows it. Thj.s isotonic
fluid enters the collecting ducts and passes down through the hypertonic
medulla; as it does so it becomes concentrated due to the passive
removal of water (and some urea) through the permeable walls of
the tubule. With ADH present some 15% of the filtrate is removed
isosmotically in the distal tubule and at least another 4 % in the
collecting ducts to produce the concentrated urine. In the absence of
ADH the distal tubule and collecting duct system remain fairly
impermeable to water and urea; as NaCI continues to be reabsorbed
actively the filtrate within those tubules becomes hypotonic and so
large amounts of dilute urine enters the renal pelvis. Even in the
absence of ADH about 8% of the filtrate is still absorbed in the distal
part of the nephron leaving about 12% (in man) to be excreted. This
amounts to some 23 litres/day of urine.
The counter-current mechanism. As we have seen, the
concentrating mechanism in the kidney depends on the establishment
of a large concentration gradient between the cortex and medulla. It
is thought that this gradient is set up because the long loops of Henle
of the juxtamedullary nephrons and the collecting ducts act with their
associated vasa recta as a counter-current system. Other examples of
such a system exist, for example the heat exchange system between
arteries and veins (venae comitantes) in the limbs. The system is
illustrated (for heat flow) in Figure 8.8. This shows that turning the
tube back on itself leads to a vastly increased temperature gradient
for the same heat input.
In the kidney an osmotic; gradient is established in the extracellular
space, in the loop of Henle, in the collecting duct and in the vasa
recta by a system which depends on the recirculation of both sodium
and urea (sometimes called the sodium and urea cycles). An essential
part of this system is that the blood flow in the vasa recta during the
production of a concentrated urine is sluggish, thus allowing the vasa
recta and interstitial space to act as a poorly stirred 'compartment'.
The movement of Na, urea and water depends on:
1. The active transport of CI- (followed by Na+) in the thick
part of the ascending loop of Henle and
2. The permeability properties in the various parts of the svstem.
THE KIDNEY AND MICfURITION
.
10ml/min
10 11' W; r--+
300 m mtn.. I
30 40
40
50
50
60
70
Figure 5.8 A simple example of a counter-c:urrent system.
153
The system is complex and not completely understood. Students
should study carefully, together with the following general outline:
1. The blood entering the outer medulla in the vasa recta
receives NaCI from the thick loops of Henle (both short and
long nephrons) and so becomes more concentrated.
2. The tubular fluid entering the thin loop loses water and so
also becomes more concentrated.
3. The fluid ascending the thin limb of the loop of Henle
exchanges NaCI for urea and then, in the thick segment,
loses more NaCI; it is thus hypotonic as it enters the distal
convoluted tubule.
4. The filtrate in the distalloop and collecting duct loses both
water and urea, thus becoming more concentrated again.
NaCI can be considered to circulate (or cycle) round the loop of
Henle and associated vasa recta, and urea to circulate round the
ascending loop of Henle, the collecting duct and the associated vasa
recta. NaCI and urea which is not excreted in the urine is removed
by ithe vasa recta back into the circulation.
It is clear that long loops of Henle are required for the system
to work, as animals with no loops of Henle (fish) or those with short
154 PHYSIOLOGY, BIOC'HEMISTRY AND BIOTECHNOLOGY
loops (for example beavers) cannot produce a concentrated urine.
Similarly, only juxtamedullary nepbrons, not cortical nephrons, are
used in this system, though the cortical nepbrons do contribute to the
system by pumping out CI-. .
5.2.8 Hormonal Control of Sodium Reabsorption
Regulation of body sodium levels occurs not only through
controlling plasma osmolality by selective reabsorption of water but
also by the hormonal control of sodium reabsorption. Sodium
deficiency results in the secretion of a steroid hormone, aldosterone,
from the adrenal cortex. Aldosterone acts primarily on the distal tubule
to promote tubular reabsorption of sodium.
Figure 5.9 Properties of the juxtamedullary nephron during the production of a
concentrated urine. The spaces between the tubules contain the vasa recta; this is
considered (together with the interstitium) to be a single compartment.
5.2.9 Urea Excretion
The isosmotic reabsorption of sodium and water in the proximal
tubule raises the concentration of the other solutes, including urea,
so that concentration gradients are established favouring passive
reabsorption. By the end of the proximal tubule about 50% of the
urea fIltered has been absorbed. During passage of the fIltrate through
THE KIDNEY AND MIcruRITION
f
.,.
I
I
I
No+ Urea
cycle cycle
I
I
I
I
+
155
Figure S.10 The Na+ and urea 'cycles' involved in the concentrating mechanism. The
movement of water from the descending limb and that of NaCI from the ascending
limb into the vasa recta acts lIS a counter-current multiplier; the movement of urea in
the collecting duct and the ascending limb act as another.
thJ loop of Henle urea diffuses from the collecting duct back into the
filtrate (the urea cycle) so that the urea concentration in it rises. The
final excretion of urea by the kidney is normally only about 13 %, so
that a total of 8 7 % of that fIltered is reabsorbed. This large
reabsorption is mainly accounted for by its movement down the
concentration gradients created by water loss from the collecting duct

5.2.10 Reabsorption of Sugars and Amino Acids
i Sugars and amino acids are completely reabsorbed in the proximal
In the case of sugars it has been shown that a stereospecific
transport system exists in the brush-border membrane of the proximal
tubular epithelial cells. Transport of sugars shows competition resulting
in different rates of reabsorption for different sugars. The relative
rates of reabsorption for some common sugars are:
Glucose > Galactose > Mannose > Fructose > Xylose > Arabinose
The transport of sugars across the brush-border membrane depends
on the existence of an electrochemical potential difference for sodium.
TQis indicates that the cellular entry of sugar is coupled to sodiwn
transport and involves a Na-K-ATPase. Net transport can be inhibited
either by oubain which blocks the Na-K-A TPase or by phlorizin which
bl9Cks sugar access to qte carrier protein. Reabsorption of glucose is
saturable and this is way the kidney can exercise a degree of
Vasa Loop of Collecting
recta Hen Ie duct
Figure 5.11 Passive reabsorption of urea. During antidiuresis around balf of the filtered
urea is removed in the proximal rubule.
course control over blood glucose levels. If glucose is present in the
blood above a certain level then the excess will be excreted.
Remember, however. that the liver is the major site for glucose
homeostasis in mammals.
Sugars leave the proximal tubule cells across the basal-lateral
membrane by facilitated diffusion down a concentration gradient
established by the sodium-dependent transport system at the brush
border. The carrier system for sugars. at the basal-lateral membrane
is not sodiumdependent and has a low sensitivity to phlorizin.
Reabsorption of different classes of amino acids OCCW"S by different
carrier mechanisms and hence they do not compete with each other.
For example, basic amino acids such as arginine and histidine do not
interfere with the reabsorption of acidic amino acids such as aspartate
and glutamic acid. The neutral amino acids also appear to have a
separate carrier mechanism although this is shared to some extent
with that for the basic amino acids. .
THE KIDNEY AND MICfURITION 157
I
I
[Glucose] in filtrate
Figure 5.12 Saturation of the glucose transport system. At a low concentration of
glucose in the filtrate total reabsorption of glucose occurs; at high concentrations a
maximum amount can be reabsorbed, the rest is excreted.
5.2.11 Tubular Secretion
Tubular secretion provides a second route by which substances
may enter the tubule from the plasma. In common with tubular
reabsorption this may be either an active or passive process. Tubular
secretion provides an important route for eliminating unwanted
metabolic breakdown products and foreign chemicals (for example,
drugs). The liver also plays an important role in the elimination of
degradative products of metabolism. Many of these substances, for
example steroids, are lipid soluble and the liver converts them to
water-soluble molecules by combining them with certain acids, for
example glucuronic acid. This also conveys specificity to the molecule
allowing the use of a tubular secretion mechanism. The most abundant
and important substances to enter the tubule by tubular secretion are
undoubtedly H+ and K+.
5.2.12 Potassium Regulation
I Almost all of the K+ in the filtrate is reabsorbed by active transport
regardless of changes in body potassiwn balance. Control over potassiwn
levels is achieved by regulating tubular secretion of K + ions back into
the distal tubule. Thus if an animal is K+ deficient there will be a
reduction in K+ secretion. Conversely, if there is an excess K+ intake
from the food there will be an increased tubular secretion and increased
excretion of K+ ions. The rate of tubular Kt secretion is controlled by
the levels of aldosterone through a different mechanism from that
~ r t i n g to conserve Na+ ions. It would appear that increased K+
Ilevels directly stimulates the aldosterone secreting cells of the adrenal
cortex resulting in enhanced tubular secretion of potassium and increased
K + loss in the urine
5.2.13 Renal Regulation of Hydrogen Ion Concentration
The daily diet of Western man produces about 60 mmoles of H l-
ion when metabolised; therefore this amount of acid (equivalent to
60 ml 0,1 N HCl) must be excreted by the kidneys each day. Almost
all of the H+ excreted in the urine enters the tubules via tubular
secretion. On a normal diet, man usually excretes a urine which is
acid (PH 6'5) relative to plasma (PH 7'4). Acidification appears to
take, place throughout the entire length of the nephron except perhaps
the loop of Henle. The amount of H+ secreted into the tubule depends
not on hormonal or nerve intermediates but on buffers present in the
epithelial cells themselves. The major mechanisms of hydrogen ion
secretion by the kidney involve: (l) HC0
3
-reabsorption; (2) titratable
acid formation; (3) ammonium secretion.
Food I ""'-
Drink '---------.1/'"
Increased K + <
elimination
in the urine r---...J
Dietary K+
1
Piasma concentration K+ [t]
1
Adrenal
cortex
l,.-[tJ
Figure 5.13 Regulation of K+ levels by wbular secretion.
Bicarbonate reabsorption. In the proximal tubule about 80-90%
of the filtered HC0
3
is reabsorbed in exchange for H + formed in
the tubular cells from the breakdown of C 3 by carbonic anhydrase.
According to this scheme, for every mole of H+ that is secreted into
the tubule lumen a mole of Na+ is reabsorbed. The H+ ion secreted
into the lwnen combines with another HC0
3
from NaHC0
3
in the
filtrate and the resulting I\C0
3
Js broken down by carbonic anhydrase
located on the brush-border membrane. The resulting C02 produced
diffuses back into the epithelial cells and hence the pH of the filtrate
remains unchanged. However. in the distal tubule and collecting duct
there is little NaHC0
3
and also carbonic anhydrase is not present in
significant amounts on the luminal surface of the tubule cells. The
maximum H + gradient against which the transport mechanism can
I
secrete in man corresponds to a pH of around 45. Thus if there were
no buffers in the tubular fluid the H+ concentration would rise so
high that there would be back diffusion into the peritubular capillary
plasma.
NaHCO;s
Jt
,- .
',;;:- - ~ ~ ~ : :'. .
~ ~ ~ . '''','
. ~ ; ~ ~ . '
',: .:---
. , "
lumen tubular
cell
Capillary
I
Tubular Renal
Figure 5.14 Reactions involved in H' secretion and bicarbonate reabsoptioD by proximal
tubular cells.
Titratable acid formation. The major titratable acid in the tubular
fluid is phosphate.
HPO!- + H+ -+ H
2
P0
4
Approximately one-third of the excess H+ produced is excreted in
this form. Other weak acids in the urine include creatinine and (3-
hydroxybutyric acid. These are usually only present in small amounts
and contribute little to titratable acid formation except in certain
pathological conditions such as diabetic acidosis.
.... :-' .

fieCO
. ....
. -. '.-: ':..

<-.', -;-:
COa+HzO
Figure 5.15 Mechanism of ammonia excretion in tubular cells. The ammonia diffuses
in the non-ionic form (as NHJ because the membrane has a low permeability to the
ionic form (NH4 +).
Ammonium secretion. The remaining two-thirds of excess H+ is
excreted in the form of anunonium. Ammonia is produced metabolically
within the tubular epithelial cells from glutamic acid. The ammonia
diffuses into the tubule where it combines with H+ to form NH
4
This
in turn reduces the ammonia concentration in the tubule causing more
diffusion of from the cell which is able to combine with still
more H+. This is a base generating process since the secretion of H+
': is accompanied by the generation of HCO
l
which diffuses into the
blood. The 'mopping' up of H+ by ammonia represents the major
mechanism whereby the kidney can secrete H+ ions under conditions
of acidosis. .
5.2.14 Mechanism of Micturition
Urination is a local spinal reflex which is influenced by higher
centres in the brain. Urine collects in the bladder by peristaltic
contractions of smooth muscle in the ureteral walls. The bladder walls
are made up of thick layers of smooth muscle supplied with stretch
receptors and a rich parasympathetic nerve supply. The afferent
neurones of the stretch receptors in the bladder wall synapse in the
spinal cord with these parasympathetic nerves. There is also a circular
THE KIDNEY AND MlCfURI110N
1 1 Stimulatory neurone
Bladder
(smooth
External urethral
sphincter
(skeletal musclel ___
Figure 5.16 Structure and nerve supply to bladder.
161
Facili tatory and
inhibitory input
from higher
brain centers J;
layer of voluntary skeletal muscle around the base of the bladder called
the external urethral sphincter. This sphincter is able to hold the urethra
closed even against strong bladder contractions. Stimulation of the
bladder stretch receptors activates the parasympathetic nerves causing
bladder contractions and simultaneously inhibits the somatic motor
nerves bmervating the external urethral sphincter resulting in urination.
In adults descending pathways from the cerebral cortex inhibit the
parasympathetics, resulting in stimulation of the motor nerves
to ;e external sphincter. This opposes the synaptic input from the
blad r stretch receptors and allows urination to be delayed at will.
Con 01 of the local spinal reflex for urination is a learned process
and must master the controls of these pathways during 'toilet
training'.
6
Reproductive Physiology
The great diversity of species in the animal kingdom is mainly a result
of the development of sexual reproduction, in which the genetic
material from two individuals is combined in their offspring.
The sex of higher animals is determined by one pair of chromos-
omes, the sex chromosomes. The nucleus of body cells of a genetic
female contains two similar sex chromosomes, the X chromosomes,
while those of the genetic male contain one X and one Y smaller
chromosome: the many aspects of sexual differentiation arise either
directly or indirectly from this chromosome difference: In the normal
human there are, in addition to the sex chromosomes, 44 non-sex
chromosomes or autosomes.
Fertilisation occurs when a male germ cell (the spermarozoa)-
containing half the normal number of chromosomes (i.e. 22X or 22Y)-
combines with a female germ cell (the ovum) again containing half
the normal number of chromosomes (i.e. 22X).
6.1 SEX DIFFERENTIATION
The male and female reproductive systems both consist of three
parts: 1. organs where the germ cells are made-the gonads (testes
in the male, ovaries in the female); 2. ducts for conveying germ cells;
and 3. the external sexual apparatus. The development of these three
parts occurs as fonows.
1. The gonads develop under genetic control from a primordial
ridge of tissue near the adrenal glawA. Initially' the primordial
gonad is divided into an outer cortical part and an inner
medullary part, identical in both Sf,xes. In the male the medulla
enlarges to form the testes, in the female the cortex becomes
the ovary. Once the testis de';elops it starts to produce the
male hormone-testosterone-which then influences the
162
REPRODU('''TIVE PHYSIOLOGY 163
subsequent development of the foetal reproductive apparatus.
2 Before the gonads differentiate to an ovary or testis, they
are connected to the body surface by two sets of ducts; after
sexual differentiation one or the other set survives, depending
on the sex of the child. In the normal male, during
development, the testis produces both testesterone and a factor
(Mullerian regression factor, MRF) which causes degeneration
of the female ducts. Together, testesterone and Mullerian
regression factor cause the development of the male duct
system.
Indifferent gonad
Epithelium
Primary sex cords
Cortex
Mesonephros

Medulla
Cortex
- _Secondary
sex cords
. Testis Ovary
6.1. Differentiation of the gonads. When first formed the embryonic gonad
iJu; the potential to develop into either a testis or an ovary; the course of development
it takes depends upon the genetic constitution of the embryo. In a normal male the
primary sex cords of the medulla develop to form the seminiferous tubules. In a normal
female the primary sex cords regress and are replaced by secondary sex cords derived
from the gonadal cortex.
3. Testosterone alone is required for male external sexual
apparatus to develop; in its absence the female ducts and
external apparatus develop. The reproductive apparatus
therefore develops through a sequence of steps. Initially the
genetic information carried in the foetal cells controls gonadal
development: in the male testicular induce sub
sequent male development. Female development takes place
in their absence. Aberrant sexual development can therefore
result from genetic or hormonal abnormalities.
6.2 FEMALE REPRODUCTIVE SYSTEM
The ovaries have the dual function of producing germ cells and
of secreting female sex hormones. The foetal ovary contains many
primordial germ cells, which divide mitotically and develop into a
Wolfflan
ducf __ -fI
MUllerian
duct --_..-t\
Bladder - - - ~ r l H H - -
Ind ifferent
No
testosterane or
~
fallopian
tube
~ H - __ Uterus
H!!!I'---Bladder
Figure 6.2 Differentiation of the genital ducts. The genital ducts of the foetus initially
have the potential to develop into those of either sex. If testosterone and Mullerian
regression factor (MRF) are both secreted by the foeta) testis. a male duct system is
formed. In the absence of these substances female ducts alone persist.
REPRODUCTIVE PHYSIOLOGY
l65
!::..
'1::"
. . ~
~ u
K
. ~
U
largt:r type of cell (oogonia). The oogonia themselves undergo further
mitotic division and fmally give rise to primary oocytes. Oocytes are
the cells from which ova will be formed by meiotic (reduction) cell
divlsion. Multiplication of female germ cells is completed before birth.
:f.
, at if they are destroyed at this stage they cannot be replaced.
. ry oocytes remain in a state of arrested development from the
. at which they are formed (before birth) until just before ovulation.
w h ~ n they divide by meiosis. Meiosis consists of two cell divisions:
one in which the number of chromosomes is halved, and a second
non-reductive division. The second meiotic division of the oocyte does
not actually take place until fusion Occurs between a sperm and the
oocyte surface. At both meiotic divisions, there is an uneven
distribution of cytoplasm to the daughter cells; those which receive
least cytoplasm are termed polar bodies and will eventually degenerate.
Although the female germ cell is usually termed an ovum after
ovulation, strictly speaking it remains an oocyte until after the second
. meiotic division at fertilisation.
Oocytes in the foetal ovary become surrrounded by a single layer
of cells termed the membrana granulosa; together, the oocyte and
the surrounding cell layer constitute a primordial follicle. At puberty
in the human, each ovary contains approximately 100,000 to 500,000
primordial follicles; this represents a great over-production, since only
a few hundred ova will ever be released ..
6.2.1 Ovarian and Uterine Cycles
Release of female germ cells is cyclical and intermittent; this is
reflected by corresponding cyclic structural and functional changes
throughout the female reproductive system. These changes are
dependent on two interrelated cycles: the ovarian cycle, and the uterine
(menstrual) cycle. Both these cycles, although variable, last
approximately 28 days. The menstrual cycle is contolled by the ovarian
cycle via sex hormones.
2.2 Ovarian Cycle
The structures which may be seen within the ovary at different
phases of the ovarian cycle. At no one time are all these stages present
together. At the beginning of each cycle several follicles start to
enlarge. Normally one continues to enlarge, while the others regress
and are said to become atretic. The number of follicles that develop
is determined by the circulating levels of pituitary gonadotrophic
hormones (see below). During the development of the follicle, the
granulosa cells proliferate and form a layer several cells thick. A
layer of flattened cells (termed the theca) then forms around the
granulosa cells. As the follicle develops further the granulosa cells
start to secrete a fluid which forms a space in the follicle-the antrum.
After further enlargement the mature (Graafian) follicle is ready to
be released from the ovary (ovulation). This normally takes place at
about 14-16 days after the start of the preceeding menstrual period.
(Both the ovarian and menstrual cycles are said to begin on the fIrst
. day of the menstrual period.)
REPRODUCl1VE PHYSIOLOGY
167

r
..
Primordial
46

Qertn cells
@
@.
I
@
Multiplication I
46

birth

OoQonia

46

BIRTH

Growth of oocyte

and follicle

PUBERTY
Folilcular

46
motunItlon
46
+ polar
First r:: body
body
em tted (may
o -'(i"j
23
divide)
(.:.} -dory
OVULATION
..... oocyte
+
Spenn pelleltation
!?
23
MeIosis 2
PronIcleate
fertilisation and eQQ
46
emission of second

palar bod,
Sec:ond polar
body
FIgure 6.4 Development of female germ cells. Before birth there is an immense
prolifpration of germ cells in the foetal ovary. By the time of birth this phase of
is complete; the germ cells remain in a state of arrested development
until before ovulation. when they undergo the first meiotic (reduction) cell
divisibn. The second meiotic division takes place after a spermatOzoon penetrates the
oocytr membrane.

follicle

tissue
:Q.. Ovulated
oocyte
follicle
- -f+---Corpus
luteum
1""Hf-----Regressing
carpus
luteum
Figure 6.5 Diagram of the ovary. At no one time are all the structures illustrated
present. At the beginning of the ovarian cycle several primordial follicles start to
develop; one of these will mature to form a Graafian follicle, wbUe other regress.
Ovulation probably occurs in the following way. The enlarging
Graafian follicle stretches the capsule surrounding the ovary, so locally
restricting blood flow through the capillaries. This causes local
degeneration and weakening ovarian tissue, which, apparently together
with enzymic tissue breakdown, causes the ripe follicle to break
through the ovary wall. The oocyte is released along with many
granulosa cells, some of which remain attached to the oocyte and
some which form a cloud of cells around the oocyte (the 'cumulus
oophorus'). After ovulation the oocyte (at this stage often loosely called
an ovum) is picked up by the finger-like projections at the mouth of
the fallopian tube. Cells of the Graafian follicle which are left in the
ovary after ovulation persist and form a folded structure (the corpus
haemorrhagicum) the cavity of which becomes tilled with blood. The
granulosa cells at this stage begin to proliferate and soon replace the
blood clot; the whole structure is then known as a corpus luteum. In
an infertile cycle the only lasts for about 10 days before
REPRODUcrIVE PHYSIOLOGY 169
it starts to regress, fmally forming an avascular scar (a c"orpus
l albicans). If pregnancy does occur the corpus luteum is maintained
for several months.
6.1.3. Uterine Cycle
The uterus has three layers: a thin layer in contact with the body
cavity, a thick muscular layer (myometrium) and a mucous membrane
bounding the uterine cavity (endometrium). During the course of the
uterine cycle the superficial layers of the endometrium undergo cyclic
stJilctural changes which can be divided into three phases: the
phase, during which time the superficial layers are lost;
the proliferative (or follicular) phase, during which the destruction
resulting from menstruation is repaired by proliferation of endometrial
cells; and the secretory (or progestational) phase, during which g1ands
in the endometrial wall become more complex in structure and start
to secrete mucus. In this last phase the uterine lining is prepared for
implantation of the fertilised ovum. The endometrial changes which
during the CO!U'Se of one cycle.
Day of cycle
Figure 6.6 Uterine (menstrual) cycle. After a menstrual period the uterine eDdomebium
rapidly tbickeDs as a result of the cellular proliferation (proliferative phase). In the
BeClOnd balf of tbt' cycle the structDre of the glands in the endometrium becomes more
and they stan to secrete mucus (secretory phase). Fmally. the endomebium
Imjaks down (menstrual phase).
, Shedding of the outer layers of the endometrium during menstru-
ation probably takes place as follows. Spiral arteries which supply the
superficial layer of the endometrium constrict and cause local ischaemia
and weakening of the capillary walls. The spiral arteries then re-open
and blood leaks out of the blood vessels, so detaching the outer layers
of the endometrium and producing the menstrual flow.
6.j.4 Ovarian Hormones
. The ovary produces two major types of steroid hormone:
oestrogens and progesterone. In the human the major oestrogen is 17
In addition to other important functions, these ovarian
Uterine cavity
steroids are of key importance in maintaining both the ovarian and
menstrual cycles. During the early part of the ovarian cycle, oestradiol
is synthesised by the developing follicle sO that the plasma level of
this hormone increases progressively and reaches a peak approximately
2 days before ovulation; oestradiol is responsible for the proliferative
(follicular) phase of endometrial development. The corpus luteum.
formed after ovulation, starts to synthesise both 17$-oestradiol and
Figure 6.8 Blood concentratioos of 17 p-oestradiol and progesterone during the course
of the ovarian cycle. The blood concentration of oestradiol rises as the secretory activity
of developing ovarian follicles increases; the level starts to fall just before ovulation.
The peaks in progeslelone and oestradiol concentratioos which occur after ovulation
result from secretion of these hormones by the active corpus luteum.
REPRODUCTIVE PHYSIOLOGY 171
progesterone. Progesterone induces the secretory (progestational) phase
of the uterine cycle; that is to say, it prepares the uterine endometrium
for implantation of the fertilised egg. To be maintained in its secretory
phase, the endometrium requires the honnonal suppon of oestrogen
and progesterone. If fertilisation does not take place, the corpus luteum
starts to regress towards the end of the uterine cycle and so secretes
decreasing amounts of oestrogen and progesterone: the circulating
levels of these hormones fall, and so the uterine endometirum is shed.
Anterior Pituitary and Hypothalamus
, tSo far we have seen that cyclic changes in the ovary hormonally
con 01 corresponding changes in the uterine endometrium. The ovarian
cyc e is, however, itself controlled by the anterior pituitary, which,
in turn, is under the influence of the hypothalamus.
The anterior pituitary produces three protein hormones (gonadotrop-
hins) which are of primary importance in reproductive function; they
are follicle stimulating hormone (FSH); leiIteinising hormone (LH)
and prolactin. The secretion of gonadotrophins is dependent on the
activity of the hypothalamus, which is one reason why female reproduc-
tive cycles and fertility can be greatly affected by emotional influences.
The hypothalamus controls the anterior pituitary by the release of
substances from the terminals of specialised neurosecretory neurones
into the hypophyseal blood portal system. Although at one time it was
thought that the secretion of FSH and LH was controlled independently
via two specific releasing factors produced by the hypothalamus, it
now seems that a single hypothalamic releasing factor controls the
release of both these gonadotrophins. Unlike FSH and LH, prolactin
secretion is controlled by prolactin inhibiting factor (PIF), which, when
from the hypothalamus, inhibits the secretion of prolactin
the pituitary. Therefore, prolactin is secreted in the absence of
PIl1.
6.Z.6 Control of the Ovarian Cycle
At the onset of the ovarian cycle the blood concentrations of both
LH and FSH are relatively low. As the cycle progresses their
concentrations start to increase. The rise in FSH is primarily responsible
for triggering follicular development and oestradiol secretion; later
stages of follicular development are dependent on both FSH and LH.
The progressive rise in oestradiol secretion which occurs during
follicular development has two effects on the anterior pituitary. First.
it iirectly inhibits further release of FSH by negative feedback loop
through the hypothalamus. Second, it produces a surge in the release
of LH by means of positive feedback via the hypothalamus; this peak
in the circulating level of LH triggers ovulation. The circulating level
of 17 Jl-oestradiol falls a little before the peak of the LH surge. After
ovulation the concentration of LH remains slightly elevated and this
hormone, probably together with prolactin, is responsible for the
formation and maintenance of the corpus luteum. However, LH
secretion is apparently inhibited towards the end of the cycle as a
result of negative feedback exerted by a rising concentration of
progesterone. The mechanism responsible for the regression of the
corpus luteum at the end of the human ovarian cycle has not been
resolved. In some animals, such as sheep, one of a group of substances
termed prostaglandins is involved; this, however, is apparently not the
Figure 6.9. Control of ovarian function. At the beginning of the ovarian cycle follicle
stimulating hormone (FSH) secreted by the anterior pituitary gland triggers follicular
development; the developing follicles start to secrete increasing amounts of oestrogens.
These oestrogens act on the hypothalamus to produce two effects on the secretion of
pituitary gonadotrophins: first. they inhibit the secretion of FSH, and second, they
enhance the secretion of luteinising hormone (LH). LH itself stimulates further secretion
of oestrogens. The peak in LH concentration which occurs is responsible for triggering
ovulation.
REPRODUCI1VE PHYSIOLOGY
Menses
I I
I
-,
I \
I \
I \'
/ \
Menses
I
I
/ \
I.. I \ I
I . \
I . / '. I
:' .,. '. \ .. 17/3-
173
... ,..... . L I . . . . . .} . oestradiol
.......... - ----/ ' ... r o J ~
Day of cycle
FIgure 6.10 Sununary diagram of the major events which take place during the course
lof the ovarian and uterine cycles (modified from Segal).
I
(a) Blood concentration of follicle-stimulating hormone (PSH) and luteinising hormone
(LH).
,(b) Blood concentrations of ovarian steroids.
:(c) Development of ovarian follicles and the corpus luteum.
(d) Changes in the uterine endometrium.
case in man. After regression of the corpus luteum. and the fall in
levels of the ovarian steroids, oestradiol and progesterone, the
concentrations of FSH gradually increase again.
The relative time course of various events associated with the
ovarian and uterine cycles.
6.2.7. Oral Contraceptives
Although it was known for some time that oestrogens and
progesterone could theoretically influence fertility, they are not
effective as contraceptive agents because they are rapidly inactivated
in the liver after oral administration. Synthetic compounds have been
developed which have the biological activity of oestrogens or
progesterone, but which are resistant to enzymic degradation. One of
the most commonly used oral contraceptive formulations is the
'combined pill', which contains synthetic oestrogen and a progestagen
(a compound with a progesteronelike action) and is taken from
approximately the fifth to the twenty-fifth day of the menstrual cycle.
This type of oral contraceptive acts by blocking the release of
gonadotrophins from the pituitary, thus preventing ovulation. It is also
thought that they have an additional anti-fertility action at the site of
the uterine cervix and endometrium.
6.2.8 Pregnancy
Fertilisation normally takes place in the widest part (ampulla) of
the Fallopian tube. After a short delay the fertilised egg (zygote) starts
dividing and forms a ball of cells termed a morula. As the cells
continue to divide they start to form a fluid-filled cavity, the
blastocoele. A single layer of cells, the trophoblast, forms around
the blastocoele: after implantation these trophoblast cells will form
the placenta and embryonic membranes. At one pole of the blastocoele
there is an aggregate of cells, the inner cell mass, which later forms
the actual foetus. At this stage the embryo is called a blastocyst.
The fertilised egg takes about 24 days to travel from the ampulla
to the uterus (as a blastocyst). It then implants into the uterus (usually
on the posterior wall) through the burrowing action of the trophoblast
layer. After implantation the cell membranes of the trophoblast cells
start to break down to form a syncytium. In the human placenta the
trophoblast tissue disrupts the walls of blood vessels in the uterine
wall to become directly bathed in maternal blood; as the placenta
enlarges the trophoblast layer develops into the chorion.
The embryo is immunologically different from the 1ll9ther and
might be expected to be rejected as would be a graft from an
REPRODUCfIVE PHYSIOLOGY
Uterus
Early
implantation
Blastoqst
Morula
2 cell
stage
I sf. cleavage
Development
of prorucJej

penetration
U IJ\ n", __ " Fimbria
t-----Ovary
175
Figure 6.11 Fertilisation of the egg and initial development of the zygote. The oocyte
is released into the body the ripe ovarian follicle: it then enters the Fallopian
tube, where it is fertilised. The zygote then starts to divide mitotically, becoming first
a morula, then into a blastocyst; during implantation the outer layer of the
blastocyst burrows imo the uterine endometrium.
incompatible donor. At the present time the way in which the embryo
. avoids rejection is unclear.
16.2.9 Hormones in Pregnancy
, One of the first problems to be overcome by the fertilised ovum
is to prevent menstrual loss of the uterine endometrium and so prevent
abortion. To maintain the uterine lining it is essential to prevent the
fall in the blood progesterone concentration which occurs at the end of
an infertile cycle as a result of luteal regression. Although the mecha-
nism is not entirely clear, the corpus luteum is probably maintained by
i the secretion of two protein hormones from the trophoblast of the
I blastocyst. These hormones are known as human chorionic
I gonadotrophin (HCG) and human chorionic somatomammotrophin
(HCS, also known as human placental lactogen). Thus, in a sense, the
, placenta takes over the role of the pituitary in maintaining hormone
(c) .
. ::".
,':
(syncytial)
_----Uterine stroma
__ Slnusalds
filled with
maternal tIIoad
__ Syncytial
:; trophoblast
__ Yolk sac
cavity
( blastOCOele)
__ Extra-embryonic
coelom
Figure 6.12 Early stages in the development of an embryo.
(a) The blastocyst is composed of the trophoblast layer and the inner cell mass; me
inner cell mass will later form the actual embryo. while the trophoblast layer forms
the embryonic membranes and the foetal pan of the placenta.
(b) AB the blastocyst implants into the uterine wall the membranes of the invading
portions of the trophoblast develop into a syncytium.
(c) Fmally, the syncytial trophoblast breaks down the walls of the blood vessels in the
uterine lining and becomes bathed in maternal blood.
The ovary is able to synthesise progesterone from acetate (the starting point for the
synthesis of all steroids) and can convert this to oestrogens; the placenta. on the other
hand. lacks some of the enzymes necessary both to convert acetate to progesterone
and progesterone to oestrogens.
177
production by the corpus luteum at the beginning of pregnancy. In
early pregnancy, as in the ovarian cycle, the corpus luteum is the
major source of progesterone and oestrogen; however, as pregnancy
progresses the placenta itself begins to take over some of the hormonal
functions of the ovary by producing oestrogen and progesterone. In
fact, by the sixth week of gestation the corpus luteum is no longer
necessary. The concentrations of HCG, HCS and progesterone during
the course of pregnancy.
g!
~
Corpus luteum
progesterone
Placental
progesterone
e Expected
-I perlad
Implantation 1 /r
~ I l
i OwIatr /. /. . HCG
J --:/'
CD /f./ Oestrogen
",.
32 36 5
'--Days--aft-er---"';''''-Ious--per-j-ad---J' ' .... --W-eeks--O""" r""pregn--ancy--J
Figure 6.13 Concentration of hormones in the maternal blood during the course of
pregnancy. If fertilisation occurs the blood concenttation of progesterone does not fall.
but is maintained mainly through the action of human chorionic gonadotrophin (RCG)
oo:the corpus luteum. By about the fifth week of gestation the placenta starts to take
over progesteIone synthesis from the corpus luteum; after the initial slages of pregnancy
the placenta is responsible for synthesising progesterone, oestrogens. BCG and human
chorionic somatomammotrophin (RCS).
~ therefore extracts the steroid cholesterol from the maternal blood
sum1y and converts it to progesterone. Some of this progesterone
passb to the foetus, where the foetal liver and adrenal cortex convert
it to precursors which, in turn, can be used by the placenta to
synthesise oestrogens. Thus, steroids are shuttled between the placenta
and the foetus allowing the biosynthesis of oestrogens; this cooperation
has given rise to the term 'foeta-placental unit'. Of course, it must
be remembered that the mother is essential in these syntheses. since
she initially supplies the necessary cholesterol. The transfer of materials
bet\\leen the mother and the 'foeta-placental unit',
I
6.2.10 Pregnancy Diagnosis
f'Aodem pregnan'i}' diagnosis depends on the detection of human
MOTHER
FOETUS t
F"JgUFe 6.14 Biosynthetic cooperation between the foet .l-placental unit and the mother.
chorionic gonadotrophin (HCG) in the UTdle of pregnant women by
an inununological method. The urine sr.mple to be tested is mixed
with antiserum containing antibodies to J ~ C G . The antiserum plus urine
is then mixed with latex particles c(Jated with HCG. If the urine
contains no HCG the antiserum will react with the HCG on the latex
particles and produce a precipitate, indicating a negative diagnosis. If
REPRODUCflVE PHYSIOLOGY 179
the urine sample contains HCG the antibodies in the serum wUl have
been used up and so cannot react with the HCG on the latex particles
so there is no precipitation; this indicates a positive diagnosis. The
general principles underlying inmmnological tests are discussed.
6.2.11. Parturition
At present the mechanism by which parturition is started is not
fully understood. Until recently the mother was thought to be entirely
responsible for the control of parturition, while the foetus played a
~ s i v role. ..
Adrenal
cortell
Maternal
hypothalarnus
and
pituitary
Sensory slqnals
(respond to dilation
of uterus, cervi& CI1d
vagina)
~ 6.15 Control of parwrition. Stinmli (probably including stress) act upon the
foetal hypothalamus and cause the release of adrenocorticotrophic hormone (ACTlI)
(rum the anterior pituitary gland; this in turo causes the secretion of corticosteroids
(mm the foetal adrenal glands. Corticosteroids lower progesterone secretion and increase
the secretion ofprostaglandins by the placenta. so inducing uterine contractions. Once
,tarted uterine contractions are reinforced both by autonomic reflexes and through the
release of oxytOCin from the posterior pituitary gland.
The onset of parblrition was thought to be almost entirely controlled
by the relative concentrations of progesterone and oestrogens in the
maternal bloodstream, since uterine contractility is depressed by
progesterone and enhanced by oestrogens.
It now seems that the foetus can control the onset of parturition.
Certain undefined stimuli (possibly stress) can act upon the foetal
hypothalablus to trigger the release of adrenocorticotrophic hormone
(ACTH) from the foetal pituitary. The ACTH, in tum, stimulates
the foetal adrenal to secrete corticosteroids. These steroids cause a
fall in the placental progesterone concentration (and hence increase
uterine contractility) and an increase in the secretion of a prostaglandin.
(Prostaglandins can induce powerful uterine contractions.) Although
thiS view of parturition is based mainly on work with sheep, it is
probably also applicable to humans.
Once parturition has begun uterine activity itself enhances uterine
contraction; as the uterus contracts it dilates the cervix and stimulates
stretch receptors in the cervical wall. Nerve impulses travel into the
spinal cord. and cause uterine contraction via autonomic reflexes.
Impulses from $lretch receptors also travel up the spinal cord to the
brain, and act on specialised hypothalamic neurones which send their
axons through the pituitary stalk to terminate in the posterior lobe of
the pituitary. When they are stimulated they release an octapeptide
hormone (oxytocin) from their terminals into the blood. Oxytocin
produces strong uterine contractions.
6.2.12 Lactation
The mammary gland consists of two types of tissue: glandular
tissue or parenchyma. and supporting tissue or stroma. The glandular
tissue consists of, small sacs or alveoli, which are lined with a single
layer of glandular epithelium. The alveoli are surroundett by a layer
of contractile myOepithelial tissue. Milk is formed by the cells of the
glandular epithelium; it is conveyed from the alveoli to the nipple by
a system of ducts.
Lactatic n has two phases: milk secretion. and milk removal. In
milk secretion r iilk is secreted by glandular cells into the lumen' of
the alveoli. This process is controlled by a number of hormones. In
the rat both prolactin and ACTH (and hence, presumably, corticos-
teroids) 'are necessary. Since both these hormones normally influence
manUnai:y development,' after parturition their action must be switched
so 'that' they ,iriitiate:milk secretion. Probably lactogen,eslsis blacked
during gestation by the direct action of progesterone on the gland
+-___ Lobules
containing
alveoli
181
Figure 6.16 Structure of the human breast. Milk is secreted by sacs (alveoli) arranged
into a number of lobules. Milk passes from the alveoli to a number of sinuses and is
finally carried to the nipple via galactophores.
cells; when the progesterone level falls at parturition, the protein
hormones are able to induce lactogenesis.
Milk removal is the transfer of milk from the alveoli to the nipple,
where it is available to the infant. This is brought about in the following
way by the milk ejection reflex. Sensory receptors in the nipple,
stimulated by the suckling infant, send impulses up the spinal cord to
the
lbe hormonal requirements for milk secretion are not known with
although it seems that prolactin and adrenocorticotrophic
ho ne (ACTH) or corticosteroids must be supplied; it also seems
that e blood progesterone concentration must be relatively low.
hypothalamus, where they trigger oxytocin release from the
posterior pituitary. In addition to its action on the uterine myometrium
oxytocin causes contraction of the myoepithelial cells surrounding the
alveoli of the breast. This forces milk from the alveoli into the ducts
and sinuses of the manunary gland and makes the milk available.
fbe hormonal control of manunary function is extremely complex
and lS dependent on hormones from the ovary; the anterior pituitary,
the Mrenal' cortex and, during pregnancy, from the placenta: the
for duct development are different" from those for
development of the lobulo-alveolar system.
182 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY,
Lobulo- alveolar
system
.... , .. l:f' ',. ':
;" .J.;;v ... ,.. ' ..
: .... ( ,
.
\
............... )
Duct development
Oestrogen
+
GH
+
Corticosteroids

'f"
4 1
}. )
/

Development at
entire system
Oestr0gen
+
GH
+
Corticosteroids
+
Progesterone
+
Prolactin
M_ilk +
}
Prolactin
A+CTH (cortlc:osteroids?)
secretion
Low progesterone
FIgure 6.17 Sununary of the bormonal requiremems for mammary growth and fimction.
Hormones from the ovaries, piblilal'y gland and adrenal glands are necessary for normal
breast function: duel development may take place in the presence of oestrogens. growth
hormone (GH) and corticosteroids; the lobule-alveolar system will develop if, in addition.
progesterone and prolactin are available.
REPRODUCfIVE PHYSIOLOGY
183
6.3 MALE REPRODUCTIVE SYSTEM
The general structure of the male reproductive system is shown
in Figure. Spermatozoa formed in the testis are mainly stored in the
epididymis. As they pass along the reproductive tract at ejaculation
the secretions of the accessory glands (ampullary glands, seminal
vesicles, prostate, urethral glands and bulbo-urethral glands) are added
to the spermatozoa to form the semen.
6.3.1 Spermatogenesis
1'he testis consists of a number of convoluted seminiferous tubules
in wip.ch spermatogenesis takes place. Germ cells in various stages
of defelopment can be seen in cross-sections of seminiferous tubules.
ClusI:s of spermatozoa are attached to Sertoli cells: although the
funcdon of Sertoli cells is not certain they give the spermatozoa
I
I
_---Bladder
___ ...,,-_ Ampulla
Seminal
vesicle
f!g---+I:--+-'--"'-,f-- Prostate

Bulbo-urethral
gland
Vos

\t\-l-__________ Epldidymis

6.18 Human male reproductive system. Spennatozoa formed in seminiferous
tubules of the testis are mainly stored in the epididymis. At ejaculation they are expelled
from the male reproductive tract in the semen. which consists of spennatozoa suspended
in the secretions of the accessory glands.
mechanical support and may nourish them. Between the seminiferous
tubules are found the Leydig (interstitial) cells which are responsible
for secretion of testosterone, the major male hormone produced
by tile testis. The formation of spermatozoa is shown diagrammatically
in A major difference between germ-cell production/ in the
malJ' and female is that in the male germ cells continue to multiply
in life while in the female this process is comelete before birth.
Spf,rmoto-
gonium
Figure 6.19 Cross section of a human teStis showing seminiferous tubules. Within the
tubules germ cells may be seen at different stages of development; Sertoli cells are
found among the germ cells. Interstitial (Leydig) cells are dispersed in the spaces
between the seminiferous tubules.
The formation of spermatozoa from spermatids takes place by a
complex process of differentiation without further cell division. This
process is termed spermiogenesis, while the structure of the mature
human spermatozoon is shown in Figure.
Formation of the spermatozoa from spennatogonia takes a number
of weeks and can only take place at a temperature slightly below that
of the body; for this reason the testes are held in the scrotwn outside
the abdominal cavity.
6.3.2 Control of Testicular Function
Although the primary function of the testis is spermatogenesis it
is also an endocrine organ, being the source of the androgenic hormone
testosterone. Testosterone itself exerts an effect upon spennatogenesis,
as we shall see later, but it is also responsible for a wide variety of
male characteristics, such as certain aspects of behaviour, increased
muscle mass, laryngeal changes which cause the voice to 'break' at
puberty and balding. Testosterone also c,uses the development of the
accessory glands of the male reproductive tract.
Testosterone secretion is apparently controlled by the amount of
LH secreted by the pituitary. LH seems to act directly upon the Leydig
cells of the testis, causing them to secrete testosterone. The circulating
concentrations of testosterone are apparently controlled by a negative
feedback loop via the hypo thalamus similar to that controlling
progesterone secretion in the female.
The control of spermatogenesis is complex since it is influenced
REPRODUCI1VB PHYSIOLOGY
Spermato-
gonia
1 spermatocytes
185
Number of
chromosomes
46
46
46
46
46
46
23
23
23
Figure 6.20 Spermatogenesis. All male germ cells are derived from spermatogonia
wbich undergo a number of mitotic divisions, finally forming primary spermatocytes;
sq:ondary sperm atocytes are formed by a reduction division or primary spermatocytes
have 23 chromosomes. Spermatids are produced by a second (non-reductive)

. tic cen division and develop into spermatozoa without further cell division. This
w le process continues throughout the reprolfuc?tive life of the male.
by testosterone in addition to both LH and PSH. LH appears to be the
dominant hormone controlling spermatogenesis. It promotes
spermatogenesis by acting at several sites: it has a direct action upon
the germinal epithelium of the seminiferous tubules and also has an
indirect effect by inducing testosterone secretion, which itself enhances
LH appears to act also upon Sertoli cells: as these

lls have a very close relationship with developing spermatozoa, LH
y influence spermatogenesis by this route. One important queStion
t has not been answered is how the rate of spermatogenesis is
to meet the requirements. Some experimental observations
Nucleus
SpermatId
4
Cap
H----Flagellum
1
3
Acrosome
I ~ _________ b m
Figure 6.21 Spermiogenesis. This is the development of spermatozoa from spermatids
and does not involve cell division. Development of the rat spermatozoon is illustrated
(after Clermont).
1. The spermatid initially bears no resemblance to a spermatozoon.
2.Ilntially a t1agellum starIs to form and a cap develops over one pole of the nucleus.
3. The t1agellum lengthens and the nucleUs takes on its characteristic shape; by this
time the nuclear cap bas developed into an acrusome. Mitochondria migrate towards
the base of the flagellum.
4.A cytoplasmic. droplet forms on one side of the sperm head and is finally cast off.
suggest that there is a mechanism for monitoring the rate of sperm
production or the number of spermatozoa in the testes at anyone time.
Although no substance has been isolated with this function, the
provisional name 'inhibin' has been given to a hypothetical substance
which signals the level of spermatogenesis. It is suggested that 'inhibin'
may be released from the testis into the blood in amounts dependent
+
Accessory gland of
male reproductive tract.
Behaviour.
Secondary sexual
characteristics.
187
FIgiJre 6.2% Control of testicular function. Luteinising hormone (LH) alone is required
to stimulate secretion of testosterone by interstitial (Leydig) cells. rn, foDicle-stinmlating
hOdnone (FSH) and testosterone all promote spermatogenesis; it may also be enhanced
by Sertoli cells. The level of spenn production may, in turn, modulate the secretion
of pituitary gouadotrophins by the release ofinhibin'.
on the level of sperm production and that it acts by inhibiting the
release of pituitary gonadotrophins.
6.3.3 Fertilisation
i During coitus semen is deposited in the vagina by the contractions
of ithe muscular walls of the male reproductive tract which occur at
orbsm. The volwne of the ejaculate is approximately 3 ml and normally
co*tains about 100 million spermatozoa per ml. The migration of
188 PHYSIOLOGY, BIOOIEMISTRY AND BIOTECHNOLOGY
spermatozoa from the vagina to the Fallopian tube is mainly brought
about by contractions of the muscles of the female genital tract.
Spermatozoa need to remain in the female genital tract for several
hours before they are capable of fertilising an ovum ('capacitation'):
thereafter they retain this ability for about another 24 hours and so
have a limited life span. When a sperm reaches the ovum changes
take place in the surface membrane of the sperm head over the
acrosome and in the adjacent acrosome membrane so that enzymes
in the acrosome can digest a channel in the layers surrounding the
ovum and aQow the sperm to come into contact with the surface
membrane of the ovum. These changes in the sperm head constitute
the 'acrosome reaction'. Further changes take place in the acrosome,
allowing fusion of the surface membranes of the ovum and the sperm.
This fusion triggers changes in the ovum surface which prevent
fertilisation by other spermatozoa. It also triggers the second mieotic
cell division of the ovum. For some time the nuclei of the ovum and
the sperm remain separate within the ovum cytoplasm. After about
12 hours, having undergone a number of changes including a marked
increase in size, the genetic components from each parent become
truly mixed.
6.4 PUBERTY .
During the period of adolescence a number of dramatic changes
take place. These include development of the gonads, genitalia and
secondary sexual characteristics and changes in body size and
proportions. The attainment of potential reproductive capacity is
reached with puberty. In females this is achieved with the onset of
menstruation (menarche).
There are two major theories on the mechanism by which the
onset of puberty is controlled. In the first theory the hypothalamus is
considered to be very sensitive to inhibition by gonadal steroid
hormones; as a result of this, gonadotrophin release by the hypotbatami
c
-
pituitary axis is inhibited by extremely low circulating concentrations
of gonadal hormones. Synthesis of sex hormones is in tum dependent
on the presence of pituitary gonadotrophins in the blood, so their
secretion is kept in check. As puberty approaches the sensitivity of the
hypothalamus decreases and allows the concentrations of gonadal
steroids to rise, producing the hormone-dependent changes seen during
adolescence. '
According to the second theory, since the hypothalamus contains
gopadotrophin releasing factors and the pituitary contains gonadotrophins
REPRODUCfIVE PHYSIOLOGY 189
before pUberty. its onset must be controlled neurally. That is to say.
before puberty the neural circuits in the hypothalamus are unable to
bring about the release of gonadotroPhin releasing factors. Thus. the
development of this capability controls the onset of puberty. Neither
theory is completely adequate and more information will be required
before we can gain a complete understanding of the mechanisms by
which puberty is controlled.
7
Endocrinology
and Metabolism
It is convenient to deal with the endocrine system separately from
the nervous system. There is, however, no sharp distinction between
them; both are control mechanisms which involve interactions between
cells; both use chemical substances as mediators between cells, and
both cause actions at a distance. On the whole, the endocrine system
deals with events slower than the nervous system and has effects which
are more generalised.
If hormones were 'visible' then the concentration of them in the
body would be seen to vary from moment to moment throughout life.
Each cell is thus bathed with very many local and general hormones
all the time; the resulting action depends on the interaction of many of
them at anyone moment. This complexity makes for difficulty in
description because some simplifying scheme must be adopted. The
one chosen here is to descn"be the hormones separately but to devote
sections to their integrative action. The first section applies to most of
the hormones, and ought to be referred to from time to time.
7.1 HORMONES IN GENERAL
Hormones are usually secreted directly into the circulation (via
the extracellular space) in extremely low concentrations and are
recognised by specific cells, which then respond in characteristic
fashion. Some hormones are secreted as precursors and then formed
in other organs or in the circulation. Typical hormone concentrations
in the blood range from 10-6 to 10-
12
moUlitre compared to (about)
10-
1
moUlitre for Na to 10-
3
moUlitre for amino acids. Measurement
of hormone concentrations is, therefore, difficult and has only recently
been achieved for most of them. Although these hormone concentrations
190
ENDOCRINOLOGY AND METABOUSM: 191
are low, 10-
12
molllitre still means that mI of plasma contains a
hundred million molecules of a hormone.
7.1.1 Factors which Determine the Blood,
Concentration of Hormones
The plasma concentration of any hormone will depend upon its
rate of secretion into and its rate of removal out of the plasma.
Normally, for any hormone, some secretion is always occurring and
inactivation by the liver or 'kidneys is always taking place. The plasma
: concentration may therefore be altered by changing either or both of
I these rates. Usually the rate of inactivation is not changed (except that
higher plasma concentrations lead to greater rates of inactivation);
I control is therefore exercised by changing the rate of secretion of the
hormone. Glands usually have only a small amount of preformed
hormone in them aim so if the demand for secretion increases, although
some may be discharged immediately, most must be made from
precursors.
Table 7.1. The principal endocrine glands of the body and their most
important secretions (from Companion).
Glands Secretions Cbemkal nature
Anterior pituitary or Growth hormone (GH) Protein, mol. wt 21,500
adenohypophysis Adrenocorticotrophic Polypeptide. mol. wt 4500
hormone (ACI'H)
Thyroid stimulating Glycoprotein. mol.
wt 33.000
hormone (fSH)
Luteinizing hormone (LH) Glycoprotein. mol.
wt 33,000
Follicle stimulating Glycoprotein
hormone (FSH)
Prolactin Protein, mol. wt 200,000
(sheep)
. Posterior pituitary or Vasopressin or antidiuretic Polypeptide of 9 amino acids
neurohypophysis hormone (ADH)
Oxytocin Polypeptide of 9 amino acids
Thyroid Thyroxine 0'4) Iodinated amino acid
(T3) Iodinated amino acid
Calcitonin Polypeptide. mol. wt 3600
I Parathyroids Parathyroid hormone Polypeptide, mol. wt 8500
Adrenals
I (Table 7.1 contd.)
Glands
Cortex
Medulla
Kidneys
Testes
Ovaries
Placenta
Pancreas
Gastrointestinal tract
Stomach
Duodenum
Small intestine
Secretions
Aldosterone
Cortisol
Corticosterone and
Chemical nature
Steroid
Steroid
many other Steroid
steroids, including
androgens and ~ g n s
in small amounts
Adrenaline
(USA epinephrine)
Noradrenaline
(USA norepinephrine)
Renin
Brythrogenin
Testosterone and other
androgens
Oestradiol and other
oestrogens
Progesterone
Chorionic gonadotrophin
(human CG)
Oestrogens '
Progesterone
Insulin
Glucagon
Gastrin
Secretin
Cholecystokinin-
pancreozymin (CPZ)
Catecholamine
Catecholamine
Protein, mol. wt 65,000
Glycoprotein
Steroid
Steroid
Steroid
Glycoprotein, mol.
wt 30.000
Steroid
Steroid
Polypeptide, mol. wt 5800
Polypeptide, a single chain,
mol. wt 3500
Polypeptides, mol.
wts 2000-7000
Polypeptide, mol.
wt about 2700
Polypeptide, mol.
wt about 2700
7.1.2 Blood Transport
Many hormones circulate in plasma largely bound to plasma
proteins; free hormone is often quite small compared to the total in
plasma and is in equilibrium with the bound fraction:
free pormone + protein p hormone - protein complex.
Only the free hormone can react with the target cells.
Hormones, are carried in the plasma in this _ way for a variety of
ENDOCRINOLOGY AND METABOUSM
o
10
~
I Free
{P
" 10
.( I
."., I {)
I
Plasma
concentration
Inactivation 67
by liver/!
and kidneys \./ ~
193
Figure 7.1 The plasma concentration of a hormone is the resultant between the rate
of productioD and the rate of inactivatiOD. The hormones may be free iD the plasma
or bound to binding proteins; if bound the total amount in the blood is often largely
determined by the amouJlt of binding proteiD present. The amount excreted is often
used as an indicatioD of blood COncentratioD.
reasons: (1) it prevents small molecules from being excreted by the
kidneys; (2) it enables molecules which are relatively insoluble in
w ~ . to be carried; (3) it enables a 'store' of hormone to exist in
the blood which buffers the supply of hormone to the tissues.
7.13 Hormone Action on Target Cells
The primary event.is a specific interaction between the hormone
and a 'receptor' in the responsive cell: the greater the hormone
concentration the greater the number of receptors occupied. Many
,cells do not recognise certain hormones and it is supposed that these
cells contain no 'feceptors' for these hormones. The present view is
~ there are two main ways in which cells recognise hormones.
I At the cell surface. Certain hormones react with specific receptors
at the cell membrane and are then thought to cause the release of a
sec(>nd messenger within the cell-usually cyclic AMP (cAMP) but

Hormone concentration (J()
Figure 7.2 Binding of a hormone to receptors, showing Michael is-Menten kineIic:s.
At low hormone concenttation only a few sites are occupied: at high c:oncentration
virtually all are oc:cupied.
perhaps cGMP-which then activates a specific kinase in the cell; this,
in tum, causes the physiological changes specific to that cell.
Hormones acting in !'his way are the catecholamines and peptide
hormones. It may be significant that these hormones are large, are
not fat soluble and would not otherwise gain easy access to the cell
interior. This kind of receptor is sometimes known as a fixed receptor.
Several different hormones act through the aiI.enyl cyclase system and
yet their actions are separate. How is this (.one? Each receptor only
recognises its 'OWO' hormone and so is fJnly triggered by it; once
cAMP is fonned it probably activates a specific kinase which then
has various cellular functions. In the cell .he adenyl cyclase mechanism
links receptors- at the cell surface with ,'arioos parts of the biochemical
machinery within the cell; just as in tIn: whole body the action potential
mechanism links different skin receptors to various parts of the CNS.
Within the cell. Steroid and thyroid hormones are small and fat
ENDOCRINOLOGY AND METABOIJSM 195
soluble and probably enter most cells by diffusion. In responsive cells
they react with a specific receptor protein in the cytoplasm. This
complex then enters the nucleus and initiates the physiological
response. This kind of receptor is sometimes known as a mobile
receptor.
7.1.4 The Way In wbich the Primary
Event Alters Cellular Functions
Hormones regulate existing functions in cells rather than initiate
new functions; their action is thus on the rates of existing processes.
T4e ways in which the various hormones do this are varied and, as
yet, not completely understood, but most fit into one of two general
These categories are: (1) changing the activity of a critical
in some pathway and (2) changing the rate of membrane
transport of a substance.
o
f\
ATP C,clic AMP
Active
enzyme QIIIIIIlea
tal

Atmred alII
function
Hormane
(flnt rnessenqer)
Inlrm:ellular or
second messenQer
Protein kinase
system
mRNA

Protein
+
Hormone
action
(b)
Cell
Nucleus
lligure 7.3 Models of hormone acting at: (a) cell membrane; and (b) cytoplasmic
n1ceptors. In each case the specificity of action depends on the specificity of the receptor
molecule.
In addition to cAMP shown, another intracellular messenger,
cGMP. has been discovered which is produced from GTP by guanyl
cyclase. Both are destroyed by a phosphodiesterase. The concentration
of each depends on their rates of production and destruction. Currently
it is thought that prostaglandins are somehow involved on the inner
sj.JIface of the cell membrane between the membrane and the cAMP
Woduction. It is likely that this simple distinction between the two
tYPes of receptors will become blurred as evidence accwnulates.
Resting state
cyclase inactive
Hormone
Receptor
(first messenger) ___ 1-
Adenyl cyclase
mechdnism
I membrane
ATP CyClic AMP
(second messenger)

Figure 7.4 One current idea is that the receptor and adenyl cyclase molecules are
separate and free to diffuse in them embrane; if the receptor molecule is activated. it
then attaches to an adenyl cyclase molecule to form a complex wbich produces the ,
cyclic nucleotide.
Changing enzyme activity. Many processes in the body are
reversible because the forward and reverse reactions occur through
different path ways:
Enzyme A

Glucose Glycogen

Enzyme 8
thus glucose is stored as glycogen through the action of a series of
ENDOCRINOLOGY AND METABOUSM 197
enzymes (containing A) and produced from glycogen via a different
series of enzymes (containing B). Hormonal control is exercised by
altering the activity of certain critical (i.e. rate limiting) enzymes in
these pathways; insulin acts on enzyme A and glucagon on enzyme
B and so, by their relative concentrations, determine the overall
direction of the process.
The activity of enzymes may be increased by increasing the
concentration of the appropriate form of the enzyme; this in turn can
be done by making more of the enzyme or by changing the shape of
the ~ from an inactive (or less active) form to a more active
form.i The general rule is specific interactions with other hormones.
(b) .Ill more complex systems tural changes in pre-existing enzymes
while I hormones which combine with the nucleus produce more of
the enzyme.
Changing membrane transport. Many hormones act by increasing
or decreasing the transport of a substance into target cells. Examples
are: the rate of glucose entry into most cells is increased by insulin
and decreased by other hormones: water and ion movements in t ~
kidney are influenced by various hormones. Such changes in membrane
transport or movement then cause secondary effects in the cells. Thus
an increase in glucose into cells leads to a higher intracellular
concentration of glucose and so (by mass action) causes a greater
formation of glycogen. As knowledge about the detailed working of
hormones accumulates these two classes may merge; for example,
increased transport of sodium foll<;>wing aldosterone treatment may
be due to the synthesis of more transport proteins. In this case
changing membrane transport is only a special case of increased
eDzyme activity.
7.1.S Levels of Complexity of Hormone Action
'Ole actions of hormones in the body can be divided somewhat
arbittkiIy into groups of increasing complexity. (a) The simplest are
those I which subserve a single, specific function with (at present) few
that hormones which produce "intracellular messengers' produce
strucone bodily function or parameter is controlled by a group of
hormones, some of which increase, somc of which decrease its
activity. (c) The most complex systems are those in which a hormone
or grbup of hormones act on almost all body cells, for example to
promte growth or the general rate of metabolism. These. hormones
are 0 en necessary for other more specific hormones to act but do
not' themselves have actions specific to these functions; such
hormones are said to have a permissive action.

L:J \
Blood
Honnane I osmotic
(ADH). pressure
/
(a)
Hormones Thyroxine
present
"
Fat cells
Glucagon
Growth hormone
Blood
__
glucose
concentration --. .......
Insulin
Adrenaline Thyroxin Adrenaline
.... ....
"
I
ct=J
I
I
..
(b)
Result No releaSe Fatty acid release Increased le)
fatty acid
release
Figure 7.5. Levels of complexity of hormone action: (a) single hormone controls action
"ia a feedback circuit; (b) net result depends on relative concentration of several
hl :-.mnes; (c) specific action of one hormone depends on the 'pennissive' presence of
another.
1.1.6 Endocrine Investigations
Classically honnone functions have been discovered by 'nature's
experiments', i.e. diseases have been recognised and correlated with
changes in a particular gland. Later investigations were concerned
with surgical removal and either transplantation of glands or with
injection back into the animal of their contents. Modem investigations
are more concerned with the cellular actions of the hormones. From
the effects of injections of hormones various bioassay methods were
developed: these are specific but oflow sensitivity. Recently, methods
have been developed with much higher sensitivity, but these may not
always be specific for the hormone active centre as they may measure
a non-specific part of the molecule.
These methods were originally devised by Berson and Yallow in
19 59; they depend on using proteins which 'recognise' a hormone
"
by binding to it. Currently concentrations down to about 10-
12
molar
can be measured, often using a standard 'kit'; such methods have
caused a revolution in endocrinology.
7.2 THE BYPOTHALAMUS AND PITUITARY
The pituitary is a small (0 5 g) endocrine gland situated above the
roof of the mouth and just below part of the brain (the hypothalamus).
It is a compound gland derived partly from a downgrowth of nervous
iii
I
-----..
Inc!8allill9 non-radioactive
hormone present

.\
D
Hormone
with
r
lsl
SS>
Figqre 7.6 Principle of saturation analysis for hormones, This involves the displacement
of labelled (hot) hormone from a specific binding protein by unlabelled (cold) hormone,
Hypothalamic
'centres'
Releasllllil
factors
Anterior
lobe
Hormone
producllllil
cell
Short
feed-
back
loop
r-
I
I
I
I
I
I
Posterior
lobe
(a)
eNS
U
+--"
I
I
I
I
I
I LOIIIiJ
_-I feed-
I back
loop
I
I
I
I
I
I
I _______ .J
Hormones
(b)
Figure 7.7 Outline of (a) structure of anterior pituitary and hypothalamic connections,
and (b) feedback pathways important in endocrine control.
tissue and partly from an upgrowtb of the mouth (Rathke's pouch).
Clinical investigators found patients with disorders of the pituitary
showed any of a range of diseases; for example they might remain as
dwarfs or grow into giants: they might have an increased or decreased
sexual development: their metabolism might be increased or decreased;
they might pass large quantities of urine. Some of these changes are
typical of other endocrine glands; and it is now clear that the pituitary
acts largely by controlling other endocrine glands, rather than acting
directly.
I
. J Growth,
development
. 4----- and
Adrenal
cortell
<0

metabolism
Thyrald various
ar;ons
and
tissue
Cortisal ThYroid
hormone
"4---Reproductlon---..
Breast
development
and milk
production
FSH LH

Gonad:
ovary or testis
(I) (2)
Germ cell Hormane
development
(} d
OestroQenarid
progesterane
Testosterone
FigUre 7.8 Main pituitary hormones. These either have direct actions on target cells
(bOxes) or cause secretion of hormones by other endocrine glands (other shapes). The
reproductive hormones have both roles.
I The anterior pituitary and hypothalamus are controlled both by
blbod-borne hormones from target organs and instructions from the
central nervous system. The outcome of this interaction determines
the amount and type of secretion from it.
Harris showed that removal of the pituitary gave the typical
deficiency signs. If the pituitary was replaced elsewhere in the body,
put back into its normal site but insulated from the brain by waxed
paper, function was not restored; direct blood vessel connection
bttween the hypothalamus and pituitary was necessary for restoration
of normal function. The explanation for this is that hormones are
secreted by the hypothalamus and carried by the blood to the pituitary
where they influence the secretion of other hormones.
i hourly feeding
16
1111111111111111111
I
.c: .... OL--------------""---
11
J
Fastino
Noon Midnitaht
FIgure.,., Human growth hormone concentrations in the plasma of a normal subject
under the different conditions shown. A single meal causes a 2-hour depression.
The anterior pituitary secretes six main hormones. To comprehend
their actions they may be thought of in two different ways: (a) whether
they act directly on tissue target cells or on other endocrine cells, or
(b) whether their general function in the body is on reproduction or
on growth, development and metabolism. The six hormones and
indicates how each fits into these two classifications. Some hormones
do not fit a category too clearly; thus prolactin causes development
of the breasts and is concerned with milk production during
reproduction; follicle stimulating hormone and luteinising hormone act
directly on the germ cells to cause them to develop, and are then
involved in causing the gonads to secrete as an endocrine organ.
7.3 GROWTH HORMONE
As its name implies, the long-term actions of growth hormone
(GB) are on growth; excess stimulates growth. lack reduces growth.
If this occurs in young animals (more specifically when their bones
can still grow) then linear growth is affected and giants or dwarfs
are produced. If an excess occurs in the adult then there is over-
growth of the ends of the long bones, of the jaws and of the soft
tissue in man; this is called acromegaly.
Production and structure. GH is produced and released by the
acidophil cells in the anterior pituitary. Growth hormones from various
species have been analysed and shown to be proteins with molecular
weights in the range of 20,000 40,000. There is recent evidence that
ENDOCRINOLOGY AND MEfABOUSM
til
(j)
Tlme--+
(lj)
l m e ~
(ji)
S
2
8.
E
J!!
Time--+
(iii)
Time--+
(iii)
203
Figure 7.10 The concept of hDl/-time. Te . The rate of cooling of a cup of tea depends
on two factors:
GH acts on cartilage via a sulphation factor (or somatomedin) produced
in tbe liver or kidney under the influence of growth hormone.
Blood concentration. The anterior pituitary contains about 1000
times more GH than the other honnones. This may be because GH
lias primary effects on body cells rather than acting as a controller
of other endocrine glands. There is a total daily release of human
GH of about 5 % of the gland content. Serial measurements of plasma
GlJ shows marked bursts of activity, reaching 10 times the basal level.
Th,e Ti of circulating growth honnone is about 25 minutes; it is
metabolised in the liver and kidneys.
Specific effects. As GH is a polypeptide it would be expected to
act at the cell membrane rather than in the cell. At present the picture
is confused but it appears that GH does have a membrane action,
'though its subsequent effects are not through cAMP. Its cellular action
may involve control of protein synthesis at a translational levei; it
also increases the transport of neutral and basic amino acids into cells.
I The effect of GH may be considered on two time scales,
cOJ;Tesponding to a long-tenn growth effect and a short-tenn effect
on: metabolism.
The long-tenn effects of GH determine the size of t h ~ animal
when developing, and the maintenance of nonnal function once it has
grown; they my be summarised as an increase in the protein content
of the body with a decrease in the fat content. Thus, these effects
are to promote protein synthesis and hence cause a positive nitrogen
and phosphorus balance; to control the size of muscle, cartilage and
bone; the size and function of kidney; the bodily retention of the
common ions Na+. K+, CI-, Mg2+ and Cal+, etc.
TIme

~
\
Growth
hormone
(a)
{
t Fat .lisatfon
l ... ___ .. .. Glucose usage
.. Insulin sensitivity
50 - ____ Acromegalic
I ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~
o 50 100
(b)
Plasma [glucose]
Figure 7.11 (a) Shon-term effects of a fall in glucose concentration on fat mobilisation
and glucose usage. Rises in plasma glucose have opposite effects. (b) Effect of plasma
glucose concentration on plasma OH concentrations in normal subjects, acromegaJies
and dwarfs (note log scale). In normal subjects a change in plasma glucose concentration
causes changes in OH concentration; in dwarfs and acromegalies the plasma OH
concentrations are fixed and unaffected by glucose concentration.
ENDOCRINOLOGY AND METABOLISM 205
The short-tenn effects can best be understood and remembered
as adjustment of starvation. In this (and indeed between meals) the
flow of food from the gut stops and bOOy food stores are metabolised.
The plasma glucose concentration drops, this switches on a release
of GH which in turn changes energy usage from carbohydrates to
fats. As a consequence the breakdown products of fats (ketones) appear
in the blood and the plasma glucose rises. Conversely a rise in plasma
glucose concentration decreases the production of GH and this
leads to a fall in the plasma glucose concentration. These
between glucose and GH are particularly important
disease; it is, however, not altogether clear if normal changes in
glucose concentration act in this way. '
Control of GR release. The release of GH from the anterior
pituitary depends on the interaction between hotmones released by
the hypothalamus and factors in the blood according to the general
scheme. The hypothalamus probably releases a growthhonnone-
releasiIig factor (GHRF) which acts to increase the release of GH. A
variety of signals act on the hypothalamus to cause release of GHRF,
for example 'stress' and sleep. As noted above, an important blood
factor acting on GH release is the glucose concentration in the plasma,
which bears a reciprocal relationship to plasma GH concentration.
Glucose
Growth hormone
Growth hormone
Ifigure 7.12 Some stimuli acting on GR release from the anterior pituitary gland. It is
certain that these stimuli act where shown. GRIF is also known as somatostatin,
The mechanism of action of glucose here is obscure, but is thought
to involve the action of metabolite on an intracellular messenger.
7.4 PROLACTIN AND THE
PITUITARY GONADOTROPIDNS
These honnones are concerned with the reproductive activities
of the body, both directly or via other endocrine organs. The pituitary
gonadotrophins are descnDed in detail in the reproductive section, and
are only outlined here; the actions of prolactin are dealt with in this
section. In brief, in the male, follicle stimulating honnone maintains
the spennatogenic epithelium and luteinising honnone causes the
secretion of testosterone output from the testes; in the female follicle
stimulating honnone is responsible for the early growth of the ovarian
follicles whereas luteinising honnone is responsible for their final
maturation and oestrogen secretion and then is responsible for their
conversion to the corpus luteum (the final stage) in the cycle and the
secretion of progesterone.
Prolo.ctin. Until recently it was difficult to separate prolactin from
human growth honnone, partly because there is very little prolactin
present in the human pituitary except during pregnancy and lactation,
and partly because OH has lactogenic effects somewhat similar to
prolactin. Immunoassay methods have now separated them. In the
female prolactin is responsible for milk secretion from the breasts
after priming by oestrogen and progesterone. In the human male the
function of prolactin is uncertain.
Control of prolactin release. The release of prolactin from the
pituitary is normally inhibited by a hypothalamic factor (prolactin
inhibiting factor-PIH); control is exercised by varying this inhibition.
Normally, in man, the prolactin release is increased by nipple
stimulation or 'stresses' of various kinds, with a diurnal rhythm in
which rises occur at night. During pregnancy the secretion increases,
rising to a peak at the time of parturition; thereafter it falls to non-
pregnant levels but is increased promptly by suckling, at least for the
first 3 months of nursing.
7.5 HORMONES OF TIlE POSTERIOR
PITUITARY GLAND .
As in the anterior pituitary this is a point of contact between
neural and homional systems. In this case two honnones, antidiuretic
honnone (ADH, or vasopressin) and oxytocin, are made in the bodies
of nerve cells in the and then-in combination with
protein carrier molecules, the neurophysins-pass slowly down the axons
of these cells to the nerve endings in the posterior pituitary gland.
Here they are stored and released, with the carrier molecules, into
the bloodstream by exocytosis; the rate of their release is comrolled
by impulses which originate, like the honnones, in the bypothalamus
and which travel down the same axons as the hormones did. The
whole complex of hypothalamic nuclei, stalk and posterior pituitary
can be considered as a functional unit, in that they make, transport,
release and control the release of these hormones; the advantage of
system is not clear.
I Environmental stimulus

Plasma volume
recaptor input

Antenor
piiuitary
Osmoreceptors
Supraoptic and ....-Increased
paraventricular osmolality and
nuclei decreased volume
of blood
"\""-4: __
Su
praoptico-
hypophyseal
tract
IFigure 7.13 Posterior pituitary. Hormones made in the bypothalamus 1l10Ve down the
,tract to the endings in the posterior pituitary and are then released ioto the circulation.
Protein carrier molecules (the neurophysins) pass down the tract with the hormones
and are also released into the blood with them. The fibres from the supra-optic nuclei
control ADH secretion, while those from the paraventricular nuclei control oxytocin
secretion.
The two hormones secreted are very similar to each other, differing
only in two amino acids in a total of nine. This led to difficulties in
separation, particularly as their functions overlap.
Antidiuretic hormone. The main action of ADH is in the control
I of water excretion; it does this by increasing the water
of the collecting ducts of the kidneys. leads to increased ,
reabsorption from the urine, which becomes more concentrated thus
conserving water. The way this occurs is unknown except that it causes
the release of cAMP in the tubular cells, which presumably mediates
the effect.
Release of ADH. The secretion of ADD depends upon the
osmolality of the extracellular space fluids; raising this by 1-2% causes
increased secretion of ADD, reducing it abolishes ADD secretion.
The osmoreceptors for this effect are situated within tissues in the
brain, supplied by the internal carotid artery; it is likely that some at
least are situated in the hypothalamus, a possible mechanism. The
capacity of this system to keep the osmolality of the extracellular
fluids constant is evident.
In addition to this simple control system based on osmolality,
ADD secretion also depends to some extent on the/volume of the
extracellular space: if this is reduced by 10-30'%" for example by
bleeding, then ADD secretion occurs no matter what the osmolality
is. This mechanism does not play much part in day to day life, but
is used in emergencies, probably the left atrial pressure receptors-
which indirectly measure blood responsible. To replace
extracellular water the body needs to replace sodium chloride and
water; the renin-angiotensin mechanism retains sodium. This
mechanism retains water both directly and indireotly once the
osmolality has increased.
Alcohol produces its characteristic diuresis by inhibiting the release
of ADD.
OSmoreceptor
cell
.. 0 Na+entry
o
""-Angiotensin n
increases size or
number of Na+
channels
Figure 7.14 Model of osmoreceptor action. Sodium entering the cell causes a
depolarisation and increased rate of firing of action potentials. The amount of sodium
entering increases as the sodium concentration of the CSF bathing the cell increases.
Angiotensin I[ increases the sodium leak hence sensitising the system to osmotic changes.
Oxytocin is involved in various aspects of reproduction. Its
primary effect is to cause certain smooth muscles to contract; thus it
causes contraction of uterine muscle during parturition and causes milk
ejection during breast feeding (by an action on the myoepithelial cells
lining the ducts). The signals for oxytocin release in these situations
is stretching of the cervix and breast stimulation respectively.
7.6 THE THYROID GLAND
General effects. The thyroid gland makes and releases molecules
which influence the growth, development and metabolic activity of
all In man an increase in circulating thyroid hormone (TO)
conpmtration increases the metabolic rate and causes an over-excited
'neJivous' type of personality. A decrease slows the metabolic rate
and causes sluggish mental activity. Reduced thyroid activity in
childhood produces dwarfs who are mentally retarded, whereas reduced
growth hormone secretion in childhood produces dwarfs of normal
intelligence.
Colloid
follicle 50 JUrI
Active J
7.15 Position of thyroid gland in the neck below larynx and across trachea,
with structure of follicles inactive and inactive states. The blood supply is
between the foDicles and accounts for I % of the cardiac output.
Synthesis of thyroid hormones. The cells in the thyroid follicles
actively accumulate iodine and amino acids from the plasma. The
amino acids are made into thyroglobulin which. together with the
iodine, is secreted into the colloid within the follicle. The iodine reacts
with part of the thyroglobulin in a series of reactions ending in the
of TH. The hormone is stored on the surface of the
thyrfglobulin until required. When required the hormone-protein
conwlex is taken into the cells. the hormone (i.e .. T
J
and T
4
) is split
off $ld then secreted into the plasma.
210 PHYSIOLOGY. BIOCHEMISTRY AND BIOfECHNOLOGY
Blood
Thyroid
cell
Colloid
Amino
acids r-

Protein
I
Thyroglobulin
Q
Thyroid
hormone
releosed
Figure 7.16. Synthesis of thyroid hormones. For simplicity the side chain is only
shown on the first step. Thyroglobulin is made by the follicle cells and exported to
the colloid. Iodide (!-'I is actively taken into the cells and converted to iodine llJ by a
peroxidase reaction at the inner surface of the cells. Release of free iodine causes
iodination of the tyrosine molecules on the surft;;P. of the thyroglobulin molecule.
forming MIT (monoiodotyrosine) or DIT These then couple
spontaneously to form T
J
(triiodothyronine) or T, (tetra odothyronine or thyroxine).
Blood transport and concentration. MOf.t of the TH in the plasma
is associated with the plasma proteins and less than 1 % is free. This
free TH is in with the bound fe'.ID and is the part responsible
for the tissue actions of the hormones. 'T he amount of binding protein
may be changed in various states (:t1creased in pregnancy) thus
increasing the capacity to store TH. If the concentration of TH per
unit of binding protein remains constant then the free TH will not be
ENDOCRlNOLOGY AND METABOUSM 211
altered. A measurement of total blood TH (or of bound iodine) does
not therefore reflect the thyroid status of the individual. Such a system
of transport means that the TH supply to the tissues must be relatively
constant, perhaps with minor and slow modifications imposed by
changes in the thyroidbinding globulin. This fits with the observation
that replacement therapy is adequate if a constant level of TH is
majnt.ained in the plasma.
10000
E
8 1000
....
J
1
100
.=:.
10
Cytoplasm
receptors
Figure 7.17 Distribution of thyroid honnones in the plasma. The free T4 and T,
concentrations are about 3 and 1 S ngl100 ml (40 and 20 pmolll) respectively.
The binding proteins are of three kinds (1) thyroxine-binding
globulin (TBG), (2) thyroxine-binding prealbumin (TBPA) and (3)
albumin. T4 binds to all these in the proportion of 60, 30 and 10%.
T3 binds rather weakly to TBG and albumin only. The free T 3 and T 4
is distributed throughout the extracellular space of the body whereas
die bound hormone is confined to the blood spaces. This means that
la larger fraction of T 3 than of T 4 is to be found in the extracellular
space.
MetaboUsm and turnover of TB. Little 111 is excreted by the
kidneys, presumably because of the low free levels in the plasma.
Some is excreted in the bile but most is metabolised by peripheral
tissues (mainly muscle and liver) and the iodine recycled. As with
other hormones the turnover of the hormone is related to its speed of
. action. Thus. T3 starts to act in about 5 hours and has a T of turnover
lof 1 day, whereas T4 takes about 2-3 days to act and has a T; of
i turnover of some 7 days. There is some evidence that T4 may act as
i a prohormone for T3 and it is possible that T3 is the active hormone
212 PHYSIOWGY, BIOCHEMISTRY AND BIOTECHNOLOGY
in the tissues. This fits with the observation that T3 has a biological
activity several times greater than T
4

Table 7.2. Thyroid effect at various levels of organisation. Most of
these changes require prior synthesis of new proteins before they occur.
Level of
organisation Hypothyroid
BehavioUr: Sluggish mentally and
physically
Sensitive to cold
Whole body: Low BMR
Organs:
CVS
GI
Muscle
TISsues:
Enzymes:
Deficient growth
Raised blood cholesterol
Cardiac output -1" rate -I,
Sluggish movements
Low glucose uptake
Weakness, hypotonia
Q02 of liver, kidney,
muscle, etc.
Nonnal Q0
2
of brain,
retina, testes
Adrenaline effects
decreased
-I, oxidative enzymes
-+ 'close coupled'
mitochondria*
Hyperthyroid
Active and restless
mentally and physically
Sensitive to heat
High BMR
Negative N2 balance
Lowered blood cholesterol
Cardiac output t, rate t
Hyperactive movements
High glucose uptake
Weakness, tremors, brisk
reflexes
Q02 of same tissues t
Nonnal QOa of brain,
etc.
Adrenaline effects
potentiated
t oxidative enzymes
-+ 'uncoupled'
mitochondria*
Effects of TH. TH enters cells, probably binds to receptors inside
them and goes to the nucleus. This is followed in about 5 hours by
the production of new mRNA, then by new nbosomal RNA, then by
new protein synthesis and an increase in oxygen consumption some 3
6 hours later. The TH concentration affects the rates at which various
processes occur in the body, excess causing an increase and a deficit
a decrease.
Control of thyroid /unction. The thyroid gland is controlled by
the thyroidstimulating hormone (TSH) released from the anterior
pituitary gland. It acts on the thyroid cell membrane producing an
intracellular messenger, cAMP; this in turn acts on all stages of TH
".
production to change the thyroid cell from a state of torpor to one of
great activity. Thus, there is increased amino acid and iodine uptake,
increased iodination, increased pinocytosis, etc. There are typical
morphological changes in the thyroid. The end result is an increased
TH secretion into the blood.
TSH production from the anterior pituitary' is' controlled by a
negativefeedback system mediated by TH in the blood (but it appears
that there is no TH effect on the hypothalamus). The anterior pituitary
in 1fD is affected by a thyrotropic-releasing factor (TRF)
in hypothalamus. The rate of release of TRF is affected by body
food intake, etc.
7.7 THE CONTROL OF GROWTH
Animals start as single cells and by repeated divisions reach a
size of many millions of cells. During this process cells differentiate
and, by an orderly sequence of changes, become grouped into tissues
and organs; this process involves an increase in the total protein and
in the length and size of the animal, not just an increase in weight.
With our limited knowledge of the way this process works it appears
to be extraordinarily complex; here we can only indicate some of the
more important factors.
e
Figure 7.18 The connol of plasma thyroid concentration. A disturbance of plasma
[thyroid hormone] (shown as an increase) is followed by the consequences shown
(arrows in other compartments) which correct the disturbance. See Figure 5.48 for an
explwtion of the convention used.
Genetic and external factors. It is supposed that genetic factors
the range of size which an animal can attain; the actual
size within this range is determined by external factors. Of these the
most important is the food supply. This must be adequate in protein.
es'sential vitamins and other factors and in calories. If th(> calorie
214 PHYSIOWGY, BIOCHEMISTRY AND BIOTECHNOLOGY
supply is too low then protein is burnt for energy: this deprives the
animal of amino acids for protein synthesis.
Growth periods. Man shows two periods of rapid growth: the
first of these is just after birth and is actually a continuation of the
rapid growth which occurs in inl ra-uterine life; the second is during
adolescence. This curve of total growth conceals differences between
various organs; thus we all know from observation that babies have
big heads (containing big brains) early in life and that the reproductive
organs do not grow to adult size until adolescence.
100
o
Age in years
Fipre 7.19 The relative size of the whole body. brain and reproductive organs in
man at various ages.
Hormonal influences. The main hormones affecting growth are
growth hormone, thyroxine and the androgens. Insulin is also necessary
during the entire growth period .
Growth
hormone-
Thyroid
hormone
Androgens
. ~
~
~
o 5
10 15 20
Ave (years)
Figure 7.20 The relative importance of various honnones in human growth at various
ages.
ENDOCRINOLOGY AND MEl' ABOUSM 215
Growth hormone is of major importance in controlling growth
from birth to adolescence. It exerts this influence by its action on
amino-acid transport into cells and by the synthesis of new protein.
The most obvious target organs during growth are the long bones,
growth of which controls the linear dimensions of the animal. The
long bones grow by converting one edge of the cartilaginous plate at
their ends to bone and simultaneously forming new cartilage at the
other edge of the plate. The cells responsible for changing cartilage
to bone are the osteoblasts. GH promotes both of these processes by
protein synthesis at both sites. This effect is probably via
which GH causes the liver to release.
". . . :. Proliferation
Cartilage
formed
here
Epiphyseal
plate
Cartilage
convertid
to bOne
here
/
'.
'. of cartilage
cells here
.' Cells Increase
In size as
""",,,,,,,,,,,...,.,.,,,0:1 they age

Bone formed
by asteoblasts
on calcified
cartilage
matrix
long bone extended as shown by arrow; the shape of the shaft is then altered by
remodelling.
Thyroxine is essential in normal amounts for growth; excess does
not produce overgrowth as with GH, but causes an increase in
catabolism of protein and other nutrients. Thyroxine at normal
concentrations thus has a permissive effect on the action of growth
hOIl1One on protein synthesis: in its absence amino acid uptake and
protjein synthesis are not much stimulated by GH. Thyroxin is
particularly necessary for the rapid early growth of the nervous system;
in its absence mental retardation occurs which cannot subsequently
be corrected (cretinism). This defect may be due to a failure of
myelination.
Androgens and oestrogens. Androgens from the adrenals in both
sexes and testosterone from the gonads in males start to be secreted
from about 10 years of age in humans, and increase progressively to
a plateau in the next 5-10 years. In females oestrogen secretion from
the ovaries increases in a similar way over this period, and causes
development of the female reproductive organs, but has little direct
effect on protein synthesis; oestrogen probably also stimulates adrenal
secretion of androgens. Androgens increase protein synthesis. causing
both an increase in the size and development of the reproductive organs
and of the general body size. They stimulate rapid growth of the
epiphyseal plates of the long bones but also, eventually, stop it by
causing complete conversion of the plate to bone. This accounts for
the spurt and then stopping of growth in children and explains why
eunuchs are often very tall; growth, though slower, goes on for longer.
7.8 ADRENAL GLAND
The adrenals are small 'cocked-hat' shaped glands closely applied
to the upper poles of the kidneys. They are really compound glands,
~
Glomerular No
4- zone --+ Aldosterone --+metabollsm
Reticular Sea
4- zone --+ Androgens --+c:toacleristics
Blood
spaces
Figure 7.22 simplified structure and function of the layers of the adrenal gland. The
cortex accounts for about 90% of the total size. The blood supply is communal. only
the medulla has a nerve supply.
both in the sense that they are derived from two types of embryonic
tissue, and in the fact that the outer part (the cortex) secretes steroid
hormones whereas the inner part (medulla) secretes catecholamines.
The main effects of the outer part are in the control of salt and organic
metabolism. If the adrenals are removed death occurs in about a week,
due mainly to sodiwn and water loss. If salt is given the animals live
longer and effects due to cortisol deficiencies on organic metabolism
become apparent. It is perhaps unlikely that two such different tissues
~
u l be so intimately connected without some functional reason; it
been suggested that as cortical secretions are necessary for some
of the synthetic reactions occurring in the medulla a common blood
supply might confer an advantage.
7.9 ADRENAL CORTEX
The hormones produced by the adrenal cortex have numerous
and diverse effects on the body, but their similar chemical structure
gives them several common features which will be discussed in the
Table 7.3. Comparison of steroid and other hormones. The
catecholamines behave like the peptide group, and thyroid hormone
like the steroid group.
Type of
hormone
Derivation
Common features
Mode of action
Form of
t;ircuJation
Peptide Steroid
Gut and related structures* Mesodermal origin
Few-except for basic Molecules all very
amino-acid similar
composition-may be
very complex
Interact with
cell-membrane
receptors
intracellular
messenger
*?structural changes
in pre-existing proteins
Water soluble and usually
free in circulation
Affect nucleus (after
combining with -
binding protein in -
the cytoplasm)-*
synthesis of new
proteins
Usually sparingly
water soluble and
circulate with binding
proteins
* They may, however, have been derived ultimately from neuroectoderm.
given table lists the ways in which the steroids are similar to each
other and contrasts their behaviour with the other large group of
hormones. the polypeptides.
The characteristic structure of the steroid hormones contains four
rings on one plane with the functional groups projecting on one or
other side of the plane of the main structure. Different steroids are
produced by attaching different functional groups (-H. =0. -CH
3
,
-08) to the basic ring structure. These groups project above (b) or
below (a) in the plane of the rings. These relatively minor changes
alter the steroids so that the reaction between the steroid and the
appropriate receptor becomes very specific. They also change the
specificity of the steroids to the transport proteins so that some are
highly bound (cortisol) while others are hardly bound at all
(aldosterone). .
The steroids are all synthesised from cholesterol by a small
number of steps. Therefore, if there is an error of metabolism in
I
2/4
8/10 8/11 13/15
3 1/5 7/9 12/14 18/17 l a
Figure 7.23 The basic steroid strucnue consists of three cyclohexane rings and one
cyclopentane ring. The carbon atoms are numbered as shown for identification of the
functional groups. The main side chain is attached to atom 17, both in cholesterol and
its derivatives. The four rings are not in a flat plane as conventionally shown in I. but
have the approximate form shown in n. in which the observer is looking a long the
plane of the strucnue. Groups are either attached above (/3) or below (a) the plane.
The numbers on 11 refer to the molecule in th at plane.
cholesterol then all are affected. There is practically no storage of
steroids. so they are made as required. The steroids have aT, in the
circulation of 20-100 minutes. They are inactivated in the liver by
removal of the functional groups, then made water soluble by
conjugation with glucuronide and sulphate and excreted in the kidneys.
The ring structure is excreted intact. .
The various groups of adrenal honnones have largely different
effects and will be discussed separately.
7.9.1 Glucocorticoids
As with most other honnones the actions of the glucocorticoids
range over several layers of organisation from whole animals to
membrane interaction. Again, as for most other hormones, no single
or 1imple explanation of the effects can be given.
In man, sheep, dog, cat and cow the main hormone in this group
is q>rtisol. In rodents it is corticosterone. In man most of the plasma
cortisol is bound to a transport protein (transcortin). There is a
circadian rhythm of plasma cortisol concentration which seems to be
related to the sleep pattern and is mediated by the pituitary. It may
be due to an inherent rhythm in the brainstem.
'i
50
Cushing'S
ii
-:8
25 e-
ll>'
OD'
Normal
. ~
S
~ 0
Midnight
Noon
Midnight
la)
Hours
Stress
......
+
S
50
. ~ -
......
Ig 25
a.g:
"8-
;
0
lb) Days
Figure 7.14 Patterns of total plasma cortisol concentration: (a) throughout one day in
the i nonnal subject and a patient with Cushing's syndrome (a disease characterised by
an abnonnally high glucocorticoid production); and (b) the effect of stress on a nonnaI
SUbject.
Specific effects. There is still uncertainty about the actions of
cortisol at normal tissue concentrations, though there is wide
acceptance of the view that cortisol exerts a 'permissive' action on
metabolic and enzyme functions, thus permitting a variety of other
hormones to exert their effect. In some cases these permissive effects
are to induce new proteins which are then acted upon by other
hormones.
The adrenals are involved in the body's response to 'stress', i.e.
exposure to noxious or potentially noxious stimuli. In short-term sudden
stress the immediate reaction is the release of catecholamines leading
to the 'fight or flight response'; the longer-term reaction is the release
of increased amounts of cortisol. Many situations in everyday life
lead to increased cortisol secretion and indeed the word 'stressful' is
often used for these situations which do lead to this response. These
effects of cortisol in stressful conditions act through their high blood
concentrations; they can be understood and remembered as follows.
Animals faced with such situations: (1) have no time to eat so the
effects of cortisol are to supplant and conserve the energy derived
from circulating glUGose; (2) might require to repair damaged tissues
and so have a need for higher free amino acid concentration in the
body. These effects are accomplished by inhibiting glucose usage in
peripheral tissues; by causing a flow of amino acids to the liver for
new glucose synthesis (gluconeogenesis); by changing muscle
metabolism from glucose to fatty acids and by mobilising fatty acids
from adipose tissues. The response is rather like the 'fasting response',
which occurs between meals, except that it leads to deposition of
glycogen as a consequence of the gluconeogenesis. This action led to
the name-glucocorticoidsused to describe this group of steroids.
Protein breakdown Tissue wasting
.
Glucose formation Glycogen formation

Insulin secretion

Fat formation (centripetol ie. face and hands)
Figure 7.25. Steps in the production of obesity caused by excess glucocorticoids.
ENOOCRINOLOGY AND METABOliSM 221
Table 7.4. Psychosocial situations shown to be associated witb
increased plasma concentration or urinary excretion of adrenal
cortical steroids.
Human beings
I Normal persons
(a) Acute situations
I Aircraft flight
2 Awaiting surgical operation
3 Final exams (college students)
4 Novel situations
5 Competitive athletics
6 Anticipation of exposure to cold
7 Decreased during weekends
8 Many job experiences
(b) Chronic life situations
I
2
Predictable personality-behaviour profile: aggressive,
ambitious, timeurgency
Discrepancy between levels of aspiration and achievement
(c) Experimental techniques
1
2
Stress' or 'shame' interview
Many motion pictures
II PsychioJric patients
(a) Acute anxiety
(b) Depression, but only when patient is aware of and involved
in a struggle with it
(c) Decreased in the manic state
High plasma cortisol concentrations pharmacological effects. If
cortisol concentrations are raised for prolonged periods-by certain
diseases or during treatment of various disorders-then: ( I) the
generalised breakdown of protein which occurs in skin, muscle, bone
and lymphoid tissue, gives wasting and/or weakness of these tissues
and a negative nitrogen balance therefore develops. This increased
protein breakdown will greatly increase the plasma glucose and
glycogen stores; this leads to an increase in insulin secretion causing
a greater conversion of glucose to fat and the obesity of Cushing's
syndrome; (2) suppression of the typical effects of injury either when
produced mechanically or by allergens. These injury effects are an
increased fluid leakage, increased leucocyte appearance and eventually
the laying down of fibrous tissue. This suppression is 'useful' or
otherwise, depending on the circumstances; in allergies it is useful,
in wound healing not. An attractive suggestion for this action of
cortisol is that it stabilises the lysosomes in cells and so does not
allew them to escape from partially damaged cells, to prolong or
increase the local damage; by this means the degradative enzymes
present in the lysosomes can be contained. Perhaps associated with
this action of cortisol is its effect on decreasing the circulating
eosinophils.
Low plasma cortisol leads to:
1. Inability to cope with physically 'stressful' conditions,
whether manmade, for example surgical operations, or the
natural hazards of living.
2. Inability to handle a water load. This is not primarily a
matter of altered kidney function but' seems to be because
of a maldistribution between the body fluid compartments.
e
I
I
Anterior pfrultory
ACTH release ..... .[ ==. l .... t-----I
Figure 7.26 The control of plasma cortisol concentration. A disturbance of plasma
cortisol (shown as an increase) is followed by the consequences shown (arrows in
other compartmentS) which correct the dislUrbance.
3. There is a fall in blood pressure, and noradrenaline no longer
has a constrictor effect on the cardiovascular system.
4. The blood glucose concentration falls, and liver glycogen
decreases. This is partly becanse of the removal of the
'permissive' eftect of cortisol on other hormones and partly
by removal of its direct effect on gluconeogenesis.
Control of cortisol secretion. ACTH (adrenocorticotrophic hormone)
released from the cells of the anterior pituitary is responsible for
maintaining normal adrenal structure and function, for the normal
release of the glucocorticoid secretion and for exerting a permissive
action in mineralocorticoid secretion (i.e. necessary, but not the main
stimulus). ACTH release in turn is increased by corticotrophic-releasing
factor (CRF) and inhibited by plasma cortisol. CRF structure has not
yet been worked out, but is probably a small (10 amino acids)
polypeptide. ACTH structure for several species has been worked out.
It consists of a polypeptide of 39 amino acids with an active part and
~ species-specific part.
I .
2
3 -------24 25 26---32 33 34-----39
~ I I biological actlylty here
All interspecies variation .. Comman ..
occurs here
Figure 7:rt Outline stnu:bJl'e of ACfH. Each number represents an amino acid. Amino
acids 1-13 are fouDd iD some species as melanocyte-stimu1ating hormone.
7.9.2 MineraIoeorticoids
Aldosterone, the main hormone in this group, is secreted at a
daily rate depending on the salt balance. There is a diurnal rhythm
of plasma aldosterone with much the same pattern as that of cortisol.
This seems to be related to posture, in that lying flat inhibits it.
Spedjic effects. Aldosterone causes the kidneys to retain Na+ and
excrete K+ and CI-, mainly by an action on the distal tubules. This
leads to a secondary retention of water (mainly via antidiuretic
hormone) and so an expansion of the extracellular volume results.
jSimilar effects on Na+, K+ and H+ handling occur in the salivary
I
glands, sweat glands and intestine and perhaps in the proximal renal
tubule.
. The way in which aldosl.erone acts on target cells is not yet clear.
It seems probable that the hormone-receptor complex enters the nucleus
of appropriate cells and that new protein synthesis occurs, but it is
likely this new protein is a 'permease' which increases the Na +
leakiness of the cells leading to a secondary increase in sodium
transport.
, Control of plasma aldosterone concentration. The aldosterone
I concentration in the blood is related to sodium metabolism. If the
'total body sodium is high-by sodium loading-then the blood aldosterone
concentration is low and sodium is excreted; if the body sodium drops-
. '.
ACTH release ..
.
: ' . ~ ~ .'
:
I
I
I
I
I
I
I
I
I
I
I
I

. .
. .
1
al cortex Kidney
'" .-... No + ",,,,,,,,,,," \
Blood
volume t
Juxtamedullary apparatus
Blood
~ Pressure'
nsln JI Renin
Ition +
14
secretionf
~
[Na]
Angio
~
Volume receptors
(chest)
'---------------
- Rate of firing t
Figure 7.28 The control of blood volume (simplified). A disturbance of blood volume (shown as an
increase) is followed by the consequences shown (arrows in other compartments) which correct the
disturbance. See Figure 5.48 for an explanation of the convention used.
~
~
::sl
I
~
~
~
-<
~
o
S
~
~

-<
by sodium deprivation-then the blood aldosterone concentration is high
and sodium is retained. To some extent potassium changes show
opposite effects.
It is difficult to imagine a system which would measure total
body sodium directly, but easier to imagine one which would measure
some indirect effect of sodium. The current view is that the size of
the extracellular fluid volmne is used as an index of the sodium loading
of the body. If the body is deprived of sodium chloride then the
osmoreceptors cause an excretion of water to correct the osmotic
hence the extracellular volmne-and the blood volmne-shrinks.
TIiere is thus a fall in the pressure in the arterioles perfusing the
kidney and this leads via a series of hormones-the renin-angiotension
system to an increase of aldosterone release. An increase in body
sodium works via the same system to reduce the plasma aldosterone
and hence causes excretion of sodium. The apparatus which detects
changes in perfusion pressure is the juxtamedullary body, situated
where the blood supply enters the glomerus, with the distal convoluted
tubule nearby. It can be thought of as a cuff around the afferent vessels
to the glomerulus.
2 3 4 567
Angiotensin II
r r-
,nI
8 i 9 10 1 :n 12----::=,.
Lung
enzyme
Figure 7.29 Production of angiotensin D. Activation consists of chopping pieces off
thl! larger plasma globulin molecule. Each number represents an amino acid.
I The situation is more complex than this in that the sodium
concentration in the distal convoluted tubule is also measured by the
juxtamedullary body and used to control aldosterone production and
release. Plasma potassium probably also has a direct effect on
aldosterone release by the adrenals. ACTH is probably permissive
rather than controlling.
A bewildering array of other information is available on the
system, most of which does not fit easily into a simple
sfheme. Three separate aspects which do will be mentioned:
1. Renin-like activity is present in many organs such as brain,
salivary glands, uterus, arterial wall, etc. Release from these
would presumably be triggered by low perfusion pressures
and thus expand the blood volume as described.
2. Angiotensin II acts on the brain to cause increased drinking.
This action seems understandable in that if sodium is retained
water also needs to be retained to expand the extracellular
volume.
Efferent
arteriole
Macula densa
round distal
convoluted .
fubuleJ
(a)
- ~ . , . . . . . . , . __ Glomerulus
.:.. _Renal nerves
(b)
Figure 7.30 The main features of the renal juxtam.duDary apparaws; (b) is an enlarged
view of the black area in (a). Pressure in the after:nt arteriole, the sodium concentration
in the disral convoluted tubule and the renal ne ves all influence its activity and hence
the renin production (after Davies).
3. Angiotensin II is a very potent constrictor substance; thus
causing a raised blood pressure by a direct action on
227
arterioles in addition to an indirect action via fluid retention.
Adrenal androgens. These have a keto group in the 17 position
of the basic ring structure and are hence known collectively as the
17-ketosteroids. Half of the plasma androgens ,are bound to a sex-
hormone-binding globulin (SHBG). Secretion is partly under the CODtroI
of ACTH from the pituitary. The role of these androgens in normal
man is unclear in that they are unable to maintain secondary sex
characteristics in castrated men or animals; it has been suggested that
they may be responsible for female libido. Their main clinical interest
is thitt inborn errors of metabolism and tumours may lead to excessive
prodUction and 'virilisation' of females or young boys.
7.10 ORGANIC METABOLISM AND
THE GLUCOSE REGULATING HORMONES
The hormones dealt with so far are those released by the discrete
organs which have a variety of effects on many systems in the body.
In the next two sections the emphasis is rather different, in that we
are concerned with the control of particular functions by several
hormones; the first of these functions is organic metabolism, the second
is calcium metabolism; neither system is much affected by anterior
pituitary secretions.
Animals eat intermittently, with periods of fasting in between,
yet their cells require a continuous supply of energy. The mechanisms
which have evolved to cope with these alternating periods of feeding
and fasting are hormonally controlled; the states themselves are often
called absorptive and post-absorptive. We shall start by considering
the events which occur during these states, and then describe the
hormones which control these changes.
Interconversion of carbohydrates, fats and proteins. In order to
understand the events during feasting and fasting it is essential to
realise that body cells-particularly those of the liver-have a remarkable
ability to convert one type of molecule into another. These conversions
allow animals to survive and grow on a very wide range of foods,
provided that certain essential items are present. It is found that not
only are molecules of one sort converted to those of another following
digestion, but that virtually all the molecules in the body are being
broken down and rebuilt all the time. So the body is in a continual
state.
! The main pathways for interconversion of protein, carbohydrate
and fat. The main difference between protein and the other two are:
228 PHYSIOLOGY, AND BIOTECHNOLOGY
(1) protein cannot be stored to any extent-it is merely used for
replacing protein molecules in cells-and it cannot be metabolised
directly but must first be converted to carbohydrate or fat; and (2)
carbohydrate and fat can be stored as such and are the main fuel to
supply the body with energy. The figures show the similarities of
handling of all three molecules; we have omitted the minor losses
which occur through the kidney and when cells are lost from the
skin. etc.
The essential items which must be in a diet are:
1. A minimal amount of protein to replace 'the nitrogen which
is excreted.
Structural
CHO and fat
______ __
fat pools
CotaboIism 10
CO
2
+
+ energy
NH:s
Derivatives
steroids
etc.
Amino acid
pools
I g=-I
'----.. (1 D
Figure 7.31 Main carbohydrate and fat metabolic pathways. Botb can be stored and
used for energy directly.
2. Certain essential amino acids, which cannot be made from
others in the body. The best source of these is animal muscle,
but an adequate mixture of plant protein can supply all our
essential amino acids. As protein cannot be stored to any
. great extent, all the essential amino acids must be eaten at
the same time. If some are missing then a negative nitrogen
balance ensues.
3. Certain vitamins and fatty acids are required, which again
cannot be made in the body. Vitamins are used as cofactors
or coenzymes for chemical reactions and not for energy.
7.10.1 Outline of Absorptive and Post-absorptive Events
Food taken into the digestive tract starts a series of events leading
to the production of the digestive enzymes which break the food down
to a mixture of smaller molecules. These are then absorbed into the
bloodeither directly or via the lymph-causing a rise in the blood
conpentrations of glucose (+galactose and fructose), amino acids and
fats. During this period glucose is used as a major source of energy
by the cells; most of the amino acids and fats, as well as the excess
carbohydrate not used for energy, are stored as fat or glycogen. A
small fraction of the ingested amino acids and fats is used to
resynthesise the body proteins and structural fat which is being
continuously degraded.
Between meals the tidal wave of newly ingested molecules
decreases, so that the direction of flow of materials is now reversed;
some ce11smain1y those in the liver and fat tissue-now export rather
than import molecules. The blood sugar is kept constant by breaking
down the glycogen stores in liver and some other cells, and by the
synthesis of new glucose; this is necessary because brain, renal medulla
and erythrocytes have no other energy source. The energy supply of
most other cells is switched from glucose to fatty acids. thus sparing
glucose; most of the energy supply for the body now comes from
fat. Protein and fat synthesis decreases, leading to a net breakdown
of these materials.
These various adjustments are made by the hormones insulin,
glucagon, GH and the catecholamines. Of these insulin is the hormone
of i absorption; it is released by the rise in blood glucose during a
meat. Glucagon, GH and the catecholamines are the hormones of
fasting; they are released by lowering the blood sugar. These four
hormones are directly involved in the control of organic metabolism;
others are, however, also involved in a permissive way, i.e. their
presence is necessary for action to occur but they are not controlling.
Thus, background concentrations of thyroid hormones and cortisol are
necessary for the proper action of most of these honnones. In keeping
with the close links between digestion and these metabolic processes,
it Js now clear that gut hormones, gastrin, secretin and pancreozymin
augment the release of insulin and glucagon in preparation for the
appearance of glucose and amino acids in the blood during a meal.
7.10.2 The Absorptive State
Following a meal monosaccharides and amino acids enter the portal
blood and go to the liver; the fats enter the lymph as chylomicrons
containing biglycerides, and so enter the general circulation. The
monosaccharides consist of galactose, fructose etc., as well as glucose
but can, for simplicity, be considered as glucose. We shall consider
separately the fate of each of these substances which enters the body
following an average meal; say carbohydrate, fat and protein in the
ratio of 65:17:18, respectively.
Gut
Urea
,/ .... -p
+ ........ AmIno--+CH
acids ....
Trlglycerldes
CHO Glucose/
I ~
Glucose
Trlglycerldes
Trlglycerides
and fatty acids
Fat cells
Figure 7.32 Metabolic pathways of absorption. Carbohydrate fonns both the glycerol
and the fatty-acid molecules for triglyceride synthesis; it also acts as the energy source
for all cells except the liver which use amino acids. Most of the food entering the
body is stored as fat and to a lesser extent as glycogen. Fat is a good way for mobile
animals to store energy. for it is light (it contains little water) and has twice the energy
yield of carbohydrate and protein per gram of material. Glucose is stored as glycogen
for osmotic reasons.
ENDOCRINOLOGY AND METABOliSM
231
Glucose. Much of the absorbed carbohydrate enters the liver cells;
here it is stored as glycogen or converted to fat which is then stored
partly in the liver but mainly elsewhere after transport in the blood.
The glucose which does not enter the liver is used by almost all body
cells as the main energy source; the remainder is stored as glycogen
(mainly in skeletal muscle), and cOnverted to fat in the adipose-tissue
cells.
Amino acids. Much of the absorbed amino acids also enters the
liver cells; here they are converted into carbohydrate (keto acids) by
removal of the ~ which is converted to urea and excreted via the
blood and kidneys. These keto acids enter the Krebs cycle to supply
most of the liver's energy during the absorptive phase. They are also
converted to fats which are then stored in the liver or elsewhere.
The amino acids which are not taken up by the liver enter other cells
(mainly muscle) only to replace that which is being continuously
broken down; excess amino acids are not stored as protein but as
carbohydrate and fat.
Triglycerides. Practically all absorbed fat enters the adipose-tissue
cells, where it is stored. During the absorptive phase adipose-tissue
cells receive triglycerides: (a) directly; (b) from those manufactured
in the liver; and (c) those made in the fat cells from glucose. A little
fat is metabolised by tissue cells to produce energy during this phase.
7.10.3 Post-absorptive or Fasting State
The main problem during this state is that the plasma glucose
concentration must be maintained (at 70 mg/lOO ml approx. 4 mmoV
litre)because the brain, renal medulla and erythrocytes require glucose-
and yet there are limited stores of glycogen available to do so. The
bcly solves this problem in two ways: (1) forming new glucose from
~ and protein (gluconeogenesis); and (2) converting the metabolism
of most cells to work on fat rather than glucose-thus sparing glucose.
7.10.3.1 Sources of plasma glucose
The glycogen in liver and muscle is broken down to supply glucose
for the plasma. The liver glycogen can be broken down directly, but
muscle lacks the necessary enzyme (glucose-6-phosphatase) to produce
glucose from glycogen; instead pyruvate and lactate are produced in
muscle, go to the liver and there are used to synthesise glucose. This
source supplies glucose for about 8 hours at average body rates (100
calIhour).
Triglycerides are broken down to glycerol and fatty acids; the
glycerol can be converted to glucose by the liver and the fatty acids
(which cannot be so converted) are used directly by the body cells as
an energy source.
r---------------------
I
J
I
J
I
J
I
J
I
I
, ....
Lactate
-and
pyrr
Glucose
Keto" '\. oc
Amino acids

Plasma glucose
1
Ic:l
L:J
BlOOd
FIgure 7.33 Metabolic pathways during die post-absorptive slate. Initially blood glucose
is supplied from liver and muscle glycogen, later glycerol from body fat is used to
make glucose. In prolonged starvation protein breakdown occurs. The main energy
soun:es for cells are fatty acids, but glucose is used by die brain, renal medulla and
erytbrocytes.
During prolonged fasting-which can go on for many days provided
water is supplied-an additional source of glucose is the body proteins,
mainly of muscle. A large fraction of this protein, which is apparently
not absolutely essential for function, can be broken down to amino
acids which are then converted to glucose. In normal life, however,
protein is not used for this purpose to any great extent.
233
1.10.3.2 Glucose sparing and Jot usoge
Almost all body cells, including the liver but excluding the nervous
system, are converted to using fat during fasting, thus sparing glucose
for the use of the nervous system. The triglycerides of adipose tissue
are catabolised to glycerol and fatty acids, these fatty acids circulate
in the blood, are taken up by tissue cells and then used in their Krebs
POSI-absorption

Glucose
--- AmIno adds
_--<-<! GI,cerol
F'atty acids
--- Glucose
Glucose
Figure 7.34 Changes of metabolism from the absorptive to post-absorptiYe smres.
cycles. In the liver, however, these fatty acids are converted to ketoDe
OOdies (for example, acetone), instead of entering the Krebs cycle,
aq<t then released into the circulation to be used by tissue cells in
their Krebs cycle. If the concentration of ketone bodies is high. as in
prplonged starvation or untreated diabetes mellitus, then it gives a
characteristic odour to the breath.
7.10.41nsuUn
Insulin is formed and stored (to a small extent) in the p-cells of
the Islets of Langerhans in the pancreas. These make up 60-90% of
the 1-2.000.000 islet cells. a-Cells. secreting glucagon. and D-cells.
which secrete gastrin. are present in small numbers. The pancreas is
derived from the gut and here the general rule that gut hormones are
proteins applies.
Pancreatic P cell
It I
Ribosornes
,l..!.Pro-lnsulin
cAMP
(
[J Golgi apparatus
,r--InSUlln
+ f::::;::?
,l.
Insulin
+
C peptide
(a)
P chain
(b)
C peptide
Figure 7.3S (a) Production of insulin in pancreatic cells; (b) Strucl\ll'e of pro-insulin.
The size and structure of the C-peptide varies with the species. that of insulin (shown
shaded) shows only Slight variations. (The numbers of amino acids are shown.)
Insulin is composed of two peptide chains joined by disuIphide
bonds. It is actually made as a pro-insulin in the noosomes of the
rough endoplasmic reticulum of p-cells and stored by the Golgi apparatus
as granules. These granules contain trypsin-like enzymes which split
off the C-peptide. forming insuIin. On receiving a signal to cause
insulin release these granules fuse with the cell membrane and release
both insu1in and the C-peptide (and perhaps some 5% pro-insuIin).
After a glucose meal there is a large (x 5) and rapid rise in the
insulin concentration in the blood, which returns to normal in 2-3
BNDOCRINOLOGY AND MEr ABOLISM 235
hours. If the plasma glucose concentration is measured in the same
subject. then this shows a roughly parallel rise and fall. The insulin is
released into blood going to the liver and about half is inactivated by
passage through the liver; this means that the liver, which is an
important site of action of insulin, receives a higher insulin concentration
than the rest of the body. Insulin probably exists in the plasma in two
components: one in the same form as in the pancreas and the other
one in a different form. The fonner acts on the tissues. Injected insulin
disappears with a T'Ii of 4 minutes, but becomes bound to the tissues
_ there much longer.
GI)QlCJen
* t
Glucose -+-.Glucose ---+ FaHy acids .. Triglyceride
\ 0""_
Krebs cycle ---+C0
2
+ H20 + -P
Amino acids -+-*-+Amino acids ---+ Protein
Blood
"'-Cell membrane
Figure 7.36 Main effects of insulin on metabolism (indicated by). lnsulin increases
g1uccse and amino--acid uptake by membrane effects (but not in the brain or liver);
this leads to increased synthesis by mass action. It also has a direct effect on the
glucose-glycogen pathway and inhibits triglyceride breakdown.
I Specific effeas of insulin. Insulin is the main hormone which
changes the body from the fasting to the absorptive phase; a large
injection of it will reproduce most of the effects of absorption, including
a profound fall in blood glucose-without. of course. uptake from the
gut. It has two primary membrane effects; it increases both the uptake
of glucose and of amino acids into most cells, thus stimulating glycogen,
fat and protein synthesis by an increase of glucose and amino-acid
concentration in the cells. It also has direct stimulating effects on the
ratelimiting steps in glycogen synthesis from glucose, causing increased
synthesis. It decreases the breakdown of triglycerides, thus causing a
net increase in stores. Insulin, therefore, promotes glucose
utilisation and decreases glucose synthesis.
236 PHYSIOLOGY. BIOCHEMrSTRY AND BIOTECHNOLOGY
In the absence of insulin these reactions are reversed. The uptake
of glucose and of amino acids by most cells is reduced: this, plus
the absence of the direct enzymic effects, causes breakdown of
glycogen, fat and protein. The raised triglycerides in the plasma causes
more to enter cells and increases the triglyceride usage for energy.
The glucose uptake by brain and liver is not much affected because
it is not dependent on insulin.
Control of insulin release. The rate of release lof insulin, and
hence its plasma concentration, is mainly controlled by the plasma
glucose concentration. Other factors interact with glucose: for example,
metabolic fuels, several peptide hormones and some autonomic
transmitters. When the fl-cells are stimulated to release insulin they
can do so immediately 1 minute) from the storage granules, but
then show a slower sustained release which requires new hormone
synthesis. There is no effect of the anterior pituitary on insulin release.
The Il-cells have an adenyl cyclase mechanism. An increase in
cAMP in the cells causes an increase in insulin secretion and
production, and a decrease inhibits secretion. It has been suggested
that glucose enters the fl-cells, is metabolised to glucose-6-phosphate,
forms A TP and stimulates cAMP production. This is an unusual way
for an increase in cAMP to occur, usually membrane receptors are
required.
7.10.4.1 The metabolic effects of gluco,gon, growth
hormone and adrenoJine
These three hormones have effects which, at an elementary level
anyway, are opposite to those of insulia. They are, therefore, the
hormones of fasting, whereas insulin is the hormone of feasting. The
precise effects of these hormones and the importance of each is not
yet clear, but the overall effect is that they raise the plasma glucose
and fatty-acid concentrations; this latter effect increases fatty-acid entry
into and usage by the body cells and thus, as a secondary consequence,
spares glucose.
The main stimulus for their release is, as expected, a decrease
in the plasma glucose concentration, but the pathway for this effect
and the detailed interactions differ with each hormone. Glucagon is a
small polypeptide released from the a-cells of the pancreas; a related
form is released from the intestinal mucosa during glucose absorption.
It has an action on the membrane of liver cells, causes the release of
cAMP which then mediates its intracellular effects. The detailed factors
acting on glucagon release are quite complex, and, like insulin, release
ENDOCRINOLOGY AND METABOLISM 237
is influenced by gut hormones. The release of growth hormone by
the nituitary and adrenaline by the adrenal medulla occurs when the
hypothalamic glucose receptors are stimulated by low plasma glucose.
Table 7.5 Summary of the main effects of hormones which oppose
those of insulin. The" glucagon action on liver is on two time scales;
a rapid one on glycogen breakdown and a slower one on gluconeog-
enesis.
QlycogenolyslS t glucose
Gluconeogenesis t glucose
Fat mobilisation t fatty acids and
glucose
Growth
Glucagon Adrenaline hormone
yes yes
yes
yes
Interactions of insulin and glucagon. The way in which insulin
and glucagon normally interact is more complex than suggested in
the above account, and cannot properly be descn1Jed as simply as we
have done. In addition to the states of feasting and fasting, the kind
of food to be metabolised alters the hormones required to do it. If a
carbohydrate meal is eaten, the blood is flooded with glucose and
other sugars, insulin is secreted and the combination of this insulin
and carbohydrate depresses glucagon production. If a protein meal is
eaten, the blood is flooded with amino acids, this causes a large rise
in plasma insulin but now there is no corresponding rise in glucose
uptake to keep up the plasma glucose. In this circumstance the plasma
glucagon concentration also rises, thus keeping up the plasma glucose
by glycogenolysis and glyconeogenesis in the liver. Normally, of
course, meals consist of mixtures of carbohydrate, fat and protein so
that the hormone response is a mixture of insulin and glucagon
It is therefore more useful to think in terms of the relative
blood of both hormones in any given situation, rather
than of each hormone separately. This can be done by considering
the molar ratios of insulin and glucagon (I/G ratio).
It has been found that if the molar ratio of insulin to glucagon
exceeds 5 then glucose is being stored in the liver, if it is less than 5
then glucose is being released by the liver. After a large carbohydrate
meal the ratio may rise as high as 30; during starvation it may fall
to 05. Thus, during energy storage the molar ratio of insulin to
glucagon is high, favouring the deposition of glycogen, protein and
fat; during energy mobilisation the molar ratio is low favouring
glycogen breakdown, protein conversion to glucose and fat breakdown.
~
~
Liver, muscle, fat cells
: ...... Food storage t t----
:
Pancreas
~ ..... Gluconeogenesis. ~
Insulin release t

Plasma

.......
Glucagon release.
! ~
[Glucose] t
...... :

General body fuel
:

i ..
Glucose t I----

.......
Free fatty acids ~
Figure 7.37 Interactions of insulin and glucagon on the control of plasma glucose concentration.
A distribution of plasma [glucose] (shown as an increase) is followed by the consequences
shown (arrows in other compartments) which correct the disturbance.
f-
~
00
~
I

I
~
i
~
ENDOCRINOLOGY AND MEfABOUSM 239
Meal
rIslet of
/ Langarhans
~
Glucagon
I , .. ----- ...
I,' ... ,
'. . ........ --
: ....................... .
I
: .. ................................... ;-
I ......... -----,'
-+D
InsulIn
Insulin
o 2 4
(a)
Hours since meal
---- Protein -Carbohydrat.
(b)
Figure 7.38 (a) Factors affecting insulin and glucagon release-all are positive except
for the dotted line (after Catt). (b) Effeds on the plasma concentrations of glucagon.
glucose and insulin of a protein and a carbohydrate meal. The ordinate is in arbitrary
units.
Both glucose and amino acids by DXJUth give much larger insulin and glucagon responses
than if given intravenously. This is thought to be due ID the augmentation by the digestive
enzymes shown. The amino acids which can be converted ID glucose (glucogenic ones,
for example, glycine) give a bigger glucagon response than the others.
7.11 BONE AND THE CALCIUM
REGULATING HORMONES
I In this section we are concerned with the control of one substance
calcium-by the action of three hormones-vitamin 0, parathormone and
calcitonin-working at a variety of sites. We will tackle this by first
considering the general properties and movements of calcium and then
the action of these hormones on this.
7.11.1 ~ i u m
Calcium is widely distributed in the body and is essential for
10st bodily functions. Examples of this are that it is required for:
.: 1. Normal penneability of cell membranes; if altered spontaneous
activity often occurs.
2. 'Messenger' activity in, for example, muscle cells, gland
secretion and transmitter release. The general rule is: external events
which produce a change in the electrical activity of the cell membrane
are coupled to intracellular events by changes in the free calcium
concentrate within the cell; external chemical events (for example.
honnones) are coupled by changes in cyclic-nucleotide Concentrations.
3. Blood coagulation.
4. Bone and teeth formation; indeed about 99% of the bodily
calcium is held within these structures.
Glucose
stored
In liver
Glucose
produced
by liver
Starvation
Insulin Iglucagon ratio
Overnight Large carbohydrate
fast meal
Figure 7.39. Effect of insulin/glucagon ratios on glucose storage or producdon by the
liver. The various conditions producing these ratios are iDdicated along the bottom of
the graph. The iDsulinIglucagon ratio is on a log scale; the liver glucose is OD an
arbitrary scale.
The distribution of calcium between the extracellular fluid and
the cells is regulated by calcium pumps at the cell membrane.
Phosphate is important for such substances as A TP, buffers, bone
and teeth. It is present mostly in blood as HPOr (85%), less as
H
2
PO 4 and organic phosphates with little bound. The actual
concentration is some 5-6 mg/IOO ml (l'S,mmolllitre of phosphorus)
in children and about half in adults. Most of tlte absorbed phosphate
is in the inorganic form. or is converted to t'.ris during digestion.
Bone is a heterogeneous material. that ~ it is made up of several
different materials combined together. fhe advantages of such
structures are that they combine the virtuf.s of each of the constituent
materials. Everyday examples of such mao.erials are reinforced concrete
and glass-reinforced plastics. These consist essentially of fibres (steel
or glass) which resist extension and a matrix (concrete or resin; a
polymer) which binds the fibres together and withstands compression.
Bone consists of a combinatioo of three substances: collagen fibres
bound together with a soft mucopolysocclu:uidl! polymer, hardened with
deposition of bone minerals on the collagen fibres. Man-made
structures and natural bone are therefore similar iD construction; in
both iDstances the orientation and density of is arranged at stress
points to give maximum strength.
- ...
ea
2
+ .... -+-_ ea
2
+
< I fU"OI/litre I mmol/litre
Ca2+
Figure 7.40 Pompneak movements of calcium across the cell membrane. The numerical
values for free calcium are approximate.
The bone minerals are laid down as small flat crystals some 20
om long by 3 DID thick, attached to the fibres in a regular way. Each
crystal is surrounded by a watery shell which allows exchanges of
the ions to occur. Because of their small size, the crystals have a
high surface to volmne ratio facilitating rapid exchanges. Bone contains
three types of cell: (1) the osteoblasts which are found where new
bone is being made, so are perhaps producing extracellular elements
collagen and mucopolysaccharides; (2) the osteocytes whose function
is unclear but may be descendants of osteoblasts; and (3), osteoclasts
which are giant multinucleated cells present where bone is being
and which perhaps ingest parts of bone and break it down.
These cells are polarised like thyroid cells with villi and phagocytic
processes absorbing bone at one side. It is probable that hormonal
control of these cells occurs on two time scales: (1) a minute-to-
minute modulation; and (2) a coordinated stimulation of the
differentiation of one cell type or another so that the bone is
predominantly proliferation or resorptive. Ordinarily these processes
of remodelling bone go on all the time.
The calcium in bone is laid down as a compound of Ca 2+. P0
4
'
and OH- called hydroxyapatite Ca(OH)2]' Bone also
cpntains Na +-, Mg + and K + combined in some unknown way; these
cpntain 40,60 and 5%, respectively of the body's stores of these
sUbstances. At present it is not clear how this deposition of calcium
I
crystals occurs; it is possible that the hydroxyapatite is precipitated
directly or via an intermediate substance which then hydrolyses. It is
known that the enzyme alkaline phosphatase, which is found in
association with the osteoblasts, is important .for this process.
Resorption of bone is a1Sd not understood. Perhaps osteoblasts
cause a local raising of the concentration of citric and lactic acids
with formation of a Ca-citric acid complex, and consequent movement
of calcium out of the bone.
7.11.2 Calcium Regulation
The plasma calcium concentration is normally kept within a range
of 3% of its normal value of 10 mg/l00 ml (2S llBIlolllitre);
approximately half of this exists as free calcium, the rest is bound. It
takes part in three main exchanges and is regulated by three hormones.
The exchanges are between the plasma and the gut, bone and kidney.
The hormones acting on these exchanges are vitamin D, parathyroid
hormone (PH) and calcitonin. The first two act to increase the plasma
calcium concentration, the last to decrease it. It seems, however. that
considerable exchanges continue in the absence of these hormones,
so that they should be considered as 'fine controls'. In the absence
of these hormones the plasma calcium settles down at a lower than
normal concentration (7 mgl 100 ml); the net hormone effect is
therefore to raise plasma calcium.
Vitamin D is the term used to descn"be a group of closely related
sterols prodUCed by the action of ultraviolet light on various precursors.
The release of the active form 1.2S-DHCC from the kidney is
increased by low plasma calcium concentration. As the active form
is the derivative of a steroid it is thought to act by a cytoplasmic
receptor and synthesis of new protein. Together with parathyroid
hormone, l,2S-DHCC increases gut absorption and reabsorption from
bone and kidney. It is also neceSsary for new bone formation. The
serum concentration of 1,2S-DHCC increases greatly during periods
of growth; an effect controlled ultimately by the pituitary.
Parathyroid honnone. PH is secreted from the chief cells of the
parathyroid glands (two pairs) which are always close to the thyroids
in the neck. The parathyroids are derived from gut and so-as in the
general rule-the hormone structure is a polypeptide; the first 27-34
amino acids contain all the activity. The release of PH is controlled
by the plasma calcium concentration acting directly on the parathyroid;
low plasma calcium concentration increases its secretion. There is
evidence that it is made as a prohormone and a part of it is cleaved
ENDocRrNOLOGY AND METABOliSM
off before secretion. The hormone acts on the surface of the target
cell producing the intracellular messenger cAMP. PH acts on bone
on two time scales; an early effect 20 minutes) leading to calcium
release and a slower effect on bone turnover and remodelling.
+ 1000mg
~
900mg
Renal
tubules
1
Urine
100mg
Free plasma
calcium
140
Bone and
teeth
20,000
--+
+--
20,000
~
~ ~ \
Ellchangeable
5000
Figure 7.41 Quantitative oudine of calcium exchanges. The figures (in milligrams)
represent the turnover per day or the total size of the various pools. The stars indicate
the reactions affected by the hormones.
Calcium exchanges also take place with other tissues and with that bound to the plasma
albumin (lOO mg); some 20-300 mglday is also lost by sweat. The sizes of the bone
calcium compartments shown are proportional to the log of the calcium contents; the
linear proportions are 1 : 2 00. Survival is possible on calcium intakes as low as 200
mg/day: in this case changes occur in the uptake mechanism from the gut.
In addition to its action on calcium movements in bone, gut and
kidney it also increases renal loss of phosphate. This action means
th;tt the product of Ca x phosphate in the blood is not increased by
its action and so precipitation does not occur.
\ J "
UV light 0 -:.

.---..
E ..... 'd_l

Kidney
Low ea
2
+
Ioigure 7.42 Steps in the production and activation of the vitamin D group of substances
(cholecalciferol or Ce). Our bodies can produce the animal fonn of vitamin D by
S!WI irradiation or from the plant form. CerIain other animals cannot use the plant
form.
Calcitonin is secreted by the C-cells of the thyroid gland. It is a
small polypeptide which probably acts on the surface of target cells
to produce an intracellular messenger. The release of calcitonin is
controlled by the plasma calcium concentration acting directly on the
producing cells. High plasma calcium concentration increases its
secretion. It acts more quickly than the other two hormones. The
main action of calcitonin is to decrease the calcium coming out of
the bone; therefore, the plasma concentration drops due to the
continued entry of calcium into bone. In adult man its role is not too
clear; it is probably an accessory fine control to PH.
Control of plasma calcium concentration. Calcitonin, PH, and
1,25-DHCC interact to control the plasma calcium concentration. The
cellular interactions of the hormones can be explained by the scheme,
C cells of thyroid Bone
~ Rote of calcitonin release'" ...... ~ Net Co2+ output ..
Kidneys GI troet
Blood
Plasma
~ Rote of 1-25 DHCC reIease+ "T".
Net eo
2
+ uptake from ..
[eo2+] .,.
f-r+
Po rat hyroids
:
Kidney tubule
r-+
Rate of PTH release +
..... i ...
Net eo
2
+ reabsorption ..
Figure 7.43 Hormonal control of plasma calcium concentration. The upper name In each box identifies
the anatomical region; the lower name the variable. Lines jolnlng boxes mean 'has an effect on'. Dotted
lines refer to hormones, solid lines to Ca2+. A disturbance of plasma [Ca2+] (shown as an increase) is
followed by the consequences shown (arrows In the other compartments) whlch correct the disturbance.
246 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLoGy
where 1,25-DHCC produces a transport protein whose action is
allowed by PH but ineffective in the presence of calcitonin. This
scheme, primarily developed for bone, is also applicable to gut and
kidney if the action of calcitonin is excluded.
7.11.3 Other Modulators of Ca
Mechanical stress on bone promotes the deposition of calcium
and its absence causes resorption. This -is a problem in prolonged
bed rest and in space flights. A secondary complication which arises
is deposition of calcium in the excretory p ~ w a y s causing renal
stones.
Bone
Calcium channels
opened by parathyroid
hormone, closed by
calcitonin
Blood
Ca
2
+
Calcium 'pump'
depends on
vitamin D
Figure 7.44 Scheme for interaction of the three hormones affecting calcium mobilisation
from bone. Vttamin D causes the production of a ttansport pmtein which pumps calcium
into the blood from the lining cell. Parathyroid increases and calcitonin decreases the
calcium permeability of the side of the cell facing bone, thus controlling the rate at
which calcium enters the cell and hence the overall rate of transport from bone to
blood.
Blood pH affects ionised plasma calcium, such that a rise in pH
decreases the ionised calcium concentration and a fall in pH increases
it. pH acts on plasma calcium by two mechanisms, both working in
the same direction. If the pH is increased then: (1) more anionic
sites appear in the plasma proteins and so more calcium is bound;
and (2) calcium deposition in bone is favoured; both mechanisms,
therefore, decrease the ionised calcium. Lowering blood pH reverses
both mechanisms.
Clinically these effects are of importance in hyperventilation,
which raises the blood pH and so lowers the plasma calcium
concentration and may give tetany.
The detailed actions of these hormones are more complex than
stated above, and still imperfectly understood.
8
Introduction to
and
Energy Metabolism
8.1 CHARACTERIZATION OF METABOLISM
The ,vital activity of any living organism is determined by the
speCific organization of biological structures, metabolic processes,
energy metabolic processes, energy metabolism, genetic information
transfer. and regulatory mechanism. Damage of any of these links
develops a pathological process and a disease in the organism. An
understanding of the molecular mechanisms involved in the vital
activity or malfunction of the organism constitutes the basis for the
search and clinical applications of biological medicinal preparations.
In 'the overall metabolism of the living organism distinguished
are: exogenous metabolism, which comprises extracellular
transformations of materials on the way to their uptake and excretion
by the cells, and intennediary metabolism, which occurs in the cells.
1Jte intermediary metabolism is conceived as the sum total of chemical
rf;actions that occur in the living cell.
Functionally, metabolism encompasses the following major
processes:
(1) accumulation of energy from decomposition of compounds
or supplied by light;
(2) utilization of energy for synthesis of essential molecular
, componentS (monoms, macromolecules) and the performance
of work (osmotic, electric, mechanical);
i' (3) decomposition of renewable structural components of the cell;
, .' ' 247
(4) synthesis and decomposition of specialized biological molecules
(hormones, mediators, hormonoids, cofactors, etc.).
The sequences of chemical reactions involved form metabolic
pathways, or cycles, each of these performing a definite function.
Conventionally, central and special metabolic pathways are
distinguished. Central pathways are common to the decomposition and
synthesis of major macromolecules. Actually, they are much alike in
all representatives of the living world. Special cycles are characteristic
of the synthesis and decomposition of individual monomers,
macromolecules, cofactors, etc. Special cycles are extremely diversified,
especially in the plant kingdOm. For this reason, the plant metabolism
is conventionally classified into primary and secondary metabolisms.
'. The primary metabolism includes the classical processes of synthesis
and deeradation of major macromolecules (proteins, carbohydrates,
lipids, nucleic acids, etc.), while the secondary metabolism ensuing
from the primary one includes the conversions of special biomolecules
(for example, alkaloids, terpenes, etc.) that perform regulatory or other
functions, or simply are metabolic end bypr'oducts.
rn the metabolism, two oppositely directed I processes, or phases,
are commonly distinguished: catabolism and anabolism. Catabolism is
the sum of degradative processes leading to the cleavage of large
molecules into smaller ones. Anabolism is the sum of metabolic
processes leading to the synthesis of complex molecules from simpler
ones. Catabolism is accompanied by a release of energy that can be
stored as energy-rich ATP. Anabolic processes proceed through
consumption of A TP and decomposition of the latter into ADP and
H.P0
4
Therefore, ATP may be said to be a coupling energetic link
between the two metabolic pathways. However, ATP is not the only
linking component shared by catabolism and anabolism. Other simple
metabolites are also formed by the catabolic pathway from
macromolecules and monomers to be used as starting materials for the
subsequent synthesis of monomers and macromolecules, i.e. in the
process of anabolism. This linking pathway, or cycle, unifying
degradatlve and synthetic routes, is called the amphibolic pathway.
This signifies that the catabolic and anabolic pathways are coupled not
only via the energetic A TP-ADP system, but also through their common
metabolites, which renders the metabolism more versatile and
economical. At need, simple intermediates can be utilized in the
biosynthesis, without the necessity of their supply from the exterior.
The amphibolic pathways are associated with the terminal, or ultimate,
system of oxidation of the materials involVed, as the latter are degraded
INTRODUCTION TO METABOUSM AND ENERGY METABOLISM 249
to the end products, CO
2
and Hp, with a release of a large amount
of energy. Apart from them, urea and uric acid, which are produced
by specific metabolic reactions of amino acids and nucleotides, are
also end products of metabolism.
Uric lEid
Cerbohydrates
Proteins. Lipids I
Polynucleotlc;les
Figure 8.1 Scheme for catabolic and anabolic pathways (shown is their interrelation
through ATP-ADP system and amphibolic metabolite cycle)
During catabolic and anabolic processes, a renovation of the
cellular components takes place. It should be emphasized
that the catabolic and anabolic pathways are independent of each other.
Be these pathways coincident and differing in the cycle direction only.
the metabolism would have been side-tracked to the so-called useless.
or futile. cycles. Such cycles anse in pathology, where a useless
turnover of metabolites may occur. To avoid this undesirable
contingency. the synthetic and degradative routes in the cell are most
commonly separated in space. For example. the oxidation of fatty
acids occurs in the mitochondria. while the synthesis thereof proceeds
extramitochondrially. in the microsomes.
8.2 ENERGY CYCLES IN ANIMATE NATURE
Nutrients and energy are supplied to the living organisms from
PHYSIOLOGY. SIOCHEMImY AND BIOTECHNOLOGy
various sources. As far as the nutritional soQrceS are concerned, living
organisms are classified into large groups. aulotrophs (from the
Greek fJUlos, self, and trophos, food) capable of ass1niiJating CO
2
as
a starting nutrient for the buildup of other materials,
and heterotrophs (heteros, other) which utilize diverse organic
compounds as synthetized by other organisms. In a sense, the autotrophs
are primary to the heterotrophs. Reduced organic compounds (for
example, glu cose) synthetized- by the autotrophs from CO
2
contain a
larger amount of energy as compared with that in the consumed carbon
dioxide. .
In regard to the energy living organisms are divided into
phototrophs, for the sun light is a source of energy, and
chemotrophs, which utilize the energy of reduction-oxidation (redox)
reactions. In redox reactions, the energy that has been acquired by the
cell is released on transport of the from' a donor to an acceptor
(or oxidant). In these processes, the donor and the acceptor act as
partners and constitUte a donor-acceptQr pair, or a redox pair. If the
redox pair is made' up of organic compounds, the living organisms
involved are called chemoorga 'notrophs, and if it consists of iqorganic
compounds, chemolithotrophs. They are further diversified by their
relationship to oxygen as an electron acceptor. The organism cells that
utilize oxygen are referred to as aerobic ones, those capable of
dispensing with oxygen are anaerobic. Most' commonly, the
cells of higher organisms and bacteria possess both types, anaerobic
and aerobic, of energetic&. For this reason, such cells and organisms
are called facultative anaerobes, although the degree of faculty, or the
dependence'on oxygen supply, in them may vary. For example, higher
organisms are incapable of subsisting without oxygen for longer periods
of time. There are obligate anaerobes, in pcuIar, microorganisms,
.do not need oxygen altogether and even ,.defy it as poisonous.
, The, green possess a combined type of energetics,
phototrophic aqd'chemoorganotrophic (respiratory and glycolytic), which
enables them to abSQrb the energy of sun light at different periods of
theiI: ,development; in the absence, of light, the plants make, use of
chemical energy. Microorganisms are especially remarkable for, the
diversity of energetics types and combinations ther:eof. The energetics
in a number of representative!!: the animate ,tumlre
are listed in Table. ' :
Owing to the diversity of nutrition ,forms and energy consumption,
the living organiSms are'in nature closely related. The'interrelation
1ableJI.l Epergetics Types Encountered. in Living Organisms. _ 52

Source of. energy
2. . Red()x reactiQils of
. ca! compoUllds(donor-ac-
-. -Donor: organtc compounds;
. acceptor: O
2
c
Donor and acceptor:
organic compounds
Donor: inorganic
acceptor: 02
Donor and acceptor:
inorganic cOmpounds
Donor: organic compoun<lS;
acceptor: of mixed type
(organic compounds and 02)
3. Combiried type: light and
redox reactions
. Light aIidredox .
reactions of organic
compounds
Light and redox reactions
of inorganic compounds
Energeties type
PhototrolJbic .
Cbemotropbic _
tosynthetic plant cells ceptCr) .
Cbemoorganotropbic
(aerobic)
Cbemoorganotropbic .
(anaerobic)
Cbemolithotropbic
(aerobic)

(anaerobic)
. Cbemoorga otropbic .
(facultativ anaerobic)
Photocbemotropl. .c
Pbotochemoorganotroph- Idem
ie
Photocbemolithotropbic
-
Representatives . 0
g
Photosynthetic organs of higber <1
plagts. algae, bacteria _ . :3
Animal aIid bacterial cells. nonpbo
Idem
Idem
Bacteria
. ldeQl .
: Cells of higher animals: bacteria
Photosynthetic plant cells at differ
ent (light and dark) pbases'
Photosynthetic bacteria
d

!
at
>

at

between nutrition and utilization of energy sources may be conceived
of from the standpoint of specific cycles operative in the ~ t
nature. Major participants of a global cycle are the Sun as a source
of energy, autophototrophs capable of acquiring the solar energy and
of synthetizing carbohydrates and other organic materials from CO
2
,
and animals which consume organic materials and oxygen generated
by phototrophs.
Energy losses associated with the vital activity of all organisms on
the Earth are being compensated for by the energy of solar radiation.
It should be noted that the cells of man and animals utilize highly
reduced, i.e. hydrogen-containing, compounds (carbohydrates, lipids,
proteins, etc.) as energetic materials. Hydrogen is an energetically
valuable material. Its energy in a transformed form is stored in the
A TP chemical bonds in the cells of heterotrophic organisms.
8.3 ENERGETICS OF BIOCHEMICAL REACTIONS
To get a deeper insight into the metabolic and energetic processes,
knowledge of general principles of chemical energetics appears to be
of help.
In the living cell, all chemical reactions contributing to the
metabolism obey the laws of energetics. The first law of energy
conservation states that the energy of a chemical reaction can be neither
annibilated nor generated from nothing, it can merely be converted
from one form into another. In terms of this law, it becomes possible
to define the energy balance of a chemical process.
Spontaneous chemical processes can proceedgonly in one direction
towards a state of equilibrium; the state of equilibrium having been
reached, the process is brought to termination. The second law of
thermodynamics (energetics) enables one to predict the direction of
biochemical processes. According to this law, any spontaneous process
takes the route corresponding to a maximum of entropy under the
given conditions until an equilibrium state for the reaction is reached.
Entropy conveys a measure of the disorder in a system. An increase
in entropy in the course of a reaction prevents the return of the reaction
to the initial state, since for that to occur a diminution in entropy
would be required. A spontaneously disordered system is never capable
of turning into an ordered one. Therefore, all reactions that proceed
with a concomitant. increase in entropy are irreversible. For their
reversal, an additional energy is needed to be spent to compensate for
the losses in entropy change, i.e. to bring the system from the disordered
into the ordered state.
INTRODUCTION TO METABOLISM AND ENERGY METABOLISM 253
However, from the practical standpoint it appears expedient to
use the so-called free energy which, in contrast to entropy, is amenable
to measurement in the course of a reaction. The free energy defines
a portion of the total energy of a system that can be converted to
work at pressure and temperature kept constant. The free energy is
denoted AG. The portion of total energy of a chemical process that
cannot be converted to work at pressure and temperature kept constant
is called the bound energy and is expressed as the product TAS. where
T is the absolute temperature and AS is the entropy change of a system
under the given chemical reaction conditions. The sum of the changes
in free and bound energies is called enthalpy (internal energy of a
system) or heat energy content of a system and is denoted OH.
Enthalpy can be measured experimentally and is equal to the amount
of heat released in the given process. The enthaply change is defined
by the equation
M/ = AG + TAS
Otherwise stated,
AG=M/- TAS
The energetic state of any system, including that of a cell and an
organism, can be defined in terms of this very important equation.
The free energy is expressed in kilojoules per mole of substance. kIf
mol.
The free energy is a very convenient parameter for defining the
spontaneity or nonspontaneity of a chemical process. For spontaneous
processes, the free energy is seen to decrease, i.e. AG has a negative
value. Such reactions are called exergonic, i.e. proceeding with a
r l ~ of energy. These reactions supply energy to the cells. The
p r ~ for which the free energy is increased (i.e. l!iJ has a positive
value) cannot proceed spontaneously. They require energy supply from
the outside. Such processes are referred to as endergonic. i.e.
proceeding with an expenditure of energy. In a state of equilibrium,
AG = O.
The free energy of chemical reactions may be estimated both
under the standard conditions and under real, or physiological,
conditions. The standard free energy, AGO, of a biochemical reaction
is defined as a free energy change under the standard conditions, i.e.
at the concentration of reactants 1 mol/litre, temperature 25C <298
X), and pH 7.
If water is involved either as a starting compound or as a reaction
product, its concentration is taken equaI to 1.0 molllitre, although the
254 BIOCHEMI$ffiY AND BIOTECHNOLOGY
true concentration of water in dilute aqueous solutions is close to 55
mQlllitre.
The standard ,free" energy is found as a dUlerence belweep the
sum val'Je$ Qf free energy for the end products and the initial reactants.
The value of A Gp for ,a biochemical rcaction proceeding under the
physiological conditions is estimated with allowance for, the actual
concentrations of' cOlIlponents involved. '
With reference to the, free energy as a characteristic of metabolism
one may say that catab(llic reactions proceed with a release of energy
and ,anabolic ones, with, a consumption of energy' The anabolic
reactions can proceed only. as closely coupled to. the catabolic reactions.
High-energy, or DUlCroergic, compounds act as energetic mediators
between these two, types of reactions.
8.4 mGHT-ENEGRY AND LOW-ENERGY
PHOSPHATES: CONSlDERATlONS
Commonly, all compounds are classified into and low-
energy ones. As a conventional borderline between these two classes.
the free energy of about 20 kJ/mol for the phosphate bond hydrolysis
has been taken. This value used for the purpose of characterization
of biochemical prooesses should not be,confused with the bond energy
which is conceived as an energy required for disruption of the bond
between two neighbouring atoms in any molecule. Cleavage of the
phosphate bond results, alongside energy release, in the formation of
inorganic phosphates.
Energetic characteristics for some of the compounds of interest
are listed in the following Table.
TABLE 8.2 Standard free Energies, AGO, for Hydrolysis' of Some
High-energy and Low-energy CoDlp(lunds, and Free for
Hydrolysis 9f Compounds Under Physiological Conditions,AGp.
- AGO, - AG..,
kJ/mol kJ/mol
High-energy compounds
Phosphoeriol pyruvate '61.7' ,66.7
1,3-Diphosphoglycerate 49.2 41.7
phosphatt;. ' ' 42,5
A TP 30.4 50.,0
Acetyl-CoA 30.4
(Table 8.2 Contd.)
\
INTRODUctION 1'0 METABOUSM AND ENERGY METABOUSM 255
Compound,
ADP
-I1GO,
kJ/mol
28.3
':
, 28.3
Pyrophosphate (H4P20,)
UDP.;.glucose 24.2
20:8
Low-energy compounds
15.8
GlUCOSe I-phosphate
Fructose 6-phosPhate
AMP
I
Glucose 6-phosphate
a-Glycerol phosphate ,
14.1
13.8
9.2
-I1G"
kJ/mol
50.0
50.0
23.8
It shoUld be emphasized that ATP, the key 'mediator in energy
metabolism, is not the most energy-rich species. A TP is found in the
middle of. energy scale.
The most frequently occurring route is cleavage of the end
from A TP:
ATP ADP + (1)
The end phosphate adds water and is transferred onto another
compound, causing thereby the phosphorylation of the latter. An
alternative route for the phosphate bond energy release is exemplified
by pyrophosphate cleavage of A TP:
ATP AMP + H
4
P
2
0? (2)
This type of reaction is less frequent in biological processes. Its
distinctive feature is the formation of pyrophosphate, which ranks
among energy-rich materials. Hydrolysis of pyrophosphate (3) releases
roughly as much energy as hydrolysis of the A TP end phosphate
bonds. In biological processes, energy-rich pyrophosphate bonds are
seldom used for synthesis of other compounds because of the heat
energy released by pyrophosphate hydrolysis.
ADP may be used as a high-energy reactant in biochemical
processes. The cleavage of the ADP end phosphate bond (4) releases
the same amount of energy as that generated by splitting the end
phosphate bond in A TP. At frrst glance, it may appear that A TP can
be adequately replaced by ADP in chemical reactions, in particular
in the phosphoryla9 ion of other compounds. However, as evidenced
by the available experimental data, such a possibility has never been
in biological processes. At least such reactions are as yet
It is known, however, that ADP can be hydrolyzed to low
energy AMP and phosphate with heat release. For a long time, there
have been difficulties in determining the free energy for the ATP
phosphate bond. The standard free energy for hydrolysis of the A TP
phosphate bond, AGo, is equal to about -30.4 kJ/mot. This value has
been derived under standard conditions, i.e. at concentrations I M
for initial reactants and end products, pH 7.0, temperature 37C, and
excess of Mgl+ ions. It stands to reason that in the cell under
physiological conditions, the concentrations of initial reacnmts, end
products, and Mg" ions are substantially inferior to the standard values;
variations in pH are also possible. Therefore, the real free energy
for hydrolysis of the end phosphate bond in ATP and ADP, and of
the phosphate bonds in pyrophosphate is close to -50.0 kJ/mot. To
be noted, the values of AGO for other compounds differ from the
standard value, but not necessarily towards higher energies.
8.5 ENERGY TRANSFER IN
BIOCHEMICAL PROCESSES
The biological activity of the cell is closely associated with the
continuous redistribution of the energy delivered by the compounds
that enter the cell. The storage of energy in the specific phosphate
bonds of A TP constitutes the basis for the energy transfer mechanism
in the living cell. The living cell is a nonequilibrated chemical system-
the circumstance that permits storing the energy, produced by catabolic
reactions of nutrients, in the A TP phosphate bonds.
The A TP energy in the cell can be converted, via tree major
routes, to energy of chemical bonds, to thermal energy. and to energy
for performing work. We now consider in general terms the transfer
of chemical bond energy. The chemical reaction is associated with
the generation of A TP energy. If the reaction is in a state of
equilibrium. the energy transfer is accomplished to a 100% efficiency.
Nonetheless. no A TP energy will accumulate, since at equilibrium,
the free energy release is zero. Therefore, in order to provide for
energy storage in the A TP phosphate bmds, the reaction must be a
nonequihorium one. This is accomplished as .1 portion of the chemical
reaction energy is lost as heat. The remaiY.ing energy is transferred
onto the ATP phosphate bonds to be Hored therein. It follows.
therefore. that biological generators of er.ergy cannot attain the 100%
efficiency.
The third route of energy convers:on leads to the performance of
work. The chemical energy of A TP phosphate bonds can be spent on
osmotic. electric. mechanical, and other types of work. In doing so,
INTRODUCTION TO METABOUSM AND ENERGY METABOUSM 257
not all of the ATP energy is used for performing work; a portion of
it is dissipated as heat. Especially Qluch heat is produced by muscle
contraction, which represents a mechanochemical mechanism for heat
generation in the living organisms. Heat is of little use for performing
work in the living system. It is only in warm-blooded animals and
man that the heat is consumed for warming and maintaining a constant
temperature of the body, equal to about 37C. .
All chemical processes in the living organism can proceed only
with the involvement of enzymes. For this reason, prior to consider
the mechanism of energy extraction and various metabolic pathways,
it appears worthwhile to discuss enzymes and their functions.
!
9
Carbohydrate
Metabolism
I I.,
Carbohydrate metabolism in the organism tissues encompasses enzymic
processes leading either to the breakdown of carbohydrates (catabolic
pathways), or to the synthesis thereof (anabolic pathways). Carbohydrate
breakdown leads to energy release or intermediary products that are
necessary for other biochemical processes. The carbohydrate synthesis
serves for replenishment of polysaccharide reserve or for renewal of
structural carbohydrates. The of various routes of
carbohydrate metabolism in tissues and organs is defined by the
availability of appropriate enzymes in them.
9.1 CARBOHYDRAGE CATABOLISM IN TISSUES
A number of routes for carbohydrate catabolism in tissues are
known. They include glycolysis and its variant, glycogenolysis, which
are auxiliary pathways to energy production, respectively, by breakdown
of glucose (or other monosaccharides) and glycogen to lactate (under
anaerobic conditions) or to CO
2
and HzO(under aerobic conditions).
The involvement of glycolysis and glycogenolysis in the energetic
function has been discussed in detail in the foregoing section
'Bioenergetics" .
'i There is known one more catabolic route for carbohydrates
commonly referred to as the pentose phosphate cycle (also called
hexose mono phosphate. shunt, or phosphogluconate pathway).
As a tribute to the biochemists who have played a decisive role
in its investigation, the pentose phosphate cycle is also referred to as
the Warburg-Dickens-Horecker pathway.
The pentose phosphate cycle represents a multienzyme system in
258
CARBOHYDRATE METABOLISM 259
which the imponant intermediates are, as the name implies, pentose
phosphates. This cycle may be regarded as a branching, or shunt, at
the glucose 6-phosphate step in the overall glycolysis.
9.1.1 Pentose Phosphate Cycle
To provide for all steps of the pentose phosphate cycle, at least
three glucose 6-phosphate molecules are required. Let us consider
separate reactions of this cycle.
1. Dehydrogenation of glucose 6-phosphate is the reaction that
directs glucose 6-phosphate via the pentose phosphate pathway; it is
catalyzed by glucose-6phosphate dehydrogenase (in the schemes below,
for a lfuller description of the cyclic process, ~ glucose 6-phosphate
moletules are used)
H ~ ~
+8!0 __ +81
3 H01-H .;;:> c::::::::: 3 Ho-y-a
Hy-oH 3NADP + 3NADPH+H+ H1-O
H
I
Hy ~
HZC-oP01Hz RzC-oPO,R
J
glucose &-phosphate 6-pholphogJucona1B ....
Glucose-6-phosphate dehydrogenase is a dimer with a molecular
mass of about 135 000. Up to eight electrophoretically separable
isoenzymes for this enzyme are known. A specific feature of the above
reaction is the formation of NADP. ~ . The reaction equilibrium is
strongly shifted to the right, since the lactone formed is liable to
hydrolysis, which is spontaneous or lactonase-assisted.
2. Hydrolysis of 6-phosphogluconate lactone to 6-phosphogl-
uconate:
I
o
C; COOH
H ~ O H I H-b-OH
I Laetonase I
3 HO-C-H 0 --3 HO-C-H
I I +3H.O I
H-C-OH H-C-OH
H-t---...: H ~ O H
I I
HaC-OPOa
H
HIC-OPOaH.
8-plloglhqluconate 8-phoaphoglueonate
JilI:tone
3. Dehydrogenation of 6-phosphogluconate to ribulose 5-
phosphate. This reaction is catalyzed by 6-phosphogluconate
dehydrogenase according to the scheme:

"roH
l "o-r-H
a--y-oH
"1-0"
Hjc-opO
l
H
2
Hzy-oH
v-o
3 H
1
-OH
H-V-o
H

&phmphagIucanaI D-ribul0l8 &-phosphate
The reaction equilibciwn is shifted to the right. 6-Phosphogluconate
dehydrogenase is a climer with a molecular mass of about 100 000.
Several isoenzymes are known for this dehydrogenase. A specific
feature of this reaction is that dehydrogenation leads to an unstable
intermediate which is immediately decarboxylated on the surface of
the enzys.Ue. This is the second oxidation reaction in the pentose
phosphate cycle that leads to NADP therefore, the conversion
of glucose 6-phosphate to ribulose 5-phosphate is commonly referred
to as the oxidative phase of the pentose phosphate cycle. The sequence
of reactions starting from ribulose 5-phosphate to the formation of
initial -glucose 6-phosphate is called the nonoxidative, or anaerobic,
phose of this cycle.
4. Intereonversion or isomerization of pentose phosphates.
Ribulose 5-phosphate is capable of a reversible isomerization to other
pentose phosphates-xylulose S-phosphate and nOOse S-phosphate. These
reactions are catalyzed by two respective enzymes, viz., pentose-
phosphate epimerase and pentose-phosphate isomerase, according to
the scheme below:
HaC-OH
I
C=O
I
2 HO-C-H
,
H-C-OH
I
lIaC-OPOaH.
UoQlvlase

HaC-OH

eplmerue I
3 H-C-OH
I
H-C-OH
I
RsC-OPo.H.
lHIbulase
&op:Iotpbate
H-C=O
I
PeDloaeophvapballl B-C-OH
. IIII-.ai I

B-C-OH
I
H-C-OH
I .
HaC-OPO.H.
I).rIJIoIe
IoPIIoBPbaIll
Two other pentose phosphates (ribose 5-phosphate and xylulose
S-phosphate), which are derived from ribulose 5-phosphate, are
important for the subsequent reaction of the cycle. Two molecules of
CARBOHYDRATE METABOUSM 261
xylulose 5-phosphate and one molecule of noose 5-phosphate are
required for this.
5. Transfer of glycolic aldehyde from xylulose 5-phospbate onto
ribose S-phosphate or the first transketo)ase reaction. The next
reaction, which is catalyzed by transketolase, involves the pentose
phosphates produced by the foregoing reaction (the transferable moiety
is shown in the box):
HsC-OH
I
c=o
I
2 BO-C-H
I
H-C-OH
I
H.C-OPOaH.
.ulaJDIIB
I\opluIIpIIate
u.c-OH
I
c=o
ep/mmA 3
I
H-C-OH
I
BaC-OPOaB.

H-C=O
I
PlDtooe-pIuIspbate B-C-OH
- I
H-C-OH 4-

u.b-Ol'OaBa
A n"bose 5-phosphate molecule and one of the two xylulose 5-
phosphate molecules are used during the first transketolase reaction.
The other xylulose 5-phosphate molecule is consumed later, in the
second transketolase reaction.
Transketolase is a dimer with a molecular mass of 140000. Its
coenzyme is thiamine bisphosphate. Mgt+ ions are required for the
reaction. Both transketolase reaction products are used as substrates
at the next step of the cycle.
6. Transfer of dihydroxyacetone moiety from sedoheptulose
'-phosphate onto glyceraldehyde 3-phosphate. This reaction is
reversible and is catalyzed by transaldolase according to the scheme:
r.-----:1

I ,
H-e-o ' e-o.J'
I ",---
H-C-OH HO-C-H
I Transke10lase I
+ H-C-OH H-e-OH +
I Mal+ I
H-C-OH H-C-OH
I I
HJC-OPOJH
z
H-C-OH
I
H,C- OPOlH,
H-C-O
I
H-C-OH
I
HzC-OPOJH
z
D-rlboIe
&-phosphate
D-sedoheptulose
7..phasphate 3-pfiosphate
Transaldolase is a dimer with a molecular mass of about 70 000.
TIle fructose 6phosphate molecule produced by this reaction enters
glycolysis, while erythrose 4-phosphate is used as a substrate at
the subsequent steps of the cycle.
7. Transfer of glycolic aldehyde from xylulose S-phosphate onto
erythrose 4-phosphate or the second transket10ase reaction. This
reaction is related to the first transketolase reaction and is catalyzed
by the same enzyme. The only distinction is that erythrose 4-phosphate
acts as an acceptor for glycolic aldehyde:
r-----'
I H,(:-OH I
I I I
I c-o I
L __ , ___
HO-C-H
I
H-C-OH
I
H,C-OPO)H,
D-xylulose
6-phosphata
Fructose 6-phosphate and glyceraldehyde 3-phosphate also enter
the glycolysis.
Thus, in the course of reactions catalyzed by the intrinsic enzymes
of the pentose phosphate cycle, two fructose 6-phosphate molecules,
one glyceraldehyde 3-phosphate molecule, and three carbon dioxide
molecules are produced from three glucose 6-phosphate molecules.
In addition. six NADP -Hz molecules are formed. The overall scheme
for the pentose phosphate cycle is:
3 Glucose 6-phosphate-6NADP+ -+ 2 Fructose 6-phosphate
+ Glyceraldehyde 3-phosphate + 3C0
2
9.1.1 Interrelation of the PentOse
Phosphate Cycle and Glycolysis
These two pathways for carbohydrate conversion are closely
related. The products of the pentose phosphate route-fructose 6-
phosphate and glyceral-dehyde 3-phosphate-are likewise glycolysis
metabolites; for this reason, they are involved in glycolysis and
undergo conversion by glycolytic enzymes. Two molecules of fructose
6-phosphate can regenerate to two glucose 6-phosphate molecules
through the agency of the glycolytic enzyme glucose-phosphate
isomerase. Here, the pentose phosphate pathway functions as a cycle.
The other product, glyceraldehyde 3-phosphate, enters the glycolysis
to be either. converted to lactate (under anaerobic conditions) or
. oxidized to CO
2
and aerobic conditions). As can easily
be estimated, the conversion of glyceraldehyde 3-phosphate to lactate
. Synthesis of nu-
cteotides and nu-
cleosldes. nucIeo-
caellzymes.
... palynucleatld8s.
and histidine
I
Figure !U Scheme for integration of pentose phosphate shunt and glycolysis.
leads to the formation of two A TP molecules, the combustion
to CO
2
and "20 produces 20 A TP molecules. _t follows therefore
'. that under physiological conditions, when the pentose phosphate
pathway for carbohydrate conversion is in the glycolysis,
the overall process of glucose 6-phosphate conversion may be
expressed via the pentose phosphate cycle. Under conditions:
3 Glucose 6-phosphate+6NADP+ + 2P
t
2 GlUcose 6-phosphate
+ Lactate + 2ATP + 6NADP . + 3C0
2
Under aerobic conditions: !
3 Glucose 6-phosphate + 6NADP+ + 20ADP + 20P
t
2 Glucose 6-phosphate + 6C0
2
I + +20ATP
. At first glance, the energetic value of this of glucose
6-phosphate via the pentose phosphate cycle appears to be inferior to
that of the aerobic glycolysis pathway, the latter providing a maximum
of 38 ATP molecules. However, it should be me in mind that a
major portion of energy is stored in NADP and 6 NADP
molecules are energetically equivalent to 18 ATP molecules.
Consequently, the energetic effect remains the same.
'jol03 The Biological Function of
the Pentose Phosphate Cycle I
The biological' function of the pentose phosphate cycle involves
the production of two compounds: NADP 'Yhich is a "reductive
force" in the synthesis of various materials, and the metabolite ribose
5-phosphate, which is used as a building material in the synthesis of
various species. The major functions of the pentose phosphate cycle
are:
(1) amphibolic function: the cycle is a route to degradation of
carbohydrates and, simultaneously, to the supply of materials
used in synthetic reactions (NADP Hz and ribose 5-
phosphate);
(2) energetic function, since the involvement of pentose phosphate
cycle products (glyreraldehyde 3-phosphate) in the glycolysis
produces energy;
(3) synthetic function, as a major function associated with the
use of NADP and ribose 5-phosphate.
NADP Hz is used:
(I) in the detoxification of drugs and poisons in the monooxy-
genase oxidation chain of the endoplasmic reticulum of the
liver;
(2) in the synthesis of fatty acids and other structural and reserve
lipids;
(3) in the synthesis of cholesterol and its derivatives-bile acids,
steroid hormones (corticosteroids, female and male sex
hormones), and vitamins D; and
(4) in the neutralization of ammonia under reductive amination.
Ribose 5-phosphate is used in the synthesis of histidine,
nucleosides and nucleotides (nucleotide mono-, di-, and triphosphates),
nucleotide coenzymes (NAD, NADP, FAD, and CoA), and polymeric
nucleotide derivatives (DNA, RNA, and short-chain oligonucleotides).
The pentose phosphate pathway for carbohydrate conversion is
primarily operative in the organs and tissues in which an intensive
utilization of NADP Hz is needed for reactions of reductive synthesis
and for reactions involving ribose 5phosphate in the synthesis of
nucleotides and nucleic acids. For this reason, a high activity of this
pathway is observed in fat tissue, liver, mammary gland tissue
(especially during lactation, since the milk fat synthesis is essential
in this case), adrenal glands, gonadal glands, marrow, and lymphoid
tissue. Relatively high is the activity of pentose phosphate shunt
dehydrogenases in the erythrocytes. A low activity of the pentose
phosphate pathway is observed in muscular tissue (heart and skeletal
muscle).
CARBOHYDRATE METABOUSM
9.2 BIOSYNTHESIS OF
CARBOHYDRATES IN TISSUES
265
In the human and animal tissues and organs, synthesis of
carbohydrates occurs. Since glucose is the starting structural unit for
producing other monosaccharides and for assembling polysaccharides,
it is expedient to consider potential routes for the glucose synthesis
in tissues and organs. Formation of glucose from noncarbohydrate
materials is attested by the fact that, under prolonged starvation (in
an extreme contingency or as applied in therapy), the polysaccharide
reserves are rapidly consumed, while the glucose level
in the circulating blood is maintained to supply tissues, especially brain,
with nergy. .
9.2.1 Gluconeogenesis
The synthesis of glucose from noncarbohydrate sources is refened
to as the ghu:oneogenesis. It is feasible only in certain organism
tissues. The major site for gluconeogenesis is the liver. To a lesser
extent, the kidneys and intestinal mucosa are involved in this process.
Mechanism for Gluconeogenesis. Since the glycolysis involves
three energetically irreversible steps at the pyruvate kinase,
phosphofructokinase, and hexokinase levels, the production of glucose
from simple noncarbohydrate materials, for example, pyruvate or
lactate, by a reversal of glycolysis ("from bottom upwards") is
impossible. Therefore, indirect reaction routes are to be sought for.
The Jirst indired route in glucose synthesis .invoives the formation
of phosphoenolpyruvate from pyruvate without the intervention of
pyruvate kinase. This route is catalyzed by two enzymes. At first,
pyruvate is converted into oxaloacetate. This reaction occurs in the
mit3hOndria as the pyruvate molecules enter them, land is catalyzed
by ruvate carboxylase according to the scheme
PrrPaie eubosJ-
CH.- -COOH+HCOi+ATP
A
JJnI .... 1e
This enzyme. similar to all CO
2
assimilating enzymes, contains
biotin for a cofactor. Oxaloacetate is released from the mitochondria
into i the cytoplasm to enter gluconeogenesis. In the cytoplasm,
converts to phosphoenolpyruvate via a reaction catalyzed
by carboxylase:
- CH.=C-COOH+GDP(ADP)+CO.
b-POaBa
~ 1 _ f I I
The reaction equilibrium is shifted to the right. The major supplier
of phosphate groups is GTP, but for this purpose, A TP may also be
available.
All of the glycolysis reactions ranging from phosphoenolpyruvate
to fructose 1,6-bisphosphate are reversible, and the phosphoenolpyruvate
molecules formed are consumed for producing fructose 1,6-bisphospbate
by making use of the same glycolysis enzymes.
The second indirect route involves the formation of fructose 6-
phosphate from fructose 1,6-bisphosphate without the intervention of
phosphofructokinase reaction. This route is catalyzed by fructose
bisphosphl1lase: .
Fructose 1,6-bisphosphate + H
2
0 FlUI:IOSeblspbospbalase
Fructose 6-phosphate + 1\ P
The reaction is irreversibly shifted to the right. Fructose 6-
phosphate is isomerized to glucose 6-phosphate by glucose-phosphate
isomerase.
The third indirect route involves the formation of free glucose
from glucose 6phosphate by circumventing the hexokinase reaction.
This route is catalyzed by
Glucose 6-phosphate + H
2
0 Gha:ase6-pbospbalase) Glucose+H
2
P
The free glucose produced by this reaction is supplied to the blood
from the tissues. As exemplified by gluconeogenesis, one may easily
envision the economical organization of these metabolic routes, since,
apart frOm four special gluconeogenesis enzymes-pyruvate carboxylase,
phosphopyruvate carboxylase, fructose bisphosphatase, and glucose 6-
phosphatase-individual glycolytic enzymes are also used in the
gluconeogenesis.
. Noncarbohydrate Sources for Gluconeogenesis. In addition to
pyruvate and lactate, which are delivered to the liver and kidneys,
other noncarbohydrate compounds serve as substrates for glucose
synthesis. In accordance with the gluconeogenesis scheme, it may be
anticipated that all materials of noncarbohydrate nature that are
. I
CARBOHYDRATE METABOLISM
t,
Q
,..
c:
n Fruetft!1IA
o
z
m
o
Q
m
z
m
i
Fig1U'e 9.2 Schematic representation of gluconeogenesis.
267
,.. ,
n
o
,..
-<
amenable to conversion to a glucolysis metabolite (first group of
materials), to pyruvate (second group), or to oxaloacetate (third group)"
E
' serve as potential sources for glucose synthesis. For example,
ycerol may be included in the first group of materials, since this
. 01, which can convert to dihydtoxyacetone phosphate, can further
take up, depending on the reaction conditions, either gluconeogenesis
route, or glycolysis route. The involvement of glycerol in
gluconeogenesis proceeds according to the scheme: '
(1)
Glycerol phasphokln_
Glycerol 7 "\ aGlycerol phosphate
ATP ADP
! a-Glprol phosphate
'(21 11 Glycerol phosphate __ de_"t.,..roge .. __ ... Dihydroxyacatone
" phosphate
NAQ+ NAD-H+H+
Subsequently. dihydroxyacetone phosphate is used in glucose
synthesis.
The Krebs cycle acids convertible to oxaloacetate (third group
materials) may also be assigned to gluconeogenesis substrates.
However, amino acids that can convert the major source for
gluconeogenesis to both pyruvate and oxaloacetate and, consequently,
to glucose are the major source for gluconeogenesis. Amino acids
involved in the gluconeogenesis are referred to as glycogenic amino
adds. They encompass all of the protein amino acids, barring leucine.
9.2.2 Biosynthesis of Glycogen (Glyeogenogenesis)
Synthesis of glycogen is carried out in all the cells of organism
(the erythrocytes, perhaps, being the only exception), but this process
is especially active in the skeletal muscles and in the liver. The reaction
of glycogen breakdown, which is catalyzed by glycogen phosphorylase,
is nearly irreversible; for this reason, this enzyme takes no part in
the synthesis of glycogen. Two routes for glycogen synthesis are
possible. One route involves a successive addition of glucose units to
the extant glycogen moiety (glycogen primer); the other one originates
in glucose molecules. The source of glucose residues during glycogen
synthesis is the active form of glucose, uridine diphosphate glucose
(UDP-glucose), which is produced from glucose I-phosphate and
uridine triphosphate (UTP) through the agency of the enzymeglucose-
I-phosphate uridyltransferase according to the scheme:
Glucose I-phosphates-UTP UDP-glucose+H
4
PP7
The next step involves a transfer of the glucose residue from
UDP-glucose onto the glycogen primer through the aid of the enzyme
glycogen synthetase:
UDP-glucose + (Glucose) -+ UDP +(Glucose)n + I
To be noted, the glycogen synthetase catalyzes the formation of a-
1 -.4-glycoside bonds only. The "branching" enzyme, amylo-(a-l,4
-a-l,6)-transglycolysase, transfers short fragments (two or three
glucose residues) from one portion of the glycogen molecule onto
another and forms a-I -.6-glycosidic bonds (branch points). The
alternating action of these two enzymes results in the lengthening of
the glycogen molecule.
If the synthesis starts from glucose molecules, then the initial step
is the transfer of glucose residues from UDP-glucose onto an
intermediary acceptor-doliclwl phosphate (membrane-bound polyprenol
phosphate). Dolichol phosphate assists in the synthesis of an
oligosaccharide which is then transferred onto the protein. The
successive addition of oligosaccharide chains to the glycogen molecule
proceeds in the same manner as in the former route. The feasibility of
the latter route is substantiated by the fact that glycogen is always
bound with some protein.
Interrelation of Glycogen Synthesis and Degradation. Glycogen
synthetase exists in two interconvemble forms. The phosphorylated,
or inactive, form is called glycogen synthetase D,' the non phosphor-
ylated, or active, form is referred to as glycogen synthetase I. The
tfalISition from one form to the other is accomplished by two enzymes.
synthetase kinase (1) and glycogen synthetase phosphatase
according to the scheme:
ATP ADP
Glycogen synthetase I qlycogen synthetase D
(nonpholphorylated 4 7 2 (phosphorylated
enzyme) , enzyme)
H,P0
4
The processes of glycogen synthesis and degradation in the cells
are controlled via phosphorylation mechanisms involving the key
enzymes of glycogen metabolismglycogen synthetase and glycogen
phosphorylase. The activation of adenyJ.ate cyclase (for example. with
adrenalin or glucagon) leads to the production of CAMP which triggers
the "cascade" mechanism of phosphorylation of glycogen synthetase
and glycogen phosphorylase. with the resultant formation of the
inactive (phosphorylated) glycogen synthetase D and active glycogen
synthetase I. This favours glycogen synthesis.
Biosynthesis of Other
, Oligosaccharides and Polysaccharides
I Homo- and heteropolysaccharides are carbohydrate components
plasmic and structural glycoproteins in the mammals. These
carbohydrates are made up of a small set of monosaccharides: galactose.
mannose. N-acetylglycosamine, N-acetylgalactosamine. fucose, and
sialic acid. The connective tissue polysaccharides contain glucuronic
acid. iduronic acid. xylose, and sulphated derivatives of Nacetyl-
glucosamine or N-acetylgalactosamine. These homo- and heteroglycans
can be assembled by activated monosaccharide forms, such as nucleoside
phosphate derivatives of monosaccharides. UDP-derivatives of glucose,
!
tose, N-acetylglucosamine. N-acetylgalactosamine. glucuronic acid
d xylose; GDP-derivatives of mannose and fucose; and CMP-
d rivatives of sialic acid are used in the synthesis. Most nucleoside
p osphate saccharides are produced through reaction of monosaccharides
or their derivatives with the corresponding nucleoside triphosphates.
Occasionally. certain monosaccharides convert to other monosaccharides
as constitutive components of nucleoside diphosphate saccharides. For
example. UUPglucuronic acid and UDP-xylose are formed from UDP-
glucose:
UDPlIlucose 7 '"
2NAD+ 2NAJ).1I,
UOP.gIucuranlc acid UDP-xylose
CO
2
GDP-fucose is formed from GDP-maDD088:
GOP-mannose GDPfucose
NADP+ NADP-II,
+ 11,0
For example. it is to be noted that UDP-glucuronic acid which
is formed in the tissues. is used not only for polysaccharide synthesis.
but also for neutralization' and removal of toxic and useless materials
or foreign compounds from the organism.
Synthesis of oligosaccharides is carried out with the participation
of specific glycosyltransjerases whose diversity and functional
specialization in each cell provide for the production of various types
of homo- and heteroglycans. In the cell. glycosyltransferases are bound
with the membranes of endoplasmic reticulum or 'Golgi apparatus.
i.e. the organelles primarily responsible for assembling oligosaccbarides.
It may be presumed that initially the assembly is made on the polyprenol
phosphate molecule onto which the monosaccharide residues from
nucleoside phosphate saccharides are successively transferred by
glycosyltransferases. Subsequently. the synthetized oligosaccharide is
transferred onto proteins to form glycoproteins. Simultaneous transfer
of glycosyl residues and assembly of proteinbased oligo- and
polysaccharides are also possible.
9.3 CARBOHYDRATE METABOLISM
CONTROL IN THE ORGANISM
The carbohydrate metabolic routes in various tissues of the organism
discussed above differ in intensity. which is defined by metabolic
features specific of each tissue and organ. However. from the standpoint
of activity of the whole organism. certain specializations of the
carbohydrate metabolic routes in individual tissues are profitably
complementary. For example. strenuous muscular exertion requires
energy which is initially supplied by the breakdown of glycogen to
lactic acid. The latter compound is excreted into the blood to be supplied
CARBOHYDRATE METABOliSM 271
to the hepatic tissue, where it is used for the synthesis of glucose
during gluconeogenesis. From the liver, glucose is delivered in the
blood to the skeletal muscles to be consumed for energy generation
and to be deposited as glycogen. This intertissue (or interorgan) cycle
in the carbohydrate metabolism is referred to as the Cori cycle (called
also glucoselactate cycle):
Liver Muscle
Glucose_Glucose_Glucose
! l __ "",,",,, __ i---
G
-
, The maintenance of a constant glucose level in the blood is of
primary importance for the organism, since glucose is the major energy
substrate for the nervous tissue. The normal glucose content in the
blood is 3.3 to 4.0 mmolllitre. An increased concentration of glucose
in blood is known as hyperglycemia . .(fhyperglycemia reaches as high
as 9 to 10 mmoll1itre, the glucose excess is released into the urine,
i.e. glucosuria sets in. On the contrary. a decreased glucose percentage
in the blood is known as hypoglycemia. Hypoglycemia as low as about
I.S mmolllitre leads to the syncopal state, while a still lower glucose
concentration results in high excitability of the nervous system and
ultimately leads to convulsions and coma.
To gain a better understanding of the mechanism that controls
the glucose level in the blood, it is important to examine processes
that contribute to an increased or lowered glucose concentration.
Processes leading to hyperglycemia:
(I) absorption of glucose from the intestine (alimentary
hyperglycemia);
: (2) breakdown of glycogen to glucose (commonly, in liver);
I (3) (in liver and kidney).
Processes leading to hypoglycemia:
'(1) transport of glucose from the blood to tissues followed by
glucose oxidation to end products;
(2) synthesis of glycogen from glucose in liver and skeletal
muscles;
(3) production of triacylglycerol from glucose in fat tissue .
. The dietary intake of carbohydrates leads to a short-term (within
1 2 hours) hyperglycemia and, occasionally, glucosuria.
I Starvation stimulates the consumption of glycogen reserves in liver
and skeletal muscles, which prevents the development of hypoglycemia,
but within the space of a few hours only. Then, under lasting starvation,
the glucose level is maintained solely owing to the gluconeogenesis,
primarily at the expense of proteinic amino acids which suffer
degradation in the tissues. In point of fact, the potential ability to
sustain starvation is determined by the protein reserves available for
glucose production.
Glycogen
breakdown
Blood
AbsIQtlon
from 1nt8stive - .....
Gluconeogenesis
Glycogen (liver,
skeletal muscle'
C
2
0+ H
2
0
(numerous tissues)
Trlacylglycerides
(adipose tissue,
The glucose level in the blood is monitored by neurohormonal
mechanisms. Excitation of the sympathetic portion of the autonomic
nervous system increases the glucose level in the blood, while excitation
of the parasympathetic portion produces a reverse effect. The only
hormone capable of reducing the glucose content isinsulin. It stimulates
all of the three processes of glucose assimilation (mtracellular transport
and degradation of glucose, synthesis of glycogen, and synthesis of
triglyceride from glucose in fat tissue). All other hormones make the
glucose level increase; for this reason, they are occasionally referred
to as contrainsular hormones. These include adrenalin, glucagon,
thyroxin and triidothyronin, somatotropin (which stimulate glycogen
degradation), and glucocorticoids (which stimulate gluconeogenesis).
I
10
Lipid
Lipids are continually renewed in the organism tissues. The major
part of lipids in the human body is represented by triacylglycerides
which occur as in-closions in most tissues; the fat lissue, which consists
nearly totally of triacyl-g1ycerides, is especially lipid-rich. Since
triacylglycerides play an important role in the energetics of the
organism, the processes of their renewaJ (the conversion haJf-time
for triacylglycerides in different organs varies from 2 to 18 days)
involve both mobilization and deposition during energy production.
Compound lipids (phospholipids, sphingolipids, g1ycolipids, and
cholesterol and its esters) that make part of the biomembrane are
subject to a less active renew-a1 as compared with triacylglycerides.
Their renewal is associated either with the restoration of an impaired
portion of the membrane, or with the replacement of a "defective"
molecule by a new one.
The renewal of tissue Iipids involves their preliminary intracellular
bydrolysis by enzymes.
DEGRADATION OF LIPIDS IN TISSUES
lO.1.1 Intracellular Hydrolysis of Lipids
Hydrolysis of triacylglycerides in tissues is effected by a tissue
enzyme, tri-acylglyceride Upase, which hydrolyzes triacylglycerides to
glycerol and free fatty acids. There are a variety of tissue lipases that
differ primarily in their optimum pH and their location in the cell.
The acidic lipase is contained in Iysosomes; the basic lipase, in
microsomes; and the neutraJ lipase, in cytoplasm. A specific feature
E
the tissue lipase is its sensitivity to hormones which, by activating
enylate cyclase, elicit the transition of the ina,?tive tissue lipase to its
ctive form via phosphorylation with protein kinase. This mechanism
273
274 PHYSIOWGY. BIOCHEMISTRY AND BIOTECHNOLOGY
bears resemblance to the activation of phosphorylase B. Lipases mobilize
triacylglycerides. This process is also known as the tissue lipolysis.
The cell membrane phosphoglycerides are hydrolyzed with phosp-
holipases A
2
, C, and D, which are located chiefly in lysosomes.
However, certain phospholi-pases also occur in other cell organelles.
Hydrolysis of phosphoglycerides yields glycerol, fatty acids, nitrogenous
alcohols, and inorganic phosphate. There are also known specific
for hydrolysis of spbingolipids and glycolipids; the enzymeS
are involved in the renewal of these lipids.
As is known, hydrolysis of intracellular lipids does not lead to a
storage of glycerol and fatty acids. This indicates that the hydrolysis
rate for the lipids is balanced against the rate of their intracellular
oxidation. In the adipose tissue, glycerol and fatty acids as produced
by triacylglyceride hydrolysis are not subject to oxidation and are
released into the blood to be supplied to other organs.
10.1.2 Oxidation of Glycerol
The glycerol metabolism is closely related to the glycolysis
involving glycerol metabolites according to the following scheme:
HZt:H G'-' ..s.-...... r- HzCH HCH
--... __ --
H -elH ? "" H H ? " I
HzA-oH ATP AOP NAO+ NAO'H+W HzC-oPO,H
z
.'G1-...",-,,-
At first, glycerol is converted to a-glycerol phosphate through the
agency of glycerol phosphokinase. a-Glycerol phosphate, by the action
of NAD-dependent a-glycerol-phosphate dehydrogenase, is converted
to dihydroxyacetone phosphate, which, as a common glycolysis
metabolite, enters glycolysis to be reduced by enzymes to lactate under
anaerobic conditions, or to CO
2
and Up under aerobic conditions.
Conversion of one glycerol molecule yields one A TP molecule under
anaerobic, and 19 A TP molecules, under aerobic conditions. Glycerol
is a profitable energy sub-strate and is used as an energy source
practically by all organs and tissues.
10.1.3 Oxidation of Fatty Acids
Oxidation of higher fatty acids was first studied in 1904 by Knoop
who fed animals with phenyl-substituted fatty acids and analyzed the
products in the urine. He showed that the fatty acid oxidation results
in the successive cleavage of two carbon moieties from the carboxyl
end. Knoop coined the fatty acid oxidation mechanism asn-oxidaJion.
As has been established by Kennedy and Lehoinger in 1948-1949,
oxidation of fatty acids occurs in the mitochondria only. Lynen and
UPID METABOLISM 275
coworkers have outlined major enzymic processes in fatty acid
oxida-tion. At the present time, the p-oxidation of fatty acids is referred
to as the Knoop-Lynen cycle.
The fatty acids, as produced by intracellular hydrolysis of
triacylglycerides or supplied to the cell from the blood, must be brought
into a state of activation. Their activation is effected in the cytoplasm
with the participation of acyl-CoA synthetase according to the scheme:
ctB.-(CHtlll-CH.-CHs-COOB+ATP+CoASB aerl-4'.DA
o
1
.. - SCoA+AMP+a.P,Ct
Since the activation process is effected extramitochondrially,
transport of acyls across the membrape into the mitochondria is
necessary.
The transport is accomplished with the participation of carnitine.
which takes up the acyl from acyl-CoA on the outer membrane side.
Acylcarnitine assisted by carnitine translocase diffuses to the inner
side of the membrane to give its acyl to the CoA located in the matrix.
The process of reversible acyl transfer between CoA and carnitine on
the outer and inner sides of the membrane is effected by the enzyme
acyl-CoA-camitine transferase.
Mltllchundliuli
y
R-C-SCM
1

Figure 10.1 Transport of fatty acids across rhe mitochondrial membrane.
The oxidation of fatty acids within the Knoop-Lynen cycle occurs
in the matrix. The Knoop-Lynen cycle includes four enzymes that act
successively on acetyl-CoA. These are: acyl-CoA dehydrogenase (FAD-
enzyme), enoyl-CoA hydratase, 3-hydroxyacyl-CoA
de"rdrogenase (NAD-dependent enzyme). and acetyl-CoA acyltrans-
ferase. Each turn, or revolution. of the "fatty acid spiral" produces
I
an acetic acid residue split from the fany acid as acetyl-CoA to yield
one FAD molecule and one NAD H2 molecule. The cycle
turns are then repeated until the fatty acid chain becomes shortened
to a four-carbon fragment, i.e. butyryl-CoA. In the last tum, butyryl-
CoA splits apart, and two, rather than one, acetyl-CoA molecules
are formed.
CHl-(CH')"YHl'HJ.-SCoA
II H
Acyl.coA dehydrogenase
FADH, Y
CH,-.'H,l,,-CH==C'II-(, - S CoA
+H'ol E-,",,",,_
seoA

011 II
'NA '3-Hydoxyacyt-coA dehydrogenase
NAD'II, Y
clI,--(cH""llll-c-sCOA
+C
SH
1 ; H --y ___ _
CII,-ICH
2
),,-C-SCoA + CH,-c-SCoA
$

y W
CH,;-l'-SCoA CH,-C -SCoA
Figure 10.2 Scheme for oxida-tion of fatty acids in the Knoop-Ly-nen cycle,
The oxidation products of an even-numbered fatty acid are acetyl-
CoA, FAD Hz and NAD Subsequently, acetyl-CoA enters
the Krebs cycle, and FAD Hz and NAD Hz are directly supplied
to the respiratory chain,
The specific behaviour of odd-numbered fatty acids under
LIPID METABOUSM 277
oxidation is that one propionyl-CoA molecule
per oxidized fatty acid mole-cule, alongside the products acetyl-CoA,
FAD and NAD (common to even-numbered fatty acids),
is formed. Propionyl-CoA converts to succinyl-CoA:
C.!tJ Y
OOH
CH,-cHzCO-5CoA ,>". CH,-cH-CO-scoA -
A'IP AIK'+' IUIIDInyI'CoA'
mathytnwlonyl-coA
Carboxylation of propionyl-CoA is accomplished by propionyl-
CoAl carboxylase (biotin, which is the carboxyl group carrier, serves
as aicoenzyme for this enzyme); the presence of ATP is also required.
The. methylmalonyl-CoA formed is converted by methylmfllonyl-CoA
mutase (whose coenzyme, deoxyadenosylcobalamin, is a derivative of
vitamin BIZ) to succinyl-CoA; the latter enters the Krebs cycle.
The specific behaviour of unsaturated fatty acids under oxidation
is determined by the position and the number of double bonds in the
fatty acid molecule. The stepwise oxidation of an unsaturated acid to
the position of a double bond in it proceeds in a manner similar to
that of saturated acid oxidation. If the double bond retains the same
configuration (trans-configuration) and position (1l.2, 3) as those of the
enoyl-CoA, which is produced during the oxidation of saturated fatty
./ acids, the subsequent oxidation proceeds via conventional route.
Otherwise, the oxidation reaction proceeds with the involvement of
an accessory enzyme, fl.3, 4-ciS-fl.
2
, J Jrans-enoyl-CoA isomerase; this
facilitates the translocation of the double bond to an appropriate
position and alters the double-bond configuration from cis totrans.
The unsaturated fatty acid oxidation proceeds at a rate higher
that for saturated acids. For example, if the oxidation rate for

stearic acid is taken as a reference value, the oxidation rate
fo oleic acid is 11 times, linolic acid, 114 times, llilolenic acid, 170
. , and arachidonic acid, nearly 200 times as high as that for stearic
acid.
In addition to p-oxidation, two other oxidation routes are known
for fatty acids, referred to as a- and ro-oxidation. However, they
exhibit a lower activity and initially involve the formation of a- and
o-hydroxy acids, with subsequent con-versions thereof. These oxidation
routes are of inferior energetic value as compared with p-oxidation;
pr6umably, they are implicated in special functions of the cell.
I Energy Balance of Fatty Acid Oxidation. The energetic value
of an even-numbered fatty acid is estimated in the following manner.
COmplete oxidation of a fatty acid composed of at carbon atoms yields
278 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOWGY
n acetyl-CoA molecules (each acetyl con-taining two carbon atoms)
and n-l FAD ~ and NAD ~ molecules (since the last tum of the
"fatty acid spiral" yields two acetyl-eoA molecules, one FAD ~
molecule and one NAD ~ molecule). Oxidation of FAD ~ gives
two ATP mole-cules, and oxidation of NAD Hz. three A TP
molecules, i.e. a total of 5 ATP mole-cules, or, in the general case.
5 (n-I) A TP molecules. As has been noted above, the complete
oxidation of one acetyl-eoA molecule results in the formation of 12
A TP molecules, while n acetyl-CoA molecules provide 12n A TP
molecules. One A TP molecule being used for the fatty acid activation,
1211-1 ATP molecules remain. Now, the ATP balance for the complete
oxidation of an even-numbered fatty acid may be expressed by the
formula
5 (n - 1) + 12n - 1 = (l7n - 6) ATP molecules
where n is equal to half the number of carbon atoms in a given fatty
acid. For example, the palmitic acid molecule, which contains 16
carbon atoms, produces 130 A TP molecules.
The energetic value of fatty acids is superior, for example, to
that of glucose. For example, the complete oxidation of capronic acid
(whose molecule contains the same number of carbon atoms as
glucose) yields 45 ATP molecules as compared with 38 molecules
which can be derived from glucose. However, the acetyl-eoA
molecules as produced by P-oxidation require a sufficient amount of
oxaloacetate tQ be degraded by the Krebs cycle. In this respect,
carbohydrates have an advantage over fatty acids, since the breakdown
of the formet: species leads to pyruvate serving as a source for both
acetyl-eoA and oxaloacetate (pyruvate-carboxylase reaction), i.e. the
acetyl-CoA conversion within the Krebs cycle is thus facilitated. It is
not without reason that in the older biochemical literature the notion
that "fats bum down in the carbohydrate flame" was popular, since
the ATP from glycol-ysis can be used for the cytoplasmic activation
of fatty acids, while the pyruvate-derived oxaloacetate facilitates the
insertion of fatty acid acetyl residues into the Krebs cycle.
Importance of Fatty Acids as Energy Substrates for Various
Organs and 'lissues. The organism tissues dif(er in the extent of
utilization of fatty acids and their intermediary oxidation products,
the so-called ketone bodies, as energy substrates. Fatty acids a,re
ktively consumed in the heart as well as in the kidneys and skeletal
muscles (under prolonged physical' exertion). In these organs, the
ketone bodies undergo oxidation to yield additional energy. In the
LIPID METABOUSM 279
nervous tissue, the con-sumed amount of fatty acids and ketone bodies
as sources of energy is insignifi-cant.
10.2 BIOSYNTHESIS OF LIPIDS IN TISSUES
10.2.1 Biosynthesis of Fatty Acids
In the organism tissues, fatty acids are continually renewed in
order to provide not only for the energy requirements, but also for
the synthesis of multicomponent lipids (triacylglycerides, phospholipids,
etc.). In the organism cells, fatty acids are resynthetized from simpler
compounds through the aid of a supramolecular multienzyme complex
r ~ to as fatty acid synthetase. At the Lynen laboratory, this
s lase was first isolated from yeast and then from the liver of
birds and mammals. Since in mammals palmitic acid in this process
is a major product, this nmltienzyme complex is also calledpalmitate
synthetase. .
Biosynthesis of fatty acids exhibits a number of specific features:
(I) fatty acid biosynthesis, as distinct from oxidation, is localized
in the endo-plasmic reticulum;
(2) the source for the synthesis is malonyl-CoA, which is
. produced from acetyl-CoA;
(3) acetyl-CoA is involved in the synthetic reactions as a primer
only;
(4) NADP . H2 is used to reduce fatty acid biosynthesis
intermediates;
(5) all the steps of malonyl-CoA fatty acid biosynthesis are cyclic
processes that occur on the surface of palmitate synthetase.
Production of Malonyl-COA for the Fatty Add Biosynthesis.
Aet;tyl-CoA serves as a substrate in the production of maJonyl-CoA.
Thqre are several routes by which acetyl-CoA is supplied to the
cyt6plasm. One route is the transfer of acetyl residues from the
mitochondrial matrix across the mitochondrial membrane into the
cyto-plasm. This process resembles a fatty acid transport and is
likewise effected with the participation of camitine and the enzyme
acetyl-CoA-camitine transferase. Another route is the production of
acetyl-CoA from citrate. Citrate is delivered from the mitochondria
and undergoes cleavage in the cytoplasm by the action of the enzyme
ATP-citrOle lyase:
I Citrate + ATP + CoA -+ Acetyl-CoA+Oxaloacetate+ADP+P]
The reaction is practically irreversible, and is shifted to the right.
280 PHYSIOLOGY, BIOCHEMISTRY ~ BIOTECHNOLOGY
The acetyl-CoA supplied to the cytoplasm via the above routes
is used for the synthesis of malonyl-CoA:
CHi COOH
6=O+HCO.+ATP B - = ~ btl + ATP+BaPO.
~ t=O
Mdl'14tA koA
maicmyloCoA
The reaction is catalyzed by the biotin,enzyme acetyl-CoA
carboxylase (E-biotin) assisted by Mg2+ ions. This enzyme is a
tetramer with a molecular mass of 400 000-500000.
Steps of Fatty Acid Biosynthesis Assisted by Palmitate
Synthetase. palmitate synthetase is composed of seven enzymes; of
these, each is assigned a definite ftmc-tion. The acyl carrier protein
(ACP) is located at the centre of the nmltienzyme complex; the other
six enzymes occupy peripheral positions. ACP acts both as an acceptor
and a distributor of acyl groups. ACP contains a covalently bound 4-
phos-phopantethein bearing a free SH group for accepting an acyl
moiety. In addition to this central SH group, palmitate synthetase has
a peripheral SH group. Both SH groups participate as acyl acceptors
in the synthesis of fatty acids on the sur-face of the nmltienzyme
complex.
The cyclic process of fatty acid synthesis may be represented by
a series of consecutive reactions (hereafter palmitate synthetase is
designated by the symbol (SH).
SH
1. Transfer of acetyl moiety from acetyl-CoA onto the synthetase:
o
II
SH CH, S - C-CH,
E/ + ~ = O _ ( +CoASH
'SH b "aH
This reaction is carried out by the first enzyme of palmitate
synthetase-acetyl-transacylase, which possesses an SH group. At this
stage of the synthesis, the acetyl acts as a primer.
2. Transfer of malonyl moiety from malonyl-eoA onto the
synthetase:
LIPID METABOUSM 28h
0 COOH 0
II

II
S-C-CH. S-C-CH.
,
+

....
, 0
+ CoASH
"
l
" II
sa SCoA
S - C-CH.-COOH
The reaction is effected by the second synthetase enzyme-
f1Illlonyltransacylase.
3. Acetyl-malonyl condensation and decarboxylation of the product
fOj: 0
I II
S-C-CB.
, 0
SH
_, 0 0 + CO.
" II
S - C-CH.-COOH
's -
The reaction is catalyzed by the third synthetase enzyme-3-ketoacyl
synthetase. An acetoacetyl, which is bound to synthetase, is formed
at this stage.
4. The first reduction of the intermediate with the involvement
of NADP eH,.:
ISH
7" '5'" V-CH' -CII-t'H
NAJllH+H+ NAJP+ 2 "' - )
The reaction is catalyzed by the fourth synthetase enzyme- P-
ketoacyl reductase, to yield intermediary hydroxybutyryl.
is. Dehydration of the intermediate:
S, 8H
./ . 0 OH -, 0 + H.O
"II I " II
S _ C-CH.-CR-CH. S - C-CH==CH-CH.
The reaction is catalyzed by the fifth synthetase enzyme-
hydroxyacyl hydratase, to produce crotonyl.
6. The second reduction of the intermediary product with the
involvement of
SJI
" 0 0
\ ....
282 PHYSIOLOOY, BIOCHEMISTRY AND BIOTECHNOLOGY
The reaction is catalyzed by the sixth synthetase enzyme-
to form an enzyme-bound butyryl. The butyryl thus
synthetized is transferred, through the mediacy of the first synthetase
enzyme. acetylttansacylase. onto the SH group (the upper one in the
Scheme) initially bound to the acetyl primer. The SH group (the lower
one in the Scheme). thus freed. accepts a new malonyl residue:
o
sa
0
S -
); _ 8-CB
1
-CHI-ca
a
-./
)a
The synthetic cycle is thus repeated.
Seven cycles are implicated in palmitic acid biosynthesis and,
accordingly. seven malonyl residues and one acetyl are required.
Acetyl is the end moiety in fatty acid biosynthesis. The palmitic acid
thus synthetized is either transferred onto the outer CoA to produce
acyl-CoA. or. more commonly, is hydrolyzed by the.speciticpalmiJale
deacylose to yield a free fatty acid.
Fatty Add Chain Elongation., The mitochondria and endop1asmic
reticulum provide the conditions for an eventual chain elongation of
the cell-synthetized or dietary fatty acids. This process is different
from the fatty acid biosynthesis in the proper sense of the term. In the
mitochondria. the chain elongation is achieved through the aid of an
enzyme complex by adding acetyl residues from acetyl-CoA. In the
endoplasmic reticulum. the chain elongation is accomplished by an
enzyme complex through making use of malonyl-CoA.
Biosynthesis of Unsaturated Fatty Adds. In the mammalian
tissues. the forma-tion of monoene fatty acids is only possible. Oleic
acid is derived from stearic acid. and palmitooleic acid. from palmitic
acid. This synthesis is carried out in the endoplasmic reticulum of
the liver cells via the monooxigenase oxidation chain. Any other
unsaturated fatty acids are not produced in the human organism and
must be supplied in vegetable food (plants are capable of generating
polyene fatty acids). Polyene fatty acids are essential food factors for
mammals.
10.2.2 Biosynthesis of Triglycerides
Triglyceride biosynthesis proceeds with the involvement of the
lipids deposited in fat tissue or in other tissues of the organism. This
process is localized in the hyaloplasm of cells.
LIPID METABOUSM 283
a-Glycerol phosphate and acyl-CoA, rather than corresponding
free glycerol and free fatty acid, are utilized in the direct synthesis of
triglycerides. a-Glycerol phosphate is produced either by phosphor-
ylating the glycerol supplied to the tissue, or by reducing dihydroxy-
acetone phosphate as an intermediary product of glycolysis.
The first step of triglyceride biosynthesis is the formation of
phosphatidic acid with the involvement of glycerophosphate. acyltrans-
ferase:
:;::::==
R-CO-SCOA
RL..cO-SCoA
y
H21-O-C-R
"". Q
2 CoA SfI
H
2
C-OP0
1
H
2
. phosphatldic: acfd
Further, the phosphatidic acid is subject to an attack by phospha-
tidate phosphatase to yield diglyceride:
o
11
'J=:=i :.
s.bopq,a.
IIIdd .u.1JCldde
The third acyl residue is transferred onto diglyceride by means
Qf diglyceride acyltronsferase
"l:t.

R"-CO-SCaA
""
Co" SH
dleCY'glycarlde

H
2
C-O-X-R
f
y
H --O-CR'
H,
trlecylglycarlde
The triacylglyceride thus synthetized is stored as fat inclusions
in the cell cyto-plasm.
10.2.3 Phospholipid Biosynthesis
Biosynthesis of phospholipids is associated with the renewal of

284 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHN0LOqY
FJgUI"e 10.3 Two pathways for \he s ~ t h s i s of certain phospholipids.
membranes. This process is accomplished in the, tissue hyaloplasm.
The first steps of phospholipid and triglyceride biosyntheses coincide;
subsequently, these routes diverge at the level of phosphatidic acid
and diglyceride.
Two routes to phospholipid biosynthesis are known; in either,
the participation of CTP is necessary. The first route involves
phosphatidic acid in pbosphoglyceride biosynthesis. Phosphatidic acid
reacts with CTP to yield CDP-diglyceride which, as a coenzyme,
can participate in the transfer of diglyceride onto serine (or inositol)
to produce phosphatidylserine (or phosphatidylinositol). Serine
phosphatides are liable to decarboxylation (pyridoxal phosphate acting
LIPID METABOliSM 285
as a coenzyme) to yield ethanolamine phosphatides. The latter species
are subject to methylation by S-adenosylmethionine (which donates
three methyl groups), tetrahydrofolic acid and methylcobalamin acting
as methyl group carriers.
The second synthetic route involves activation of an alcohol (for
example, choline) to produce CDP-choline. The latter participates in
the transfer of choline onto diglyceride to form phosphatidylcholine.
The phospholipids thus obtained are transported by lipid-carrier
cflOplasmic proteins to the membranes (cellular or intracellular) to
rj lace the used or jmpaired phospholipid molecules.
Because of the competition between the phospholipid and
triglyceride synthetic routes for common substrates, all substances that
favour the former route impede the tissue deposition of triglycerides.
Such substances are referred to as lipotropic factors. They include:
choline, inositol, and serine, as structural components of phospholipids;
pyridoxal phosphate, as an agent facilitating the decarboxylation of
s,mne phosphatides; methionine, as a donor of methyl groups; and
folic acid and cyanocobalamin, involved in the formation of methyl
group transfer coenzymes (tetrahydrofolic acid and methylcobalamin).
They may be used as drugs preven-tins excessive deposition of
. triglycerides in tissues (the so-called fatty infiltra-tion).
10.2.4 Biosynthesis of Ketone Bodies
Three compounds: acetoacetate, P-hydroxybutyrate, and acetone,
are known as ketone bodies. They are suboxidized metabolic
intermediates, chiefly those of fatty acids and of the carbon skeletons
of the so-called ketogenic amino aGids (leucine, isoleucine, lysine,
.,henylalanine, tyrosine, and tryptophan). The ketone body production,
~ ketogenesis, is effected in the hepatic mitochondria (in other tissues,
ketogenesis is inoperative). Two pathways are possible for ketogenesis.
he more active of the two is the hydroxymethyl glutarate cycle which
is named after the key intermediate involved in this cycle. The other
one is the deacylase cycle. In activity, this cycle is inferior to the
former one. Acetyl-CoA is the starting compound for the biosynthesis
of ketone bodies.
Hydroxymethyl Glutarate Cycle. At the first step of this cycle,
condensation of two acetyl-CoA molecules takes place, with the
participation of acetyl CoA acetyltransferase:
!CH.-C-SCoA+CH.-C-SCoA - CH.-C-CH.-C-SCoA+CoASH
i & g & &
I aeetYloCoA -'7I-coA IIIletoaat710C0A
286 PHYSIOLOGY, BIOCHEMISTRY AND BIOTECHNOLOGY .
Further, acetoacetyl-CoA becomes coupled once more to an acetyl-
CoA molecule through the assistance of hydroxymethylglutaryl-CoA
synthase:
ClIo
CH.-C-CH.-C - SCoA+CB.-C - SCoA -
A A g -0 bH 0
__
/3-Hydroxy-f3-methylglutaryl-CoA is split by hydroxymethylglutaryl-
CoA lyase into acetyl-CoA and acetoacetate:
CH.
I
HOOC-CHa-C-CH.-C - SCoA - CH.-C_ SCoA+HOOC-CH.-C-CH.
6H A A g
Acetyl-CoA is again used at the first step and closes thereby the
whole process into a -cycle. Acetoacetate, as a representative of the
ketone body family, is the end product of the hydroxymethyl- glutarate
cycle.
The other ketone bodies are derived from acetoacetate: P-
hydroxybutyrate, by reduction with the involvement of NAD-dependent
hydroxybutyrate dehydrogenase, and acetone, by decarboxylation of
acetoacetate with the participation of aceto-acetate decarboxylase:
V . fH
HOOC--<:HzC-CH
J
o. HOOC-CH
2
-CH-CH
J
AD=;;:-. ,-hyclroxybutyrate
The Deacylase Pathway for Ketogenesis is feasible after the
formation of acetoace-tyl-CoA which is subject to hydrolysis to
acetoacetate in the liver with the involvement of acetoacetyl-CoA
hydrolase, or deacylase.
In the liver, the ketone bodies suffer no transformation, and are
excreted into the blood. The normal contents of ketone bodies (as
acetoacetate or /3-hydroxy-butyrate) amount to mere 0.1-0.6 mmol/
litre). Other tissues and organs (heart, lung, kidney, muscle, and
nervous tissue), as distinct from the liver, utilize the ketone bodies
as energy substrates. In the ceUs of these tissues, acetoacetate and 1-
hydroxybutyrate enter ultimately the Krebs cycle and "burn down"
. to CO
2
and H
2
0 to release energy.
LIPID METABOLISM 287
10.2.5 Biosynthesis of Cholesterol
In the experiments with acetic acid labelled radioisotopically and
fed to ani-mals, it has been established that the cholesterol carbon
framework is made up entirely of the acetic acid carbon.
Biosynthesis of cholesterol from acetyl-CoA proceeds, assisted
by the enzymes of endoplasmic reticulum and hyaloplasm, in many
tissues and organs. This pro-cess is especially active in the liver of
adult humans.
Cholesterol biosynthesis is a multistage process; in general, it
may be divided into three steps:
I (1) production of mevalonic acid from acetyl-CoA;
(2) synthesis of an "active isoprene" from mevalonic acid
followed by the con-densation of the former to squalene;
(3) conversion of squalene to cholesterol.
The initial reactions in the first step, prior to the formation of
P,:,hydroxy-p-methylglutaryl-CoA from acetyl-CoA, resemble those
involved in ketogenesis witb the only distinction that ketogenesis occurs
in the mitochondria, while cho-Iesterol biosynthesis is carried out
extramitochondrially:
2 Acetyl-CoA Acetoacetyl-CoA + Acetyl-CoA

Further, l3-hydroxy-l3-methylglutaryl-CoA is converted with
hydroxymethylgluta-ryl-CoA reductase to mevalonic acid:
?" "">P'
i 1R A aNADPoJI+a+ a +
eH,
I
HOOC-CHrC-CHa-tHaOH +CaASH
I
OR
I This reaction irreversible and is a rate-limiting stage of the
cholesterol biosynthesis.
An alternative route to mevalonic acid is also possible, which
differs from the former one in that the formation of I3-hydroxy-13-
methylglutaryl residue occurs on the surface of an acyl carrier protein
(like in fatty acid biosynthesis). The intermediary product in this route,
P-hydroxy-p-methylglutaryl-S-ACP, is re-duced by another enzyme to
mevalonic acid.
During the second step, mevalonic acid is implicated in a number
of enzymic reactions involving A TP, and is converted to isopentyl
pyrophosphate and to its isomer 3,3-dimethylallyl pyrophosphate.
Actually, the two compounds constitute the "active isoprene", which
is consumed in the production of squalene. During the third step.
cholesterol is generated from squalene: ,
Squalene -+ Lanosterol -+ Cholesterol
The steroid ring hydroxylation proceeds with the involvement of
the monooxygen-ase chain of endoplasmic reticulum membranes.
Cholesterol esters are produced by transferring an acyl moiety
from acyl-CoA or from phosphatidylcholine onto the cholesterol
hydroxyl group. The latter process is catalyzed byphosphatidylchoUne
cholesterol acyltransjerase:
Phosphatidylcholine + Cholesterol-+
Lysophosphatidylcholine + Cholesterol ester
Cholesterol esters are produced especially actively in the intestinal
mucosa and iIi the liver.
Thus, the tissue cholesterol can be syntbetized from any materials
whose break-down leads to acetyl-CoA. These include carbohydrates,
amino acids, fatty acids, and glycerol.
The liver plays a decisive role in the cholesterol metabolism. The
liver accounts for 90% of the overall endogenic cholesterol and its
esters; the liver is also impli-cated in the biliary secretion of cholesterol
and in the distribution of cholesterol among other organs, since the
liver is responsible for the synthesis of apoproteins' for r ~
lipoproteins, a-lipoproteins, and j3-lipoproteins which transport the
secreted cholesterol in the blood. In pan, cholesterol is decomposed
by intestinal micro-flora; however, its major pan is reduced to'
coprostanol and cholestanol which, together with a small amount of
nonconvened cholesterol, are excreted in the feces.
Cholesterol, mostly esterified, is utilized in the buildup of cell
biomembranes. BeSides, cholesterol is a precursor to biologically
important steroid compounds: bile acids (in liver), steroid hormones
(in adrenal cortex, male and female sexual glands, and placenta), and
vitamin D
3
, or cholecalciferol (in skin).
10.3 REGULATION OF LIPID
METABOLISM IN THE ORGh.NISM
The rate of lipid metabolism in the organism tissues is dependent
on the dietary supply of lipids and on tt.e neurohormonal regulation.
An excessive intake ofbigb-calory food (.;arbohydrates and triglycerides)
impedes the consumption of endogenic triglyceride reserves stored in
the fat tissues. Moreover, carbohydrates provide a very favourable
basis for the neogenesis of various lipids; for this reason, a large
LIPID METABOLISM
I
dietary intake of carbohydrate-rich food exerts a significant influence -
on the production of b'iglycerides and cholesterol in the organism.
Synthesis of endogenic cholesterol is also controlled by exogenous
cholesterol supplied in food: the more dietary cholesterol is digested,
the less endogenic cho-Iesterol is produced in the liver. Exogenous
cholesterol inlnbits the activity of hydroxymethylglutaryl-CoA reductase
and the cyclization of squalene to lanosterol.
The dietary ratio of various lipids plays an bnportant role in the
lipid metab-olism in the organism. The available amounts of polyene
fatty acids and phospho-lipids acting as solvents for fat-soluble vitamins
affect not only the absorption of the latter species, but also the solubility
mid stability of cholesterol in the organism fluids (blood plasma and
lymph) and biliary ducts. Vegetable oils with a high percentage of
phospholipids and polyene fatty acids impede an excessive accnmulation
of cholesterol and .its deposition in blood vessels and other tissues, and
facilitate the removal of cholesterol excess from the organism. These
processes are most markedly affected by com oil, safflower oil,
cottonseed oil, and sunflower oil. The consumption of unsaturated -
fatty acids contained in vegetable oils pro-duces a favourable effect on
the synthesis of endogenic phospholipids (for which these acids are
substrates); polyene fatty acids are aIs<; needed in the production of
materials, for example, prostaglandins. Unsaturated fatty acids
ac as uncouplers for the oxidative phosphorylation and thus accelerate
o dation processes in the mitochondria and control thereby an excessive
tri yceride deposition in the tissues.
The lipotropic factors exercise a marked effect on the biosynthesis
and triglycerides. As has been mentioned above,
the facilitate the phospho-lipid synthesis. The dietary deficiency of
Ii tropic factors favours the triglyceride production in the organism.
I Starvation elicits mobilization of triglycerides from the adipose
tissUe and inhibits the endogenic cholesterol synthesis owing to the
low activity of hydroxy-methylglutaryl-CoA reductase. The latter
process provides the possibility. for the- active production of ketone
bodies in the liver.
I
I The neurohormonal control of lipid metabolism chiefly affects the
and synthesis of triglycerides in the fat tissue. The
lipolysis in tissues is dependent upon the activity of triglyceride lipase.
All the regulators that favour the conversion of the inactive
(non phosphorylated) lipase to the active (phosphorylated) one, stimulate
the ,lipolysis and the release of fatty acids into the blood. Adrenalin
and noradrenalin (secreted in the sympathetic nerve endings), hormones
(glucagon, adrenalin, thyroxin, triiodothyronine, somatotropin, 3-
lipotropin, corticotropin, etc.), tissue hormones, including biogenic
amines (histamine, serotonin, etc.) act as stimulators for this process.
Insulin, on the contrary, inhibits the adenylate cyclase activity,
preventing thereby the formation of active lipase in the fat tissue,
i.e. retards the lipolysis. In addition, insulin favours the neogenesis
of triacyl-glycerides from carbohydrates, which, on the whole, provides
for lipid deposition in the fat tissues as well as for the cholesterol
production in other tissues. The thyroid hormones thyroxin and
triiodothyronine assist in the oxidation of the cholesterol side chain
and in the biliary excretion of cholesterol in the intestine.
10.4 PATHOLOGY OF LIPID METABOLISM
Most commonly, the lipid metabolism pathology is manifest as
hyperlipenaia (elevated concentration of lipids in blood) and tissue
lipidoses (excessive lipid de-position in tissues). Normally, the lipid
contents in the blood plasma are: total lipids, 4-8 gllitre; triglycerides,
0.5-2.1 mmolllitre; total phospholipids, 2.0-3.5 mmoillitre; total
cholesterol, 4.0-10.0 mmolllitre (esterified cholesterol accounts for
2/3 of total cholesterol).
Hyperlipemia may manifest itself by an increased concentration
of lipids, or certain groups thereof. For example, hypercholesterolemia
and hypertriglyceri-demia may be mentioned in this connection. Since
practically all the blood plasma lipids make part of lipoproteins,
hyperlipemias may be reduced to one of the hyper-lipoproteinemia
forms which differ in the varied ratios of plasma lipoproteins of
different groups.
Distinguished are exogenous, or alimentary, hyperlipemias which
are actually associated with a normally increased blood lipid
concentration after the intake of a food high in fat. and endogenic
hyperlipemias caused by impaired lipid metab-olism. The endogenic
hyperlipemia may be due to a primary hereditary defect in apoproteins
or in a lipid metabolism enzyme. However, of more frequent
occur-rence are hyperlipemias attributable to secondary causes, for
example, to regulatory disturbances of the lipid metabolism or to unfav-
ourable environmental factors. Five types of primary hyperlipopro-
teinemias are distinguished.
Hyperlipoproteinemia, Type I, is characterized by the enhanced
content of chylo-microns in the blood plasma; simultaneously, the
percentage of a- and P-Iipopro-teins may be lowered. The triglyceride
LIPID MET ABOLISM
291
content is 8-10 times above the norm, while cholesterol does not
exceed the normal level. Presumably, Type I is associated with a
defective lipoprotein lipase that destroys chylomicrons.
Hyperlipoproteinemia, Type II, is characterized by an increased
I-lipoprotein content in the blood plasma and, respectively, by a 1.5-
2-fold higher, against the norm, cholesterol concentration. The familial
form of hyperlipoproteinemia, Type Ila, is also known, wHich
manifests itself in the occurrence of a defective apoprotein for 13-
lipoproteins, and in a slower breakdown of these materials in the
tissues.
!Hyperlipoproteinemia, Type III, is a rare hereditary disease (also
familial dysbetalipoproteinemia) manifested by the occurrence
of an uncommon P-lipo-protein form. Cholesterol and triglyceride
contents in the patients may occasion-ally be 2-5 times superior to
the norm.
Hyperlipoproteinemia, Type IV, is characterized by increased
contents of pre-J3-lipoproteins and triglycerides (2-5 fold) in the blood
plasma. Its incidence rate is higher in aged patients. Hereditary forms
of this disease (called also familial hyperprebetalipoproteinemia) have
been described.
-Hyperlipoproteinemia, Type V. This pathology is manifested by
increased con-tents of chylomicrons, pre-l3-lipoproteins, triglycerides,
and cholesterol in the patients' blood plasma.
Secondary hyperlipoproteinemias, which arise from a disordered
lipid tissue metabolism or its impaired control, are observed in diabetes
thyroid gland hypofunction, alcoholism, etc.
Lipidoses. Hyperlipoproteinemias may lead to tissue
Lipidoses can also arise from hereditary defects of the
involved in the synthesis and breakdown of lipids in the
tissues. We now discuss certain instances of tissue lipidoses.
Atherosclerosis is a wide-spread pathology, manifested chiefly by
the deposition of cholesterol in arterial walls, which results in the
formation of lipid plaques (atheromas). Lipid plaques are specific
foreign bodies around which the connective tissue develops abnormally
(this process is called sclerosis). This leads to the cal-cification of
the impaired site of a blood vessel. The blood vessels become inelastic
and compact, the blood supply through the vessels is impeded, and
thj' plaques may develop into thrombi. -
Atherosclerosis results from hyperlipoproteinemia. All of the
lippproteins, ex-cepting chylomicrons, are capable of penetrating the
vessel wall. However, are rich in proteins and
phospholipids, are liable to an easy break-down within the vessel wall,
or are apt to leave it because of their small size. Jl-Lipoproteins and,
pardy, pre-Jl-lipoproteins containing much cholesterol exhibit
atherogenic properties. Elevated concentrations of lipids of these groups
and an increased vessel wall permeability are conducive to deposition
of atherogenic lipoproteins within the walls, with the subsequent
development of atherosclerosis.
Fatty infiltration of the liver. In this pathology, the triglyceride
concentration in the liver is 10-foid superior to the norm. The
accumulation of fat in the cyto-plasm of hepatic cells leads. to an
impaired liver function. The causes of this pathol-ogy are numerous;
one of these may be a deficiency in lipotropic factors and the
associated therewith synthesis of excess triglycerides.
Ketosis is a pathologic state produced by an excess of ketone
bodies in the organism. However, ketosis may be regarded as a lipid
metabolism pathology with a certain reserve, since excessive
biosynthesis of ketone bodies in the liver is sequent upon an intensive
hepatic oxidation not only of fatty acids, but also of keto-genic amino
acids. The breakdown of the carbon frameworks of these amino acids
leads to the formation of acetyl-CoA and acetoacetyl-CoA, which are
used in
ketogenesis. The ketosis is accompanied by ketonemia and
ketonuria, which is manifested by the increased concentration of ketone
bodies in blood and their ex-cretion in the urine. In an aggravated
form of ketosis, the ketone body concentra-tion in blood may be as
high as 10-20 mmolllitre. The ketone bodies are normally present in
the daily urine in trace amounts, while in pathology, 1 to 10 g (or
even more) of ketone bodies per day is excreted in the urine. Most
commonly, ketonemia and ketonuria are observed in diabetes mellitus
(the manifest ketosis symptoms are dependent on the extent of diabetes
mellitus), as well as in prolonged starvation or in "steroid" diabetes.
10.5 APPLICATIONS OF LIPIDS AND THEIR
COMPONENTS IN PHARMACOTHERAPY
Fat-emulgated preparations for parenteral administration have been
elaborated for clinical applications. Since these are administered to
the patients intravenously, the size of fat emulsion particles should
not exceed the size of the largest naturally occurring lipoproteins-
chylomicrons, i.e. about I /Lm. Fat emulsions on the basis of com
oil (preparation lipomaize), cottonseed oil (lipofundin, lipomol),
~ P I METABOUSM 293
soybean oil (intralipid) have been proposed. These preparations are
composed of lipids (10 to 20%), emulsifying agents (phosphatides and
other materials) and, occasion-ally, glycerol. They are prescribed to
asthenic patients for increasing energy re-sourl;es of the organism. In
addition, lipotropic preparations (methionine, choline, and inositol),
which make part of natural phospholipids, are used in the prophy-Iaxis
of fatty infiltration of the . liver .

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