1.

Biological Molecules

Carbon is central to the living world.

• Carbon is central element to life because most biological molecule are built on a
carbon framework.
• The complexity of living things is facilitated by carbon's linkage capacity.
• Carbon has the great bonding capacity due to its structure.
• Carbon's outer shell has only four of the eight electrons necessary for maximum
stability in most elements.
• Carbon atoms are thus able to form stable, covalent bonds with a wide variety of
atoms including other carbon atoms.

Functional Groups:
• Groups of atoms known as functional groups can confer special properties on
carbon-based molecules.
• Carbon is a central element to life because most biological molecules are built on
carbon framework.
• For example, the addition of an -OH group to a hydrocarbon molecule always
results in the formation of an alcohol.
Sr No. Groups Structural Found in
Formula

1. Carboxyl (-COOH) Fatty acids,

Amino acids.
2. Hydroxyl (-OH) Alcohols,
Carbohydrates
3. Amino (-NH2) Amino acids

4. Phosphate (-PO4) DNA, ATP

5. Carbonyl (-CO) Aldehyde,

Ketone

1
• Functional Groups often impart on electrical charges or polarity onto molecules
thus affecting their bonding capacity.

Dehydration synthesis: Formation of large molecules by the removal of water.

- monomers are joined to form polymers (also known as condensation
polymerisation).

Hydrolysis: Breakdown of large molecules by the addition of water - polymers

are broken down to monomers.

Dehydration synthesis Hydrolysis

Polymers and Macromolecules:

• Macromolecules are giant molecules.
• 3 types of macromolecules : Polysaccharides, proteins and nucleic acid.
• Macromolecules - many repeating sub-units →Polymers.
• Polymerisation → condensation of monomers.
• Polysaccharides – monosaccharides.
• Proteins - Amino acids.
• Nucleic acid – nucleotides.
• Natural polymers - cellulose and rubber.

2
A macromolecule which is not a polymer is fat.

Carbohydrates :
• Carbon, hydrogen and oxygen
General formula: Cx(H2O)y

3 main group: (CH2O)n

- Monosaccharides
- Disaccharides
- Polysaccharides

Monosaccharides:
• Sugar
• They are 3, 4, 5 and 6 carbon atom
• Called trios, tetrose, pentose, hexose, respectively
• They undergo isomerism.
• Isomers have different structural formula but same molecular formula.
• Various types of isomers
- Aldose and ketose
- Open chain and ring form
- α - isomers and β - isomers
• Either Aldehyde group (-CHO) or Ketone group (-CO)
Eg. Aldose: Glyceraldehyde, ribose, glucose
Ketose: Dihydroxyacetate, ribulose, fructose

3
 Fructose Glucose Galactose

Open chain and ring form:

• Pentose or hexose when dissolve in water: long chain bond rounds up, bringing the
carboxyl group close to reduce one hydroxyl group.
• Hexose sugar - C-1 combines with oxygen atom on carbon atom 5; six-membered
ring - pyranose ring.

α and β isomers:
• Hydroxyl group - carbon atom in glucose can be below or above the plane of ring.
• Alpha - isomer: hydroxyl group below the plane of ring.
• Beta - isomer: hydroxyl group above the plane of ring.
• β and α isomers of glucose - Interconvertable in aqueous solution.

Role of monosaccharaides in living organism:

Two major function
1. Cellular respiration- energy (ATP)
2. Building blocks for larger molecules.

4
Carbohydrates
Disaccharides:

• 2 monosaccharaides linked together by dehydration synthesis.

eg. Glucose + Galactose Lactose + water
Glucose + Glucose Maltose + Water
Glucose + Fructose Sucrose + Water
• Reaction – 2 monosaccharaides condensation reaction.
• Condensation reaction- removing water- bond formed- 2 monosaccharaides –
glycosidic bond.
• Monosaccharide units- residue
• Glycosidic bond in a disaccharides broken by hydrolysis reaction.

Source of Disaccharides:
• Lactose is found in milk.
• Maltose- germinating seeds
• Sucrose- Transported in plant,
Abundantly found in sugarcane.

5
Disaccharides and the glycosidic bond:
Two monosaccharides joined together by condensation reaction

Properties of Disaccharides:
• They are sweet.
• Crystalline in nature.
• Readily soluble in water.

Polysaccharides:
• Long chain of sugar.
• Used for energy storage.
• Plants used starch; animal used glycogen.
• Used for structural support.
• Plants used Cellulose; animal used chitin.
• Polymers of monosaccharides.
• Base function- starch and glycogen.
• Structural polysaccharides- cellulose.

Starch:
• Mixture of two substance- Amylose and Amylopectin.
• Both contain α-glucose.
• Amylose component– Unbranched.
• Glucose residue- Linked by α (1, 4) glycosidic bond.
• Amylose chain coiled into helix-with 6 glucose molecules.
• Hydroxyl group of glucose –Projected interior.
• No crosslink- amylose chain.
• Amylopectin- branched.
• Glucose residue- linked by α (1, 4) glycosidic bond.

6
 • Branch point- formed by α (1, 4) glycosidic bond.
• Amylopectin- coiled into a helix.
• This is no cross linking between amylopectin chain
• Amylopectin chain – red violet colour- iodine.

α-(1, 6) glycosidic bond (Amylopectin)

• Starch is present in all plant.

• Starch is hydrolysed by chemical method.
• Incubating with dilute acid at 100°C.

Enzyme Method:
• Incubating with enzyme at room temperature. eg- amylose.

7
 • Starch- maltose

Glycogen:
• Glycogen is present in animals.
• Glycogen is like that of animals.
• Chain 14 linkage α glucose molecules and 1,6 linkage forming branches.
• Glycogen molecules bind together- granules.
• Stored in liver, Nerve, Cells, Skeletal muscles of vertebrate of animals and fungi.

Starch and glycogen- good storage molecules because of following reasons:

1. Starch and Glycogen- Insoluble in water.

2. Stored in large quantities without having great effect on water potential of cells.
3. Indivisible chain of starch of glycogen- fold into a compact shape, large amount of
starch or glycogen- stored within a cells.
4. Easily hydrolysed to monosaccharides (no cross- linked).

Cellulose:
• Abundant organic molecule.
• Plant cell wall.

8
 • Slow rate of breakdown.
• Cellulose – Polymers – β glucose.
• Glycosidic bond with C-4.
• Here, -OH Below-ring-glucose molecule must be upside down that is rotated 180°.
• Re-emit strong molecule.
• Because – Hydrogen atom of OH group attracted oxygen atom some cellulose
molecules.
• Hydrogen bond – weak alone, strong when several formed together.
• 70 cellulose molecules tightly cross linked from bundle (micro fibrils).
• Micro fibrils combines to form macro fibrils.
• Fibers - run – different directions.
• Cellulose 20–40 % average cell wall.
• Cellulose fibers – very high tensile strength – almost equals to steel.
• Plant Cell – can withstand greater pressure.
• Determine the shape of cell, gives strength.
• Cellulose – freely permeable – water and solutes.

Lipids:
• Made up of C, H, and O.

9
• Lipids are insoluble in water.
• Soluble in organic solvents such as – Ethanol, Ethene and Trichloroethane
(Chloroform).
• Lipids are in solid state at 20°C - fats.
• Lipids are in liquid state when it is above 20°C – oil.

Classification of lipids:
→ Simple lipid
→ Compound lipid
→ Steroid and Sterols

Simple Lipids:
• Formed by joining fatty acid to alcohol (Glycerol).
• Bond formed by ester linkage.
• Two types of simple lipids
1) Fats
2) Waxes
• Fats are formed by joining fatty acid and glycerol.
• Waxes – forming fatty acids – high molecular weight alcohol.

Compound lipids:
Lipid + Non lipids

10
Structure of lipids:
• Fatty acid + Glycerol.
• Fatty acid consists of hydrocarbon chain and carboxylic group.
• Fatty acids are two types.
1) Saturated fatty acids – RCOOH (R- Hydrocarbon chain).

Steroid and Sterol

Palmatic acid:

2) Unsaturated fatty acid:

• Hydrocarbon chain – C=C
• Oleic acid – 18 carbon atom 4 contains one C=C.

11
Glycerol:

Glycerol / Propane 1,2,3-triol.

Fats:

• Fats and Glycerol are the subunits of fats.

• 3 fatty acids + 1 Glycerol = Triglycerides.

• Hydroxyl group has glycerol and carbonyl of fatty acid.

• Water is removed.
• Bond is formed – ester bond ( -COO-)
• Reaction is condensation or dehydration.
• Fatty acid + Glycerol = Monoglyceride + Water (H2O).
• 2 Fatty acid + Glycerol = Diglycerides + Water (2H2O).
• 3 Fatty acid + Glycerol = Triglycerides + Water (3H2O).

Function of lipids:
• Triglyceride – storage form of energy.

12
 • Found around delicate organs eg. Kidney.
• Suburaneous fat – Skin.
• Triglyceride- large molecule and unchanged insoluble in water stored in large
quantities.
• Triglyceride- great no carbon atom/ gram /one mole 38 kj.
• Triglyceride -water-oxidised than glucose.
• Triglyceride-good thermal insulator.
• Long term energy store.
• Important to hibernating animal.
• Less dense than water-aquatic organism can broyaney eg. whale.

Proteins:
• Made up of C, H, O, N
• S is also present build up amino acid

• Various amino acid differ R group

Neutral amino acid:

• Have one carboxyl group and one amine group eg. Alanine.

Basic amino acid:

• More than one amine group.
• R group contains amine group.
Eg. Lycine

13
Acidic Amino Acid:
• More carboxyl group than amino group.
• R-group contain carboxyl group.

-SH: R group

14
-OH: R Group

Hydrophobic: R Group

Properties of amino acids

• Amino acids are soluble in water, but insoluble in organic solvent.
• Dissolve in water to form ions.
• The carboxyl group can lose a hydrogen ion to become negative charged.
• The amino group can accept hydrogen ion to become positive charge.
• Neutral aqueous solution contains amino acids which are amphoteric (act as buffer).

Formation of peptide bond:

• Amino acid join together to form polypeptides.
• It involves a combination of amino acids.
• Condensation reaction takes place.
15
 • Water molecule is removed to form peptide bond.
• Reaction occurs between -COOH & NH2.

Linkage of Polypeptide:
• Polypeptide has specific three dimension shape called conformation.
• There are four different bond between amino acide.
1) Di sulphate
2) Ionic bond (electro covalent Bond)
3) Hydrogen bond
4) Hydrophobic Interaction

Disulphate Bond:
• When two cystine residues are added they oxidised and a disulphate bond is
formed.

16
Ionic Bond:
• Amino acids are zwitter ions.
• Acidic amino acid + basic amino acids give an ionic bond.
• Availability of NH3 & COOH group in acidic & basic amino acids.

Hydrogen Bond:
• Formed between hydrogen atoms and the element N OR O to which it is attracted.
• H atom has a small positive charge.
• N and O are higher electronegative than hydrogen atom.
• Oppositely charged atoms, i.e. between H and N OR with O attracted hydrogen bond.

Hydrophobic Interaction:
• Interaction between hydrophobic R groups of amino acid residue.
• Polypeptide ford- Shields hydrophobic R group from aqueous environment.

Structure of a protein:

4 leaves of structure:
• Primary structure
• Secondary structure

17
 • Tertiary structure
• Quaternary structure

Primary Structure:
• Sequences of amino acid- Polypeptide chain.
• Substitution of a simple amino acid causes a major attraction of function in a
protein. eg. Sickle cell anaemia.

Secondary Structure:
• Alpha helix, beta pleated sheet, triple helix.
• Secondary structure is stabilised by hydrogen bond.

Alpha Helix:
• Form a spiral spring.
• 3-6 amino acid residues in one complex form.
• Alpha helix are stabilised by hydrogen bond.
• Protein keratin are component of hair (it has alpha helix structure).

Beta Pleated Sheet:

• Forms a sheet
• Number of adjacent polypeptide chain.
• If it turns in same direction, it is called a parallel beta pleated sheet.
• If it turns in opposite direction, it is called anAnti-parallel beta pleated sheet.
• Sheet is stabilised by hydrogen bond between -CO group of one polypeptide and -H
group of adjacent polypeptide.

Triple Helix:

• Saturated unit of collagen which is tropocollagen.

• Collagen is a compound in the skin, bones, blood, vessels and teeth..

18
 • Tropocollagens are triple helix structure.
• It consists of 3 polypeptide chains.
• Each chain contains 1000 amino acid.
• Their helical strands wind around each other with a hydrogen bond.

Tertiary Structure:
• Refers to relationship of amino acid residues that are far apart and are linear
sequences.
• Due to the bending and folding of α- helix structure into a globular shape.
• Four types of bond are present/ formed.
1. Disulphate Bond.
2. Ionic Bond.
3. Hydrophobic Interaction.
4. Hydrogen Bond.

Quaternary Structure:
• Protein consisting of more than one polypeptide chain which display on the addition
structure.
• Number of polypeptide chains is associated with non-protein molecules.
• Subunits are held together by hydrogen bond, hydrophobic interaction, ionic bond.
eg. Haemoglobia.

Classification of protein according to structure:

Fibrous Protein:
• Don’t have tertiary structure
• Secondary structure is the most important.
• Polypeptides have cross linked intervals to from a long fibre or sheet.
• Insoluble in water due to large molecules of hydrophobic R groups of amino acid
residues are exterior of their molecules.
• Length of polypeptide chain may vary as 2 simples are of the same fibrous protein.
19
 • Amino acids sequence of fibrous protein is remarkably the same.
• Fibrous proteins perform structural function.
• Amino acids sequence may vary slightly between two samples of the same fibrous
protein.

Globular protein:
• Have a tertiary structure
• Quaternary structure may or may not be present
• Peptide chains are tightly folded to form a spherical shape
• They dissolve in water to form colloidal solution. This is due to the hydrophobic R
group of amino acids residues, which are placed outwards.
• Amino acid sequence is highly specific, and they never vary between two samples of
the same globular protein.
• Length of polypeptide is always identical in 2 samples of the same globular proteins.
• Globular proteins rarely exhibit regularities of amino acid.
• They perform metabolic activities. Eg. Enzymes and Haemoglobin.

Classification of proteins according to components:

It has two groups.

1. Simple proteins: It will have only amino acids and they form the structure of simple
proteins. Ex: Albumin.

2. Conjugated protein: Amino acids are combined with non-protein compounds called
the prosthetic group. Eg: Haemoglobin.

Relating structure of haemoglobin and collagen to their functions.

• Haemoglobin eg: Globular protein.

• It is required to transport oxygen.
• It is a quaternary structure.
• It consists of 2 alpha chain sub unit and beta sub units.
• I t is good for oxygen transportation for two reasons
1) Globular shape, packed-RBC for transport of oxygen
2) Four sub nits are capable to binding with oxygen.

Haemoglobin is Allosteric protein:

20
• It means binding of one oxygen to one haem group, facilitates or fastens the binding
of oxygen of another haem group in same haemoglobin molecule.
• Each sub unit consists of protein and non-protein component.
• Structure of globulin is globular except for deep hydrophobic cleft which is a haem
binding site.
• Haem group has porphyrin and iron ring.
• Iron placed in the centre of the polar porphyrin ring.
• Haem is oriented with Fe2+ one face with the complex amino acid leaving another face
accessible for oxygen binding.
• Oxygen combines reversibly with Fe2+.
• Haemoglobin has both hydrophilic and hydrophobic residues.
• Hydrophobic amino acids are buried in the interior of the globular structure while
hydrophilic amino acids found outside.
• Haemoglobin is soluble in aqueous medium and hence it is a good transport protein
for O2 in blood.
• Collagen is a fibrous protein.
• Skin, bones, blood vessels and teeth have some components of collagen.
• Tropocollagen has three polypeptide chains with about 1000 amino acids.
• It has three helical stands which wind around each other. Its called as triple helix.
• Collagen is rich in glycine and proline (Amino acids). Insoluble in water.
• They are large molecules in size-Tropocollagen.
• Consist largely of glycerine and proline residues which are hydrophobic in nature.
• Collagen has great tensile strength.
• Tropocollagen has three polypeptide chains and are cross linked to form of collagen
fibre.

Functions of proteins:
• Proteins functions as a buffer, they have both acidic and basic properties.
• Proteins act as a buffer when stabilizing PH of blood plasma.
• Globular proteins have a composition of crystal in the cell.
• Hormones have protein in nature.
• Enzymes are proteins in nature.
• Proteins serve in transport function.
• Proteins serve in shortage function. Eg. Myoglobin, stores O2 in muscle.
• Proteins also participated in body defence mechanism.
• Structural proteins eg: Collagen, Keratin.
• Source of energy during saturation.

Water:
• All living cells contains 50% of water by mass.

21
• Water consists of an Oxygen atom covalently bonded with two hydrogen atoms.
• Water is polar.
• Oxygen atom has high electronegativity. Due to this oxygen pulls the bonded
electrons of hydrogen. This arises partial negative charge on oxygen atom and partial
positive charge on hydrogen atom.
• Attractive forces between small negative charge on the oxygen atom and the small
positive charge on hydrogen atom constitutes H-bond.
• Properties of water and its importance to living organism.

Water as a Solvent:
• Many solutes are dissolved in water (Universal solvent) or biological fluids.
• Hydrophilic compounds are soluble.
• Solutes dissociates in to an ions and molecules, that is why its soluble.
• This property is important for transportation and secretion.

Thermal properties:
• Has high heat capacity.
• Lot of heat is required to increase water temperature by 1 oC.
• Water can act as a thermal buffer and resist large change in temperature.
• It will have s a high heat of vaporization.
• Lot of heat can be lost for negligible loss of water.
• It is important for regulating body temperature.
• High heat of fusion.
• Lot of heat must be lost before water freezes.

Density and freezing properties:

• Dense at 4 oC.
• Density starts to decrease from 4 oC- 0 oC.
• Water in ponds freezes from the surface to the bottom to prevent complete
solidification.

Cohesion and Surface tension:

• Cohesion is a force of attraction between same as molecules.
• Cohesion in water molecule has a hydrogen bond between them.
• Water molecules pulled inwardly towards each other, forming spherical drops which
forces skin like layer and is called surface tension.
• Surface tensions allows insects to walk of water surface.
• Cohesion of water molecules leads to ascent of sap.
• Cohesion forces leads water molecule to support the structure of organism.
• Hydrostatic skeleton of earthworm.

22
• Turgidity for example herbaceous plant.
• Eye shape is maintained by aqueous humour and virtuous humour.
• Protection of amniotic fluid to foetus to prevent it from mechanical shock.
• Adhesive force is the attraction between un like molecules.
• Water molecules are attracted by the cellulose of Xylem vessel in plants.
• Water is transparent to make visible light penetrable for photosynthetic plants in an
aquatic ecosystem.
• Low viscosity property makes useful lubricants.
• Mucus secreted externally to aid the movement of animals.
• Oesophagus helps in the movement of food.
• Movement of sperm in oviduct is done through semen.
• Synovial fluid lubricates the movement in vertebral joints.
• Pericardial fluid lubricates the movement of the heart.
• Metabolic role of water
Hydrolysis.
Biochemical reactions.
• Water is needed for diffusion in materials across the surface.
• Water is a substrate of photosynthesis.

Role of Inorganic Ions:

Calcium:
• Calcium phosphate provides hard, strong. Insoluble material bones and teeth in
mammals.
• Required for blood clotting in animals.
• Calcium pectate in plant cells forms a material in which cellulose fibres lie.
• It involved in the transmission of action potential from one neuron to another and in
muscle contraction.

Sodium:
Constantly pumped out of the cell by active transport in exchange for potassium ions,
providing a positive charge out side the cell which is important in the transmission of
nerve impulse.

Potassium: constantly pumped in to the cell by the active transport in exchange for
sodium ions, important in the transmission of nerve impulse.

Magnesium: Forms part of the chlorophyll molecule in plant, important for

absorption of light energy to drive the reaction of photo synthesis.

23
 Chloride: Moved out of the cells lining the lungs and digestive system provide a low
water potential outside the cell, causing water to follow. So that mucus is not too thick
and stiff (failure of this cause of cystic fibrosis).

Nitrate: Used in the production of Amino acids (and therefore proteins) and also
carbohydrate synthesis (In plants by photosynthesis).

Phosphate:
• Production of nucleic acids in cell.
• Production of ATP.
• In production of phospholipids, essentials in cell membrane.

2. Cell

Cell structure:
• Cells are the basic units of living organisms.
• Microscope in cell studies.
• Detailed structure of typical animal and plant cells, as seen under the electronic micro
scope.
• Outline functions organelles is plant and animal cells.
• Characteristics of Prokaryotic and Eukaryotic cells.

Microscopy:
• Microscope is very useful in study of living cells.
• Most of living organism are too small to be seen in any detail with the human eye,
cells and their organelles can only be seen with the aid of a microscope.
• Cells were first seen in 1665 by Robert Hooke (Who named them after Monk’s cells
in a monastery), and were studied in more detail by leeuwehoek using primitive
microscope.

Units of measurement:
meter m 1m
millimetre mm 10-3 m
micrometre μ 10-6 m
nanometre nm 10-9 m
picometre pm 10-12 m
Angstrom Ao 10-10 m (obsolete)

24
Magnification and resolution:
• By using high power lenses, one can magnify the cell structure, but it does not mean
that detailed structure can be seen.
• The magnification depends on the resolving power of a microscope, which is the
smallest separation at which two separate objects can be distinguished (or
resolved).
• The resolving power of a microscope depends on the wave length of light (400-600
nm for visible light).
• To improve the resolving power of a shorter wave length microscopes with blue
filters can be used.
• Magnification=objective lens x Eye piece lens.

Resolution

• Resolution is the ability to distinguish between two points of an image.

• The resolution of an image is limited by the wavelength of radiation used to view the
sample.
• The wavelength of light is much larger than the wavelength of electrons, so the
resolution of the light microscope is much lower.
• Using a microscope with a more powerful magnification will not increase this
resolution any further, it will increase the size of the image, but objects closer than
200nm will still only be seen as one point.

Light microscope Electron microscope

Low cost
Easy to operate
Small and portable
Simple and easy sample preparation
Material rarely distorted by
preparation.
Natural colour of the cell is maintained.
Magnifies objects up to 2000 times.
Specimens can be living or Dead
Stains are often needed to make the
cells visible.
High cost
It involves production of electron
beam, so it becomes expensive.
Large and requires special room.
Lengthy and complex sample
preparation
Preparation distorts material.
The cells can be seen only in white and
black colour.
Magnifies objects up to 500,000 times.
Specimens are dead, as they must be
fixed in plastic and viewed in a vacuum.
The electron beam can damage
specimens and they must be stained
with an electron-dense chemical

25
 (usually heavy metals like Osmium,
lead and Gold).

• A molecule = 1 nm
• Cell membrane thickness = 7.5 nm
• Virus = 100 nm (20-200 nm)
• Bacteria < 10 μm
• Organelles < 100 μm
• Eukaryotic cells < 100 μm

Surface area to Volume ratio:

• The rate of metabolism of a cell is a function of its mass/volume.
• The rate of material exchange in and out of a cell is a function of its surface
area.
• As the cell grows, volume increase faster than surface area (leading to a
decreased surface area/volume ratio).
• It is metabolic ratio is greater than the rate of exchange of vital materials and
wastes, the cell will eventually die.
• Here the cell must consequently divide in order to restore a viable SA: vol
ratio and survive.
• Cells and tissues specialised for gas or material exchange (ex. alvroli) will
increase this surface area to optimise the transfer of materials.

The Cell

• Cell Theory- Schleiden and Schwann- 1938-39

• The cell is the basic unit of life.
• All organisms are made up of cells
• All the cells arise from pre-existing cells by division (R Vischow’s 1955)
• Basis of cellular organisation-organisms are divided into 2 categories.
1. Prokaryote- “before carrier bag” i.e. without a nucleus
2. Eukaryote- “good carrier bag” i.e. with a nucleus

Prokaryotic Cell:

• Cell Wall: A rigid outer layer made of polypeptide glycon that maintains shape
and protects the cell from damage or bursting if internal pressure is high.
• Cell Membrane: Selectively permeable-barrier that controls the entry and exit
of substances.

26
• Cytoplasm: Fluid component which contains the enzymes needed for all
metabolic reactions.
• Nucleoid: Region of the cytoplasm which contains the genotype (the
prokaryotic DNA)
• Plasmid: Additional DNA molecule that can exist and replicate independently
of the gonophore-it can be transmitted between bacterial species.
• Ribosomes: Complexes of RNA and protein that are responsible for
polypeptide synthesis (prokaryotic ribosomes are similar than eukaryotes-
70s).
• Slime Capsules: A thick polysaccharide layer used for protection against
desiccation (dry out).
• Flagella (Singular flagellum): Long slender projection containing a motor
protein which spins the flagella like a propeller, enabling movement.
• Pili (singular pillus): Hair-like extensions found on bacteria which can serve
two roles.
• Attachment Pilli: shorter in length, they allow bacteria to adhere to one
another or to available surface.
• Sex pili: longer in length, they allow for the exchange of genetic material
(plasmids) via a process called bacterial conjugation.

Prokaryotic cells Eukaryotic cells

• Small cells • Longer cells
• Always unicellular • Often multicellular
• No nucleus or any • Always have nucleus and
membrane Bound other membrane bound
organelles such as organelles.
mitochondria
• DNA is circular, without • DNA is linear and associated
proteins. with proteins to form
chromatin.
• Ribosomes are small (70 s) • Ribosomes are large (80 s)
• No cytoskeleton • Always has a cytoskeleton.
• Motility by rigid rotating • Motility by flexible having
flagellum made of flagellin. underlipodium, made of
tublin.
• Cell division is by binary • Cell division by mitosis or
fission. meiosis.
• Reproduction is always • Reproduction is always
asexual. asexual or sexual.

27
 • Huge variety of metabolic • Common metabolic
pathways. pathways.

Eukaryotic cell (consists to 3 parts)

1. Contains plasma membrane or cell surface membrane
2. Nucleus
3. Cytoplasm

Cytoplasm consists of
1.cytosol:
Aqueous solution of ions, organic compounds.
Eg. Sugars and amino acids, proteins.
2.organelles:
Includes ribosomes, endoplasmic reticulum,
Golgi apparatus, lysosomes,
3.cytoskeleton:
Consist of network of microtubules, intermediate filaments and
microfilaments.
4.Cell wall:
Found only in plant cell, absent in animal cell.
5.Nucleus:
• Longest organelle-eukaryotic cell.
• Observed -light microscope.
• Spherical or ovoid.
• Diameter- 10 μm to 20 μm

Nuclear envelope:
• Double membrane.
• Outer membrane -nuclear envelope -continuous with endoplasmic reticular.
• Pores are present on the nuclear envelope.
• Nuclear pores have channels through ten, through which molecules move
between the nucleus and the cytoplasm.

Nucleoplasm
• Semi fluid matrix fill the nucleus.

Chromosome.
28
• In cells, the DNA located in the nucleus, and is organised along with protein
(histone) into chromosomes.
• When the cell is not dividing the chromatin, which consists of the genetic
material, is too dispersed and entangled.
• During nuclear division, the chromatin becomes visible as the chromosome.
• The number of chromosomes vary according to the individual species.

The Nucleolus:
• Most of the visible structure within the non-dividing nucleus.
• One or more found in nucleus.
• Responsible for the synthesis of r-RNA.
• r-RNA is the component of ribosome.

Ribosome:
• Eukaryotic ribosome has a diameter of 20 nm.
• It has a sedimentation co-efficient of 80 s.
• It consists of 2 sub units
1) 40 s (Smaller sub unit) 2) 60 s (Larger subunit)
• Ribosome is composed of protein and r-RNA in roughly equivalent
quantities.
• Ribosome is allotted to the membranous network of the endoplasmic
reticulum.
• The ribosomes are suspended in the cystol.
• Ribosomes occur in cluster of polyribosomes or polysomes.

Functions: site for the protein synthesis.

Endoplasmic Reticulum:
• It consists of a network of membranous tubules and sac-cisternal.
• Originated from the outer membrane of the nuclear envelope.
• There are two regions of the endoplasmic reticulum.
• They differ in structure and function
1) Rough endoplasmic reticulum- presence of Ribosomes on its surface.
2) Smooth endoplasmic reticulum- absence of Ribosomes on its surface.

Functions:
1. The rough endoplasmic reticulum is the site for protein synthesis and
transport.

29
2. Proteins are transported either through cisternal of packed in vesicles.
3. Smooth endoplasmic reticulum synthesises lipids including membrane
phospholipids and steroid hormones.

Galgi apparatus:
1. It was identified by the Italian scientist Camillo Golgi in 1898.
2. Its appeared as stack of flattened Sac.
3. Galgi bodies are formed from the rough endoplasmic reticulum.
4. Proteins are made and stored in the rough endoplasmic reticulum, and are
transported to the galgi bodies.
5. These proteins are modified and combined with carbohydrates and fats.
6. Modified proteins are carried in small vesicles and are secreted.
7. Sometimes the vesicles that are pinched off from the Golgi bodies becomes
lysosomes.

Main functions:
1. Assembling glycoprotein such as mucin by combining carbohydrates and
protein.
2. Transporting and storing lipid.
3. Formation of lysosomes.
4. Production of digestive enzymes.
5. Secretion of carbohydrates for the formation of plant cell wall insect cuticle.

Lysosome structure:
• Small and spherical shape.
• 0.2 μm to 0.5 μm.
• Surrounded by a single membrane and contains hydrolytic enzymes.
• Hydrolytic enzymes are acidic, and the enzymes have a low optimum pH.
• Lysosomes formed from the Golgi body.

Functions:
1. Autophagy is a process by which worn out organelles are engulfed digested
within the lysosomes.
2. Amoeba digests food materials in to food vacuoles.
3. Phagocytosis is the self-digestion of a cell by releasing the content of
lysosomes.

Mitochondria:

30
1. Cylindrical shape or rod shape.
2. Width range from 0.5 μm-1.5 μm and length from 3 μm-10 μm.
3. Bounded by double membrane.
4. Outer and inner membrane are separated by intermembrane space.
5. Inner membrane is extensively folded from partitions called cristae.
6. Cristae is projected in to the semi fluid matrix.
7. Circular DNA molecule and 70 s ribosomes are present.
8. Endosymbiont theory.

Functions of mitochondria:
1. Involved in cellular respiration.
2. Series of bio chemical reaction result in formation of ATP.
3. Often known as the power house of the cell.
4. More than a 1000 mitochondria are found in a metabolically active cell.

Chloroplast:
1. Large organelles.
2. Length: 5 μm-10 μm.
3. Bounded to the double membrane.
4. The inner membrane give rise to the membranes lamellae or thylakoids.
5. The interior of the chloroplast is filled with a gel like matrix called stroma.
6. Within the stroma, a group of thylakoids stalk together to form grana.
7. Stalks of grana are joined together by integral membrane.
8. Grana and intergranal lamellae are photosynthetic pigments.
9. The stoma of the chloroplast contains circular DNA Molecule and 70 s
ribosome.

Functions:
1. The chromoplast is the site of photosynthesis.
2. Light dependent reaction of photosynthesis occurs in the lamellae.
3. All light dependent reactions occur in stroma.

Vacuole:
1. Fluid filled sac- bounded by a single membrane.
2. Vacuoles in animal cells are smaller and more in number.
3. Young plant cells- numerous small vacuole, later merge to become large
central vacuole.

31
4. Vacuole is surrounded by a single membrane known as the tonoplast.
5. Plant vacuole contains a fluid called cell sop.
6. Cell sop is a solution consisting of mineral salts, sugar, organic acid, pigment
and waste products.

Functions:
1. Vacuoles is a protoctist (Amoeba) for osmoregulation.
2. Vacuoles is a protoctist (paramecium) for osmoregulation.
3. Plant Vacuole – keeps cell turgid and it is important for cell expansion and
cell growth.
4. Plant Vacuole contains pigments such as anthocyamins.
5. Dissolves substances and uses it as a food resource.
6. Act as a store for waste products. Eg. Tannins.

Microvilli: (Singular Microrillus)

1. Finger like extensions on the cell surface membrane.
2. Typically-certain epithelial cells.
3. Increases the surface area of the cell surface membrane.
4. Used by absorption in the gut.
5. Reabsorption: Proximal convoluted tubules.

Microtubules:
1. Components of cytoskeleton.
2. Long, Hollow, cylinder-found cytoplasm.
3. 25 nm in diameter.
4. Microfilament + Actin filament and intermediate filament makes
cytoskeleton.
5. Cytoskeleton is structural component of cells.
6. Determines the cell shape.
7. Microtubules are proteins called tubulin.
8. There are two forms of tubulin
1) Alpha tubulin
2) Beta tubulin
9. Alpha and Beta tubulins combined to form dimer form. (Double Molecule).
10. Dimers joints end to end to form long proto filaments.
11. 13 protofilaments line up alongside.
12. This forms a ring that is a hollow cylinder.
Centrioles:
1. Found in animal cells and lower plant cells.

32
 2. Absent in higher plant cells.
3. Exist in a pair, with rod-like structure.
4. Are positioned right angled to each other, next to the nucleus.
5. In higher animal cells the centrioles from the mitotic poles.
6. The centrioles function as the microtubule amazing centre, it is an
important event in major cellular process. That is cell division and flagella
formation.
7. The centriole pair duplicates within a cell and the two pairs migrates to the
opposite ends of the cell to organise the mitotic spindle. During the
transition phase of GI/sec of the interphase stage. The existing centrioles
disengage from each other.
8. Each centriole gives rise to a new centriole. The centriole that are newly
formed remain tightly attached to the parent centriole and it is elongates
during the S and G2 phases.
9. In the prophase stage, the centriole pair start moving towards the opposite
poles of the cell, forming the spindle simultaneously.
10. The migration and the positioning of the centrioles determines the
orientation of the spindle. It is also influences the chromosomes attachment
to the spindle fibres.

The spindle fibres are responsible for two segregation of chromosomes into the daughter
cells

At end of each cell cycle the cell has two centrioles-one mother centriole and other newly
formed centriole (known as the daughter centriole)

After segregation the centrioles determines the position of the nucleus and also influence
the cellular organs in the newly formed daughter cells

The centrides may produce flagua or cilia

The fibre of the tail of sperms also arises from the centrioles

The disfunctioning of centrosome is also responsible for the development of certain

cancers

Cell wall
Laid down during development of a cell

Starts as a thin layer of pectin which forms in the middle lamella

The cell wall is fully permeable

33
Made of cellulose fibres

Consists of gaps/spaces/holes/between fibres/other cell wall components)

Plasmodesmata
Allows transport of water, sucrose. aminoacids, organic substances and also helps the ATP
molecules to move from cell to cell without crossing the membrane or without going
through the protein ethanols

The movement of water through the plasmodesmata is known as symplast pathway

Plasmodesmate allows the sucrose molecules to move from companion cell of the phloem
to the sieve tube elements

Plasmodesmata also helps in communication or signalling between the cell

Plant cell Animal cell

Cell wall present Cell wall absent
Pits plasmodesmata present in cell wall Absent
Chloroplast present Absent
Mature cell normally have a single large Vacuoles if present are small and
central vacuole with cell sap scattered through the cell
Tonoplast present Absent
Starch grains present Glycogen granules present
Cilia and flagella absent in higher plant Cilia and flagella sometimes present
cell
Microvilli absent Microvilli may be present

All biological membrane similar in structure

Membranes are composed of lipids and proteins

Their proportion may vary depending on their function and location

1972-singles and Nicolson-fluid mosaic model

Membranes differs in thickness (5nm to 10nm)

Phospholipids forms a bilayer in the cell membrane

Phospholipids have a hydrophilic head and hydrophobic tail

Changes groups (head)-faces outward

Interact with aqueous environment on either side

34
Hydrocarbon chain-fatty acids-faces inward

Structure

2 types of proteins
Peripheral proteins/extrinsic protein

Integnal protein/intrinsic protein

Peripheral protein
Lies on the surface of the phospholipid layer

Protein is bound to the membrane near the hydrophilic layer

Integral protein
Penetrates partially or completely through the phospholipid layer

Protein maintains the position by the cell membrane by the hydrophobic and hydrophilic
bonds

Cell surface membranealso contains glycolipids and glycoproteins

Cell membrane contains cholesterol, which is found between phospholipid molecules.

Role of carbohydrate
Glycolipids and glycoproteins are include is cell to cell recognition

Cell to cell adhesion

Roll of lipids
Increases the fluidity of cell membrane

Role of protein
Facilitate diffusion,osmosis,active transport

35
Membrane protein function as enzymes

Membrane proteins function as a receptor site for hormones

Function of membrane
Plasmamembrane forms a binary between the contents of the cell and external
environment

Plasmamembrane helps the cell to maintain structural relationship with neighbouring cell

It maintain a constant internal environment

Regulate the passage of substances into and out of the cell

Enables the cells communicate with the external environment

Cell membrane enables separate compartment, which is formed within the cell with
specific biochemical pathways

Transport in and out of cell

Transport across cell surface membrane is vital for many reasons

It allows the entry of glucose for respiration which provides energy

Allows the excretion of waste products

Allows the secretion of extracellular enzymes

Maintain a stable ph and iconic contraction within a cell for enzyme activity

Ionic gradient which is essential for action potential

Passive transport
Does not regular or input of energy

2 types a passive transport:- 1 simple diffusion, 2 facilitate diffusion

Simple diffusion

36
Define net movement of particles from a region where they are at a relatively high
contraction to a region where they are at a lower concentration

Net movement of particles down a concentration gradient

Energy is provided by the kinetic energy of particles

Simple diffusion does not required the expenditure energy by a cell

Simple diffusion always proceeds to a concentration gradient and will continue until
eventually particles are uniformly system

Simple diffusing does not always involve a passage of substances through a membrane

Membrane present must be permeable in the molecule

The rate of diffusion

Concentration gradient
Greater the difference to concentration between two region, like greater the rate of
diffusion

Distance over which diffusion occurs

Shorter the distances over which diffusion occurs, the greater the rate of diffusion across it

Area over which diffusion occurs

Larger the area, the greater the rate of diffusion

Size and nature of the molecule

Smaller molecules diffuse faster than the larger molecule

Eg:oxygen molecule diffuse more quickly than carbon dioxide molecule

Molecules that are soluble in the substance of diffuse faster through

Eg: lipid soluble substance diffuse faster across plasma membrane than water soluble
substance

Facilitate diffusion
Charged particles and hydrophilic molecules don’t rapidly pass through the plasma
membrane because they are relatively insoluble in lipids

37
Plasma membrane contain proteins which assist such particles (hydrophilic and charged
particles)to diffuse in and out of the wall

Facilitate diffusion is similar to simple diffusion in two ways:

1. Net movement of particles is always down the concentration gradient

2. Energy from ATP is not required

Facilitate diffusion differs from simple diffusion by protein molecule being embedded in
the plasma membrane which is needed to carry the particles of a substance across the
membrane

Facilitate diffusion consists of transport protein which can transport substances in either
direction,depending on the concentration gradient

Transport proteins have same properties of enzyme

Transport proteins can transport only one substance across the plasma membrane

2 types of transport protein

1 channel protein

2 carrier protein

Channel protein
Configuration of the proteins molecule is such that it forms a water filled membrane

Unlike the interior of the phospholipid bilayer of the plasma membrane of the channel is
hydrophilic so water soluble substance pass through it

Channel proteins are partially consed with transporting ion into and out of the cell

Channel protein have gates which open and closed

Gated ion channel-open when they receive appropriate signals

Signals mechanical distance or a change in the voltage across the membrane

Only some channel proteins have gates

Carrier proteins
Carrier proteins exist in 2 forms;

38
1 binding site for the diffusion substances which are exposed(outside the wall)

2 some site exposed inside the cell

Changes it forms because it can be trigged by the binding and release of the transport
molecule

Osmosis
Is the net movement of water molecule from a region of higher water potential to a region
of lower water potential through a partially permeable membrane

Osmosis is considered to be a form of facilitated diffusion as water is flowing down

concentration gradient

Osmotic pressure
Osmotic pressure is measured in a osmometer

Osmometer contain officially partially permeable membrane

Bottom filter funnel is covered with a membrane which is permeable to water but not salt.

Filter funnel is filled with salt solution and immersed in pure water to the level of solution

Salt solution rises in the filter funnel while the level of water in the containers outside cells

Piston is used to exert pressure or identical salt solution

Osmotic pressure is pressure that must be applied to prevent entry of water into salt
solution

Opposite force is a tendency of salt solution to gain the water from pure water across the
membrane

Osmotic pressure is directly proportional to the concentration of solute particles in the

solution

For eg; 1% glucose solution would generate about twice the osmotic pressure as 1%
sucrose solution

Concept of solute potential

Solute potential is new name of osmotic potential

Solute potential is the potential of the solution to gain water

39
It is always regulate and has a magnitude equal to the osmotic potential of a solution

Solute potential is numerically equal to osmotic potential but is given negative sign

The more concentrated the greater its osmotic potential and the lower its solute potential

Water moves from a region of higher solute potential to a region of lower solute potential
down the solute potential gradient

Water flows from a region of higher water potential to a region of lower potential

Water potential of a solution is defined as the tendency for water molecule which is leave
the solution by osmosis

Measured in the unit of pressure:-SI unit – pascal (pal)

At standard temperature and pressure pure water has a negative water potential

Water molecule in pure water are free to move

Presence of solute particles lower the water potential

The more concentrated solution is the lower is its water potential and the smaller tendency
for water to leave the solution

In a concentrated solution most of the water molecules are associated with solute
molecules and are therefore not free to move

Higher the water potential implies and greater tendency for water to leave a solution

Osmosis in animal cells

RBC is placed in a solution where the water potential is the same as it is in the cell(isotonic
solution)

Cells neigher shrink not swell

Cells do not ecperience a not gain or loss of water

RBC is placed in a solution where the water potential is higher than that of the
cell(hypotonic solution)

Their not gain of water by endo osmosis which causes the cells to swell

Large amount of water entres the cell wall it cannot prevent the cell from bursting

There will be a net loss of water-exosmosis

40
This causes the cell to shrink

The RBC become generated

Osmosis in plant cell

Hypotonic solution
Effect the plant cell when the solution with a higher water potential than cells

Water molecules move through the plasma membrane and tonoplast into the vacuole by
endosmosis which results the cell to swell

Unlike animal cells, plant cells do not burst

Because the cell wall is stretched and develops a tension expansion of the cell

Water enters the plant cell and the cell contents starts to push against if form of inside of
the cell wall

In turn cell wall pushes back on the expanding cell this is known as pressure potential

Pressure potential opposes the continuous uptake of water into the cell by endosmosis

When pressure potential reaches the maximum than the cell cannot take in any more water

41
At this point the cell is described to the fully turgid

Hypertonic solution
Effect of placing the plant cell in a solution of lower water potential volume of the cell
decreases so thw water flows out of the vacuole by exosmosis

The plasma membrane starts to pull away from the cell wall

Space between the cell wall and plasma membrane will be filled by the outside solution

With of the plasma membrane from the cell wall is called plasmolysis

Plasmolysis is reached when the plasma membrane has completely with from the cell wall

Active transport
Defined as a movement of ions or molecules across a membrane from a region of lower
concentration to a region of higher concentration

Against the concentration gradient by the means of specific transport proteins

Expenditure of energy from the cell

Energy is usually from the hydrolysis of ATP

Active transport differs from facilitate diffusion in the following ways;

1 carrier proteins for active transport can bind with their molecule or ions on one side of
the membrane only

2 conformational changes of the carrier protein input of energy

Importance of active transport

It allows the cells to obtain nutrient even when the concentration outside the cell is lower
than that the concentration inside the cell

It enables the cells to get of waste products when the concentration outside the cell are
higher, even than that of the cell

Evidence that ATP is required for active transport is as follows;-

The factor influcing the rate of respiration will influence the rate of active transport

The chemical that interfears with the synthesis of ATP

42
The cells carry out active transport on large scale have an exceptionally large number of
mitochondria

In active transport the carrier proteins pump the molecules and ions from one side of the
membrane to other

These membranes proteins called protein pump

Examples of protein pump in active transport;

Na, K pump

Proton pump

Na, K pump
When compared with the surrounding of the animal cell it has a much higher concentration
of k ions and a much lower concentration of na ions

This is due to the Na,K pump that is in each complex

This process energy from the hydrolysis of ATP molecule

Na,K pump contributes for the resting potential of the nerve cell

Proton pump
Found in inner mitochondrial membrane

Pumps H+ from mitochondrial matrix to inner membrane spaces

Energy for the operation of proton pump is derived from the transport of the electrons
through the electrone carries with progressively a lower energy level

The uptake of glucose and Na from the lumen of proximal convoluting trouble of the active
transport

Bulk transport
Certain particles are either too large too pass through the small pores of the membrane or
they are too hydrophilic to diffuse through the plasma membrane

These materials are transported into or out of the cell by bulk transport

Bulk transport is defined as the transport of materials into and out of the cell by enclosing
within vesicles

43
Vacuoles are fluid filled membrane bound vesicles are small

In bulk transport materials are released from the cell,by exocytose or taken into the cell by
endocytosis

Both exocytosis and endocytosis are active processes that requires an expenditure of
energy

Exocytosis
Larger molecules released from the cell

Eg; secretion of hormones,enzymes,anti-bodies

This vesicles containing these materials are from the golgi apparatus and more towards the
surface of the cell

The vesicles fuse with the plasma membrane and open extrinsic and its contents leave the
cells

Endocytosis
It provides a mean of transport for the large molecule to enter the cell

there are two types of endocytosis

1 phagocytosis
2 pinocytosis

Phagocytosis
Cells in the liver the war out erythrocytes and bacteria

WBC (neutrofils & macrophages) engulfs the bacteria

Amoeba engulf the food particles

Phagocytes are specific

The process of phagocytosis is as follows;

The particles to be contact with the cell surface membrane

The cell membrane investigates to from a flask like depression around the particles

The cell membrane extends outward forming around the particles

44
Subsequentally the react of the flask closed seals of the investigation forming a separate
phagocytic vesicles

Pencytosis is non selective is substance which its transpoits

Cell signalling molecules

Cell to cell recognition

Although cells can act as self contain units they don’t exist in isolation

Even a unicellular organism must detect and respond to outside the cell

In a multicellular organisms of tissues and system=ms brings more complexity

Communication involves tramissing and recieing information

A signalling cell a single and is reci0eved by a target cell

If a change in the form of a single is required it is called a signal transduction

Communication system
Endocrine Secretion of hormone into blood stream
for disporsa the signally cell and target
cell can be very slow method eg; insulin
Parocrine Secretion of a local mediator
This affects cells in the immediate area of
the signalling cell eg; histamine
Neuronal Nerves cells response by the release of
neurotransmitter at synapsecan signal
over very long distances via nerve cells
eg; GABA
Contact dependant Signal molecule in the plasma membrane
of the signal cell interact with membrane
bound receptor the target cell these
signals are therefore connected to cells
which are in direct contact

Extracellular hydrophobic signaling molecules

Some small hydrophobic molecule can cross the plasma membrane and enter the cell by
diffusion

45
Best classes are the steroids hormones eg; costisol and testosterone and the thyroid
hormone eg; thyroxin

The hormones can diffuse across the plasma membrane and bind to receptor proteins that
are located either in the cytosol or in the nucleus

They work by activating gene regulatory protein in the cell which stimulate transcription of
prticles of gene in the molecules

Extacellular hydrophilic signalling molecules

In contrast to hydrophobic signals,the majority of signalling molecules are either too large
or too hydrophilic too cross the plasma membrane

The receptor proteins for these signals must therefore present a binding site to the
extracellular environment and response in the cytosol

These are main 3 classes of these cell surface transmembrane receptor all of which bind
extracellular signal molecule s bur generates intracellular responses in different ways

1 Ion-channel linked receptor:-

These are also known as chemically-gated ion channel

They open pores through the protein in response to binding a signal molecule

Ion flows through this gate generally are electrical effect

This types of receptor is found in excitable cells such as nerve and muscle cells

2 Enzyme linked receptor:-

Found in all type of cells

Generate or enzyme activity on the cytoplasmic end of the protein

This activity causes the phosphorylation of other intracellular proteins, thereby activating
them

3 G-protein linked receptor:-

Activate a binding a protein that sets off a chain of events in the cell

This group of receptors is the largest known and many different signals and responses can
be associated with G-protein activity

46
All have to same structure agreement within the membrane known as a seven pass
transmembrane protein

On binding the signals the G-protein is activated by binding og ATP

This activated proteins diffuses away from the receptor protein site and activates its target
proteins

This may be an ion channel protein or enzyme such as cyclase or phospholipase c

This enzymes catalyse the formation of small molecules known as secondary messenger
which trigger the intracellular response to the original transduction event to the cell
surface

The cyclic AMP signal transduction pathway

Adenylate cyclase activity generates cyclic AMP

Generates ionositol triphosphate(IP3)

Second messenger are important parts of the signal transduction pathway and can have
many different effects

An outline of the c-AMP is shown respectively

Signal transduction
Very complex area !

Signals can be of many different types and can act either by diffusing across the plasma
membrane or bt interacting with a receptor protein on the cell surface

The variety of signals receptors and responses means that the system of signal reception
and transduction can generate very specific effects in different types of cells

The response of cell to signal can involve ion flow activate of specific proteins or changes in
gene expression

These effects can be short linked as in the case of the action potentials as they may be
permanent alteration that control the developmental fats of the cell

It is therefore clear that the idea of a cell as a self contained unit is in fact very far from the
reality of the situation cells are constantly engaged in the exchange of information in the
form of molecular signals and it is this that enables cells in multicellular system to function
in an integrated way.

47
 ENZYMES

• Also known as biocatalyst

• It increases the rate of reaction without being changed themselves at the end of the
reaction
• Enzymes are named by attaching the suffix(ase)
• Eg. carbohydrate- carbohydrase, protein-protease, lipid- lipase etc
• All the enzymes are protein but not all protein are enzymes
• In 1982-sidney Attman Thomas each discovered RNA enzymes which are made up
of RNA
• They achieved a Nobel prize in 1989 for its chemistry

STRUCTURE OF ENZYMES MOLECULES

• Enzymes are globular proteins, and are larger than the substrate they act on.
• The small protein of the enzyme molecular comes to a direct contact with the
substrate.
• The region where substrate is in contact with the enzymes is known as the active
site.
• The amino acid of the active site of the enzyme can be divided into groups:

1. CONTACT RESIDUE

2. CATALYTIC RESIDUE

• The contact residue is responsible for the specificity of the enzyme to form a
shape which is complimentary to the shape of the substrate.

• Catalytic residue are responsible for the ability of the enzyme to catalyse a
particular chemical reaction, they act on bonds of the substrate to break
them down

48
 • The remaining amino acid residues comprise the bulk on an enzyme to
maintain the globular shape, which is responsible for the optical functioning
of the active site.

ACTIVATION ENERGY

• Consider the mixture of ethanol and oxygen in room temperature.

• The reaction between the two substance in thermodynamically possible until
and unless energy is supplied.

• The energy to required to make these two substance to react a known as

activation energy.

49
• If activation energy increase its slows the rate of reaction at any particular
temperature and vice versa.
• Presence of enzymes lowers the activation energy.
Enzymes speed up to the rate of reaction without altering to any great extent, to the
temperature at which the reaction occurs.

• Collision between the substrate and enzyme at correct orientation causes the
substance molecules to bind to the enzyme active site to form an enzyme- substrate
complex.

• Within the E-S complex the chances of the reaction occurring will be greatly
enhanced.

• This is because the enzyme molecular holds the substrate molecules the
arrangement of their forces then to be together in a correct orientation.

• As the reaction occurs the complex breakup to give the products and the unchanged
enzyme.

• The unchanged enzyme molecule is then available to catalyse another cycle of

reaction.

50
• Enzymes are another way to decreases the activation energy.

• The substrate binds to active site of the enzyme; certain bonds of the
substrate are placed under physical stress.
• The physical stress increases the breakage of the bonds.
• The R-group of the amino in the active site of the enzymes can change the
charge on the substrate.
• The disturbance in the electrons, which are within the bonds of the
substrates, overall causes other changes that increase the reactivity of the
substrate.

51
 ENZYMES SPECIFICITY

Two hypotheses have been put toward to explain the enzyme specificity:

1. Lock and key hypothesis

• Fischer in 1890, Enzymes operate on lock and key mechanism, where

complimentary shape of substrate fits accurately to the specific shape of the active
site of and enzyme molecules.
• Binding of the two molecules from a temporary structure called the enzymes
substrate complex.
• The products formed from the reaction may have a different shape of the substrate.
• So, the product leave the active site, as they no longer fit in the specific shape of the
active site.
• This enables the active site of the enzyme to pick up and attach itself to another
substrate molecule.
• THIS HYPOTHISIS IS CONSIDERED TO BE INCORRECT

2. INDUCED FIT HYPOTHESIS.

52
• Induced fit hypothesis was suggested by koshland.
• If the lock and key model was true, one enzyme would only catalyse one reaction in
actually; some enzymes can catalyse multiple reactions.
• The initial shape of the active site of an enzyme might not be complimentary to the
shape of substrate.
• Binding of the substrate to the active site of enzymes induced the conformational
change in the shape of an enzyme.
• This enables the substrate to fit more strongly in to the active site.
• This stresses the substrate, reducing the activation energy of the reaction.

The rate of an enzyme-catalysed reaction:

1. The rate of product formation:

The study of the rate of which the enzymes work is known as enzyme kinetics.

2. Measuring the rate of substrate

• Catalyse is an example of an enzyme

• Catalyse catalysis the hydrogen peroxide to water and oxygen
• 2H2O2 → 2H2O +O2

1. Molecule of catalyse can break 40 million molecules of hydrogen peroxide in each

second.
Amylase is an example which catalyses starch to maltose by hydrolysis.

The rate of reaction is determined by disappearance of starch (blue-black to reddish

brown)

53
Properties of enzymes:-

• Enzymes are effectively in small amount.

• It remains chemically unaltered at the end of the reaction.
• Extremely efficient to the process the reaction 103-108 times faster than an
unanalyzed reaction.
• Enzymes have a high turnover in number.
• Enzymes show a high degree of specificity.
• Activity of enzyme is affected by change in temperature substrate
concentration and enzyme concentration.
• The activity of the enzyme can be altered by the inhibits or activators.

54
Factor affecting the rate of enzymes-catalyzed reaction:
Effect of varying temperature:-

• As heat increases, the molecular motion increases.

• The molecule of substrate and enzymes more and more quickly as the
temperature increases (kinetic energy).
• This increases the chances of bumping of the substrate and enzyme together.
• It results in a grater probability for the reaction to occur.
• The temperature that promotes the maximum activity of enzyme is referred
to as the optimum temperature.
• It the temperature increases beyond the optimal level, and then the rate of
the enzyme-catalyzed reaction decreases rapidly.

55
 • (*the optimal temperature might differ for different enzymes)
• This is because enzymes denture at very high temperatures by losing its
secondary and tertiary structure.
• This results in the unfold of an enzyme & the precise structure of an active sit
is gradually lost.
• The bonds which are highly sensitive to the temperature (hydrogen bond &
hydrophobic interaction) are changed.
• If the temperature decreases to 0.c, then the enzymes are inactive, and its
enzyme activity will be very low.
• Enzymes will retain its catalytic activity as the temperature increases.
• The effect of temperature on an enzyme catalyzed reaction is expressed as
temperature co-efficient (Q10).
rate of enzyme catalysed reaction at x.c+10
Q10 = -----------------------------------------------------------
Rate of enzyme catalysed reaction at x.

Effect of varying catalysed reaction.

56
• The optimum PH is the PH at which the rate of an enzyme catalysed reaction
is maximum.
• At optimum PH, the intermolecular bonds are maintained, so secondary and
tertiary structure of an enzyme is not disturbed.
• If the PH is higher or lower than the optimum, then the concentration of H+
ion will change or alter the changes on the R groups of the amino acid
residues of an enzyme molecule.
• The ionic bonds which help to maintain the conformational shape of an
enzyme molecule is disturbed and the binding of the substrate is affected.
• The enzymes work within a narrow range of pH.
• If the PH is altered by a small extent form the optimum PH, then their effects
are normally reversible.
• If the PH is restored to optimum, then then the maximum activity of the
enzyme is restored.

57
 • If the PH is altered to a larger extent, the conformation shape of the enzyme
molecule is severely affected, and then the denatured enzyme would be
irreversible.

Effect of enzyme activity on varying substrate concentration;

• In a fixed enzymes concentration, the rate or reaction increases with an

increase in substrate concentration.
• An increase in substrate molecules results in an increases in substrate
molecules.
• The rate of reaction will continue to increase with an increase in substrate
concentration, to a point.
• Further increase in the substrate molecule, they no longer produce
significant change in the rate of reaction.

58
 • This is because the active site of all the enzyme molecules is saturated with
substrate molecules.
• The extra substrate molecules have to wait for the enzymes substrate
complex to be free.
• At that point the rate of reaction limited by enzyme concentration.

Enzyme can be inhibited:

• End product inhibition.
• The metabolic pathway involves a series of reaction catalysed by enzymes.
• When the end product begins to accumulate, it acts as on inhibiter.
The final product has the ability to switch of its own production as it builds
up.
• It is self regulatory because as the end product is used up the inhibition is
lifted and production is switched back once again.
• Allostatic enzymes are the ones whose activity is attained by the molecule
acting at the site. Other than the active site.
• This site where the molecule binds in an enzyme is known as the allosatic
site.
• The binding of the regulatory molecule to the allostatic site changes the of
shape of the enzyme active site.

Non competitive inhibition:

• Non competitive inhibitors bind to an allosteric site on the enzyme. The

active site is altered and the substrate cannot attach and react.

59
 • As the concentration of the inhibiter increases, the rate of reaction decreases.
This is because there are fewer functional active sites available for reaction.

• The
maximum rate of
reaction also
reduced-with fewer functional active sites, the enzyme has reduced ability to
process the substrate, even if substrate concentration is increased.

Competitive inhibition:

• A competitive inhibitor blocks the active site, preventing the substrate

entering. Higher the concentration of inhibiter, the slower the rate of
reaction.

60
• Even with competitive inhibition, the same maximum rate of reaction will be
achieved if more substrate is added-because we have not changed the number of
enzymes available.

61
ENZYME KINETICS AND INHIBITION

• The relationship between velocity and the substrate concentration of an enzyme

catalyzed reaction.
• The irreversible and reversible enzyme inhibition

1. Competitive inhibition

2. Allosteric inhibitions

• The word kinetic is derived from Greek word kinetos, which means, ’moving’.
• Using mathematical tools, we can describe as an enzyme’s catalytic power, its
substrate affinity and its responses to the inhibitors etc.
• The study of this analytic part of the enzyme is known as enzyme kinetics.
• An enzymes catalyzed reaction of substrate to product can be written as:

S→P

• Actually the enzyme and substrate must combine and E recycled after the reaction is
finished, just like any catalyst.

62
 • Because the enzymes actually binds with the substrate, the reaction can be written
as:
E + S ↔ Es → P+E
• The simplest reaction is a single substrate going to a single product.

RATE OF VELOCITY OF THE REACTION DEPENDS ON THE FORMATIONB

OF Es

• THE P REVERSIBLE Es is ignored

• The equilibrium constant (Keq) is based on the idea that the reaction is limited to
the formation of the ES complex and that only K1 and K-1 are involved because the
thermodynamics of the reverse of K2 causes it to be minimal.

Keq=k1/k-1

• How fast an enzyme catalysis in reaction is its rate.

• The rate of the reaction is in the number of moles of product produced per second.

Rate (v) =d(p)/dt=K2[ES]

• The relation between the concentration of a substrate and the rate of an enzymatic
reaction is described by looking at the concentration of S and V.
• When the reaction is first order- the rate is dependent on (s).
• When the reaction is zero order- there is no relationship between V and S,
• A second order is between 1st and 0 order- where the relationship between V and
(S) is not proportional to (S).
• To study enzyme; first order kinetics must be followed.
• Think of the graph of (S) vs V in this way:
1. The velocity increases as the substrate concentration is increased up to a
point where the enzyme is (saturated) with the substrate.

63
 2. At this point, the rate of reaction (v) reaches a maximal value and is
unaffected by further increases in substrate because all of the enzyme
active sites are bound to a substrate.

CONDITION FOR MICHAELIS –MENTEN:

• Two assumptions must be met for the michaelis - menten equation:
• EQUILIBRIUM: The association and dissociation of the substrate and enzyme is
assumed to be a rapid equilibrium and Ks is the enzyme substrate dissociation
constant.
• STEADY RATE: the enzyme substrate complex (ES) is at a constant value. That is
the ES is formed as fast as it releases the product. For this to happen the
concentration of substrate has to be much higher than the enzymes concentration.
That is why we only study the initial velocity. Cater in the reaction lower and the
rate of product starts to be limited by diffusion and not the mechanism of the
enzyme.

GRAPHICAL MODEL OF THE REPRESENTATION OF THE MICHAEL’S

– Menta Equation

64
 • Reaction velocity (v) vs concentration of the substrate (s).
• As (s) increases, velocity increases and eventually levels off v max.
• 1st order vs zero order rates of reaction = back to the two assumptions.
• There are two important values for each enzyme that are described by the
M&M equation Vmax & Km(M&M constant).
Graphically these are shown as 1\α V max =Km can not reach real V max so
• Km is a measure of affinity of the enzyme for its substrate and also
informs about the rate of reaction. The binding constant is approximated
by km.
Mathematical model of the representation of mem equation.
E+S ↔ ES → P+E

1. The Michael’s constant km is:

Km =k-1+K2/K1

Think of what this means in terms of the equilibrium large vs a small Km

2. When investigation the initial rate (Vo)the Michael’s – menta eq.is:
Vo =V max +(S)/(S)+Km

GRAPHICAL REPRESANTATION IS A HYPERBOLA:

65
 • When [S]< Km, the velocity is dependent on [S]
• When [S]> Km, the initial velocity is dependant of [S]
• When [S]=Km, the Vo = ½ Vmax

Line Weaver-Burt (double reciprocal plot)

• V max and Km are not likely to do determined by increasing [S].

• Instead the [S]Vs-Vo data are transformed to a plot of their reciprocal of each value.
• 1/[S] VS 1/Vo

V0 = Vmax[s]/[s]+Km → 1/Vo = Km+[S] / Vmax [S]

And this can be simplified to:

1/Vo = (km/V max). 1/[S] +1/Vmax

This is the equation for a straight line:

Y = mx +b

Y = 1/Vo & X = 1/[s]

• Km related to affinity, Vmax relates to efficiency

• Km tells us how much substrate to use in an assay
• If more than one enzyme shares the same substrate then Km will also determine how
to decide which pathway the substrate will take.
• Vmax tells us about the pathways.
• Rate limiting enzymes in pathway:
Km and Vmax can be used to determine effectiveness of inhibitors and activators for
enzymes studies and clinical applications.

66
Mixed Inhibition

67
[1] – inhibitor

• The result will decrease in Vmax and either result in an increase or decreases in km
• The effect of a non- competitive inhibits can only be partially overcome by high
concentrations of the substrate.

Non competitive

68
Immobilized enzymes are widely used in industry

1. Detergent: contain proteases &lipases to help breakdown protein and fat stains.

• Enzymes are used to break down the starch grains into bio-fuels that can be
combusted.
• In the textiles industry, enzymes help in the processing of fibers
Eg; polishing clothes to make it appear more shiny
• Paper production uses enzymes to help in the pulping of wood.
• In the brewing industry, enzymes help a number of processes including the
clarification of beer.
• In medicine and biotechnology, enzymes are widely used in everything from
diagnostic tests to contact lens cleaners, and cutting DNA in genetic engineering.
• Enzymes widely used in food industry
• Eg: fruit juices pectin to increase juice yield from fruit fructose is used as a
sweetener, it is converted from glucose by isomerise.
• Resin is used to help in cheese production.
• Enzymes are used in industry are usually immobilised. They are attached to a
material so that their movement is restricted.
Common ways of doing this:
• A solution of sodium alegnateis prepared by dispersing 2g of Alegnatein
100Cm3 of distilled water- of a temperature of 40°C
• The alegnate and enzyme solution is prepared by stirring 2Cm3 of enzyme
concentration in 40Cm3 of cooled alegnate solution.
• Palates are already produced by filling the syringe with alegnate solution and
arranging it to drop into a beaker of calcium chloride. Insoluble palates are
separated with a nylon or plastic siev.
• These palates are washed in distilled water and are allowed to harden.
• The palates are placed in a narrow tube.

69
Advantages of Enzymes:

• With whole cell immobilized, a number of enzymes can act together at the same
time.
• Substrate can be exposed to a higher enzyme concentration which increase the rate
of reaction.
• Recycled enzymes can be used many times, immobilized enzymes are easy to
separate from the reaction mixture, resulting in a lower cost altogether.
• Separation of products is easier (this also means reaction can be stopped at the
correct time).
• Stability of the enzyme to changes in temperature and pH is increased reducing the
rate of degradation, again resulting in a lower cost altogether.

Disadvantages of Enzymes:

• An immobilized mechanism does not alter the shape or the catalystic ability of the
enzyme.
• Reaction is stable, hard and palate are hard and expensive that is inevitably
reflected in the cost of the industry.
• If the enzyme is detached and combines with the product in the solution, which is
unnoticeable.

END of CHAPTER

70
 Nucleic acid

Genetic control of protein structure and function

There are two types of nucleic acids formed in living cells

1. DNA
2. RNA
DNA: Deoxyribonucleic acid
• Carries the genetic information in cells and consists of thousands of genes
• Genes serve as a recipe for building protein molecules
• Proteins are the building blocks of cells and perform functions similar to that of an
enzyme, hormones, receptor proteins, transport proteins and gene regulatory
proteins.
• Nucleic acid is made up of basic units called nucleotides.
• DNA and RNA are made up of 3 components
1. Pentose (5C)
2. Phosphoric acid
3. Nitrogen base

DNA Nucleotides

5C Pentose

• The sugar present in nucleotides is the pentose -5C atoms

71
 • DNA and RNA are efficient types of pentose sugars.
• Deoxyribose sugar is present in DNA
• Ribose sugar is present in RNA
• Deoxyribose sugar differs from ribose sugar – in which the hydrogen atom at
carbon-2 in deoxyribose sugar is replaced by a hydroxyl group (OH) in the ribose
sugar.

Nitrogen base

• Nucleic acid contains 4 different types of nitrogen bases

• Categorized into purines and pyrimidines
• Purines are of two rings
• Pyrimidines are of one ring
• In DNA and RNA, purines are of adenine and guanine
• In DNA, pyrimidines are of cytosine and Thymine
• In RNA, pyrimidines are of cytosine and uracil (in place of thymine)
• Bases are commonly represented by the initials i. e. A, G, C, T and U.

Nitrogenous bases

Purines

72
 • Adenine (A)
• Guanine (G)

Pyrimidine

• Thymine (T)
• Cytosine ©

73
 Formation of nucleotide

• 3 components are linked to form a nucleotide condensation reaction

• Nitrogen base are joined to carbon atom 1 of sugar molecule.
• Phosphoric acid is attached to sugar molecule at carbon 5
• In total reaction, 2 molecules of water is removed
• Adjacent nucleotides are linked to each other by the phosphodiester bond.
• 5’ end of polynucleotide ends with phosphate group attached to carbon atoms in
sugar molecules
• 3’ end of polynucleotide ends with the –OH group of carbon atom 3 in sugar
molecule
• Sugar and phosphate groups – polynucleotide chains are identical.
• Difference between polynucleotides lies in sequence at nitrogen case

74
 • Sequences of bases carriers the information which controls the organism’s
appearance, behavior and development.

History of DNA

History of the DNA double helix structure

• Fredrick Griffith discovers that a factor in diseased bacteria can transfer harmless
bacteria into deadly bacteria (1928)
• Rosalind Franklin – X-ray photos of DNA (1952)
• Watson and Crick – Describe the DNA molecules from Franklin’s x-ray photo (1953)

Watson and Crick’s Proposal

• DNA had specific pairing between the nitrogen bases

Adenine – Thymine

Cytosine – Guanine

• DNA was made of two long strands of nucleotides arranged in a specific way called
the ‘complementary rule’

A=T

C=G

Structure of DNA

• Consists of α polypeptide chains

• Twisted around each other in a double helix
• 2 strands run is opposite direction (anti parallel)
• One strand – 5’ 3’ directional
• Other strands – 3’ 5’ directional
• Two strands are held together by hydrogen bonds between nitrogen bases
• Purines always pair with pyrimidines

75
e.g., A=T, G=C

• Complementary base pairing

• Width between two sugar phosphate back bone of DNA molecules constant and
equal to width of purine and pyrimidines.
• Within a DNA molecule, the number of nucleotides are equal i. e. adenine = thymine;
cytosine = guanine
• One complete turn of DNA double helix has 10 base pairs
• Sugar phosphate backbone of both the strands lie outside and nitrogen base in the
center of DNA molecules

Structure of RNA

• Single strand except some RNA virus

• Similar to single strand DNA
• Sugar ribose is found in the nucleotides of RNA instead of deoxyribose of DNA
Nitrogen base in RNA adenine, cytosine and uracil. (instead of thymine)

There are 3 types of RNA

1. m RNA
2. r RNA
3. t RNA

These are involved in protein synthesis

Difference between DNA and RNA

1. The basic unit of DNA is deoxyribonucleic acid

2. The basic unit of RNA is Ribunucleic acid
3. Sugar in DNA is deoxyribose
4. Sugar in RNA is Ribose
5. Nitrogen based in DNA is AGCT
6. Nitrogen based in RNA IS AGCU

76
 7. Structure of RNA – double helix
8. Structure of RNA – single strand
9. DNA is only of one form
10. RNA is of 3 forms (mRNA, rRNA, tRNA
11. DNA is huge molecule
12. RNA is a small molecule
13. DNA is found only in the nucleus and small amounts in mitochondria and
chloroplast
14. RNA is synthesized in the nucleus but founf in the cytoplasm
15. Amount of DNA is constant for a cell of a species (except suring interphase in
gametes)
16. Amount of RNA varies from cell to cell and even within a cell (protein synthesis)

The importance of base pairing and hydrogen bonding in DNA molecules

In DNA replication

• In order to maintain the hereditary material, the structure of DNA is allowed to for
its own replication
• The daughter nuclei after nuclear division have identical copies of DNA
• Double helical structure of DNA enables the semi conservative replication
• During replication, α strands can unwind and separate
• Each strand acts as a template
• Complimentary set of nucleotide will attach to form base pairing and hydrogen
bonding
• Each original DNA molecule will give rise to 2 copies of DNA with identical structure
and base sequence

In DNA repair

1. DNA is constantly being subjected to environmental favour which leads to mutation

2. In such cases complimentary strands can be used as a template to guide the repair.

77
3. Repair mechanism ensures the integrity of base sequence of DNA molecules to
remain intact

Instability of DNA molecules

1. Bases are held together by hydrogen bonds – DNA double helix

2. 2 hydrogen bonds between A and T
4. Hydrogen bonds between G and C
5. Hydrogen bonds and hydrophobic interactions which stabilize the double helix
structure
6. Adjacent nucleotides within each strand are held together with strong covalent
phosphodiester bonds.
Packing of DNA into chromosomes
• In eukaryotic cells, DNA is packed into a structure called a chromosome
• DNA is associated with proteins called histones
• DNA wind around the outside gap of 8 histones are positively charged. As a result,
they form an ionic bond
• Nucleosomes are linked to form the chromatin to chromosomes
• Chromatin fibers coil itself to form solenoid (6 nucleosomes in one turn of a helix)
• Coiling of solenoid gives rise to chromatin
• Condensation of packing makes chromatin to chromosomes
• Each DNA molecule has been packed to chromosome i. e. 50,000 times shorter than
its extended length.
What are telomeres?
Telomeres are:
• Repetitive DNA sequences at the ends of all human chromosomes.
• They contain thousands of repeats of six-nucleotide sequence, TTAGGG
What do telomeres do?
• They protect the chromosomes
• They separate one chromosome from another in the DNA sequence

78
• Without telomeres, the ends of the chromosomes would be ‘repaired’, leading to
chromosome fusion and massive genomic instability
• Telomeres are thought to be the ‘clock’ that regulates how many times an individual
cell can divide.
• Telomeric sequences shorten each time the DNA replicated
• Telomeres effectively ‘cap’ the end of each chromosome in a manner similar to the
way the plastic on the ends of our shoelaces ‘caps’ and protects our showlace from
unraveling. (Geron Corporation)

Telomere and aging

• Once the telomeres shrink to a certain level, the cell cal no longer divide. Its
metabolism slows down, it ages, it dies.
• Healthy human cells are mortal because they can divide only a finite number of
times, growing older each time they divide. Thus, cells in an elderly person are much
older than the cells in an infant.
• It has been proposed that telomere shortening may be a molecular clock mechanism
that counts the number of times a cell has divided and when telomeres are short,
cellular senescence (growth arrest) occurs.
• It is believed that shortened telomeres in mitotic (dividing) cells may be responsible
for some of the changes we associate with normal aging.
• Yeron corporation links the telomere and aging condition to this:
• For the cell, having a long telomere can be compared to having a full tank of gas in
your automobile; having a short telomere is like running on an empty tank, Each
time a cell divides, the telomere become a little shorter until the cell simply can no
longer divide (eg. it runs out of fuel).
• "After a certain number of cell divisions, the telomere would be so short at to
somehow prevent the cell from further proliferation - putting it in a state called
secescence. in other words, he proposed that telomeres length offered a clock for
telling a cells longevity" -Scientific American
Replication of DNA

79
In 1950, three hypotheses were proposed:
1. Conservative hypothesis:
• “Both strands of the DNA molecules act like a template for synthesizing an entirely
new DNA molecule.
• “Parental DNA is intact and goes into 1 daughter cell and new DNA molecule goes
into the other daughter cell.

Dispersive Hypothesis
➢ Parental DNA molecule breaks up into short fragments.
➢ Short fragments acts as a template for synthesizing DNA.
➢ Later segments are joined together.
➢ Two molecules with section of both new and old strands interspeeds along each
side.

80
 Semi Conservative Hypothesis
➢ Strands of DNA molecule separates and acts as a template and synthesizes two new
strands.
➢ Hydrogen bonds forms between bases of one old strand and one new strand.
➢ Each daughter cell inherits a DNA molecule. This is said to be hybrid. i.e. one old
strand and one new strand.

81
 Evidence for semiconservative hypothesis
➢ Melson and Stahl (1950).
➢ Used two isotopes of Nitrogen.
➢ N14 : Ordinary isotope of Nitrogen.
➢ N15 : Heavy isotope of Nitrogen.
➢ Parental DNA was labeled with N15.
➢ Labeling of DNA was done by growing Bacteria (E.coli) for many generations in a
medium which contained 15NH4Cl.

➢ Later, bacteria are transferred to a medium of 14NH4Cl.

➢ One sample of bacterial cells was allowed to divide once to provide first generation.
➢ Another sample of cells were allowed to divide twice to obtain second generation.
➢ DNA was extracted from these cells and their densities were determined by
centrifugation in Cesium Chloride solution.
➢ DNA containing N15 is denser than DNA containing N14 .
➢ Hybrid DNA consists of one N14 and one N15 strand.
82
➢ Hybrid DNA forms a band half way between the N14 DNA and N15 DNA band.

Mechanism of semiconservative DNA replication

➢ Before DNA strands starts replication – free Deoxyribonucleotides are synthesized
in cytoplasm and are transported to the nucleoplasm.
➢ Replication begins at specific sites – it is the origin of replication.
➢ HELICASE breaks the hydrogen bond between the Nitrogen bases- results DNA
strands separate.
➢ Single Stranded DNA Binding (SSB Proteins) binds to the separated parental DNA
strand.
➢ Single Stranded DNA Binding Proteins stabilizes the single strand to unwind.
➢ Single DNA strand acts as a template.
➢ Enzyme PRIMASE synthesizes short RNA chain, which is complimentary to one of
the DNA templates.
➢ Short RNA chain-PRIMER.
➢ PRIMER is the beginning of all new DNA chains.

83
➢ [Enzymes responsible for synthesizing DNA i.e. DNA polymerase III cannot start
from scratch.]
➢ DNA polymerase III recognizes bases and selects the free nucleotides that are
complimentary to the parental strand, i.e., A pairs with T, G pairs with C .
➢ DNA polymerase III catalyzes the formation of the phophodiester bond only if bases
are complimentary.
➢ DNA polymerase III has a proof reading or editing function.
➢ RNA portion is hydrolyzed - DNA polymerase I.
➢ Gap is filled by catalyzing- DNA polymerase I.
➢ Synthesis of new DNA strand from 5’ to 3’ direction.
➢ One strand of DNA is known as LAGGING strand. (DNA is synthesized in form of
short fragments.)
➢ One strand of DNA is known as LEADING strand.(DNA is synthesized continuously.)
➢ Short fragments- OKAZAKI fragments.
➢ DNA ligase requires a free Hydroxyl group to function.
➢ At the end of the replication, both the parental and daughter strands unwind to
form a double helix.
➢ The resultant double helix of replication consists of one parental strand and one
new daughter strand by the process of semi conservative replication.

Mechanism of DNA Replication

➢ Prokaryotes have a single origin of replication.
➢ Bidirectional replication with two forks – single terminus.

Summary of DNA Replication

➢ HELICASE unwinds DNA double helix and breaks the hydrogen bonds between the
bases.
➢ Single Stranded Binding proteins stabilizes single strand DNA by the action of
helicase.
➢ Primase synthesizes RNA primer.
➢ DNA polymerase III synthesizes DNA.

84
 ➢ DNA polymerase I erases RNA primer and fills the gap.
➢ DNA LIGASE joins the end of two DNA chains. i.e. OKAZAKI fragments.

The Gene and its Genetic Code

➢ Genetic information is stored in the form of sequences of bases in DNA molecules.
➢ This code helps in the production of protein molecules. i.e. sequence of bases of DNA
molecules will code for amino acid sequence of polypeptides.

Concept of Genes

• A Gene is a specific sequence of nucleotides along a DNA molecule which codes for
specific sequence of amino acids in polypeptide chains.

Central Dogma of Molecular Biology

• Flow of Genetic information from DNA to RNA and to protein is known as the central
Dogma of molecular Biology.
• Central Dogma involves the following process:
1. Replication – genetic information involved in DNA direct the DNA synthesis.
2. Transcription – genetic information expressed in the cell, flows
unidirectionally from DNA to RNA.
3. Translation- from RNA – protein synthesis.

Ammendments to Central Dogma of Molecular Biology

85
 • Significant ammendments are made to the central Dogma since it was formulated in
the 1950s
• Viruses do not use DNA to store genetic information.
• Retro viruses store the information in RNA.
• RNA is used as a template for synthesizing DNA.
• It represents the back flow of genetic information from RNA to DNA.
• Enzymes involved is reverse transcriptase and the process is known as reverse
transcription.
• In eukaryotes, the sequence of bases in DNA and the sequence of amino acids in
protein is not entirely collinear.
• Eukaryotic genes – Split genes.
• Split genes – coding sequence (exon) and non coding sequence (intron)
• During transcription, the coding and noncoding sequences of split genes are copied
on to the primary RNA transcript.
• Non coding sequence excised from primary RNA transcript.
• Coding sequence are joined together to form mRNA.
• mRNA is then transported from the nucleus to the cytoplasm for translation.

Genetic Code
• Codes for the sequence of amino acids.

86
 • Code is stored in a sequence of nucleotides of mRNA - transcription.
• Code is then translated to a sequence of amino acids.- translation

Features of Genetic Code

• Codes are found in triplets
• Every amino acid of a protein is coded by a sequence of 3 bases.
• DNA code is first copied to mRNA.
• Triplet base of mRNA – codes.
• Codes are complimentary to the triplet bonds formed in the DNA template.
• Codes are degenerative (more than 1 codon can code for the same amino acid.)
• 20 amino acids are found in a structure of protein.
• 18 amino acids are designated by more than 1 Codon.
• Codes are non overlapping (triplet codes are used only once)
• ‘AUG’ is the starting codon and it signals the initiation of translation mRNA.
• 3 codons; ‘UGA’, ‘UAG’ and ‘UAA’ are stop codons
- Signals the termination of translation.
• Codons are universal – same triplets of bases codes for same amino acid in all
organisms.

Second Letter
T C A G
TTT –Phe TCT – Ser TAT –Tyr TGT –Cys T
TTC –Phe TCC –Ser TCA –Tyr TGC –Cys C
T
TTA –Leu TCA –Ser TAA -STOP TGA - STOP A
First Letter

TTG –Leu TCG –Ser TAG -STOP TGG – Trp G

CTT –Leu CCT – Pro CAT –His CGT –Arg T

C CTC –Leu CCC –Pro CAC – His CGC –Arg C
CTA – Leu CCA –Pro CAA –Gln CGA –Arg A

87
 CTG –Leu CCG -Pro CAG -Gln CGG- Arg G

ATT –Ile ACT –Thr AAT –Asn AGT –Ser T

ATC –Ile ACC –Thr AAC –Asn AGC –Ser C
A
ATA –Ile ACA –Thr AAA –Lys AGA –Arg A
ATG –Met ACG -Thr AAG -Lys AGG –Arg G

GTT –Val GCT –Ala GAT –Asp GGT –Gly T

GTC –Val GCC –Ala GAC –Asp GGC – Gly C
G
GTA –Val GCA –Ala GAA –Glu GGA –Gly A
GTG –Val GCG -Ala GAG -Glu GGG –Gly G

Protein Synthesis

• Genetic information in a gene in transcribed mRNA in the nucleus.

• The mRNA carries the genetic information to the ribosomes (which is the organelle
for translation).
• Ribosomes will translate the sequence – mRNA into polypeptide.

Requirements for Protein Synthesis

Transcription:

• RNA polymerase catalyses the process of transcription.

• It also catalyzes the formation of mRNA molecules from free ribonucleotides.
• Free ribonucleotides are the basic unit of RNA molecule.
A in DNA pairs with U
T in DNA pairs with A
G in DNA pairs with C
C in DNA pairs with B

88
 • RNA polymerase binds to promoter.
• Promoter – sequence that is at the beginning of a gene that attract the RNA
polymerase.
• RNA polymerase transcribes the anti-sense DNA strands in the 3’ to 5’ direction –
complimentary base pairing.
• The anti-sense strand acts as a template for transcription.
• RNA transcript produced contains an identical sequence at the sense DNA strand.
• Except this as the U are replaced with T in the mRNA.
• The RNA transcript is formed until the RNA polymerase reaches the end of the gene
terminator.
• The RNA transcript is then ready for modification.
• In the gene, some sequences – the introns – do not code for protein.
• Sequence in a gene code for proteins – the exons.
• After transcription, RNA transcript contains exons as well as introns.
• Introns are removed and exons are joined – this process is known as splicing.
• This kind of post transcriptional modification form mature mRNA.
• Splicing occurs in the nucleus.
• mRNA produced in the nucleus contains information for protein synthesis.
• The genetic code on mRNA – triplet code (every 3 bases – one amino acid) –codon.
• A mRNA binds to ribosome.
• t RNA – 3 bases – anticodon (complimentary to codon)
• t RNA carries the amino acids to ribosomes and amino acids are joined by
condensation reaction.
• Ribosoms move along the mRNA – translate the mRNA sequence.
• A translation will stop when the ribosomes reach the last codon (stop codon) in the
mRNA.

Ribosomes:

• Made up of rRNA and protein.

• 2 subunits: a small subunit & a large subunit

89
 • There is an mRNA binding site and 3 t RNA binding site (E, P, A)

tRNA:

• Specific molecule, implying that it contains a specific anticodon and amino acid.
• There are 20 amino acids; there are different types of tRNA in the cytoplasm.
• tRNA – clovered structure.
• Structure is maintained by hydrogen bonds between the complimentary bases.
• At the 3’ ends tRNA, there are three bases, ‘CCA’, which allows tRNA to bind to
amino acids.

Translation:
• 3 stages
• Initiation, Elongation & Termination
• Start Codon, ‘AUG’, is located at the 5’ end of the mRNA.
• The start codon and second codon – mRNA binds to the P site and A site of
ribosomes respectively.
• tRNA – corresponding anticodons bind to the codon accordingly to the
complimentary base pairing rule.
• The amino acid in the P site is transferred - binds to the amino acid in the A site and
peptide bind is formed between these two.
• The enzyme peptidyl transferase forms a peptide bond between the two amino
acids.
• The ribosomes move from 5’ to 3’ of the mRNA.
• The result is that the first codon corresponding to the tRNA will be in the E site,
while the second codon, with the tRNA, carries the dipeptide will be in the P site.
• A site in empty and this allows another tRNA with correct matching to bring another
amino acid to ribosome to build a polypeptide.
• Ribosoms read the codon one by one from 5’ to 3’end, until it reaches the stop
codon.
• In mRNA can be translated by more than one ribosome at a time.

90
 • Structure that contains an mRNA with many ribosomes are called polysomes.

Difference between Prokaryotes and Eukaryotes in steps leading from

genes to polypeptides.
• In prokaryotes both transcription and translation occur in the same compartment of
the cell.
• In eukaryotes, transcription occurs in nucleus and translate occurs in the cytoplasm.
• In prokaryotes, the sequence of bases along the gene is transcribed directly into
mRNA.

91
• In coding sequence (exon) and non-coding sequence (introns) called split genes are
transcribed to primary RNA transcription.
• In non-coding sequences, are excised from the primary RNA transcription and the
coding sequence is simultaneously ligated by the process called RNA splicing to
form mRNA.
• In eukaryotes, the sequences of bases in DNA and sequence of amino acid in protein
is not entirely collinear.
• Eukaryotic genes – split genes
• Split genes – coding sequence (exon) and non-coding sequence (intron)
• During transcription – coding and non-coding sequences of split genes are copied
onto primary RNA transcript.
• Non-coding sequence – excised from primary RNA transcript.
• Coding sequence are joint together to form mRNA.
• mRNA is then transported from the nucleus to the cytoplasm for translation.

92
End of Chapter

93
 Cell Division

Introduction:

• Cell Division – consist of:

➢ Karyokinesis (division of nucleus)
➢ Cytokinesis (division of cytoplasm)
• 1879 – Bouri & Flemming – described event occurs within – nucleus leading to the
production of 2 identical cells.
• 1887 – Weismann suggested specialized form of division occurs in gametes.
• Two types of division:
1. Mitosis
2. Meiosis

Chromosome:
• Chromo – Colour / Some – body
• Important structure in cell during cell division
• Responsible for the transmission of heriditary information from one generation to
another.
• Contains DNA – the molecule of inheritance
• During cell division – DNA replicates into two identical molecules.
• The two parts are reffered to as chromatids.
• Each chromatid of a pair contains one of the two identical DNA molecules.
• Beginning of the cell division, chromosomes are loosely coiled, long, thin thread
spreads throughout the nucleus – chromatin
• Before cell division, the chromatin coil becomes short, thick and recognizable
structure – chromosome.

94
• Each chromosome contains chromatids.
• Chromatids are held together by a point called the centromere.
• It occurs anywhere in the length.
• Each species have characteristic numbers of chromosomes in each cell.
• 2 sets of chromosomes:
1. Autosomes
2. Allosomes
• Cell – 2 set of chromosome – diploid
➢ Symbol – 2n
➢ eg. Somatic cell
• Few simple organism have only one set of chromosomes and are referred to as
haploid cell.
➢ Symbol – n
➢ eg. Sperm and egg cell (gametes)
• Organism including plants have 3 or more sets and are referred to as polyploidy.

Advantages of possessing 2 sets of Chromosomes:

• Genetic variation increases
• Gene on one chromosome of a pair is faulty; the second chromosome may provide a
normal back up.
• Homologous chromosome:
➢ The identical male and female chromosome occurs in pairs and are known as
homologous chromosome.
• Gene:
➢ The fundamental physical and functional unit of hereditary, which carries
information from one generation to the next.
• Allele:
➢ One of the two or more forms that can exist at a single gene locus
distinguished by their different effects of the phenotype.
• Genotype:

95
 ➢ The genetic make up or constitution of an individual with reference to the
traits under consideration usually expressed in symbol.
• Phenotype:
➢ The appearance or discernible character of an individual which is dependant
on its genetic make up usually expressed in words.

Factors influencing the cell division

• In multi-celled organisms (like humans) cells specialize for specific functions thus
the original cells must divide to produce different kinds of cells.
• Cells can only take in nutrients and excrete waste products over the surface of the
membrane that surrounds them. The surface to volume ratio decrease with the
square of the volume (unless special accommodations are made).
• Nucleocytoplasmic:
➢ Cell increases beyond a certain size the cell divide to restore a fundamental
Nucleocytoplasmic ratio so that the nucleus efficiently direct and control the
activities in the cytoplasm
• Mechanical factor:
➢ Cell on hand and feet – caused by repeated cell division
➢ Abrasion removes cells from there surfaces and stimulates the cell beneath
to divide more rapidly than normal.
• Hormones:
➢ Auxine in plants initiates cell division and determines the rate of cell division.
• Wound Surface:
➢ It produce substance – causes the cell to multiply and grow over an injured
area.

The Cell Cycle

It can be defined as the entire sequence of events happening from the end of one
nuclear division to the beginning of the next.

96
Cell cycle is divided into 3 parts.
1. Interphase
Divided into 3 pases:
A. ‘G 1’ Phase
▪ Immediately after the cytokinesis and lasts for an hour to a month, or
even a year.
▪ Resting phase and called first gap phase.
▪ Cell – manufacturing protein such as histones, ribosoma proteins and
tubulin.
▪ Synthesis of cell organelles
▪ Cell – builds up a large store of energy.

B. ‘S’ Phase:
▪ Replication of DNA and synthesis of histone protein
▪ New histones are required – Masssive amount immediately at the
beginning of S phase provide new DNA – nucleosomes.
▪ End of the S phase, each chromosome has 2 DNA molecules and a
duplicate set of genes.
▪ 2n – 4n

C. ‘G 2’ Phase:
▪ Synthesis of RNA and protein which is required for cell growth.

97
 ▪ Cell continues to build up energy.
▪ Manufacture protein and cyto plasmic organelles
Genetic material in the nucleus is thread like in structure i.e.
uncondensed form called chromatin.
2. Nuclear Division (by Mitosis)
3. Cell Division (Cytokinesis)

The Cell Life Cycle

G1: Gap 1 – Doubling of all size Regular cellular activities.
Transcription and Translation, etc.

S : Synthesis of DNA – Regular cell activities cell activities cease and a copy of
all nuclear DNA is made.

G2: Final preparation for division

M: Mitosis – Cell Division

Mitosis – 4 Phases
1. Prophase:

98
 • Appearance of thin thread like condensed chromosome Marks the first phase of
Mitosis.
• Cell becomes spheroid, more retractile and viscous
• Each chromosome contains two chromatids.
• With condensation – Chromatids show centromere
• Nucleolus disintegrates and disappears
• Cytoplasm – the spindle is formed between the aster and move towards the poles
• Microtubule forms – spindle is also called the spindle fiber.
• 2 types of spindle fibres run from one pole to another
• Another type runs from the pole to centromere of each chromosome.
• Higher plants have no centriole – still forms spindle and undergo normal mitosis.
2. Metaphase:
• Beginning of Metaphase – NE disintegrates and mixing of nucleoplasm with the
cytoplasm occurs.
• Chromosomes – attached to the microtubules of the spindle and are oriented at the
equatorial plate.
3. Anaphase:
• Centromere splite and spindle fibre contracts or shorten faster.
• It causes 2 chromatids – each chromosome to separate and migrate to opposite pole.
• Shortening of the spindle fiber due to progressive removal of the tubulin molecule of
which they are made.
• Energy for this phase – provided by the mitochondria, which are observed to collect
around the spindle fibre.

4. Telophase:
• During this phase, chromatids reach – respective phase, new NE form around each
group.
• Chromatids uncoil and lengthen and become invisible.
• Spindle fibres disintegrates and a nucleolus reforms.

99
Cytokinesis:
• After nuclear division, the cytoplasm is divided into 2 parts.
• Animal cell – furrow develops into cell membrane.
• Cell membrane furrow eventually joins up and completely separate 2 daughter
nuclei
• Plant cell – series of Golgi vesicles line up in the middle of the parent cell.
• Golgi vesicle contribute – cell plate
• Cell plate eventually fuses with the parental cell wall and cell membrane, separating
– 2 daughter cell.

What is a Kinetochore?

• Kinetochores play an important role in cells, especially during cell division.

• A Kinetochore is a protein structure that forms on a chromatid during cell division
and allows it to attach to a spindle fibre on a chromosome.
• A chromatid is one of two strands that form when a chromosome replicates.
• During the replication process, the two chromatids unite by way of a centromere, or
the part of the chromosome that is connected to the spindle fiber.
• The purpose of the kinetochore is to pull the chromatids apart.
• Kinetochores also help during cell division by making sure that each new cell has
one chromatid from each pair.

100
Kinetochore Structure:
• Kinetochores consist of three regions: an inner and outer region as well as a fibrous
corona
• Each region works in its own particular way to aid in the separation of the sister
chromatids.
• Each chromatid recieves its own kinetochore.
• The inner plate of the kinetochore is connected with the centromere, where it works
closely with centromeric DNA.
• The centromere is where the sister chromatids connect and form a
chromosome, an important source of genetic information located in the
nucleus of almost all living things.
• The outer plate of the kinetochore is attached to, and works in conjugation
with, the microtubules, which are connected to the spindles at either end of
the cell poles.
• The fibrous corona is created from a network of permanent and temporary
proteins and helps regulate the attachment of the microtubules to the outer
plate.

101
 • Due to their location and organization, all three regions of the kinetochore
share an equal role in making sure that the structure is doing its job.
• Their activities and relationships only occur during all division and are
essential in that they help to pull the chromatide.

Significance of Mitosis
• Helps the cells in maintaining proper size
• Helps in the maintenance of an equilibrium in the amount of DNA and RNA in the
cell.
• Provides an opportunity for growth and development to organs and the body of an
organism.
• Old decaying and dead cells of body are replaced.
• Certain organisms are involved in asexual reproduction.
• Cleavage of egg during embryogenesis & division of blastema blastogenesis in
mitosis.

Role of Mitosis

The length of the cell depends on the type of cell and the external factors – nutrients,
Oxygen supply and temperature.

Meiosis

• 2 cells division
• Starts with 2 copies of each chromosome (homologous) each with 2 chromatids.
• In meiosis 1, crossing over in prophase mixes allels between the homologues.
• In metaphase of meiosis 1, homologous pair up and in anaphase, the homologues
are separated into 2 cells.

102
 • Meiosis 2 is just like mitosis. The centromeres divide in anaphase giving rise to a
total of 4 cells, each with 1 copy of each chromosome, and each chromosome with
only one chromatid.

Summary

MITOSIS
• An equational division that
seperates sistes chromatids.
• One division per cycle i.e., one
cytoplasmic division (cytokinesis)
per equational chromosomal
division.
• Homologous chromosomes do not
synapse; no chiasmata form
• Genetic exchange between
homologous chromosomes does not
occur.
• Two daughter cells produced per
cycle
• Genetic content of mitotic daughter
cells is identical to mother cells.
• Chromosome number of daughter
cells is the same as that of the
mother cell.
MEIOSIS
• Meiosis 1 is a reductional division
that separates homologous
chromosome sister chromatids
separate during meiosis 2.
• Two cytoplasmic divisions: one
following reductional
chromosomal division meiosis 1
and one following equational
chromosomal division meiosis 2.
• Chromosomes synapse and form
chiasma
• Genetic exchange occurs between
homologous chromosomes.
• Four daughter cells, called gametes
produced.
• Genetic content of mitotic
daughter cell is different from each
other and the mother cells.
• Chromosome number of daughter
cells is half that of the mother cells.

103
• Mitotic products are usually capable
of undergoing additional mitotic
division.
• Normally occurs in almost all
somatic cells through the life of the
organism.
• Meiotic products cannot undergo
additional meiotic divisions,
although they may undergo
subsequent mitotic division.
• Occurs only in specialised cells of
the gem line in the mature
organism.

CANCER
• Involves the uncontrolled division of cells.
• Certain genes are responsible for normal cell growth and division
• Cancer caused by mutation
- Spontaneous
- Induced
• Normal cells have the gene – proto-oncogene.
• Promotes normal growth and division of cell
• Cancer cells have the gene – oncogene.
• Mutated form of proto-oncogene.
- When the oncogene is switched on, the cell divides excessively.
- Normal cells have tumour suppressor gene.
- Function – prevent or suppress cell growth.
- Mutation occurs in these genes – it loses this ability and cells can start growing and
divide in an uncontrolled manner.
- BRCA1 and BRCA2 are well known tumour suppressor genes.
- Mutated form of BRCA1 and BRCA2 are believed to be responsible for about half a
case of inherited breast cancer.
- Tumour cells divide normally with respect to normal cell.
- Form a mass of cell – cancerous cell or tumour.

104
- Some form of cancer migrates to other parts of the body through blood vessels or
lymphatic system.
- Migrated cells give rise to cancer, some distance from the original one.
- Tumour spread in this way is described as Malignant.
- Process by which cancerous cells are released from a tumour and spread to other
parts of the body is called Metastasis.

NORMAL CELL CANCEROUS CELL

Normal cells have proto-oncogene, Cancerous cells have oncogene which

whose function is to promote the are mutated form of proto-oncogene
normal growth and division of cells. when oncogenes are switched on, cells
divide excessively.

Cancerous cells do not show

programmed cell death as they can
Normal cells show programmed cell divide indefinitely.
death they divide for a certain number
of times and then stop dividing.
Cancerous cells do not show contact
inhibition.
Normal cells show contact inhibition –
this means that they do not divide
further when in contact with other cells.

Normal cells differentiate properly to

become specialized cells.
Cancerous cells fail to differentiate
properly.

105
 Causes of Cancer

➢ Age:
• Chances of developing cancer increases with age.

➢ Chemicals:
• Numbers of chemicals are known to cause cancer-carcinogen.
• E.g. tar in cigarette smoke causes lung cancer.
• Low fiber intake increases the chance of colon cancer.
• Ethidium Bromide and Carbon Tetra Chloro Methane.

➢ Radiation:
• Exposure to too much UV radiation in sunlight causes skin cancer.
• X rays cause cancer.

➢ Viruses:
• Human papilloma virus causes cervix cancer.

➢ Genetic Factor:
• Certain cancers eg. colon cancer runs in families.

106
 CIRCULATORY SYSTEM

The circulatory system is comprised of the heart, veins, capillaries, arteries, lymph vessels and lymph
glands, which work together to supply the body tissues with nourishment and collect waste
materials.

Functions of the Circulatory System:

- Distribute nutrients
- Transport and exchange O2 and CO2
- Remove waste materials
- Distribute secretions of endocrine glands
- Prevent excessive bleeding
- Prevent infection
- Regulate body temperature.

The Need for Circulatory System:

- Organisms need to exchange materials within themselves and the environment.

- The respiratory gases and the raw materials for growth must pass into an organism
and waste products must be removed.
- In small organisms there is a lack of specific system for transport and distribution of
materials.
- In small organisms, they possess a sufficient larger surface area to volume ratio to
allow efficient exchange of most of the materials over the surface of the body through
diffusion.
- The distance for materials to travel is small enough.

107
 - As the organism increases in size and complexity, there is a need for development of
internal transport system.
- An increase in size and metabolic rate of organism, there is a corresponding increase
in requirement for food, oxygen, as well as there is an increase in waste material.
- Increase in the size reduces the surface area to volume ratio. This means the exchange
of material through the body surface through simple diffusion is inadequate.
- The material has to travel within the body increases.
- An increase in size and specializations of an organism’s tissue and organ is also
increasingly dependent on one another.
- This leads to the development of internal transport system and facilitates the
exchange of materials between the tissue and organ.

Mammalian Circulatory System:

Circulatory system consists of:

- Blood: fluid that is transported throughout the system.

- Blood vessel: tubes that carry blood.
- Heart: to pump blood through the network of blood vessel.

Circulatory System – 2 types

➢ Open blood system

- Blood doesn’t flow through the blood vessels.
- Pumped out of the heart into larger spaces in the body cavity.
- Blood comes in close contact with the tissues and the exchange of materials takes
place.
- Then blood returns to the heart.

➢ Closed Circulatory System

Eg. Mammals.

108
 Blood flows in blood vessels.

Closed circulatory system once again divides into two:

1. Single Circulation
2. Double Circulation

Single Circulatory System:

- Eg. fish
- Here the blood flows from the heart to the gills.
- The oxygenated blood is then carried to the tissue.
- From the tissue, the deoxygenated blood is returned to the heart.
- Blood passes through the heart only once in one circulation of the body.

Double Circulatory System:

The blood passes through the heart twice in one circulation of the body.

Advantages: -

- Sends the blood to the lungs to pick up the oxygen and then to return to the heart
before travelling around the body.
- When blood passes through the capillaries in the lungs, it loses its pressure- this
circulation makes the pressure to boost and restores it before the blood is circulated
around the body.
- Double circulatory is possible as the heart is divided into two.
- One half pumps deoxygenated blood to the lungs and the other half pumps
oxygenated blood to the rest of the body.
- Beginning of double circulatory system is seen in amphibians, mammals and reptiles.
- The organs are arranged in parallel instead of series.

109
 - If the organs were arranged in series, the blood will start losing the pressure. This
would damage the blood vessels linking the two organs, which interrupts the whole
circulation.

Blood Vessels:

110
The blood is circulated around the body and passes through a series of arteries, capillaries and veins.

- Arteries and veins have three layers:

1. Endothelium (tunica interna)
• It is an inner lining made up of squamosal epithelium
2. Tunica Media
• It is a middle layer made up of smooth voluntary muscles and elastic fibres.
3. Tunica Externa
• It is the external layer which is mainly made up of inelastic fibres.
• White fibre (collagen fibre).

Arteries:

- Arteries transports the blood away from the heart

- Tunica Media is thick and composed of elastic smooth muscles.
- There is no semilunar valve except where it leaves the heart.
- Pressure of blood is high as pulse flows rapidly with low blood volume.

111
 - The blood is oxygenated except for the pulmonary artery.
- The larger arteries which are nearer to the heart (Aorta, Subclavian) are thick and are
made up of elastic fibres to withstand pressure.
- This characteristic enables them to dilate but not rupture when heart forces blood at
high pressure.
- In between, the heartbeat, the arteries undergo elastic recoil and contract tending to
the smooth outflow of the blood along their length.
- Arteries are supplied with sympathetic nervous system.
- The stimulation from the sympathetic nervous system makes the arteries to dilate
and control the flow of blood to different parts of the body.

Arterioles:

- Arterioles consist of only endothelium wrapped around by a few muscle fibres at

intervals.
- Arterioles possess sphincter muscles at the point of entry of capillaries.
- Sphincter muscles are circular fibre muscle, they prevent the blood from flowing into
the capillaries.
- In certain regions of the body, there are cross connection or shunt vessels.
- It acts like a short cut between arterioles and venules.
- Shunt vessels serve to regulate the quantity of the blood that flows to the capillary
bud, according to the needs of the body.

Capillaries:

- It links arteries and veins, site of exchange of material between blood and tissue.
- No tunica media but only tissue present is squamous epithelium.
- Capillaries do not have elastic fibres and semi lunar valves.
- The pressure of the blood will be falling in capillaries so no pulse is detected.
- Blood flow is slowing with high blood volume.

112
 - There will be a region of mix-oxygenated and de-oxygenated blood.
- The endothelium wall of capillaries is only permeable to water and dissolved
minerals.

Venules:

- The blood from capillaries is drained into venules.

- The wall of the venules consists of thin collagen fibres which are tough and inelastic.
- Venules passes the blood to veins and eventually carries back the blood to the heart.

Veins:

- Possess less muscle and elastic fibre in its middle layer.

- The diameter of the lumen is greater than arteries.
- The semilunar valves are present.
- The valves are formed from the folds of the inner wall of the vein.
- Function is to prevent the backflow of blood.
- Number of veins is located between the larger muscles of the body.
- When larger muscles contract, they exert pressure on the veins and squeezes them
flat.
- This helps the flow of blood to the heart.

Characteristic features of Vein:

- Relatively thin and only slightly muscular.

- Semi lunar valves are present at an interval along the length to prevent the back flow
of the blood.
- The pressure of the blood is low so no pulse is detected.
- Much higher blood volumes are present in vein than in capillaries or arteries.

113
 - The blood in the veins is deoxygenated except pulmonary vein.

Composition of Blood:

There are two portions that are - blood plasma and cellular components.

Blood plasma:

- Makes 55% of the total blood.

- Watery liquid that is left when the cellular constituent of blood are removed.
- It contains dissolved substances and acts as a medium of transport.
- Substances carried by plasma are plasma proteins like albumin, mostly globulins –
immune globulins.
- Proteins play an important role in maintaining osmotic pressure.
- Proteins are also an important factor for the formation of tissue fluid.
- Serum refers to the blood plasma without blood clotting factors.
- Other dissolved substances are nutrients, glucose, amino acids, fatty acids, and
creatinins and mineral salts.
- The waste products are NH4 (ammonium), urea and uracil.
- Gases like O2 and CO2.
- Enzymes and hormones are also transported.

RBC’s (Erythrocytes):

- It makes 99% of the cellular components in the blood.

- Main function is to carry oxygen from lungs to tissue and CO2 from tissue to lungs.
- Structure:
o Biconcave
o No nucleus
o The cytoplasm of the RBC contains haemoglobin

114
 o The shape of the RBC provides larger surface area and also enables the RBC to
squeeze through the blood capillaries.
o It has limited life span (120 days)
o Graveyard for RBC’s liver spleen.
o They are synthesized in adult bone marrow.
o Size of RBC is 7.5 micro meter.
o Number of RBC’s per ml is 5.1 x 108 ml in male and 4.7 x 106 in female.

WBC’s (Leukocytes):

- Number of WBC’s per ml of blood is 6000-9000

- In WBC nucleus is present and haemoglobin is absent
- WBC’s are divided into two types:
1. Granulocytes
2. Agranulocytes
- Granulocytes are of three types:
1. Eosinophils
2. Basophils
3. Neutrophils
o These cells have conspicuous granules in the cytoplasm and lobed nucleus.
- A granulocytes are of two types:
1. Monocytes
2. Lymphocytes
o Do not have granules
o Nucleus is spherical and bean shaped
o Lymphocytes – 6 to 8 µm – 20% to 80%
o Neutrophils – 12 to 15 µm – 60% to 70%
o Eosinophils – 12 to 15 µm – 2% to 4%
o Basophils – 12 to 15 µm – 20% to 30%
o Monocytes – 12 to 20 µm – 3% to 8%

115
Platelets:

- Also called thrombocytes

- They are cell fragments formed from the cytoplasm of large cells
- Sparsely distributed in the plasma
- Involved in the process of blood clotting

116
Different Types of Cells

117
Transport of Oxygen:

- Solubility of oxygen in the blood is low.

- Mammals have protein to combine with oxygen called respiratory pigment.
- The respiratory pigment combines with O2 where the concentration is high; and
releases where the concentration is low.
- The respiratory pigment is known as haemoglobin.
- Haemoglobin has 4 subunits:
- Two α subunits
- Two β subunits
- The polypeptide chain is associated with a prosthetic group i.e. haem group.
- Haem group contains ferrous ions (Fe2+).
- Haemoglobin molecule combines with 4 oxygen molecules to form oxy-
haemoglobin.
- An oxygen molecule reversibly attaches to the haem portion of the haemoglobin.
- The haem unit contains Fe2+ which provides the positive force.
4O2 + Hb ⇌ HbO

Oxygen Dissociation Curve:

- Oxygen capacity – The maximum quantity of oxygen that will combine chemically
with the haemoglobin is a unit volume of blood.
o Normally it amounts to 1.34 ml of O2 per gram of Hb or 20 ml of O2 per 100 ml
of blood.
- Oxygen content – how much oxygen is in the blood.
- Oxygen saturation – A measure of how much oxygen the blood is carrying as a
percentage of the maximum it could carry.
- Respiratory pigments have a high affinity for oxygen when its concentration is high.
- The affinity decreases when O2 concentration is low.
- Oxygen concentration is measured in terms of partial pressure and its known as
oxygen tension.

118
 - In a mixture of gas, the partial pressure of each component is proportional to the
molar percentage of the mixture.
- Oxygen tension reflects the proportional of oxygen present in the atmosphere.
- The affinity of respiratory pigment for oxygen is measured experimentally by
determining the percentage saturation with oxygen, of sample of blood.
- After exposure to the air containing different partial pressures, the relation between
the oxygen and the saturation of respiratory pigment is called oxygen dissociation
curve.

Oxygen Dissociation Curve for Haemoglobin:

- Oxygen Dissociation curve is sigmoid or ‘S’ shaped.

- The steep rise part of the curve will correspond to the range of oxygen partial
pressure found in the tissue.
- The small increase in partial pressure is that oxygen brings about a large change in
percentage saturation in haemoglobin with oxygen and vice-versa.
- The partial pressure of oxygen decreases as the result of tissue utilizing the oxygen.
- Oxy haemoglobin gives up oxygen rapidly.
- The horizontal part of the oxygen dissociation curve corresponds to the situation in
lungs.
- Haemoglobin will remain highly saturated with oxygen despite the change in oxygen
temperature.
- High pressure of haemoglobin is maintained even if partial pressure of oxygen
decreases slightly.
- This favours loading of haemoglobin with oxygen in the lungs.
- Oxygen dissociation curve is ‘S’ shaped because of the phenomenon of Allostery
(allosteric property).
- Oxygen molecule binds with Fe2+ of haem group and changes the shape of
haemoglobin.

119
- The change in the haemoglobin molecule occurs when oxygen molecules attach to the
second and third haem group.
- Each facilitates quicker binding of oxygen as compared with preceding one.
- Fourth haem group will readily pick up the oxygen molecule and it does it several 100
times faster than the first.
- The reverse will occur when oxy haemoglobin is exposed to the region of low oxygen
tension that is actively respiring tissue.
- The first oxygen molecule releases to the tissue very rapidly and subsequently oxygen
molecules are given up much less rapidly.
- In oxygen dissociation curve, the partial pressure in which respiratory pigment is
50% saturated with oxygen, then it is known as uploading tension.

120
Bohr’s Effect: The effect of CO2 on Haemoglobin

121
- Higher concentration of CO2 in tissue cell.
- Curve shifts to the right.
- Always at a valve of lower percentage saturation of haemoglobin than the curve on
the left.
- Haemoglobin has a low affinity to O2.
- In actively respiring tissue cells, O2 is more easily released by haemoglobin.
- Lower concentration of CO2 in the lungs.
- Curve shifts to the left.
- Haemoglobin has a higher affinity to O2.
- In well ventilated alveolus, O2 is more easily taken up by haemoglobin.
- The region where there is an increase in partial pressure of CO2 makes the oxygen
dissociation curve shift to the right.
- This makes the haemoglobin to release the oxygen more readily to the oxygen.
- In lungs, the partial pressure of CO2 is so low that the oxygen dissociation curve shifts
to the left.
- Haemoglobin picks up the oxygen more readily.

122
Myoglobin
• This is another respiratory pigment in the vertebrates.
• It consists of singlE polypeptide chain and a single heam group.
• O.D.C for myoglobin is displaced to left that for the adult haemoglobin.
• Myoglobin has a greater affinity for oxygen than haemoglobin.
• Myoglobin is present in skeleton muscle.
• The function of myoglobin is to store oxygen in resting muscles.
• It supplies oxygen to active muscles when blood supplies inadequate of oxygen.
• Myoglobin replenishes the stored oxygen from oxy haemoglobin in the blood.
• High levels of myoglobin are present in the muscles of diving animals and flight
muscles.

123
 Oxygen dissociation curve Heamoglobin Vs Myoglobin

Heart
- Made up of cardiac muscle.
- Cardiac muscles are made up of interconnecting muscle cells.
- Each muscle cell is joined by inter calary disk .
- Inter calary disk allows the rapid spread of impulses through the tissue from cell to
cell.
- Heart is described as myogenic.
- Myogenic means that the heart rhythmically contracts and relaxes on its own.
- Heart pumps blood side by side.
- Right side of the heart pumps deoxygenated blood to the lungs.
- Left side pumps oxygenated blood to the rest of the body.
- Each side of the heart has two chambers.
- Right atrium receives blood from the vena cava.
- Left atrium receives oxygenated blood from the pulmonary vein.
- When atria contracts, blood flows into the lower chambers called ventricles.
- When right ventricles contract, deoxygenated blood is pumped into the pulmonary
artery.
- When left ventricles contract, it pumps the oxygenated blood through the aorta.
- The thickness of the wall of each chamber of the heart is related to the distance the
blood vessels need to be pumped.
- The two atria pump the blood to a shorter distance that is ventricle.
- The ventricle have thicker walls to develop enough pressure so to force the blood
further.
- Left ventricle has more muscular walls than the right ventricle.

124
 - Right ventricle forces deoxygenated blood only to lungs whereas left ventricle forces
oxygenated blood to the whole body.
- There are two sets of valves.
- The valves ensure the blood flows unidirectional.
- Atrioventricular valves are formed on each side between the atria and the ventricle.
- A.V valve prevents the backflow of blood to the atrium when the ventricles
contracts.
- The right side of the A.V valve is called tricuspid valve, which has 3 valves.
- The left side of the A.V valve is called the bicuspid valves, which has two valves.
- There are semilunar valves fount at the base of pulmonary artery and aorta.
- The S.L valves will prevent the back flow of the blood back into the ventricles.
- The septum divides the heart into two:
1. Interventricular septum
2. Inter artrial septum
- Valves are held by cardiac tendons.

Cardiac Cycle:
- It describes the sequence of events in one heartbeat.
- It is described in terms of alternate contraction: Systole and relaxation-diastole
- The cardiac cycle will last for 60/75 =0.85 seconds
- The valves of the heart respond to the pressure change during a cardiac cycle.
- The noise by the blood, when valves open and closes, make a sound called “lub dub”.
- There are two major stages of the cardiac cycle.

Atrial systole:
-In the heart, the atrium is full of blood and the ventricles are relaxed.

-Both atria contracts and the blood passes to the ventricles.

-Atrioventricules valves open as pressure of the blood is against them.

-The 70% of the blood flow passively down the ventricles.

Ventricular systole:
-Atria relaxes
-The thick muscular wall of the ventricular contracts, forcing the blood out of the heart
through the pulmonary artery and aorta.
-The pressure of the blood is against the atrioventricules valves, so it causes the valves to
shut to prevent the backflow of blood.
-This produces the first part of the sound, “lub”.

125
-Pressure of the blood is against semilunar valves so it opens.
-Pulmonary artery carries deoxygenated blood to lungs, aorta carries oxygenated blood to
the body.

Atrioventricular Diastole
-The pressure inside the ventricles drop below arteries so blood under high pressure
in the arteries causes the semilunar valves to shut.
-This prevents the blood from the arteries to go back to the ventricles, which produces
the second part of the sound. “dub”.
-During the diastole, the muscle in the heart relaxes.
-The blood from the vana cava and pulmonary vein enter the atria.
- The whole cycle repeats again.

Control of the heartbeat:

-Mammal heart is known to be myogenic.
-It means the heartbeat is initialized within the heart rather than nerve impulses
outside the heart.
-Stimulus to the heartbeat is originated from the Sino Atrial Node (S.A Node).
-It consists of a group of specialized cardiac muscle.
-S.A Node is located in the right atrium.
-That is near to the superior and inferior vena cava, which joins to the heart.
-The S.A Node determines the basic rate of the heartbeat.
-It is known as the pace maker.
-The action potential spreads from the S.A. Node across the atria.
-This causes the contraction of the atrium at the same time.
-This constitutes the atrial systole.
-The action potential from the S. A. Node moves to the atrioventricular node (A.V.
Node).
-It consists of a special group of cardiac muscle cells.
-A.V Node is located between the two atria.
-In order to force blood upward, towards the aorta and pulmonary artery.
-Ventricles contract from apex to upward.
-The action potential from the A.V Node transmitted along a bundle of hill.
-The bundle of hills are strands of cardiac fibres which run in an interventricular
septum.
-Bundle of hill divides into two – that is one for each ventricles.
-It is branched to form purkinje fibre and are distributed over the ventricular wall.
-The action potential spreads from the purkinje fibre causing contraction of the
muscles of the wall of the ventricles (from apex to upward)
-The blood is therefore forced out of the ventricles into the arteries.

126
-The constitutes the ventricular systole.

Note: The stimulus spread from the S.A Node reaches to the A.V node is delayed by
0.1-0.2 seconds because of the presence of the narrowing of non-conducting fibres
at the base of the atrium.
This helps in ventricular filling.

Starling’s law

The power of cardiac contraction is proportional to the degree of stretching.

Cardiac output
Is adjusted to meet varying needs of the body.
Cardiac output = heartbeat X stroke volume

Stroke volume:
Volume of the blood force out of the heart by each muscular contraction.

Carbon dioxide transport in the blood:

The blood carries CO2 partly as undissociated CO2 in solution in the plasma, partly
as hydrogencarbonates ions in solution in the plasma, and partly combined with
haemoglobin in the red blood cells.

127
End of Chapter….

128
 Gaseous Exchange

The human gas exchange system links the circulatory system with the atmosphere.
It is adapted to:
-Clean and warm the air that enters during breathing.
-Maximize the surface area for diffusion of O2 and CO2 between the blood and the
atmosphere.
-Minimize the distance for diffusion.
-Maintains adequate gradients for diffusion.

- Most organisms need a supply of oxygen for respiration.

-In single celled organisms the oxygen simply diffuses from the fluid outside the cell,
through the cell surface membrane and into the cytoplasm.

-In a multicellular organism, such as a human, however, most of the cells are a considerable
distance away from the external environment from which the oxygen is obtained.

- Multicellualar organisms therefore usually have a specialized gas exchange surface where
oxygen from the external environment can diffuse into the body, and carbon dioxide can
diffuse out.

-In humans, the gas exchange surface is the alveoli (Singluar: alveolus) in the lungs.

-Although each individual alveolus is tiny, the alveoli collectively have a huge surface area,
probably totalling around 70 m2 in an adult.

-This means that a large number if oxygen and carbon dioxide molecules can diffuse
through the surface at any one moment to give us a high rate of gas exchange.

Lungs:
− The lungs are in the thoracic (chest) cavity surrounded by the pleural membranes
which enclose an airtight space.
− This space contains a small quantity of fluid to allow friction free movement as the
lungs are ventilated by the movement of the diaphragm and the ribs.

129
 Trachea, Bronchi and Bronchioles:
− The lungs are ventilated with the air that passes through a branching system of
airways.
− Leading from the throat the lungs is the trachea.
− At the base of the trachea are the two bronchi (singular; bronchus), which subdivide
and branch extensively forming a bronchial ‘tree’ in each lung.
− Each bronchus divides many times to form smaller bronchioles.
− Terminal bronchioles divide to form even narrower respiratory bronchioles that
supply the alveoli with air.
− Cartilage in the trachea and the bronchi keeps these airways open and air resistance
low, & prevents them from collapsing or bursting as the air pressure changes during
breathing.
− In the trachea, there is a regular arrangement of c-shaped rings of cartilage.
− In the bronchi, there are irregular blocks of cartilage.
− Bronchioles are surrounded by smooth muscles, which can contract or relax to
adjust the diameter of these tiny airways.
− During exercise, the muscles relax to allow a greater flow of air to the alveoli.
− The absence of cartilage makes these adjustments possible.

The Structure of the Airways from the Trachea to the Alveoli

Airway Number Approx. Cartilage Goblet Smooth Cilia Site of

Diameter Cells Muscle Gas
Exchange
Trachea 1 1.8 cm Yes Yes Yes Yes No
Bronchus 2 1.2 cm Yes Yes Yes Yes No
Terminal 48,000 1.0 mm No No Yes Yes No
Bronchi
Respiratory 3,00,000 0.5 mm No No No A Few No
Bronchi
Alveolar Duct 9 X 106 400 µm No No No No Yes
Alveoli 3 X 109 250 µm No No No No Yes

− The lining is comprised of ciliated epithelium which rests on a basement membrane

made of protein fibres.
− In between the ciliated cells are goblet cells.

130
− Beneath the epithelium is an area of loose tissue with blood vessels and mucous
glands.
− The trachea as a whole is supported by c-shaped rings of cartilage, a portion of
which appears as the thick layer running across the bottom.
− There are fewer goblet cells per cm2 than in the trachea, and the epithelial cells are
not as tall. Beneath the epithelium, there are elastic fibres. Blocks of cartilage, not
rings, support the bronchus.

Warming & Cleaning the Air:

− As air flows through the nose and the trachea, it is warmed to body temperature
and moistened by evaporation from the lining, so protecting the delicate
surfaces inside the lungs from desiccation (drying out).
− Protection is also needed against the suspended matter carried in the air, which
may include dust, sand, pollen, fungal spores, bacteria, and viruses.
− All are potential threats to the proper functioning of the lungs.
− Particles larger than about 5-10 µm are caught on the hairs inside the nose and
the mucus lining the nasal passages and other airways.
− In the trachea and the bronchi, the mucus is produced by the goblet cells of the
ciliated epithelium.
− The upper part of each goblet cell is swollen with mucin droplets which have
been secreted by the cells.
− Mucus is a slimy solution of glycoproteins with many carbohydrates chains that
make them sticky and able to trap inhaled particles.
− The rest of the goblet cell which contains the nucleus is quite slender like the
stem of a goblet.
− Mucus is also made by mucous glands beneath the epithelium. Some chemical
pollutants, such as sulphur dioxide and nitrogen dioxide, can dissolve in mucus
to form an acidic solution that irritate the lining of the airways.
− Between the goblet cells are the ciliated cells.
− The continual beating of their cilia carries the carpet of mucus upward towards
the larynx at a speed of about 1 cm/min.
− When mucus reaches the top of the trachea, it is usually swallowed, so that
pathogens are destroyed by the acids in the stomach.
− Phagocytic white blood cells known as macrophages patrol the surface of the
airways scavenging small particles such as, bacteria and fine dust particles.
− During an infection, the macrophages are joined by other phagocytic cells which
are joined by other phagocytic cells which leave the capillaries to help remove
pathogens.

131
 Alveoli:

− At the end of the pathway between the atmosphere and the bloodstream are
the alveoli.
− Alveolar walls contain certain elastic fibres which stretch during inspiration
and recoil during exhalation to help force air out.
− This elasticity allows alveoli to expand according to the volume of air
breathed in.
− When the alveoli are fully expanded during exercise, the surface area
available for diffusion increases and the air is expelled efficiently when the
elastic fibres recoil.
− The alveoli have extremely thin walls, each consisting of a single layer of
epithelium cells no more than 0.5 µm thick.
− Pressed closely against the alveolar walls are the blood capillaries also with
very thin single-celled walls.
− Oxygen and carbon dioxide molecules diffuse quickly between the air and the
blood because the distance is very small.
− Diffusion is the net movement of molecules or ions down a concentration
gradient.
− So, for gas exchange to take place rapidly, a steep concentration gradient
must be maintained.
− This is done by breathing and by the movement of the blood.
− Breathing brings supply of fresh air into the lungs with a relatively high
oxygen concentration and a relatively low carbon dioxide concentration.
− Blood is brought to the lungs with a lower concentration of oxygen and a
higher concentration of carbon dioxide from the air in the alveoli.
− Oxygen therefore diffuses down its concentration gradient from the air in the
alveoli to the blood and CO2 diffuses down to concentration gradient in the
opposite direction.
− The blood is constantly flowing through and out of the lungs, so, as the
oxygenated blood leaves, more deoxygenated blood enters to maintain the
concentration gradient with each new breath.

Smoking:

(The WHO – World Health Organisation considers smoking to be a disease.)

− Tobacco smoke is composed of ‘mainstream’ smoke (from the filter or mouth end)
and ‘side stream’ (from the burning tip).
− When a person smokes, about 85% of the smoke that is released, is side stream
smoke.

132
 − Many of the toxic ingredients are in a higher concentration in side steam than in
mainstream smoke, and any other people in vicinity are also exposed to them.
− Breathing in someone else’s cigarette smoke is called passive smoking.
− The main components of cigarette smoke pose a threat to human health:
▪ Tar (which contains carcinogens)
▪ Carbon monoxide
▪ Nicotine
− In general, tar a carcinogen damages the gas exchange system, carbon monoxide and
nicotine damage the cardiovascular system.
− Tar is a mixture of compounds that settles on the lining of the airways in the lungs
and stimulates a series of change that may lead to obstructive lung diseases and
lung cancer.
− Carcinogens are cancer causing compounds. These cause mutations in the genes that
control cell division.

Lung Diseases:

− Lung diseases are a major cause of illness and death worldwide.

− Air pollution, smoking and allergic reactions are the cause of almost all cases.
− The gas exchange system is naturally efficient and adaptable.
− Healthy people breathe with little conscious effort; for people with lung diseases,
every breath may be a struggle.
− The lungs’ large surface area of the alveolar tissue is constantly exposed to moving
streams of air that may carry potentially harmful gases and particles.
− Despite the filtering system in the airways, very small particles (less than 2 µm in
diameter) can reach the alveoli and stay there.
− The air that flows down the trachea with each breath fills the huge volume of tiny
airways.
− The small particle settles out easily because the air flow in the depths of the lungs is
very slow.
− Such deposits make the lungs susceptible to airborne infections such as influenza,
pneumonia and in some people can cause allergic reactions leading to asthma.

Pneumonia:

• Persistent dry coughs that often gets worse at night

• Low grade fever
• Shortage of breath
• Fatigue or tiredness

133
 • Chest pain that gets worse when you take a deep breath or cough
• Loss of appetite

− Every disease has a characteristic set of signs and symptoms.

− Signs are the visible expression of a disease which a doctor could find my examining
a patient – for e.g.: a high temperature.
− Symptoms can not be detected by an examination and can only be reported by the
patient – for e.g.: headache or nausea.
− Often, however, the word ‘symptom’ is used more loosely to cover both signs and
symptoms.
− Chronic (long-term) obstructive pulmonary disease (COPD) such as asthma, chronic
bronchitis and emphysema are now common in many countries.
− Atmospheric pollution from vehicle and industrial emissions and tobacco smoke are
linked with these diseases.
Chronic Bronchitis:

− Tar in cigarette smoke stimulates goblet cells and mucous glands to enlarge and
secrete more mucus.
− Tar also inhibits the cleaning action of the ciliated epithelium that lines the
airways.
− It destroys many cilia and weakens the sweeping action of those that remain.
− As a result, mucus accumulates in the bronchioles and the smaller of these may
become obstructed.
− As mucus is not moved or at best only moved slowly dirt, bacteria and viruses
collect and block the bronchioles.
− This stimulate ‘smokers’ cough’ which is an attempt to move the mucus up the
airways.
− With time, the damaged epithelia are replaced by scar tissue and the smooth
muscle surrounding the bronchioles and bronchi become thicker.
− This thickening of the airways causes them to narrow and make it difficult to
breathe.
− Infections such as pneumonia easily develop in the accumulated mucus.
− When there is an infection in the lungs, the lining become inflamed and this
further narrows the airways.
− This damage and obstruction of the airways or chronic bronchitis.
− Sufferers have a severe cough, producing large quantities of phlegm, which is a
mixture of mucus, bacteria and some white blood cells.

Emphysema:

134
 − The inflammation of the constantly infected lungs causes phagocytes to leave the
blood and line the airways.
− Phagocytes are white blood cells that remove bacteria from the body.
− To reach the lining of the lungs, from the capillaries, phagocyte release the protein
digesting enzyme elastase.
− This enzyme destroys elastin in the walls of the alveoli, so making a pathway for the
phagocyte to reach the surface and remove the bacteria.
− Elastin is responsible for the recoil of the alveoli when we breathe out.
− With much smaller quantities of elastin in the alveolar walls, the alveoli do not
stretch and recoil when breathing in and out.
− As a result, the bronchioles collapse during expiration, trapping air in the alveoli,
which often burst.
− Large spaces appear where the alveoli have burst, and this reduces the surface area
for gaseous exchange.
− The number of capillaries also decreases, so less oxygen is absorbed into the blood.
− This condition is called emphysema.
− The loss of elastin makes it difficult to move air out of the lungs.
− Non-smokers can force out about 4 dm3 of air after taking a deep breath.
− Someone with emphysema may manage to force out only 1.3 dm3 of air.
− The air remains in the lungs and is not refreshed during ventilation.
− Together with the reduced surface area for gaseous exchange, this means that many
people with emphysema do not oxygenate their blood very well and have a rapid
breathing rate.
− As the disease progresses, the blood vessels in the lungs become more resistant to
the flow of blood.
− To compensates for this increased resistance, the blood pressure in the pulmonary
artery increases and over time, the right side of the heart enlarges.
− As lung function deteriorates, wheezing occurs and breathlessness become
progressively worse.
− It may become so bad in some people that they can not get out of bed.
− People with severe emphysema often need a continuous supply of oxygen through a face
mask to stay alive.
− Chronic bronchitis and emphysema often occur together and constitute a serious risk to
health.
− The gradual onset of breathlessness only becomes troublesome when about half of the
lungs is destroyed.
− Only in very rare circumstances is it reversible.
− If smoking is given up when still young, lung function can improve.

135
− In older people, recovery from COPD is not possible.
− Over 60 million people worldwide have COPD.
− The WHO predicts that COPD will become the third leading cause of death worldwide by
2030.
− There are legal controls in many countries on emissions of pollutants from industrial,
domestic and transport fuels and on conditions in work places.
− Many countries have also banned smoking in public places, such as shops, restaurants and
on public transport.

Lung cancer:
− Tar in tobacco smoke contains several substances that are carcinogens.
− These react, directly or via breakdown products, with DNA in epithelial cells to produce
mutations, which are the first in a series of changes that lead to the development of a mass
of cells, known as a tumor.
− As the cancer develops, it spreads through the bronchial epithelium and enters the
lymphatic tissues in the lung.
− Cells may break away and spread to other organs so that secondary tumors become
established.
− Lung cancer takes 20–30 years to develop.
− Most of the growth of a tumor occurs before there are any symptoms.
− The most common symptom of lung cancer is coughing up blood, as a result of tissue
damage.
− People with lung cancer also have chest pain and find it difficult to breathe.
− It is rare for a cancer to be diagnosed before it reaches 1 cm in diameter.
− A tumor like this is known as Maligant tumor.
− Tumors in the lungs are located by one of 3 methods:
1. Bronchoscopy, using an endoscope to allow a direct view of the lining of the bronchi
2. Chest X-ray
3. CT scan.

136
A Computerized tomography (CT) scan combines a series of X-ray images taken from
different angles and uses computer processing to create cross sectional images of slices.
− By the time most lung cancers are discovered, they are well advanced.
− Treatment involves surgery, radiotherapy and chemotherapy and is dependent on the
type of lung cancer, how far it has developed and whether it has spread into other areas
of the body (whether there is metastasis).
− If the cancer is small and in one lung, then either a part or all of the lung is removed.
− However, metastasis has usually happened by the time of the diagnosis so, if there are
secondary tumours, surgery will not cure the disease.
− This is why smoking is linked to so many different cancers.
− Chemotherapy with anti-cancer drugs or radiotherapy with X-rays is used.

Nicotine:
− Nicotine is the drug in tobacco.
− It is absorbed very readily by the blood and travels to the brain within a few seconds.
− It stimulates the nervous system to reduce the diameter of the arterioles and to release
the hormone ‘Adrenaline’ from the adrenal glands.
− As a result, heart rate and blood pressure increase and there is a decrease in blood supply
to the extremities of the body, such as hands and feet, reducing their supply of oxygen.
− Nicotine also increases the risk of blood clotting.
− Nicotine is a highly addictive drug that influences reward centers in the brain.
− It stimulates nerve endings in the brain to release the transmitter substance dopamine,
which is associated with reinforcing pleasurable experiences.
− This makes it very hard to give up smoking

Carbon Monoxide:
− Carbon monoxide diffuses across the walls of the alveoli and into the blood in the lungs.
− It diffuses into red blood cells where it combines with haemoglobin to form the stable
compound ‘carboxy haemoglobin’.

137
− This means that haemoglobin does not become fully oxygenated.
− The quantity of oxygen transported in the blood may be 5–10% less in a smoker than in
a non-smoker.
− Less oxygen is supplied to the heart muscle, putting a strain on it especially when the
heart rate increases during exercise.
− Carbon monoxide may also damage the lining of the arteries.
− These short-term effects are readily reversible in people who have not smoked for very
long.
− Long-term smokers put the health of their cardiovascular system at risk.
− Damage to the walls of arteries may lead to the build-up of fatty tissue and the reduction
of blood flow.
− Coronary heart disease (CHD) and stroke may be the result. These diseases are a major
cause of death and disability.
− They are responsible for 20% of all deaths worldwide and up to 50% of deaths in
developed countries.
− Cardiovascular diseases are multifactorial, meaning that many factors contribute to the
development of these diseases.
− Smoking is just one among several risk factors that increase the chances of developing
one of the cardiovascular diseases, such as CHD and stroke.

Coronary Heart Disease:

− Arteries supplying blood to the heart becomes narrowed and the blood supply to cardiac
tissues is reduced.
− Heart has to work harder to force blood through narrowed vessels/blood pressure
increases.
− Angia:
• Chest pain due to severe shortage of blood to the heart muscle cells do not die
• Pain only occurs during activity but not at rest

138
 • Caused by narrowing of coronary arteries (arthrosclerosis)
− Heart attack (Myocardial infarction):
• When a coronary artery is totally blocked by a thrombus/embolus
• No blood supply to heart muscle and cells die-often total.
− Heart Failure:
• Blockage leads to damage of heart muscles and to gradual weakening of muscle and to
gradual weakening of muscle
• Less efficient pumping
• Often accumulation of blood on right side leads to the enlargement of the heart.

Causes of coronary Heart Disease:

− The cause is atherosclerosis
− Often called hardening of the arteries
− Inner layer of artery wall thickens with deposits of
• Cholesterol
• Fibrous (Scar) tissues)
• Dead Muscle cells
• Blood platelets
− Fats in the form of lipoproteins, accumulate beneath the endothelium
• Form plague on the walls of the arteries, also called as a theroma.
• Arteries become less elastic and partially narrowed.
• Accelerated by high blood pressure
• Arterial endothelium damages are more likely but also lead to further weakening.
− Aneurys: Weak walls may burst leading to sever loss of blood (haemorrhaging)
• Brain aneurysin is called stroke
− Atheroma increases the risk of blood clotting.
• Thrombus (blood clot) may break away and lodge elsewhere in the circulation (e.g.,
brain, heart)
• Circulating thrombus is called an embolus.
− Cholesterol and Atherosclerosis:

139
− Cholesterol has important functions and is need for:
• Vitamin D production in skin
• Sex hormone production in gonads and adrenal glands.
• Making Cell membrane
• Making bile acid (salts)
− Cholesterol is an alcohol, not a fat but has properties similar to fats-soft waxy and
insoluble (difficult to remove if deposits form)
− Cholesterol is transported in the blood stream from the liver to tissues.
• Safe transport is needed due to its insolubility
• This is achieved by lipoprotein, which are soluble fatty proteins
• These are wrapped around the cholesterol
• Usually, only small amounts of free cholesterol escape.
− Fatty streaks adhere to wall of arteries
− Atheroma/atherosclerosis/plague forms
• Narrow lumen of artery
• Damages endothelium
• Can lead to formation of thrombus/blood clot

Smoking:
− Reduces levels of antitoxidants (vitamins), more damage due to release of free radials by
phagocytes
− Nicotine constricts arteries causes platelets to stick together
→ Vasoconstriction → heart must work harder to force blood through → increases
blood pressure
− Raises conc. of fibrinogen (in blood) which increases the risk of clotting
− Higher blood pressure causes damage to blood vessel lining/endothelium/collagen

140
 • Leads to rise on blood platelets and makes them more sticky/form a plug/adhere to
collagen fibres
• Release of thromboplastin/thrombokinase
• Fibrinogen converted to insoluble fibrin
• Platelet plug trapped by fibrin mesh
− Raises blood cholesterol by causing a rise in LDLs in blood
− Carbon monoxide reduces the efficiency of the blood in terms of carrying oxygen
• Haemoglobin combines with CO more readily than with oxygen → forms
carboxyheamoglobin.
• Associated with plaque formation
− Principle CHD = heart muscle receives inadequate amount of blood or
oxygen/(coronary) blood supply reduced

Treatment:
− Lower blood pressure
• Drugs which regulate heart rate/beat - prevent abnormal rhythms (beta blockers)
• Drugs which prevent blood clotting making thrombosis less likely (warfarin)
− Heart by-pass
• Vein from the leg is used to by-pass the blocked region of the coronary artery
• Involves open heart surgery
− Angioplasty
• Deflated balloon-like device is passed up to the heart via the aorta
• Guided into damaged coronary artery and inflated to stretch the artery
− Transplant
• Need to find a suitable donor
• Need to prevent rejection → drugs that suppress immune system needed for rest of life

Prevention is more cost effective then treatment

1. Screen the population for
• High blood pressure

141
• High cholesterol
• Stop smoking, healthier diet, more exercises
• Men over 35 are at highest risk
2. Monitor the behaviour of the heart during exercise
• Difficult but encouraging the population to adopt a more healthy lifestyle from an early
age is important
• Often leads to changes in diet and weight management
3. Giving up smoking and reducing alcohol intake
• Reduces blood pressure
• Many people do not heed the advice until it is too late
• Coronary heart disease is a long-term degenerative disease, starts at birth

End of Chapter
IMMUNITY
The immune response:
− Immunity: “Free from burden”, Ability of an organism to recognize and defend itself
against specific pathogens or antigens.
− The latin term ‘Immunis’ means ‘Exempt’, referring to protection against foreign agents
− Definition -The integrated body system of organs, tissues cells and cell products that
differentiates self from non self and neutralizes potentially pathogenic organisms.
− Immune Response: Third line of defense.
142
− Involves the production of antibodies and generation of specialized lymphocytes against
specific antigens.
− Antigen: Molecules from a pathogen or foreign organism that provokes a specific
immune response.
Cells of the Immune system

The immune system consists of

1. Innate Immunity –Primary Response
2. Acquired Immunity-Secondary Response
Innate or genetic immunity: Immunity an organism is born with
− Genetically determined
− May be due to lack of receptors or other molecules required for infection
• Innate human immunity to canine distemper
• Immunity of mies to poliovirus

143
Acquired Immunity: Immunity that an organism develops during lifetime
− Not genetically determined
− May be acquired naturally or artificially.
• Development of immunity to measles in response to infection or vaccination.

Types of Acquired Immunity

Naturally Acquired immunity: Obtained in the course of daily life
A. Naturally Acquired active immunity
− Antigens or pathogens enters body naturally
− Body generates an immune response to antigen
− Immunity may be lifelong (chicken pox or mumps) Or temporary (influenza or intestinal
infection)

B. Naturally acquired Passive immunity

− Antibodies pass from mother to fetus via placenta or breast feeding (colostrum)
− No immune response to antigens
− Immunity is usually short lived (weeks to months)
− Protection until child immune system develops

Phagocytosis
− If cells are under attack, they release histamine
− Histamine plus chemicals from pathogens mean neutrophils are attracted to the site of
the attack.
− Pathogens are attached to antibodies and neutrophils have antibody receptors.
− Endocytosis of neutrophil membrane makes a phagocytic vacuoles
− Lysosomes attach to phagocytosis vacuole and the pathogen is digested by the
proteases.

144
Neutrophils:
− 60% of WBCs.
− Patrol tissue as they squeeze out of the capillaries
− Large number are released during infections
− Short lived-die after digesting bacteria.
− Dead neutrophils make up a large proportion of pass.

Macrophage:
− Larger than neutrophils
− Found in the organs, not the blood
− Made in bone marrow as monocytes called marcrophages once they reach organs
− Long lived
− Initiate immune responses as they display antigens from the pathogens to the
lymphocytes.

Lymphocytes:
− Produce antibodies
− B cell mature in bone marrow then concentrate in lymph nodes and spleen
− T-cells mature in thymus
− B and T cells mature then circulate in the blood and lymph
− Circulation ensures they come to contact with pathogens and each others.

T-Lymphocytes:
− Matures T cells have T cell receptor which have a very small structure to antibodies and
are specific to one antigen.
− They are activated when the receptor come to contact with the antigen with another
host cell (e.g., on a macrophage membrane or an invaded body cell)
− After activation, the cell divides to forms:

145
 • T helper cells: Secrete cytokines
− Help B cells divide
− Stimulates macrophages
• Cytotoxic T cells (Killer T-cells)
− Kill body cells displaying antigen
• Memory T Cells
− Remains in body

T cells and Mediated Immunity

Types of T cell
1. T Helper (TH) cells: Central role in immune response
− Recognize antigen on the surface of antigen presenting cells (e.g. Macrophage)
− Activates Macrophages
− Induce formation of cytotoxic T cells
− Stimulate B cells to produce antibodies

2. Cytotoxic T cells (Tc): Destroy target cells

− Recognize antigens on the surface of all cells
− Kill host cells that are infected with viruses or bacteria
− Recognize and kill cancer cells
− Recognize and destroy transplanted tissue
− Release protein called perforin which forms a pore in target cells causing lysis of
infected cells
− Undergoes apoptosis whe stimulating antigen is gone.

3. Delayed hypersensitivity (TD) cells:

− Mostly T helper and a few cytotoxic T cells that are involved in some allergic reactions
(poison ivy) and rejection of transplanted tissue.

4. T Suppressor (Ts) cells: May shut down immune response.

146
Non specific cellular components:
1. Activated macrophages: (Stimulated phagocytosis)
− Stimulated by ingestion of antigen
− Larger and more effective phagocytes
− Enhanced ability to eliminate intracellular bacteria, virus –infected and cancerous cells

2. Natural Killer (NK) cells

− Lymphocytes that destroy virus infected and tumor cells
− Not Specific. Don’t require antigen stimulation
− Not phagocytic but must contact cell in order to lyses it.

B-Lymphocytes
− At the clone stage, antibodies do not leave the B cell
− The antibiotics are embedded in the plasma membrane if the cell and are called antibody
receptor
− When the receptors in the membrane recognizes and the antigen on the surface of the
pathogen of the cell divides rapidly.
− The antigens are presented to the B cells by macrophages
− Some activated B cells form plasma cells. These produce lots of antibodies < 1000/sec
− The antibodies travel to the blood, lymph, lining of gut and lungs.
− The number of plasma cells goes down after a few weeks.
− Antibodies stay in the blood longer but eventually their numbers fo down too.
− Some activated B cells form memory cells
− Memory cells divide rapidly as soon as the antigen is reintroduced.
− There are many more memory cells than there were alone cells
− When the pathogen/infection infects again it is destroyed before any symptoms show.

147
 Assembled Antibody Molecule

− Also known as immunoglobulins

− Globular proteins
− The heavy and light chains are polypeptides
− The chains are held together by disulphide bridges
− Each antibody has 2 identical antigen binding sites
− They are known as Variable regions
− The order of amino acids in the variable region determine the shapes of the binding site

Now Antibodies work

− Some act as labels to identify antigen for phagocytes
− Some work as antitoxin i.e., they block toxins for those causing diphtheria and tetanus
− Some attach to the bacteria flagella making them less active and easier for phagocytes to
engulf

148
− Some case agglutination (lumping together) of bacteria making them less likely to
spread.

Consequences of antigen-antibody binding

Antigen-antibody complex: Formed when an antibody binds to an antigen it recognizes

Affinity: A measure of binding strength

1. Agglutination: Antibodies causes antigens (microbes) to clump together.
• IgM (decavalent) is more effective than 1 g (bivalent)
• Hemagglutination: Agglutination of red blood cells
Used to determine ABO blood types and to detect influenza and Measle viruses.

2. Opsonization: Antigen (Microbe) is covered with antibodies that enhances its ingestion
and lysis by phagocytic cells

Antibody Response after Exposure to Antigen

Different Immunoglobins:
Type # of Site of action Function
antigen
binding
sites

149
 IgG 2 Blood Increase Macrophages activity
Tissue Fluid Antioxins
Can Cross placenta Agglutination
Ig M 10 Blood Agglutination
Tissue Fluid
Ig A 2 or 4 Secretions (saliva, Stops bacteria from adhering to host
tears, small intestine, cells
vaginal prostate, Prevents bacteria forming colonies or
nasal, breast milk) mucous membranes
IG E 2 Tissue Activates mast cells
− HISTAMINE worm response

Antigens
− Most are protein or large polysaccharides from a foreign organism.
• Microbes: Capsule, cell wall, toxin, viral capsids, flagella etc.
• Non microbes: Pollen, egg white, red blood cell surface molecules from transplanted
tissues.
− Lipids and nucleic acids are only antigenic when combined with proteins and
polysaccharides.
− Molecular weight of 10000 or higher
• Hapten: Small foreign molecule that is not antigen. Must be coupled to a carrier
molecule to be antigenic. One antibodies are formed they will recognize hapten.
Epitope:

• Small part of an antigen that interacts with an antibody.

• Any given antigen may have several epitopes.
• Each epitope is recognized by a different anti body.

Vaccination

1. Injection into vein or muscle.

150
2. Oral.

Why are not they always effective?

• Natural infection persists within the body for a long time so the immune system has
time to develop an effective response, vaccination from dead mice do not do this.
Less effective vaccines need booster injections to stimulate secondary responses.

Smallpox

Symptoms:

• Red spots containing transparent fluid all over the body.

• Spots fill with pus.
• Eyelids swell and become glued together.

Mortality:

• 12-30% died.
• Survivors often left blind and disfigured with scabs.

Eradication Programme:

• Started by WHO in 1956.

• Aimed to rid world of smallpox by 1977.
• Involved vaccination and surveillance.
• Over 80% of population at risk of the disease was vaccinated.
• After any reported case, everyone in the household and 30 surrounding households were
vaccinated. This is called RING VACCINATION.
• Last case of smallpox was reported in Somalia in 1977.
• World declared free of smallpox in 1980.

Eradication programme –Why was it successful?

• Variola virus was stable because it was cheap and everyone used the same vaccine.
151
• Vaccine was made from harmless strain of similar virus (vaccine).
• Vaccine could be used at high temperatures.
• Easy to identify infected people.
• Smallpox does not lie dormant in body.

Eradication programme –Why do not all work?

• Political instability
• Poor infrastructure
• Unstable

Measles

• Caused by airborne virus.

• 9th leading cause of death worldwide.
• Causes rash and fever.
• Can have fatal complications.
• Passive immunity from mothers in infants under 8 months.
• Now quite a rare disease in developed countries due to vaccination.
• Transmitted easily in overcrowded and insanitary conditions.
• Mainly affects malnourished infants with Vitamin A deficiencies.
• Responsible for many cases of childhood blindness and can cause severe brain damage.
• Herd immunity of 93-95% needed to prevent transmission within a population.

Allergies

• When the immune system responds to harmless substances.

• Allergens – antigenic substances which do not real harm.
• Allergens include house dust, animal skin, pollen, mite and its feces.
• Histamine causes blood vessels to widen and become leaky.
• Fluid and white blood cells leave capillaries.
• The area of leakage becomes hot, red and inflamed.

152
 Asthma
• Vaccines are being developed to make allergic responses less severe.
• Designed to desensitize people so they do not produce antibodies.
• Genetic tests may be used to screen children and then a vaccine could be given to prevent
them from developing asthma.

Overview of the immune response

HUMORAL (antibody-mediated) CELL-MEDIATED IMMUNE RESPONSE

IMMUNE RESONSE

153
 Defend against extracellular pathogens Defend against intracellular pathogens
by binding to antigen and making and cancer by binding to and lysing the
them easier targets for phagocytes infected cells or cancer cells
and complement

Monoclonal Antibodies
• Mouse is exposed to non-self material that carries antigen against which an antibody is
required.
• A different B-lymphocyte within the mouse produces a mixture of antibodies & cells are
excreted from the spleen of the mouse.
• To enable these B-lymphocytes to divide outside the body, they are mixed with cells that
divide readily outside the body.
• A fusogen is added to the mixture, to allow the surface membrane of both these cells to fuse
together.

154
• These fused cells are called hybridoma cells.
• The hybridoma cells are separated out and each single cell is grown into a group of
identical cells from the single ancestor hybridoma cells (clone).
• Each clone is tested to see if it is producing the required antibody.
• Any group producing the required antibody is grown in a large scale and the antibody is
excreted from the growing medium.
• The antibody then comes from cloned cell from a hybridoma cell, they are monoclonal
antibodies.
• As a mouse was used as the host organism to produce the B-lymphocytes, the monoclonal
antibodies, have to be modified to make them human before they can be used. This process
is known as humanization.

Uses of Monoclonal Antibodies:

• Antibodies are very specific to particular proteins (antigens). They can be used to target
specific substances, e.g. Specific cells.
• Monoclonal antibodies can be used to treat cancer, if they are specific to an antigen on a
cancer cell.
• The antibodies attach to the receptors on the cancer cell, block the chemical signals that
stimulate uncontrolled growth.
• The advantage of monoclonal antibody therapy is that since the antibody are host to mice,
they lead to fever as a side effect that other forms of therapy.
• Since a B-lymphocyte recognizes a specific antigen, monoclonal antibodies can be used to
distinguish between different strains of a disease causing organism.
• Detection of HIV in a blood sample.
• Blood typing helps a blood transfusion. For e. g. A, AB, B or O and Rhesus negative or
positive group.
• Tissue typing before tissue transplant operation.
• Identification of the particular type of leukemia.
• Location of tumors.

155
 End of Chapter

Transport in Plants

Why transport systems are needed in plants?

• To move substances from where they are absorbed to where they are needed. For example-
water and mineral ions are absorbed by roots and transported in the xylem to other parts
of the plant.
• To move substances from where they are produced to where they are needed for
metabolism.
• For example- sugars are produces in leaves, but glucose is needed by all parts of the plant
for respiration and for converting to cellulose for making cell walls in areas of growth.
• Glucose can be moved in a phloem as part of the sucrose molecule.
• To move substances to a different part of the plant for storage-for example, to move sugars
into a potato tuber for storage in the form of starch.
• Unlike animals, plant do not have systems for transporting carbon dioxide and oxygen
• Instead these gases diffuse through air spaces within stems, roots and leaves.
• Carbon dioxide: photosynthetic plants require a supply of carbon dioxide during daylight.
• Most photosynthetic tissue is in the leaves & most plants have evolved thin , flat leaves with
a large surface area ideal for absorbing as much as carbon dioxide as possible.
• They obtain this by diffusion from the air.
• Oxygen – All plants require a supply of oxygen for respiration, but cells which are actively
photosynthesizing produce more than enough oxygen for their own needs because oxygen
is a waste product of photosynthesis.
• Cells which are not photosynthesizing need to take in oxygen from their environment.
• Plants have much lower demands than animals, so they respire at much lower rates.

156
• They therefore do not need such a rapid supply of oxygen.
• The branching shape of plants and a network of air spaces in the plant body provide a large
enough surface area for effective absorption of oxygen by diffusion.

TWO SYSTEMS:
XYLEM AND PHLOEM
Plants have two transport systems: xylem and phloem
Vascular Tissue: Trees
Vascular tissue is located on the outer layers of the tree.

• Xylem carries mainly water and inorganic ions (mineral salts) from roots to the parts above
the ground.
• The xylem sap contained in the xylem can move in only one direction, from roots to the rest
of the plant.
• Phloem carries substances mode by photosynthesis from the leaves to the areas of the plant.
• At any one time, phloem sap can be moving in different direction in the different plants of
the phloem.
• In neither of these systems do fluids move as rapidly as flood does in a mammal, nor is there
an obvious pump such as the heart. Neither plant transport system carries CO2 or O2 which
travel to and from cells and their environment by different alone.

157
 Xylem Phloem
Made of Dead cell Living cells
Cell wall thickness Thick Thin
Cell wall material Lignin (rigid) Cellulose
Permeability Impermeable Permeable
Cytoplasm None Cytoplasm lining
Transports Water and minerals Food
Carried to Leaves Growing parts and
storage organs
Direction of flow Upwards Up and down
Tissue also has Fibers Companion cells

Tissue Process What is moved Structure

• Xylem • Transpiration • Moves water and• Columns of
minerals from hollow dead
roots to leaves reinforced cells
• Phloem • Translocation • Moves food • Columns of living
substances from
leaves to the rest
of the plant

Flowering plants (angiosperms) may be:

• Monocotyledons ( monocots) or dicotyledons (dicots)

Monocotyledons: Plants, such as grasses, typically have long narrow leaves.

Dicotyledons: Plants typically have leaves with blades and stalks (petioles).

158
 The Mechanism of transport through both types of plants are the same, but there are
differences in the distribution of xylem and phloem in their roots, stems and leaves.

Monocotyledons & Dicotyledons

• Flowering plants (angiosperms) may be monocots or dicots.

• Monocotyledonous plants such as grasses typically have long, narrow leaves.
• Dicotyledonous plants typically have leaves with blades and stalks (petioles).

Parenchyma

• Parenchyma is made up of thin – walled cells used as packing tissue.

• The cells are very metabolically active and may be used for many functions.
• They may be used for storage of foods like starch.
• Where they are turgid (fully inflated with water) they help to support the plant, preventing
it from wilting.
• Air spaces between the cells allow gas exchange.
• Water and mineral salts are transported to through the walls and through the living
contents of cells.
• Parenchyma forms the cortex in roots and stems and the pith in stems.
• The cortex is an outer region of cell.
• The pith is made up of similar cells that is the name given to the central region of stems.
• Parenchyma contains chloroplasts in leaves where it is motif to form to form the palisade
and spongy mesophyll.

Collenchyma:
• Collenchyma is a modified form of parenchyma with extra cellulose deposited at the
corners of the cells.
• This provides extra strength
• The midrib of leaves contain collenchymas.

159
 Endodermis:
• The endodermis, like the epidermis, is one cell thick.
• It surrounds the vascular tissue in stems and roots.

Mesophyll:
• ‘Meso’ means middle and ‘phyll’ means leaf.
• The mesophyll is made up of specialised parenchyma cells found between the lower and
upper epidermis of the leaf.
• They are specialised for photosynthesis and therefore contain chloroplasts.
• There are two types: palisade mesophyll and spongy mesophyll.
• Palisade means column – shaped.
• Spongy mesophyll is so called because in there dimension, it has many large spaces
between the cells.
• Palisade mesophyll cells are near the upper surface of the leaf where they receive more
sunlight. They contain more chloroplast than spongy mesophyll cells.

Pericycle:
• This is a large of cells one to several cells thick just inside the endodermis and next to the
vascular tissue
• In root, it is one cell thick and new roots can grow from this layer
• In stems, it is joined from a tissue called sclerenchyma
• This has dead lignified cells for extra strength

Vascular tissue:
• Xylem and phloem both contain more than one type of cell and together make the vascular
tissue.
• Vascular means having tubes for transporting fluids.
• Xylem contains tubes called vessels made from dead cells called xylem vessel elements.
• The walls of the cells are rain forced with a strong waterproof material lignin.

160
• Xylem allows long distance transport of water and mineral salts. It also provides
mechanical support for strength.
• In roots, it is at the center and has series of arms between which the phloem found.
• In stems, the xylem and phloem are found in bundles called vascular bundles.
• The outside of these bundles have caps made of sclerenchyma fibres which provide extra
support for the stem.
• Sclerenchyma fibres are long dead empty cells with lignified walls. They provide
mechanical support.
• The distribution of the strengthening tissues xylem and sclerenchyma is different in roots
and stems.
• This reflects the fact that these organs are subjected to different stresses and strains.
• Stems for example, need to be supported in air whereas roots are usually spreading through
soil and are subjected to pulling strains from the parts above the ground.
• In the roots and stems of trees and shrubs, extra xylem is made forming wood.
• Phloem contains tubes called sieve tubes made from living cells called sieve tube elements.
• These allow long distance transport of organic compounds, particularly the sugar, sucrose.

The transport of water

• The movement of water is passive as it is driven by evaporation from the leaves.
• The process starts in the leaves.
• The energy of the sun causes water to evaporate from the leaves, a process called
TRANSPIRATION.
• This reduces the water potential in the leaf and sets up a water potential gradient throughout
the plant.
• Water moves down this gradient from the soil into the plant- for example: through its root
hairs. Water then moves across the root into the xylem tissue in the centre.
• Once inside the xylem vessels, the water moves upwards through the root to the stem and
from there into the leaves.
From leaves to atmosphere- transpiration

161
• The air in the internal spaces of the leaf has direct contact with the air outside the leaf
through small pores called stomata.
• If there is a water potential gradient between the air inside the leaf (high water potential)
and the air outside (lower water potential), then water vapour will diffuse out of the leaf
down this gradient.
• Some of the water in the leaf will be used in photosynthesis and some evaporates and
diffuses out of the leaf by the process of transpiration.
*TRANSPIRATION IS THE LOST OF WATER VAPOUR FROM A PLANT TO ITS ENVIRONMENT
BY DIFFUSION DOWN A WATER POTENTIAL GRADIENT. MOST TRANSPIRATION TAKES
PLACE THROUGH THE STOMATA IN THE LEAVES.

WATER MOVEMENT THROUGH A LEAF

Water is, in effect, being pulled through the plant as a result of transpiration and evaporation.
Movement of water through the plant is therefore known as the transpiration stream.
1. Water vapour diffuses from an air space through open stomata, a process called
transpiration. It is carried away from the leaf surface by air movements. This reduces the
water potential inside the leaf.
2. Water evaporates from a mesophyll cell wall into the air space.
3. Water moves through the mesophyll cell wall or out of mesophyll cytoplasm into the cell
wall.
4. Water leaves a xylem vessel through our non-lignified area such as a pit. It may enter the
cytoplasm or cell wall of a mesophyll cell.
5. Water moves up the xylem vessels to replace the water loss from the leaf.

Factors Affecting Transpiration

HUMIDITY: if the water potential gradient between the air spaces in the leaf and the air
outside becomes steeper, the rate of transpiration will increase.
• In conditions of low humidity, the gradient is steep so transpiration takes place more quickly
than in high humidity.

162
 WIND SPEED AND TEMPERATURE: Transpiration may also be increases by an increase in
wind speed or rise in temperature.

LIGHT INTENSITY: In most plants, stomata open during the day and close at night. Most
transpiration takes place through the stomata (although a little water vapour can escape
through the epidermis if the cuticle is thin), so the rate of transpiration is almost zero at
night. Stomata must be open during the day to allow carbon dioxide to diffuse into the leaf
for photosynthesis. This inevitably increases the rate of transpiration. Closing at night, when
photosynthesis is impossible, reduces unnecessary water loss.

VERY DRY CONDITIONS: In especially dry conditions when the water potential gradient
between the internal air spaces and the external air is steep, a plant may have to compromise
by partially or completely closing its stomata to prevent its leaves from drying out, even if
this means reducing the rate of photosynthesis.
• In hot conditions, transpiration plays an important role in cooling the leaves. As water
evaporates from the cell wall inside the leaf, it absorbs heat energy from these cells thus
reducing the temperature.
• If the rate at which water vapour is lost by transpiration exceeds the rate at which a plant
can take up water from the soil, then the amount of water in its cells decreases. The cells
become less turgid and the plant wilts as the soft parts such as the leaves lose the support
provided by turgid cells. In this situation, the plant will also close its stomata.

Xerophytes
Xerophytes (or xerophytic plants) are plants that live in places where water is in short
supply. Many xerophytes have evolved special adaptations of their leaves that keep water
loss down to a minimum.
Marram grass (Ammophilia arenaria)
• This grass grows on sand dunes where conditions are very dry.

163
• The leaves can roll up due to shrinkage of special hinge cells exposing a thick waterproof
cuticle to the air outside the leaf.
• The cuticle contains a fatty relatively waterproof substance called cutin.
• The stomata are found only in the upper epidermis and therefore open into the enclosed
humid space in the middle of the roll.
• Hairs help to trap a layer of moist air close to the leaf surface, reducing the steepness of the
diffusion gradient for the water vapour.

Opuntia:
• It is a cactus with flattened photosynthetic stems that store water.
• The leaves are reduced to spines which lessens the surface area from which transpiration
can take place and protects the plant from being eaten by animals.

Sitka spruce:
• This is a large tree native to Canada and Alaska. Its leaves are in the form of needles, greatly
reducing the surface area available for water loss. In addition, they are covered in a layer of
waterproof wax and have sunken stomata.

Phlomis italica:
• Small shrub that lives in dry habitats.
• On the leaves there are tiny hairs like structures that act as a physical barrier to the loss of
water.

Euphorbia canariensis:
• This shrub grows in dry areas of Tenerife.
• It has swollen; succulent stems that store water and photosynthesis.
• The stems are coated with wax, which cuts down water loss.
• The leaves are extremely small.

164
 Sunken Stomata:
• Sunken stomata is a stomata in a small pit which protects the escaping water vapour from
air currents, decreasing water loss from the leaf. Sunken stomata are a feature of many plants
in deserts and other dry environments.

From xylem across the leaf:

• As water evaporates from the cell walls of mesophyll cells, more water is drawn into the
walls to replace it.
• This water comes from the xylem vessels in the leaf.
• Water constantly moves out of these vessels through the unliginfied parts of the xylem
vessels walls.
• The water then moves down a water potential gradient from cell to cell in the leaf along two
possible pathways.
1. In one possible pathway, known as the symplastic pathway, water moves from cell to cell via
the plasmodesmata.
2. In the other pathway, known as the apoplastic pathway, water moves through the cell walls.

Xylem tissues:
• The xylem tissue is made from cells joined end to end for tubes.
• The cells are dead.
• The walls of the cells are thickened with a hard strong material called lignin.
• The xylem has two functions:
1. Support
2. Transport
• Contains several different types of cell.
• In flowering plants xylem tissue contains vessels elements, tracheids, fibers and parenchyma
cells.

Vessel elements and tracheids are the cells that are involved with the transport.

165
• Flowering plants rely mostly on the vessel elements for their water transport.
• Sclerenchyma fibers are elongated cells with lignified walls that help support the plant.
• They are dead cells; they have no living contents at all.

Xylem vessels and vessel element

- Xylem vessels are made up of many elongated cells called vessels elements,
arranged end to end.
- Each vessels element begins life as a normal plant cells, whose lignin is laid down.
- Lignin is a very hard, strong substance, which is impermeable to water.
(wood is made up of xylem and gets its strength from lignin)
- As lignin builds up around the cell, the contents of the cell die, leaving a completely
empty space or lumen inside.
- However, in those parts or the original cell walls, where groups of plasmodesmata
are found, no lignin is laid down.
- These non-lignified areas can be seen as ‘gaps’ in the thick walls of xylem vessel and
are called pits.
- Pits are not open pores, they are crossed by permeable, unthickened cellulose cell
wall.
- The pits in one cell link with those in the neighboring cells so water can pass freely
from one cell to the next.
- The end walls of neighboring vessel elements break down completely to form a
continuous tube rather like a drainpipe running through the plant.
- This long non-living tube is a xylem vessel.
- It may be up to several meters long. The structural features of xylem vessels are
closely related to their function.

166
Annular or Ring-Like Thickening

- The deposition of linin takes place in the form of rings on the inner surface of the
cell wall.
- These lignified rings are placed one above the other like coins, leaving sufficient
space in between each other.
- The gaps of the walls remain unthickened. Such thickenings are commonly found in
the vessels and tracheids.

Spiral Thickening

- In such case, the deposition of thickening material (lignin) takes place in the form of
complete spiral bands.
- The number of such bands may be one or more than one.
- This type of thickening is commonly found in the vessels or tracheae of
angiosperms.

From Root to Stem and Leaf in the Xylem

- The removal of water from xylem vessels in the leaf reduces the hydrostatic presser
in the xylem vessels.
- Hydrostatic pressure is pressure exerted by a liquid.
- The hydrostatic pressure at the top of the xylem vessel becomes lower than the
pressure at bottom.
- This pressure difference causes water to move up the xylem vessels in continuous
columns.
- (it is just like sucking up water in straw)

167
 - When you suck a straw, you reduce the pressure at the top of the straw causing a
pressure difference between the top and bottom.
- The higher pressure at the bottom pushes water up the straw.
- The lower the hydrostatic pressure, the lower the water potential, so hydrostatic
pressure gradient is also a water potential gradient.
- The water in xylem vessels, like the liquid in a ‘sucked’ straw, is under tension.
- If you suck hard on a straw, its walls may collapse inward as a result of the pressure
difference.
- Xylem vessels have strong, lignified walls to stop them from collapsing in this way.
- The movement of water of xylem vessel is by mass flow.
- This means that all the water molecules (and any dissolved solutes) move together,
as a body of liquid, like water in a river.
- This is helped by the fact that water molecules are attracted to each other by
hydrogen bonding. This attraction is called COHESION.
- They are also attracted to the cellulose and lignin in the walls of the xylem vessels,
and this attraction is called ADHESION.

Cohesion and adhesion help to keep the water in a xylem vessel moving as a
continuous colum.

- These vessels are full of water. The fact that the cells are dead is an advantage,
because it means there is no protoplasm to get in the way of transport.
- If an air bubble forms in the column the column of water break and the difference in
the pressure between the water at the bottom cannot be transmitted through the
vessel.
- We say there is an air lock.
- The water stops moving upwards. the smalldiameter of xylem vessels helps to
prevent such break from occurring.
- Also, the pits in the vessel walls allow water to move out into neighbouring vessels
and so bypass such an air lock.

168
 - Air bubbles cannot pass through pits.
- Pits are also important because they allow water to move out of the xylem vessels to
surrounding living cells.
- The xylem tissue in dicotyledonous stem is arranged in a series of rods around the
center of the stem. These strong rods help to support the stem.

Root Pressure:

- Transpiration reduces the water (hydrostatic) pressure at the top of a xylem vessel
compared with the pressure at the base, so causing the water to flow up the vessels.
Plants may also increase the pressure difference at the base of the vessels.
- The pressure is raised by the active secretion of solutes, for example: mineral ions
into the water in the xylem vessels use energy to pump solutes across their
membranes and into the xylem by active transport. The presence of the solutes
lowers the water potential of the surrounding root cells. This influx of water
increase the water pressure at the base of the xylem vessel.
- Although root pressure may help in moving water up xylem vessels, it is not
essential and is probably not significant in causing water to move up xylem in most
plants.
- Water can continue to move up through xylem if the plant is dead.
- Water transport in plant is largely a passive process driven by transpiration from
the leaves. The water simply moves down a continuous water potential gradient
from the soil to the air.

From Root Hair to Xylem:

- The xylem vessels are in the center of the root, unlike the arrangement in stems,
where they are arranged in a ring, and are nearer to outside.
- Water taken up by root hair crosses the cortex of the root and enters the xylem in
the center of the root. It does this because the water potential inside the xylem
vessels is lower than the water potential in the root hair. Therefore, water moves
down this water potential gradient across the root.
- The water takes two routes through the cortex

169
 - Individual molecules can switch from one route to the other at any time. The cells of
the cortex like all plant cells are surrounded by cell walls made of several layers of
cellulose fibres crisscrossing one another. Water can soak into these walls rather as
it would soak into blotting paper and can seep across the root from cell wall to cell
wall without ever entering the cytoplasm of the cortical cells.
- This is called the apoplastic pathway.

Appoplastic pathway:

1. Water enters the cell wall.

2. Water moves through the cell wall.
3. Water may move from cell wall to cell wall through the intercellular spaces.
4. Water may move directly from cell wall to cell wall.
- Another possibility is for the water to move into the cytoplasm or vacuole of a
cortical cell by osmosis and then into adjacent cells through the interconnecting
plasmodesmata.
- This is the symplastic pathway.
- The relative importance of these two pathways varies from plant to plant and in
different conditions.
- Normally, it is probable that the symplastic pathway is more important, but when
transpiration rate are especially high, more water travels by apoplastic pathway.
- Once the water reaches the endodermis, the apoplastic pathway is abruptly blocked.
The cells in the endodermis have a thick waterproof waxy band of suberin in their
cell walls.
- This band, called the Casparian Strip, forms an impenetrable barrier to water in the
walls of the endodermis cells.

170
 Water moves into the root hair by osmosis down a water potential gradient.

- Although soil water contains some inorganic ions in a solution, it is a relatively

dilute solution and so has a relatively high water potential.
- However, the cytoplasm and cell sap inside the root hair have considerable
quantities of inorganic ions and organic substances such as proteins and sugars
dissolved in them and so have a relatively low water potential.
- Water therefore diffuses down this water potential gradient through the partially
permeable cell surface membrane and into the cytoplasm and vacuoles of the root
hair cell.
- The large number of very fi ne root hairs provides a large surface area in contact
with the soil surrounding the root, thus increasing the rate at which water can be
absorbed.
- However, these root hairs are very delicate and often only function for a few days
before being replaced by new ones as the root grows.

171
 - Root hairs are also important for the absorption of mineral ions such as nitrate and
magnesium.
- Many plants, especially trees, have fungi located in or on their roots, forming
associations called mycorrhizas, which serve a similar function to root hairs.
- The mycorrhizas act like a mass of fi ne roots which absorb water and nutrients,
especially phosphate, from the soil and transport them into the plant.
- Some trees, if growing on poor soils, are unable to survive without these fungi.
- In return, the fungi receive organic nutrients from the plant.
- The name given to a relationship such as this, in which two organisms of different
species both benefit, is mutualism.

Transport of mineral ions

- Apart from the carbohydrates made in photosynthesis, plants need a supply of
mineral ions to complete their nutrition.
Ex: nitrate, phosphate, sulfate, potassium, magnesium and calcium.
- Mineral ions in solution are absorbed along with water by the roots, particularly by
the root hair cells.
- Their route through the plant is the same as that for water, crossing the root by
apoplastic and symplastic pathways before moving in the mass flow of xylem sap up
the xylem to therest of the plant.
- From the xylem they enter the apoplastic and symplastic pathways again.
- As well as moving by mass flow through the apoplastic pathway and xylem, mineral
ions can also move by diffusion and active transport.
Eg. they can diffuse into the apoplastic pathway of the root from the soil and once
In the apoplastic pathway can diffuse in any direction according to concentration
gradients.
- They can also enter cells by diffusion, facilitated diffusion and active transport.
- Facilitated diffusion and active transport allow cells to control what ions enter or
leave cells.
- One important control point is the root endodermis, where the Casparianstrip
forces ions to pass through living cells before theycan enter the xylem.
172
Translocation
Translocation is the movement of organic solutes (e.g. sucrose) from a source to a sink
through the phloem by means of mass flow.

PHLOEM
SOURCE SINK
(eg. Palisode cells) (eg. root cells)

- The term translocationcan be applied to transport inboth xylem and phloem

- It means literally moving from one place to another.
- It tends to be used more commonly to describe the transport of soluble organic
substances within a plant.
- These are substances which the plant itself has made – for eg. sugars which are
made by photosynthesis in the leaves.
- These substances are sometimes called ASSIMILATES.
- Assimilates are transported in sieve elements.
- Sieve elements are found in phloem tissue, along with several other types of cells
including companion cells, parenchyma and fibres.

Sieve tubes and sieve elements

- Phloem contains unique tube-like structures called sieve tubes. Unlike xylem
vessels, sieve tubes are made of living cells.
- A sieve tube is made up of many elongated sieve elements (also known as sieve tube
elements), joined end to end vertically to form a continuous tube.
- Each sieve element is a living cell.
- Like a normal plant cell, a sieve element has a cellulose cell wall, a cell surface
membrane and cytoplasm containing endoplasmic reticulum and mitochondria.
- However, the amount of cytoplasm is very small and only forms a thin layer lining
the inside of the wall of the cell.

173
 - There is no nucleus, nor are there any ribosomes.
- Where the end walls of two sieve elements meet, a sieve plate is formed.
- This is made up of the walls of both elements, perforated by large pores.
- These pores are easily visible with a good light microscope.
- In living phloem, the pores are open, presenting little barrier to the free flow of
liquids through them.
- It is not easy to collect enough phloem sap to analyse its contents.
- The contents of sieve tubes are under very high pressure.
- When a sieve tube is cut, the release of pressure inside the tube causes a surge of its
contents towards the cut. When the contents come up against a sieve plate, they may
block it.
- This helps to prevent escape of the contents of the sieve tube. Then, within minutes,
the sieve plate is properly sealed with a carbohydrate called callose, a process
sometimes called clotting.
- However, castor oil plants are unusual in that their phloem sap does continue to
flow from a cut for some time, making it relatively easy to collect.
- In other plants, aphids may be used to sample sap.
- Aphids such as greenfly feed using tubular mouthparts called stylets. They insert
these through the surface of the plant’s stem or leaves, into the phloem.
- Phloem sap flows through the stylet into the aphid.
- If the stylet is cut near the aphid’s head, the sap continues to flow, it seems that the
small diameter of the stylet does not allow sap to flow out rapidly enough to trigger
the plant’s phloem clotting mechanism.

How translocation occurs

- Phloem transport is an active process, in contrast to the passive transport in xylem.
- In leaves, sugar is synthesized in mesophyll cells (the middle layer of leaf) and is
then actively pumped into the phloem, using metabolic energy.
- Any area of a plant in which sucrose is loaded into the phloem is called a source.
- This is usually a photosynthesising leaf or a storage organ.

174
 - Any area where sucrose is taken out of the phloem is called a sink – for example the
roots.
- Loading a high concentration of sucrose into a sieve element greatly decreases the
water potential in the sap inside it.
- Therefore, water enters the sieve element, moving down a water potential gradient
by osmosis.
- This causes a correspondingly high build up in pressure (equivalent to about six
times atmospheric pressure).
- The pressure is referred to as hydrostatic pressure, turgor pressure or pressure
potential.
- A pressure difference is therefore created between the source and the sink.
- This pressure difference causes a mass flow of water and dissolved solutes through
the sieve tubes, from the high pressure area to the low pressure area.
- At the sink, sucrose may be removed and used, causing the water to follow by
osmosis, and thus maintaining the pressure gradient.
- Sinks can be anywhere in the plant, both above and below the photosynthesising
leaves.
- Thus, sap flows both upwards and downwards in phloem (in contrast to xylem, in
which flow is always upwards).
- Within any vascular bundle, phloem sap may be flowing upwards in some sieve
tubes and downwards in others, but it can only flow one way in any particular sieve
tube at any one time.

Loading sucrose into phloem

- In leaf mesophyll cells, photosynthesis in chloroplasts produces triose sugars, some
of which are converted into sucrose.
- The sucrose, in solution, then moves from the mesophyll cell, across the leaf to the
phloem tissue.
- It may move by the symplastic pathway, moving from cell to cell via plasmodesmata.
- Alternatively, it may move by the apoplastic pathway, travelling along cell walls.

175
 - At the moment, little is known about how the sucrose crosses the cell surface
membrane of the mesophyll cell to enter the apoplastic pathway.
- Which of these routes – the symplastic route or the apoplastic route – is more
important depends on the species.
- It is now known that the companion cells and sieve elements work together.
- Sucrose is loaded into a companion cell or directly into a sieve element by active transport.

-
- Hydrogen ions (H+) are pumped out of the companion cell into its cell wall, using
ATP as an energy source.
- This creates a large excess of hydrogen ions in the apoplastic pathway outside the
companion cell.
- The hydrogen ions can move back into the cell down their concentration gradient,
through a protein which acts as a carrier for both hydrogen ions and sucrose at the
same time.
- The sucrose molecules are carried through this co-transporter molecule into the
companion cell, against the concentration gradient for sucrose.
- The sucrose molecules can then move from the companion cell into the sieve tube,
through the plasmodesmata which connect them (the symplastic pathway).

176
Unloading sucrose from phloem
- Unloading occurs into any tissue which requires sucrose.
- It is probable that sucrose moves out of the phloem into these tissues using both
symplastic and apoplastic routes, as with loading.
- Phloem unloading requires energy, and similar methods to those used for loading
are probably used.
- Once in the tissue, the sucrose is converted into something else by enzymes, so
decreasing its concentration and maintaining a concentration gradient.
- One such enzyme is invertase, which hydrolyses sucrose to glucose and fructose.

Differences between sieve tubes and xylem vessels

177
 - Unlike water transport through xylem, which occurs through dead xylem vessels,
translocation through phloem sieve tubes involves active loading of sucrose at
sources and unloading at sinks, thus requiring living cells.
- Xylem vessels have lignified cell walls, whereas phloem sieve tubes do not.
- This is an advantage, because it means the cells can be dead and therefore entirely
empty. Water can therefore flow unimpeded, and the dead xylem vessels with their
strong walls also support the plant.
- Sieve tubes also reduce resistance to flow by having only a thin layer of cytoplasm
and no nuclei, but whereas the end walls of xylem elements disappear completely,
those of phloem sieve elements form sieve plates.

End of chapter

Photosynthesis
Introduction:

178
2 main Types of Nutrition
Autotrophic Nutrition
Chemoautotroph
Photoautotroph

Heterotrophs

Discovery of Photosynthesis
- C.B. Van Niel, 1930
- Proposed a general formula:
- CO2 + H2O + light energy → CH2O + H2O + 2O
- Where H2A is the electron donor.
- Van Niel identified water as the source of O2 released from photosynthesis.
- Robin Hill (in the 1950) confirmed Van Niel’s proposal that energy from the light
reaction tents carbon fixation (making glucose from CO2)
- The process by which organism use carbon dioxide and water to manufacture food.
- The energy needed for synthesis is supplied by light which is absorbed by the
organisms and subsequently converted by them into chemical energy in the
presence of chloroplast.

Structure of chloroplast
- Biconvex shape
- 3-10 micro meter in diameter
- Double membrane

179
 - Internal membrane- flattened fluid filled sacs called thylakoid
- Pigments- chlorophyll- held – thylakoid membrane
- Cluster – pigment with primary pigment – surrounded by accessory pigment
- Enzymes – ATP synthase – found – membrane of grana
- Fluid substances surrounding grana – called stroma
- Stroma – lipids and starch grains
- Stroma – contains enzymes – involved – in the calvin cycle
- Carbon dioxide – fixed and starch – made in stroma
- Stroma – 70s ribosome & double stranded circular DNA
- Shape of chloroplast – varies between different species.

Adaptation of Palisade mesophyll cells for photosynthesis

- Closely packed, so absorbs more incident light
- Cells are found near the surface of leaf to maximise light interception
- Cells are arranged at right angles to the leaf to reduce the number of light absorbing
walls.
- Cylinder in shape to produce air spaces between cells.
- Intercellular air spaces act as reservoir for carbon dioxide.
- Provide large surface area for gas exchange
- Cell walls are relatively thin for short diffusion pathway
- Large vacuole pushes chlorophyll in the chloroplast, so it can absorb more light
efficiently.
- Large numbers of chloroplasts maximise the amount of light absorbed.
- Chloroplasts also move within the cell towards the light
- Higher light intensity, chloroplast move away from light to avoid any damage.

See figure on page no. 197

Photosynthetic Pigments
Chlorophyll a – Primary pigment in plants and cyanobacteria (Porphyrin Mg)
- Absorbs violet – blue and red light.
180
Chlorophyll b – secondary pigment absorbing light wavelength that chlorophyll a does not
absorb.
Accessory Pigments- Secondary pigments absorbing light wavelength other than those
absorbed by chlorophyll a
- Increase the range of light wavelength that can be used in photosynthesis.
- Includes chlorophyll b, carotenoids, phycobiloproteins
- Carotenoids also act as antioxidants.

Photo system:

• Pigment molecules in chloroplast are organized into photosystems.

• A photosystem consist of:

181
 1) An antennae complex (Light harvesting complex) of hundreds of accessary
pigment molecules that gather photons and feed energy to reaction center.
2) A reaction center one or more chlorophyll-A molecule pass electrons out of the
photosystem (photochemical reactions).
• There are 2 types of photosystems:
➢ Photosystem-I has an absorption pear of 700 nm.
➢ Photosystem-II has an absorption pear of 680 nm.

Absorption and action spectrum:

• Absorption spectrum of pigment is a record of amount of light absorbed at each

wavelength.
• Chlorophyll-a and Chlorophyll-b absorbs red and blue/ violet parts of the spectrum.
• Xanthophyll and Carotene absorbs light from other parts.
• This effectively increases the range of wavelength from which plants can obtain
energy.
• Action spectrum is a record of amount of photosynthesis occurring at each
wavelength of light.
• There is a close similarity between the absorption spectra of the major
photosynthetic pigments and the action spectrum for photosynthesis.

Chemistry of photosynthesis:

• Photosynthesis consist of 2 Stages-

1) Light dependent reaction (takes place in grana) Hills Reaction.
2) Light independent reaction (takes place in stroma) Calvin Cycle.

1) Light Dependent reaction:

• It consists of light harvesting and electron transport.
• ATP synthesized from ADP + P
• NADH is reduced to NADPH.
• O2 is formed from water.

182
 • ATP is synthesized by using light energy = Photophosphorylation.

There are two ways in which ATP can be synthesised by Photophosphorylation:

1. Non cyclic photophosphorylation

2. Cyclic photophosphorylation

Diagram/Flow chart of noncyclic photophosphorylation

Non-cyclic photophosphorylation:
183
 • Non cyclic photophosphorylation involves both Photosystem I and Photosystem II
• Light is absorbed by photosystem II and passed on to chlorophyll a
• The irradiated chlorophyll a molecules emits two electrons.
• The electron acceptor passes the electrons along the chain of electrons carriers to
photosystem I
• The energy released from the electron is used to make ATP from ADP and P+
• Light is absorbed by Photosystem I and passes on to chlorophyll a
• The energised electrons rise to a high energy level and are picked up by a second
electron acceptor
• Since both chlorophyll have now lost electrons, they will both be positive and
unstable
• The two electrons released from the chlorophyll a Photosystem II go to replace the
two that have been lost by chlorophyll a of photosystem I
• Chlorophyll a of photosystem II receives its replacement electron from the saplings
of water (Photolysis)
• During photolysis, water molecule dissociates in to electrons, hydrogen iron &
oxygen.
• Electrons go to photosystem II. The Oxygen is released as a waste gas
• The H+ IONS combine with e- held by the second acceptor to give NADPH ( Reduce
NADP)
• This passes to the reaction of the light independent stage
• So, the product of the light dependent stage are NADP, ATP & O2 (..)

Cyclic photophosphorylation:

• Involves Photosystem I only

• Light is absorbed by photosystem I and passed on to chlorophyll a
• This causes the chlorophyll a molecules to emit an electron
• The energised electron is raised to a higher energy level and is picked up by an
electron acceptor.

184
 • The electron is passed along a chain of electron carries before it is returned to the
chlorophyll-a molecule.
• As the electron passes along the electron carries chain, enough energy is released to
make ATP from ADP and P +. This ATP is needed for the light independent stage.
• No NADPH is made during cyclic photophosphorylation.

Diagram/ flowchart of cyclic Photophosphorylation:

Chemiosmosis powers ATP synthesis in the light reaction:

• The electron transport chains are arranged with the photosystem in the thylakoid
membranes and pumps H+ back through the membrane.
• The flow of H+ back through the membrane is harnessed by ATP synthase to make
ATP.
185
• In the stroma, the H+ ions combine with NADP+ to form NADPH.

186
The production of ATP by chemiosmosis in photosynthesis:

Distinguish between Non-cyclic and cyclic photophosphorylation:

Features Non-cyclic Cyclic

photophosphorylation photophosphorylation
Conditions under which When plant require ATP 2 When plant require only
process occurs NADPH ATP
Pathway of electrons Non-cyclic Cyclic
For electron donor Water Photosystem-I
(Source)

187
Last e- acceptor NADP+
(destination)
Establishing protons High H+ conc. In the
gradient for the synthesis thylakoid space is due to
of ATP photolysis of H2O and active
transport of H+ from the
stroma, across the thylakoid
membrane and into the
thylakoid space.
Products ATP, NADPH and oxygen
Photosystem-I
High H+ conc. In the
thylakoid space is due to
active transport of H+ ions
from the stroma, across the
thylakoid membrane into
the thylakoid space.
Only ATP.

2) The light independent reactions ( Calvin Cycle):

• This cycle pathway is sometimes called the Calvin cycle
• CO2 combines with a 5C compound, ribulose biphosphate (RuBP).
• The reaction is catalyzes by the enzyme RuBP carboxylase, the most common
enzyme in the world.
• The product is an unstable 6C compound that breaks down to form 2 molecules
of 3 carbon (Phosphate glycerates (3C)).
• ATP is used to phosphorylates the 2 molecules of phosphoglycerates to form two
molecules of glycerate bisphosphonate of 3C.
• The next stage involves the use of NADPH to reduce each molecules of glycerate
biphosphate to glyceraldehyde 3-phosphate (GALP).
• For every six molecules of GALP formed five are used in a series of reactions to
generate ribulose biphosphate, which can then combine with more CO2.
• One of the six GALP molecule is converted to glucose and other carbohydrates,
amino acids and lipids.

Role of NADP in photosynthesis:

188
 • NADP are called Co-enzyme.
• It serves as electron acceptor in the electron transport system during
photophosphorylation.
• Electrons are released in excited chlorophyll in photosystem-I and they travel to
electron receptors.
• From electron acceptor, they pass down a chain called electron transport chain and
ends with NADP.
• During photolysis, photons from splitting water molecules to reduce NADP to
NADPH.
• Reduced NADP passes along the electrons on the Calvin cycles.
• During cycle Glyceraldehyde-3 phosphate is reduced to 3C sugar called triose
phosphate by reduced NADP.
• NADP then reused to light dependent stage to pick up another H+ ion.

A) photosystem consists of a light- harvesting complex and a reaction

center:

Photosystem-I Photosystem-II
Located at the outer surface of the grana Located at the inner surface of grana
thylakoid membrane thylakoid membrane
The photo center is P700 The photo center is P680
Pigment absorb longer λ of light (> 680 Pigment absorbs shorter λ of light
nm). (<680nm).
Participates in cyclic as well as non- Participates only in non-cyclic
cyclic photophosphorylation. photophosphorylation.
It is not associative with photosynthesis It is associated with photosynthesis of
of water water
Main function is ATP synthesis. Main function are ATP synthesis and
photolysis of water.

189
Factors necessary for photosynthesis:

• The presence of a suitable photosynthetic pigment, a supply of CO2, H2O and light
energy are required for photosynthesis.
• The main external factors affecting the rate of photosynthesis are light intensity and
wavelength, temperature and CO2 concentration.

Light and temperature

• In the early 1900s. F.F. Backman investigated the effects of light intensity and
temperature on the rate of photosynthesis
• At higher, light intensities. This relationship varies with the light intensity, initially
increasing as the light intensity increases
• At higher light intensities, this relationship no longer holds and the rate of
photosynthesis reaches a plateau
• At higher light intensity, the rate of photosynthesis increases as the temperature is
increased over limited energy
• At low light intensity, increasing the temperature has little effects on the rate of
photosynthesis
• These two experiments illustrate two important points -firstly from other research,
We know that photochemical reactions are not generally affected by temperature.
• However, those experiments clearly shows that temperature effects rate of
photosynthesis, so there must be two set of reaction in the full process.
• These are the light dependent photochemical stage and a light independent
temperature dependent stage.
• Secondly, Backman’s experiment illustrate the concept of limiting factors

Limiting factors:

• The rate of any process which is dependent on a series of reactions is limited by

the slowest reaction in the series

190
 • In Biochemistry, if a process is affected by more than one factor, the rate will be
limiting by the factor which is nearest its lowest value.
• At low light intensities, the limiting factor governing the rate of photosynthesis
in the light intensity, one or more other factors must be limiting such as
temperature or carbon dioxide supply
• Not all wavelengths can be used in photosynthesis
• This mean that the wavelength of light reach a plants leaves may limit its rate of
photosynthesis.
• At constant light intensity and temperature, rate of photosynthesis initially
increases with an increasing concentration of C02, but then again, reaches a
plateau at higher concentrations.
• A graph of rate of photosynthesis at different concentration of Co2 has the same
shape as that for different light intensities
• At lower concentration of Co2, the supply of C02 is the rate of limiting factors
• At higher concentration of Co2, other factors are rate limiting (e.g. Light density
of temperature )
• The effects of these limiting factors on the rate of photosynthesis are easily
investigated by using an aquatic plant such as elodea or calomba in a sample
apparatus
• The number of bubbles of gas (mostly o2) produced in unit time from a cut stem
can be counted in different condition
• Alternately, the gas can be collected and the volume produced in unit time can be
measured.
• This procedure depends on the facts that the rate of production of o2 is a
measure of the rate of photosynthesis

Carbon dioxide:

• Air contains 0.0.3 percent of Co2. It is released by respiration, construction of fossil

fuel and microbe decomposition

191
 • During early morning hours and evening hours, Co2 is released in respiration is
sufficient for photosynthesis. At this stage, there is no exchange of gases between
the plant and environment. This is called compensation point
• An increase in the concentration of c02 up to 0.1 percent increase the rate of
photosynthesis
• Higher concentration of Co2 decreases the rate of photosynthesis.

Water:

• Plant absorb water and mineral salts through root hairs and pass it to the leaves
through xylem
• If there is less availability of H2o, then stomata closes (to reduce the water loss by
transpiration) and there is decreases of Co2 absorption and sunlight absorption
• Therefore, the rate of photosynthesis decreases

Chlorophyll :

Only cells having chlorophyll are photosynthetic. There is no proportionality between rate
of photosynthesis and amount of chlorophyll

Growing plants in protected environment

• An understanding of effect of environmental factors on the rate of environmental

factors on the rate of photosynthesis their management when crops are grown in
protected environment such as glass houses.
• The aim is to increase the yield of crop concerned
• For example many hectares of tomato plants are grown in glass houses.
• In the most sophisticated of these, sensor monitor, the light intensity, the humidity
of atmosphere and conc. Of co2 around the plants
• The plants grow hydroponically i.e. with the roots in a nutrient solutions whose
nutrient content can be varied at different shape of plants grown
• All of these factors are managed by a computer to maximise the yield of the crop

192
 • Such glass house-grown crops have added advantage that insects pets and fungal
disease are more easily controlled than is possible with field grown crops, further
improving yield

C4 Plants

• C4 Plants are often called tropical or warm season plants

• Corn sugarcane, sorghum and millets, as well as the switchgrass which has been
utilized as a source of biofuel are some examples
• In light dependent stage of photosynthesis, Co2 combine with RUBP to form a 6C
compound which immediately splits to form 2 3C molecules. Plants that do this are
called C3 plants, which are temperate or cool season pants
• However maize and sorghum plants and most other tropical plants grasses do
something different. The first compound that is produced in light independent
reaction contains 4C atoms, they are therefore called C4 plants
• All the plants make energy during the Calvin cycle, It is the process where plants
take up C02 and turn it into sugar and energy
• However, in hot summer , sunny dry climate, C4 plants are much more efficient than
C3 plants, which are most common type.

Avoiding photorespiration:

• Photorespiration is a wasteful pathway that occurs when the Calvin cycle enzyme,
RUBISCO, acts on O2 rather than Co2
• Rubisco catalyses two different reactions
1. Avoiding C02 to Ribulose biphosphate for the carboxylase activity
2. Avoiding O2 to ribulose biphosphate for oxygenase activity

Which one predominates depends on the relative concentration of o2 and Co2 with high
C02 and low o2 favouring the carboxylase reaction

High O2 and low Co2 favours the oxygenase reaction

193
The light reaction for the photosynthesis liberates oxygen and more oxygen dissolves in the
cytosol of the cell at higher temperature.

• Therefore, high light intensities and high temperature (above 30°C) favors the
second reaction.
• Tropical gases such as maize, sorghum and sugar have evolved a method of avoiding
photorespiration.
• They keep RuBP and Rubisco well away from high O2 concentrations.
• The cells that contain RuBP and Rubisco are arranged around the vascular bundles,
and are called bundle sheath cells.
• They have no direct contact with the air inside the leaf.
• CO2 is absorbed by another group of cells (mesophyll cells) which are in contact
with air.
• The mesophyll cells contain an enzyme called PEP carboxylase, which catalyzes the
combination of CO2 from the air with a 3C substances called phosphoenolpyruvate
(PEP).
• The compound formed from this reaction is oxaloacetate.

Ring of mesophyll cells:

• This tight ring of specialized mesophyll cells excludes air from the cell inside the
ring. The cytoplasm fixes CO2 the chloroplast capture light and carry out the light
dependent reactions, but not the Calvin cycle.

Bundle sheath cells:

• The bundle sheath cells carry out the Calvin cycle but not the light dependent
reactions. No air gets to these cells and they get CO2 from the mesophyll cells.
• Still inside the mesophyll cells, the oxaloacetate is converted to malate and this is
passed on to the bundle sheath cells.
• Now the CO2 is removed from the malate molecules and delivered to RuBP by
Rubisco in the normal way.

194
 • The light independent reaction then proceeds as usual.
• Enzymes in C4 plants generally have higher optimum temperature than those in C3
Plants.
• This is an adaptation to growing in hot climates.
• For examples, in one study, it was found that in Amarnath, which is a C4 plant, the
optimum temperature for activity of PEP carboxylase is around 45°C.
• If the temperature drops to 15°C, the enzyme loses around 70% of its activity.
• By contrast, the same enzyme in peas, which are C3 plants, was found to have an
optimum temperature of around 30°C and could continue to work at much lower
temperatures than Amarnath.

Comparison of photosynthesis in C3 and C4:

C3 Plants C4 Plants
Calvin cycle used Yes Yes
Primary CO2 acceptor RuBP PEP
CO2 fixing enzyme Rubisco PEP carboxylase
First product of CO2 fixation 3PG (3C) Oxaloacetate (4C)
Affinity of carboxylase for Moderate High
CO2
Photosynthetic cells of leaf Mesophyll cells Mesophyll and bundle
sheath
Photorespiration Extensive Minimal

• You can easily extract chloroplast pigments from a leaf too see how many pigments
are present by using paper chromatography.
• You can calculate the Rf value for each pigment using the equation,

𝑅𝑓 = 𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑝𝑖𝑔𝑚𝑒𝑛𝑡𝑠/𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑛𝑡

195
Regulating stomatal opening: The potassium ion pump- Hypothesis:

• ATP lowered proton pump actively transport H+ out of the guard cells.
• Low H+ conc. And negative (-ve) charge inside cell causes K+ channels to open.
• K+ diffuses into guard cells down an electrochemical gradient.
• Increased conc. of K+ in guard cells.
• Lower Ψ in guard cells, so H2O moves in by osmosis, down Ψ gradient.
• Entry of H2O increases vol. in guard cells.
• Thin outer wall of guard cells expands most, so the cells curve apart

RF value

RF ( retention factor) value in chromatography is the distance travelled by a given

component divided by the distance travelled by the solvent front.

This will vary depending on the solvent used but in general carotenoids have RF values
close to 1

Chlorophyll b has much lower RF values

Chlorophyll a has RF values between those of carotenoids and chlorophyll b

End of chapter

196
 Respiration

The need for energy in living organisms:

• All living organism require a continuous supply of energy to stay alive either
from absorption of light energy or from chemical potential energy (energy
stored in nutrient molecules)
• The process of photosynthesis transfers light energy to chemical potential
energy and so almost all life on earth depends on photosynthesis either directly
or indirectly
• Photosynthesis supplies living organism with two essential requirements
1. An energy supply
2. Unstable carbon compound
• All biological molecules such as carbohydrates, lipids and proteins and nucleic
acids contain carbon.
• All living organism therefore need source of carbon
• Organism that can use inorganic carbon source in the form of co2 are called
autotrophs
• Those feeding a ready made organic supply of carbon are heterotrophs
• An organic molecule is a compound including carbon and hydrogen
• The term originally meant a molecule derived from an organism but now
includes all compounds of carbon and hydrogen, ever they do not occur that
naturally
• Organic molecule can be used by living organism in two ways
1. They can serve as building brick for making other organic molecule that are
essential to the organism
2. They can represent chemical potential energy that can be released by
breaking down molecule in respiration
3. This energy can be used for all form of work
4. Heterotrophs depends on autotrophs for both material and energy

197
 5. The cycling of matter and the flow of energy within the ecosystem occur
through the interactions among different organism and physical
environment
All living systems needs matter and energy
• Matter feels the energy – releasing chemical reactions that provide energy for life
functions and provides the material for growth and repair of tissues.
• Energy from light is needed by plants because the chemical reactions that produces
plant matters from air and water requires an energy input to occur.
• Animals acquire matter from food, that is, from plants or other animals.

• Transfer of materials.
• Transfer of energy.

198
Working in a living organism includes:

• The synthesis of complex substances from simpler ones (anabolic reactions) such as
the synthesis of polysaccharides from monosaccharide, lipid from glycerol and fatty
acids, polypeptides from amino acids and nucleic acids from nucleotides.
• The active transport of substances against a diffusion gradient, such as the activity
of the sodium – potassium pump.
• Mechanical work such as a muscle contraction and other cellular movements for eg.
The movement of cilia and flagella, amoeboid movements and the movement of
vesicles through the cytoplasm.
• In a few organism bioluminescence and electrical discharge
• Mammals and birds used thermal energy, released from metabolic reaction to
maintain constant body temperature
• Most animals are ectotherms
• The thermal energy that wants them comes from outside their bodies

Ectotherms

These organisms control their body temperature through external means

They depend on environmental sources

Their body temperature rises and fall with their environment

Endotherms

Mammals and birds are endotherms, releasing enough thermal energy within their bodies
to maintain them above the temperature of their surrounding when necessary

They also maintain a constant body temperature through negative feedback loops

For a living organism to do work, energy- requiring reactions must be linked to those that
yield energy

199
In the complete oxidation of glucose (C6H12 O6) In aerobic condition, a large quantity of
energy is made possible

C6H12O6 + 6O2 6CO2+ 6H2O + 2870 KJ

- Reactions take place in a series of small steps, each releasing a small quantity of the
total available energy.
- Multi-step reactions allow precise control via feedback mechanisms.
- Moreover, the cell could not usefully control and make use of the total available
energy if all of it were made available at one instant.
- Although the complete oxidation of glucose to CO2 and H2O has a very high energy
yield, the reaction does not happen easily.
- Glucose is actually quite stable, because of the activation energy that has to be added
before any reaction takes place.
- In living organisms, the activation energy is overcome by lowering it by using
enzymes, and by also raising the energy level of the glucose by photophosphorylation.
- Photophosphorylation: A biochemical process that involves the addition of phosphate
to an organic compound.
- Examples include the addition of phosphate to glucose to produce glucose
monophosphate and the addition of phosphate to adenosine diphosphate (ADP) to
form ATP.
- Activation energy is the least amount of energy required to activate atoms or
molecules to a state in which they can undergo a chemical reaction.

200
- Theoretically, the energy released from each step of respiration could be controlled
and used directly to some form of work in the cell.
- However, a much more flexible system actually occurs in which energy yielding
reactions in all organisms are used to make an intermediate molecule ATP.

ATP as energy currency

- When a phosphate group is removed from ATP, ADP is formed and 30.5 kJmol−1 of
energy is released.
- Removal of a second phosphate produces adenosine monophosphate (AMP), and 30.5
kJ mol−1 of energy is again released.
- Removal of the last phosphate, leaving adenosine, releases only 14.2 kJ mol−1.
- In the past, the bonds attaching the two other phosphate groups have been called
high-energy bonds, because more energy is released when they are broken than when
the phosphate is removed.
- This description is misleading and should be avoided, since the energy does not come
simply from breaking those bonds, but rather from changes in chemical potential
energy of all parts of the system.

201
 Hydrolysis of ATP (P1 is inorganic phosphate - H3PO4)
- These reactions are all reversible. It is the inter conversion of ATP and ADP that is all-
important in providing energy for the cell:

- The rate of inter conversion, or turnover, is enormous.

- It is estimated that a resting human uses about 40 kg of ATP in 24 hours, but at any
one time contains only about 5 g of ATP.
- During strenuous exercise, ATP breakdown may be as much as 0.5 kg per minute.
- The cells energy-yielding reactions are linked to ATP synthesis.
- The ATP is then used by the cell in all forms of work.
- ATP is the universal intermediary molecule between energy-yielding and energy-
requiring reactions used in a cell, whatever the type of cell.
- In the words, ATP is the ‘energy currency’ of the cell.

- The cell ‘trades’ in ATP, rather than making use of a number of different
intermediates.
- ATP is a highly suitable molecule for this role.
- Not only it is readily hydrolysed to release energy, it is also small and water soluble.

202
- This allows it to be easily transported around the cell.
- Energy transfers are in efficient
- Some energy is converted to thermal energy whenever energy is transferred
- At the different stages in a multistep reaction such as respiration, the energy made
available may not perfectly correspond with energy needed to synthesize ATP.
- Any excess energy is converted to thermal energy
- Also many energy requiring reactions in cells use less energy than that released by
hydrolysis of ATP to ADP
- Again any extra energy will be released as thermal energy.

The difference between energy currency and energy storage.

- Be careful to distinguish between molecules used as energy currency and energy

storage.
- An electron currency molecule as the immediate donor of energy to the cells - energy
requiring reactions.
- An energy molecule is a short-term (glucose or sucrose) or long term (glycogen,
starch or triglyceride) store of chemical potential energy.

Synthesis of ATP

- Energy for ATP synthesis can become available in two ways.

- In respiration,
1. Energy released by reorganizing chemical bonds (chemical potential energy)
during Glycolysis and the Krebs cycle is used to make some ATP.
2. However, most ATP in cells is generated using electrical potential energy.
- This energy is from the transfer of electrons by electron carriers in mitochondria and
chloroplasts.
- It is stored as a difference in proton (H+) concentration across some phospholipids
membranes in mitochondria and chloroplasts, which are essentially impermeable to
protons.

203
- Protons are then allowed to flow down their concentration gradient (by facilitated
diffusion) through a protein that spans the phospholipids bilayer.
- Part of this protein acts as an enzyme that synthesizes ATP and is called ATP synthase.
- The transfer of 3 protons allows the production of one ATP molecule, provided that
ADP and an inorganic phosphate group (Pi) are available inside the organelle.
- This process occurs in both chloroplasts and mitochondria.
- The process was first proposed by Peter Mitchell in 1961 and is called chemiosmosis.
*H H + + e-

- Chemiosmosis involves the pumping of protons though special channels in the

membranes of mitochondria from the inner to the outer compartment.
- The pumping establishes of proton (H+) gradient.
- After the gradient is established, protons diffuse down the gradient throw transport
protein called ATP synthase.
- The flow of hydrogen catalyzes the pairing of a phosphate with ADP, forming ATP.
- ATP synthase has 3 binding sites and a part of the molecule that rotates and hydrogen
ion (H+) pass.
- This produces structural changes in the binding sites and allows them to pass
sequentially through three phases.
1. Binding ADP and Pi
2. Forming tightly bound ATP
3. Releasing ATP.

The role of ATP in Active Transport

- Active transport is the movement of molecules or ions across a partially permeable

membrane against a concentration gradient.
- Energy is needed, from ATP, to counteract the tendency of these particles to move by
diffusion down the gradient.
- All cells show differences in the concentration of ions, in particular sodium and
potassium ions, inside the cell with respect to the surrounding solution.

204
- Most cells seem to have sodium pumps in the cell surface membrane that pump
sodium ions out of the cell.
- This is usually coupled with the ability to pump potassium ions from the surrounding
solution, into the cell.
- The sodium–potassium pump is a protein that spans the cell surface membrane.
- It has binding sites for sodium ions (Na+) and potassium ions (K+) ions on the outer
side.
- The protein acts as an ATPase and catalyses the hydrolysis of ATP to ADP and
inorganic phosphate, releasing energy to drive the pump.
- Changes in the shape of the protein moves sodium and potassium ions across the
membrane in opposite directions.
- For each ATP used, two potassium ions move into the cell and 3 Na+ move out of the
cell.
- As only two potassium ions are added to the cell contents for every 3 Na+ removed, a
potential difference is created across the membrane that is negative inside with
respect to the outside.
- Both sodium and potassium ions leak back across the membrane, down their
diffusion gradients.
- However, cell surface membranes are much less permeable to sodium ions than
potassium ions, so their diffusion actually increases the potential difference across
the membrane.
- This potential difference is most clearly seen as the resting potential of a nerve cell.
- One of the specializations of a nerve cell is an exaggeration of the potential differences
across the cell surface membrane as a result of the activity of the Na+ - K+ pump.
- The importance of active transport in ion movement into and out of cells should not
be underestimated.
- About 50% of the ATP used by a resting mammal is devoted to maintaining the ionic
content of cells.

Structure of mitochondria

205
 - It is cylindrical in shape or rod shape.
- Has width range from 0.5 micrometer and length from 3-10 micrometer.
- It is bounded by double membrane.
- Outer and inner membrane is separated by intermembrane shape.
- Intermembrane is extensively folded to form protein called cristae.
- Cristae is projected into a semi fluid called matrix.
- Matrix contains circular DNA molecule and 70 s ribosome.
- Endosymbiont theory

Function of mitochondria
- It is involved in cellular respiration.
- Series of biochemical reaction can result in formation of ATP.
- It is often known as power station of the cell.
- More than 1000 mitochondria are found in metabolically active cells.
Cell respiration can be divided into four parts:

1. Glycolysis
2. Oxidation of Pyruvate/ Transition reaction
3. Kreb’s cycle
4. The electron transport chain and chemiosmotic phosphorylation

Where do the 4 parts of the Cellular Respiration Take Place?

- Glycolysis – Cytosol/cytoplasm
- Oxidation of pyruvate: Matrix
- The Kreb’s cycle: Matrix
- The electron transport chain and chemiosmotic phosphorylation : Cristae

1. Glycolysis:

- ‘Glyco’ means glucose and ‘lysis’ means breakdown

- It involves the membrane of enzyme controlled reactions

206
- It takes place in the cytoplasm of the cell.
- It does not require O2.
- Common for both aerobic and anaerobic respiration.

- Glucose is phosphorylated by ATP to glucose-6-phosphate.

- Phosphorylated glucose is no longer recognized by the glucose transport system,
therefore it is trapped inside the cell.
- Enzyme involved in this is kinase.
207
 - Glucose-6–phosphate is isomerised to fructose–6–phosphate.
- Enzyme involved in this is isomerase.
- Fructose–6–phosphate is phosphorylated by ATP to fructose 1,6 phosphate.
- Enzyme involved in this is kinase.
- Fructose–1,6–phosphate splits to glycerate 3 phosphate.
- Glycerate 3 - phosphate converts to pyruvate.
- Glycerate 3- phosphate, when converted to pyruvate, also forms NADH and ATP.
End product of Glycolysis:

- 2 molecules of ATP (4 molecules are produced, but 2 molecules are used up)
- 2 molecules of NADH
- 2 molecules of pyruvate

2. Oxidation of pyruvate/ Transiton Reaction

How does it take place?

- Pyruvate in matrix of mitochondria from cytoplasm

- Pyruvate is decarboxylated (remove of carbon in the form of CO2)
- Pyruvate is dehydrogenated (removal of hydrogen)
- Hydrogen is transferred to hydrogen acceptor.
NaD+ → NADH
- Pyruvate is converted to Acetate
- Acetate combines with co-enzyme A to form Acetyl Coenzyme A.

208
End products of Transition Reaction/Oxidation of Pyruvate

- CO2 → 2 molecules
- 2 molecules of NADH
- 2 molecules of Acetyl Coenzyme A

3. Kreb’s Cycle

How does it take place?

- Discovered by Sir Haan Krebs in 1937.
- Citric Acid Cycle / Tricarboxylic acid (TCA).
- Occurs in matrix of mitochondria
- Occurs only in aerobic respiration
- Acetyl Coenzyme A (2C) + Oxalate (4C) → Citrate
- These reaction is condensation reaction
- Enzyme involved here is citrate synthetase.
- Citrate isomerizes to isocitrate (6C)
- Isocitrate undergoes oxidative decarboxylation to give α- ketoglutarate (5C)
- CO2 is produced by decarboxylation
- NAD+ is a hydrogen acceptor and NADH is formed
- H+ is released from isocitrate (6C) through dehydrogenation
- The enzymes involved are isocitrate decarboxylase and isocitrate dehydrogenase
- α- ketoglutarate (5C) undergoes oxidative decarboxylation and dehydrogenation
gives Succinyl coenzyme A (COA) (UC)
- CO2 is produced and NADH is formed
- Enzymes used are α- Ketoglutarate dehydrogenase and α-Ketoglutarate
decarboxylase
- Succinyl CoA (UC) Succidade
- ATP is formed from ADP and Pi
- Enzyme used is succinyl COA synthetase
- Succinate undergoes dehydrogenation and gives Fumarate (4C)
- FAP (Flavine adenine- dinucleotide) is a hydrogen acceptor and forms FADH2

209
 - Enzyme used is Succiccate dehydrogenase
- Fumarate undergoes hydrogenation and give oxaloacetate (4C)
- (NAD+) + (H+) gives NADH
- Enzyme used is Malato dehydrogenase

End Products of the Kreb’s Cycle:

- 6 molecules of NADH
- 2 molecules of FADH
- 2 molecules of ATP
- 4 molecules of CO2

4. The Electro Transport Chain

- O2 is required during the final stage of aerobic reaction.

- Oxidative-phosphorylation is process by which ATP is formed as electrons are
transferred from NADH and FADH to oxygen via series of electron carriers.
- Location for this to happen is in the inner membrane of the mitochondria
- Electron transport chain involves chain of electron carrier molecule.
- Electron from NADH and FADH2 are transferred to oxygen.
- Series of reactions are known as Redox Reactions.
- Hydrogen atoms split to hydrogen ion and e-.
- Transfer of electrons along the chain releases sufficient energy to make ATP.
- Electron and hydrogen ion form hydrogen atom.
- Then hydrogen is passed on to oxygen to form water.

Chemiosmotic Theory

- Energy from electron transport chain is linked to pumping H+ from matrix to space
between two membranes.
- Results in higher concentration of H+ in inner membranal space and electro chemical
gradient is set up.

210
 - Hydrogen ion passes into the matrix through stalk granules.
- Electrical potential energy is used to make ATP from ADP+Pi
- ATP synthetase catalyzes the reaction.
- NADH and FADH2 is formed during the glycolysis and Kreb’s cycle are passed to ETC.
- ETC – present – inner membranal space and consists of cytochromes.
- NADH and FADH2 are oxidized and H2 is released.
- H2 now splits into H+ and e-.
- e– pass along the electron carrier chain and transferred to oxygen.
- Protons (H+) are actively pumped from the matrix to the outer compartment i.e. inter-
membranal space.
- A proton gradient is created between the outer compartment and inner matrix.
- H+ cannot diffuse through the cristae membrane.
- H+ only flow down the gradient- matrix through ATP synthase channels.
- This is known as chemiosmosis.
- Protons flow – ATP synthase channel (F1 Channels) they generate energy to photo
phosphorylate ADP to ATP in the presence of enzyme ATP synthase.
- Later, proton combines with oxygen to form water.
1
2𝑒 − + 2𝐻 + 𝑂2 → 𝐻2 𝑂
2
Summary

- Glycolysis – 2 ATP
- Kreb’s cycle - 2 ATP (one per turn)
- Glycolysis - 2NADH
- Link reaction - 2 NADH
- Kreb’s cycle - 6 NADH (3 per turn)
- Kreb’s cycle - (one per turn) 2 FADH2
- 1 NADP - 3 ATP
- 1 FAD - 2 ATP

ANAEROBIC RESPIRATION

211
212
213
214
215
216
Chapter 10
Infectious diseases
Learning outcome
• Explain what is meant by infectious and non-infectious diseases.
• Name the organisms that cause some infectious diseases.
• Explain how cholera, measles, malaria, TB HIV/AIDS are transmitted.
• Discuss ways in which biological, social and economic factors influence the prevention
and control of cholera, measles, malaria, TB, HIV/AIDS.
• Discuss the factors that influence global patterns of malaria, TB and HIV/AIDS.
• Outline how penicillin acts on bacteria and why antibiotics do not affect viruses.
• Explain how bacteria become resistant to antibiotics.
• Discuss the consequences of antibiotic resistance and how we can reduce its impact.

What is a disease?

• An abnormal condition affecting an organism, which reduces the effectiveness of the

functions of the organism.
• The word ‘disease’ implies something very serious, yet many conditions that make us feel
ill, such as the common cold.
• ‘Disease’ is a difficult word to define satisfactorily as it covers a wide range of human
conditions.
• A disease is an illness or disorder of the body or mind that leads to poor heath; each
disease is associated with a set of signs and symptoms.
• The study of disease is called pathology.
• See the picture in this section given on pg no. 234
Infectious and non-infectious
• Infectious diseases are diseases that are caused by organisms known as pathogen.
• They are sometimes called communicable disease as they passed from infected to
uninfected people.
• Some also affect animals and are passed from animals to humans.

217
• See the figure given in this section pg no. 234
• A disease that is not contagious is called a non-infectious disease. These diseases are not
caused by pathogens.
• Instead, they are likely to have causes such as life style factors, environmental toxins, or
gene mutations.
• Common types of non-infectious diseases include cancer, diabetes, COPD, and immune
system disease.
• These are long-term degenerative disease.
• Inherited or genetic diseases, such as cystic fibrosis and sickle cell anemia, are another
group of non-infectious diseases.
• There are other categories of non-infectious disease, including deficiency diseases that
are caused by malnutrition and mental diseases.
• See the image in pg no. 234 in this section.
• Many infectious diseases, such as the, cold, measles and influenza, only affect us for a
short period of time.
• Others such as tuberculosis (TB) may last a much longer time.
• Indeed, in the case of HIV/AIDS, there is yet no cure and treatments must be taken for
the whole of a person’s life.
• Some infectious disease can only spread from one person to another by direct contact,
because the pathogenic cannot survive outside the human body.
• Other pathogens can survive in water, human food, faces or animals (including insects),
and so are transmitted indirectly from person to person.
• Some people may spread a pathogen even though they do not have the disease
themselves.
• People like this who lake of symptoms are called carriers and it can be very difficult to
trace them as the source of an infection.
• The way in which a pathogen passes from one host to another is called the transmission
cycle.
• Control methods attempt to break transmission cycles by removing the conditions that
favors the spread of the pathogens.

218
• Control is only possible once the cause of disease and its method of transmission are
known and understood.
• Vaccination is a major control measure for many infectious diseases;
• It works by making us immune so that pathogens do not live and reproduce within us
and do not then spread to others.

Worldwide importance of infectious diseases

• Five infectious diseases of worldwide importance are discussed in detail in this chapter;
• CHOLERA, MALARIA, HIV/AIDS, TUBERCULOSIS (TB), MEASLES
• Small pox is the only infectious disease that has so far been eradicated.
• The number of people infected with cholera, malaria, tuberculosis, HIV/AIDS, measles,
particularly children and young adults, remains very high, and these diseases pose
serious public health problems now and for the foreseeable future.
• This is particularly true in parts of the world without the efficient health services
available in affluent countries

Causative agents of diseases

Disease Causative agent (Pathogen) Types of organism

Cholera Vibrio cholerae Bacterium
Malaria Four species of Plasmodium Protoclist
HIV/AIDS Human immuno deficiency Virus
virus (HIV)
Tuberculosis (TB) Mycobacterium tuberculosis Bacteria
and M. bovis
Measles 8 species of Marbilivirus Virus
Small pox (Irradicated) Variola virus virus

Endemic, epidemic and pandemic

219
• An ENDEMIC disease is a disease that is always present in a certain population or region.
Malaria is endemic to parts of Africa.
• An EPIDEMIC, refers to an OUTBREAK of a disease. An epidemic occurs when a disease
is spreading through one or more populations.
• A PANDEMIC, is the worldwide spread of a new disease or global disease outbreak.
• An Influenza pandemic occurs when a new influenza virus emerges and spreads around
the world, and most people do not have immunity.
• The death rate from different diseases is referred to as MORTALITY.

Cholera and its transmission

• Cholera is caused by the bacterium Vibrio cholerae.

• The feces of an infected person are full of these bacteria, each with its distinctive
flagellum.
• [see the figure pg no. 235 given in this section]

Pathogen Vibrio cholerae

Methods of transmission Food-borne, water-borne
Global distribution Asia, Africa, Latin America,
Incubation period Two hours to five days
Site of action of pathogen Wall of small intestine
Clinical features Severe diarrhea (‘rice water’), loss of water
and salts, dehydration, weakness
Method of diagnosis Microscopical analysis of faeces
Annual incidence worldwide 3-5 million
Annual mortality worldwide 1,00,000-1,20,000

• As a disease is water-borne, cholera occurs where people do not have access to proper
sanitation and uncontaminated food.
• Infected people, three quarters of whom may be symptom less carrier, pass out large
number of bacteria in their feces.

220
• If these contaminated water supply, or if infected people handle food or cooking utensils
without washing their hands, then bacteria are transmitted to uninfected people.
• [See the image given in the section pg no. 235]
• To reach their site of action in the small intestine, the bacteria have to pass through
stomach.
• If the contents are sufficiently acidic (pH less than 4.5), the bacteria are unlikely to
survive.
• However, if the bacteria do not reach the small instestine, they multiply and secrete a
toxin, CHOLERAGEN, which disrupts the function of the epithelial lining the intestine, so
that salts and water leave the blood
• These causes severe diarrhea and the loss of fluid can be fatal if not treated within 24
hours

[This is the similar figure given in this section pg no.236]

Treating cholera

• Almost all people with cholera who are treated make a quick recovery
• a death from cholera is an AVOIDABLE DEATH.
• The disease can be controlled quite cheaply by a solution of salts and glucose given
intravenously to rehydrate the body
• If people can drink, they are given oral rehydration theory
• Glucose is effective, because it is absorbed into the blood and takes ions (for example,
sodium and potassium ions) with it.
• It is important to make sure that a patient’s fluid intake equals fluid losses in urine and
feces and to maintain the osmotic balance of the blood and tissue fluids

221
Preventing cholera

• In developing countries, large cities that have grown considerably in recent years, but
as yet have no sewage treatment or clean water, create perfect condition for the spread
of disease
• Increasing quantity of untreated feces from a growing population fever cholera survival
• Countries that have huge debates do not have the financial resources to tackle large
municipal project such as providing drainage and a clean water supply to large areas of
standard housing.
• In many countries, the use of raw human sewage to irrigate vegetables is a common
cause of disease, as are inadequate cooking, or washing in contaminated water.
• Areas of the World where cholera is endemic are West and East Africa and Afghanistan.
• Cholera is now almost unknown in developed World, as a result of sewage treatment
and provision of clean piped water, which is chlorinated to kill bacteria.
• The transmission cycle has been broken.
• Health authorities always fear outbreaks of cholera and other diarrheal diseases
following natural disasters.
• In Haiti in 2010, a cholera epidemic broke out several months after the earthquake that
destroyed large parts of the country.
• Travelers from areas free of cholera to those where cholera is endemic used to be
advised to be vaccinated, although the vaccine only provided short-term protection.
• This recommendation has now been dropped.

Strains of cholera

• There are many different strains of V. cholerae.

• Until the 1990s, only the strain known as O1 caused cholera.
• Between 1817 and 1923, there were six pandemics of cholera.
• Each originated in what is now Bangladesh and was caused by the ‘classical’ strain of
cholera O1.

222
• A seventh pandemic began in 1961 when a variety of O1 named ‘E1 Tor’ originated in
Indonesia.
• E1 Tor soon spread to India, then to Italy in 1973, reaching South America in January
1991, where it caused in epidemic in Peru.
• The discharge of a ship’s sewage into the sea may have been responsible.
• Within days of the start of epidemic, the disease had spread 2000 km along the coast,
and within four weeks had moved inland.
• In February and March of 1991, an averages of 2550 cases a day were being reported.
• People in neighboring countries were soon infected.
• In Peru, many sewers discharge straight onto shellfish beds.
• Organisms eaten as seafood, especially filter-feeders such as oysters and mussels
become contaminated because they concentrate cholera bacteria when sewage is
pumped into the sea.
• Fish and shellfish are often eaten raw.
• Because the epidemic developed so rapidly in Peru, it is thought that disease probablt
spread through contaminated seafood.
• A new strain known as V. cholera O139, originated in Chennai (then called Madras) in
October 1992 and has spread to other parts of India and Bangladesh.
• This strain threatens to be responsible for an eighth pandemic.
• It took EI Tor two years to displace the ‘classical’ strain in India O139 replace EI Tor in
less than two months, suggesting that may be more virulent.
• Many adult cases have been reported, suggesting that previous exposure to EI Tor has
not given immunity to O139.
• [ See the photograph on pg no.237]

223
Most cases of malaria are caused by one of four species of the protoctist Plasmodium. Genetic
analysis of infection shows that some species of Plasmodium that cause malaria in monkeys
also affect humans.

Pathogen Plasmodium falciparum,P. vivax, P. ovale, P.

malariae.
Methods of transmission Insect vector: female Anopheles mosquito
Global distribution Throughout the tropics and sub-tropics
(endemic in 106 countries)
Incubation period From a week to a year
Site of action of pathogen Liver, red blood cells, brain
Clinical features Fever, anemia, nausea, headaches, muscle
pain, shivering, sweating, enlarge spleen
Method of diagnosis Microscopic examination of blood(Figure):
dip stick test for malaria antigens in blood
Annual incidence worldwide About 207 million cases of malaria in 2012
(about 80% are in Africa)
Annual mortality worldwide About 6,30,000 deaths in 2012 (about 90%
are in Africa)

• See the images of mosquitoes in this section pg no. 237.

• Female Anopheles mosquitoes feed on human blood to obtain the protein they need to
develop their eggs.

224
• If the person they bite is infected with Plasmodium, they will take up some of the
pathogen’s gametes with the blood meal.
• Male and female gametes fuse in mosquito’s gut and develop to form infective stages,
which move to mosquito’s salivary glands.
• When mosquito feeds again, she injects an anticoagulant from her salivary glands that
prevents the blood meal from clotting, so that it flows out of the host into mosquito.
• The infective stages pass from the mosquito’s salivary glands in to the human’s blood
together with the anticoagulant in the saliva.
• The parasites enter the red blood cells, where they multiply.
• The female Anopheles mosquito is therefore, a vector of malaria and she transmits the
disease when she passes the infective stages into uninfected person.
• Malaria may also be transmitted during blood transfusion and when unsterile needles
are re-used.
• Plasmodium can also pass the placenta from mother to fetus.
• A vector is an organism which carries a disease from one person to another or from
animal to a human. Do not confuse it with the causative agent, which in this case is
Plasmodium.

• The parasite has two hosts: the sexual stage occurs in mosquitoes, theasexual stage in
humans. The time between infection and appearance of parasites inside red blood cells
is 7-30 days in P. falciparum. Longer in other species.

225
• [see the figure in this section pg no. 237]
• Plasmodium multiplies in both hosts, the human and the mosquito; at each stage there
is huge increase in the number of parasites, and this improves the chances of infecting
another mosquito or human host.
• If people are continually re-infected by different strains of malaria they become immune.
• However, this only happens if they survive the first five years of life, when mortality is
very high.
• The immunity only lasts as long as people are in contact with the disease.
• This explains why epidemics in places where malaria is not endemic can be very serious,
and why malaria is more dangerous in those areas where it only occurs during and after
rainy season.
• This often coincides with the time of maximum agricultural activity, so the disease has a
disastrous effect on the economy; people can cultivate the land when they are sick.

•
• Transmission electron micrograph of a section through a red blood cells packed tightly
with malarial parasites. Plasmodium multiples inside red blood cells; this cell will soon
burst, releasing parasites which will infect other red blood cells.
• [See the figure in pg no.238]
• Red blood cells infected with Plasmodium falciparum. Notice the characteristics single
ring appearance of the parasite inside the red blood cells (X1300)

MALARIA

• Anti-malarial drugs such as quinine and chloroquine are used to treat infected people.

226
• They are also used as prophylactic (preventative) drugs, stopping an infection occurring
if a person is bitten by an infected mosquito.
• Prophylactic drugs are taken before, during and after visiting an area where malaria is
endemic.
• Chloroquine inhibits protein synthesis and prevents the parasitespreading within the
body.
• Another prophylactic, proguanil, has the added advantage of inhibiting the sexual
reproduction of Plasmodium inside the biting mosquito.
• Where anti-malarial drugs have been used widely, there are strains of drug-resistant
Plasmodium-the drug is no longer effective against the pathogen.
• Chloroquine resistance is widespread in parts of South America, Africa and New Guinea.
• However, mefloquine is expensive and sometimes causes unpleasant side-effects such
asrestlessness, dizziness, vomiting and disturbed sleep.
• Resistance to meflloquine has developed in some areas, notably the border region of
Thailand.
• The antibiotic doxycycline is also used s prophylactic drug.
• The drug artesunate, derived from the plant compound artemisin, is used in
combination with mefloquine to treat infections of P. falciperum.
• People from non-malarial countries visiting many parts many parts of the tropics are at
great risk of contracting malaria.
• Doctors in developed countries, who see very few cases of malaria, often misdiagnose it
as influenza, since the initial symptoms are similar.
• Many of these cases are among settled immigrants who have been visiting relatives in
Africa and India.
• These people do not take prophylactic drugs, because they do not realize that they have
lost their immunity.

Preventing malaria
• There are three main ways to control malaria:
1. Reduce the number of mosquitoes
2. Avoid by being bitten by mosquitoes.

227
 3. Use drugs to prevent the parasite infecting people.

[See other images in the page no. 238]

• It is possible to kill the insect vector and break the transmission cycle.
• Mosquitoes lay their eggs in water.
• Larvae hatch and develop in water but breath air by coming to the surface.
• Oil can be spread over the surface of water to make it impossible for mosquito larvae
and pupae to breath.
• Marshes can be drained and vegetation cleared.
• Two biological control measures that can be used are :
1. Stocking ponds, irrigation and drainage ditches and other permanent bodies of water
with fish which feed on mosquito larvae.
2. Spraying a preparation containing the bacterium Bacillus thuringiensis, which kills
mosquito larvae but is not toxic to other forms of life.
• However, mosquitoes will lay their eggs in any small puddle or pool, which makes it
impossible to completely eradicate breeding sites, especially in the rainy season.
• The best protection against malaria is to avoid being bitten.

228
 • People are advised to sleep beneath mosquito nets and use insect repellents.
• Soaking mosquito nets in insecticide every six months has been shown to reduce
mortality from malaria.
• People should not expose their skin when mosquitoes are active at dusk.

WORLDWIDE CONTROL OF MALARIA

• In the 1950s, the World Health Organization (WHO) coordinated a worldwide

eradication programme.
• Although malaria was cleared from some countries, the programme was not generally
successful.
• There are two main reasons for it
1. Plasmodium became resistant to the drugs used to control it.
2. Mosquitoes became resistant to DDT and the other insecticides that were used at the
time, such as dieldrin.
• This programme was also hugely expensive and often unpopular. People living in
areas where malaria was temporarily eradicated by the programme lost their
immunity and suffered considerably, even dying, when the disease returned.
• Some villagers in South-East Asia lost the roofs of their houses because dieldrin killed
a parasitic wasp that controlled the numbers of thatcheating caterpillars.
• Some spray teams were set upon and killed by angry villagers in New Guinea.
• The programme could have been more successful if it had been tackled more
sensitively with more involvement of local people.
• In 1970s, war and civil unrest destroyed much of the infrastructure throughout Africa
and South-East Asia, making it impossible for mosquito control teams to work
effectively.
• [See the photo given I this section 239]
• The reasons for worldwide concern over the spread of malaria are:
1. An increase in drug-resistant forms of Plasmodium.
2. An increase in the proportion of cases caused by P. falciperum, the form that causes
severe, often fatal malaria.

229
 3. Difficulties in developing vaccines against malaria.
4. An increase in the number of epidemics, because of climatic and environmental
changes that favor the spread of mosquitoes.
5. The migration of people from areas where malaria is andemic, for economic and
political reasons.
• Malaria is still one of the World’s biggest threats to health: 40% of the World’s
population lives in areas where there is risk of malaria.
• Between 2000 and 2011, control measures have achieved a decrease in mortality
rates of about 25% across the World, and 33% in the WHO’s African region.
• Control methods now concentrate on working within the health system to improve
diagnosis, improve the supply of effective drugs and promote appropriate methods
to prevent transmission.
• Several recent advances give hope that malaria may one day controlled. The
introduction of simple dip stick tests for diagnosing malaria means that diagnosis can
be done quickly without the need for laboratories.
• The whole genome of Plasmodium has been sequenced, and this may be lead to the
development of effective vaccines. Several vaccines are being trialed, but it is not
likely that a successful vaccine will be available for some time.
• Drugs are used in combination to reduce the chances of drug resistance arising.
• Three factors may lead to improvements in the control of malaria:
1. Use of modern techniques in gene sequencing and drug design.
2. Development of vaccines targeted against different stages of the parasite’s life cycle.
3. A renewed international will to remove the burden of disease from poorest parts of
the World, allied to generous donations from wealthy individuals and foundations.

Acquired immune deficiency syndrome (AIDS)

• HIV stands for human immunodeficiency virus.

• If left untreated, HIV can lead to the disease AIDS (acquired immunodeficiency
syndrome)
• Unlike some other viruses, the human body can’t get rid of HIV completely.

230
 • So once you have HIV, you have it for life.

Pathogen Human immunodeficiency virus

Methods of transmission In semen and vaginal fluids during sexual
intercourse, infected blood or blood
products, contaminated hypodermic
syrighes, mother to fetus across placeta, at
birth mother to infant in breast milk
Global distribution Worldwide, especially in sub Saharan Africa
and South-East Asia.
Incubation period Initial incubation a few weeks, but up to ten
years or more before symptoms of AIDS
may develop.
Site of action of pathogen T helper lymphocytes, macrophages, brain
cells
Clinical features HIV infection-flu like symptoms and then
symptomless.
AIDS-opportunistic infections including, TB
and cancers; weight loss, diarrhea, fever,
sweating, dementia

231
Methods of diagnosis Testing blood, saliva or urine for the
presence of antibodies produced against
HIV.
Estimated total number of people infected 35.5 million (89% of these in sub-Saharn
with HIV worldwide in 2012 Africa)
Estimated number of new cases of HIV 2.3 million
infection worldwide in 2012

Estimated number of deaths from AIDS- 1.6 million (UNAIDS estimate)

related disease worldwide in 2012

HIV AND AIDS

• Human immunodeficiency virus (HIV)

• The outer envelope contains glycoproteins.
• The protein core contains genetic material(RNA) and two enzymes; a protease and
reverse transcriptase. Reverse transcriptase uses the RNA as a template to produce
DNA once the virus is inside the host.
• [see the figure in this section pg no.240]

232
 • A series of transmission electron micrograph showing HIV budding from the surface
of an infected lymphocyte and becoming surrounded by membrane derived from the
cell surface membrane of the host cell (X176000)
a. The viral particle first appears as a bump.
b. Which then buds out.
c. Is eventually cut off.
d. The outer shell of dense material and less dense core are visible in the released virus.

What is retro virus

• HIV is a retrovirus, which means that its genetic material is RNA, not DNA.
• Once inside a host, the viral RNA is converted ‘back’ to DNA (hence ‘retro’) to be
incorporated in to human chromosomes.
• The virus infects and destroys cells of the body’s immune system so that their
numbers gradually decrase.
• These cells, known as helper T cells, control the immune system’s response to
infection.
• When the numbers of these cells are low, the body is unable to defend itself against
infection, so allowing a range of pathogens to cause a variety of opportunistic
pathogen.

233
 • AIDS is not a disease; it is a collection of these opportunistic diseases associated with
immuno deficiency caused by HIV infection.
• Since HIV is an infective agent, AIDS called an ACQUIRED IMMUNE DEFICIENCY to
distinguish it form other types- for example, an inherited form.
• [See the Image in pg no. 240]
• [See the Image in pg no. 241 (HHMI)]

Transmission of HIV

• The WHO estimated that by 2010 over 25 million people had died of HIV/AIDS.
• HIV is a virus that is SPREAD BY INTIMATE HUMAN CONTACT; their space IS NO
VECTOR (unlike in malaria) and the virus is unable to survive outside the human body
(unlike choleras or malaria pathogens).
• Transmission is only possible by direct exchange of body fluids.
• HIV transmission
1. Unsafe sex
2. From an infected mother to her child
3. Contaminated needles
4. Blood products.
• In practice, this means that HIV is spread most easily through sexual intercourse,
blood donation and the sharing of needles used by intravenous drug users.
• HIV is also transmitted from mother to child across the placenta and, more often,
through the mixing of blood during birth.
• An HIV + pregnant women can transmit to her baby 3 WAYS;
1. During pregnancy
2. During vaginal child birth
3. Through breast feeding
• The initial epidemic in North America and Europe was amongst male homosexuals
who practices anal intercourse and had many sex partners, two forms of behavior
that put them at risk

234
 • The mucous lining of the rectum is not as thick as that of the vagina, and there is less
natural lubrication.
• As a result, the rectal lining is easily damaged during intercourse and the virus can
pass from semen to blood.
• Heaving multiple partners, both homosexual and heterosexual, allows the virus to
spread more widely.
• Also at high risk of infection were hemophiliacs who were treated with a clotting
substance (factor 8) isolated from blood pooled from many donors.
• Much of the transmission of HIV has been by heterosexual intercourse.
• This is particularly rapid in some African states, where equal numbers of males and
females are now HIV positive (HIV+).

THE GLOBAL DISTRIBUTION OF HIV/AIDS

• 70% of the World’s deaths from AIDS occur in Africa.

• In 2007 it was estimated that 15 million people had died of HIV/AIDS in sub-Saharan
Africa since the beginning of the pandemic.
• 25% of the adult population of Botswana is infected with HIV.
• Between 15% and 25% of people aged 15-49 in Botswana and Zimbabwe are infected
with HIV.
• Over 16 million children are estimated to have lost one or both parents to AIDS; in
some places this is 25% of the population under 15.

235
• _______________________________Or people affected by HIV has been estimated between 4,
30,000 and 1.5 million, somewhere below or around 0.1% of the population.
• The prevalence of HIV among women attending antenatal clinics in Zimbabwe was
around 20% in 2012.
• A large proportion of women in Rwanda are HIV positive following the use of rape as
a genocidal weapon in the civil war of the early 1990s.
• The average life expectancy in South Africa dropped from 65-55 during 1195-1999.
• HIV is spread through direct contact with certain body fluids from someone who has
HIV.
Blood
Semen and pre-seminal fluid
Rectal fluids
Vaginal fluids
Breast milk
• HIV is a slow virus and, after infection, there may not be any symptoms until years
later.
• Some people who have the virus even appear not to develop any initial symptoms,
although there often flu-like symptoms for several weeks after becoming infected,
• At this stage, a person is HIV positive but does not have AIDS.
• 1.8 MILLION CHILDREN worldwide are living with HIV. Most of these children were
infected by their HIV positive mothers during pregnancy, child birth or breastfeeding.
• The infections that can opportunistically develop to create AIDS tend to be
characteristic of the condition.
• Two of these are caused by fungi:
1. Oral thrush caused by Candida albicans
2. A rear term of pneumonia caused by Pneumocystis jiroveci
• Phenumocystis pneumonia is not commonly found in the lungs of healthy people.
• But being a source of opportunistic infection, it can cause a lung infection in people
with a weak immune system.

236
 • Phenumocystis pneumonia is especially seen in people with cancer undergoing
chemotherapy, HIV/AIDS and the use of medications that suppress the immune
system.
• [see the diseased images in this section pg no. 242]
• As and when the immune system collapses further, it becomes less effective in finding
and destroying cancers.
• A rare form of skin cancer, Kaposi’s sarcoma, caused by a herpes-like virus, is
associated with AIDS.
• Kaposi’s sarcoma, and cancers of internal organs are now the most likely causes of
death of people with AIDS in developed countries, along with degenerative disease of
the brain, such of dementias.
• [see the diseased images in this section pg no. 242].
• At about the same time that AIDS was first reported on the West Coast of the USA and
Europe, doctors in central Africa reported seeing people with similar opportunistic
infection.
• We have seen that HIV/AIDS is now wide spread throughout sub-Saharan Africa from
Uganda to South Africa.
• It is serious public health problem here because HIV infection means people more
vulnerable to existing disease such as malnutrition, TB and malaria.
• AIDS is having an adverse effect on the economic development of countries in the
region, as it affects sexually active people in their 20s and 30s who are also potentially
the most economically productive, and the purchase of expensive drugs drains
government funds.
• The World Bank estimated that AIDS had reversed 10-15 years of economic growth
for some African states by the end of the 20th century.

TREATING HIV/AIDS

237
• There is as yet NO CURE FOR AIDS and NO VACCINE FOR HIV.
• No-one knows how many people with HIV will progress to developing full- blown
AIDS
• Some people think it is 100%, although a tiny minority of HIV-positive people do
appear to have immunity and can leave as entirely symptomless carriers.
• Drug therapy can down the onset of AIDS quite dramatically, so much so that some
HIV-positive people in developed countries are adjusting a suddenly increased life
expectancy.
• However, the drugs are expensive and have a variety of side-effects ranging from the
mild and temporary (rashes, headaches, diarrhea) to the severe and permanent
(nerve damge, abnormal fat distribution).
• If used in combination, two or more drugs which prevent the replication of the virus
inside host cells can prolong life, but they do not offer a cure.
• The drugs are similar to DNA nucleotides (for example, ZIDOVUDINE is similar to the
nucleotide that contains the base thymine).
• ZIDOVUDINE binds to the viral enzyme reverse transcriptase and blocks its action.
• This stops the replication of the viral genetic material and leads to an increase in some
of the body’s lymphocytes.

238
 • A course of combination therapy (taking several drugs) can be very complicated to
follow.
• The pattern and timing of medication through the day must be strictly followed.
• People who are unable to keep to such a regimen can become susceptible to strains
of HIV that have developed resistance to the drugs.

NRTI/NtRTI NNRTI Protease inhibitor Fusion inhibitor

Zidovudine (ZDV) Nevirapine (NVP) Indinavir (IDV) Enfuvirtide (T-20)
Didanosine (ddI) Efavirens (EFV) Ritonavir (RTV)
Stavudine (d4T) Nelfinavir (NFV)
Lamividine (3TC) Lopinavir/r (LPV/r)
Tenofovir (TDF) Fosamprenavir
Combivir (COM)
Abacavir (ABC)
NRTI- Nucleoside reverse transcriptase inhibitor; NtRTI- Nucleotide reverse transcriptase
inhibitor; NNRTI- Nonnucleoside reverse transcriptase inhibitor

PREVENTING HIV/AIDS

• The spread of HIV/AIDS is difficult to control.

• The virus’s long latent stage means it can be transmitted by people who are HIV positive
but who show no symptoms of AIDS and do not know they are infected.
• The virus changes its surface proteins, which makes it hard for the body’s immune
system to recognize it.

239
• This also makes the development of a vaccine very difficult.
• For the present, public health measures are the only way to stop the spread of HIV.
• People can be educated about the spread of the infection and encouraged to change
their behavior so as to protect themselves and others.
• CONDOMS, FEMIDOMS and DENTAL DAMS are the only effective methods of reducing
the risk of infection during intercourse, as they form a barrier between body fluids,
reducing the chances of transmission of the virus.
• A dental dam is a small sheet of rectangular latex that acts as a protective barrier
between your mouth and the vagina or anus.
• [See the images in the pg no.243]
• Some countries have promoted the use of condoms as well as other measures.
• As a result, infection rates in these countries have slowed and the number of new
cases reported each year has either decreased or remained the same year on year.
• It is estimated that the rate of HIV infection across the world decreased by 25%
between 2001 and 2009.
• Contact tracing is an important part of controlling the spread of HIV.
• If a person who is diagnosed as HIV positive is willing and able to identify the people
whom he or she has put at risk of infection by sexual intercourse or needle sharing,
then these people will be offered an HIV test.
• This test identifies the presence of antibodies to HIV, although these only appear
several weeks after the initial infection.
• Injecting drug users are advised to give up their habit, stop sharing needles or take
the drugs in some other way.
• Needle-exchange schemes operate in some places to exchange used needles for sterile
ones to reduce chances of infection with HIV and other blood-borne diseases.
• Blood collected from blood donors is screened for HIV and heat- treated to kill any
viruses.
• People who think they may have been exposed to the virus are not strongly
discouraged from donating blood.
• In some low income countries, not all donated blood is tested.

240
 • Anyone concerned about becoming infected by blood transfusion during and
operation may donate their own blood before the operation to be used instead of
blood from blood bank.
• Widespread testing of population to find people who are HIV positive is not expesive,
but governments are reluctant ti introduce such testing because of the infringement
of personal freedom.
• In the developed World, HIV testing is promoted most strongly to people in high risk
groups, such as male homo sexuals, prostitutes, injecting drug users and their sexual
partners.
• If tested positive, they can be given the medical and psychological support they need.
• In Africa and South-East Asia, the epidemic is not restricted to such easily identifiable
groups and widespread testing is not feasible due to difficulty of reaching the majority
of the population and organizing testing.
• People of these regions find out that they are HIV positive when they develop the
symptoms of AIDS.
• Both VIRAL PARTICLES and INFECTED LYMPHOCYTES are found in BREAST MILK.
• Mother-to-child transmission is reduced by treating HIV-positive women and their
babies with drugs.
• However, HIV-positive women in high-income countries are advised not to breatfeed
their babies, because of risk of transmission even if they have secure supply of drugs
during this period.
• In contrast, HIV-positive women in low- and middle income countries are advised to
breastfeed, especially if they have secure supply of drugs during this period, as the
protection this gives against other diseases and lack of clean water to make up
formula milk may outweight therisk of transmitting HIV.

TUBERCULOSIS

• TB is caused by either of TWO bacteria, Mycobacterium tuberculosis and

mycobacterium bovis.
• These are pathogens that live inside human cells, particularly in lungs.

241
 • This is the first site of infection, but the bacteria can spread throughout the whole
body and even infect the bone tissue.
• [See the figure on pg no. 244]

Pathogen Mycobacterium tuberculosis

Mycobacterium bovis
Methods of transmission Air borne droplets (M tuberculosis via
undercooked meat and un pasteurized milk)
Global distribution Worldwide
Incubation period few weeks or up to several years
Site of action of pathogen Primary infection in lungs; secondary
infection in lymph nodes, bones and gut
Clinical features Paching cough, coughing blood, chest pain,
shortness of breath, fever, sweating, eeight
loss
Methods of diagnosis Microscopical examination of sputum for
bacteria, chest X-ray
Annual incidence worldwide in 2012 8.6 million
Annual mortality worldwide in 2012 1.3 million (including 320000 deaths of
people who were HIV+)

Transmission electron micrograph of Mycobacterium tuberculosis dividing in to two, it may

multiply like this inside the lungs and then spread throughout the body as the dormant,
becoming active many years later

242
 • Some people become infected and develop TB quite quickly, while in others the
bacteria remain inactive for many years
• It is estimated that about 30% of the world’s population is infected with TB without
sowing any symptoms of the infection; people with these inactive infection do not
spread the disease to others
• However, the bacteria can later become active, and this is most likely to happen when
people are wreaked by other disease, suffer from malnutrition or become infected
with HIV
• Those who have the active form of TB often suffer from debilitating illness for a long
time
• They have a persistent cough and, as part of the defense, cells release hormone like
compounds, which cause fever and suppress the appetite
• As a result, people with TB loss were and often look emaciated.
• [see the image on pg no. 244]
• TB is often the first opportunistic infection to strike HIV-positive people
• HIV infection may reactivate dormant infection of M. tuberculosis which may have
been present from childhood or, if people are uninfected, make them susceptible to
infection.
• TB is now the leading cause of death among HIV-positive people.
• The HIV pandemic has been followed very closely by a TB pandemic.

Transmission of TB

• TB is spread when infected people with the active form of the illness cough or sneeze
and the bacteria are carried in the air in tiny droplets of liquid.
• Transmission occurs when people who are uninfected inhale the droplets.
• TB spreads most rapidly among people living in overcrowded conditions.
• People who sleep close together in large numbers are particularly at risk.

243
• The disease primarily attacks the homeless and people who live in poor, substandard
housing; those with low immunity, because of malnutrition or being HIV positive, are
also particularly vulnerable.

Fig. 1 on page no. 245 (transmission of TB)

• The form of TB caused by M. Bovis also occurs in cattle and is spread to humans in
meat and milk.
• It is estimated that there were about 800000 deaths in UK between 1850 and 1950
as a result of TB transmitted from cattle.
• Meat and milk still remain a source of infection in some developing countries.
• The incidence of TB in the UK decreased steeply well before the introduction of a
vaccine in the 1950s, because of improvements in housing conditions and diet.
• The antibiotic streptomycin was introduced in the 1940s, and this hastened the
decrease in the incidence of TB.
• This pattern was repeated throughout the developed world.
• Fig.2 on page no. 245 (TB in cattle)
• TB is on the increase.
• There are high rates of incidence all across the developing world and in Russia and
surrounding countries.
• This increase is due in part to the following factors:
• Some strains of TB bacteria are resistant to drugs
• The HIV/AIDS pandemic
• Poor housing inner cities and homelessness
• The breakdown of TB control programmes; partial treatment for TB increases the
chance of drug resistance in Mycobacterium.

244
 Treating TB
• When a doctor first sees a person with the likely symptoms of TB, samples of the
sputum (mucus and pus) from their lungs are collected for analysis.
• The identification of TB bacteria can be done very quickly by microscopy.
• If TB is confirmed, then patients should be isolated while they are in the most
infectious stage (which is at two to four weeks).

• The treatment involves using several drugs to ensure that all the bacteria are killed.
• If not killed, drug-resistant forms remain to continue the infection.
• The treatment is a long one (six to nine months, or longer), because it takes a long
time to kill the bacteria, which are slow growing and are not very sensitive to the
drugs used.
• Unfortunately, many people do not complete their course of drugs, because they think
that when they feel better, they are cured.
• People who do not complete their treatment may be harboring drug resistant bacteria
and may spread these to others if the bacteria become active.

245
 • Fig. 1 on page no. 246 (global distribution of TB in 2010)

Drug-resistant TB

• Strains of drug-resistant M. tuberculosis were identified when treatment

with antibiotics began in the 1950s.
• Antibiotics act as selective agents killing drug-sensitive strains and
leaving resistant ones behind.
• Drug resistance occurs as a result of mutation in the bacterial DNA.
• Mutation is a random event and occurs with a frequency of about one in
every thousand bacteria.
• If three drugs are used in treatment, then the chance of resistance arising
to all three of them by mutation is reduced to one in a thousand million.
• If four drugs are used, the chance is reduced to one in a billion.
• If TB is not treated or the person stops the treatment before the bacteria
are completely eliminated, the bacteria spread throughout the body,
increasing the likelihood that mutations will arise, as the bacteria survive
for a long time and multiply.
• Stopping treatment early can mean that M. Tuberculosis develops
resistance to all the drugs being used.
• People who do not complete a course of treatment are highly likely to
infect others ith drug-resistant forms of TB.
• It is 10 to 15 others, especially if the person lives in overcrowded
conditions.

246
DOTS

• The WHO promotes a scheme to ensure that patients complete their courses of
drugs.
• DOTS- DIRECT OBSERVATION TREATMENT, SHORT COURSE, involves health
workers or responsible family members making sure that patients take their
medicine regularly for six to eight months.
• The drugs widely used are ISONIAZID AND RIFAMPCIN, often in combination with
others.
• This drug therapy cures 95% of all patients, and is twice as effective as other
strategies.
• Multiple-drug-resistant forms of TB (MDR-TB) now exist.
• MDR-TB strains of TB are resistant to at least the two main drugs used to treat
TB-isoniazid and rifampicin – which are known as first-line drugs.
• In 1995, an HIV unit in London reported an outbreak of MDR-TB with form of M.
tuberculosis that was resistant to five of the major drugs used to treat the disease,
including isoniazid, which the most successful drug.
• Extensively (or extremely) drug-resistant TB (XDR-TB) has also emerged as a very
serious threat to health, especially for those people who are HIV-positive.

247
 • XDR-TB strains are resistant to first line drugs and to the drugs used to treat MDR-
TB.
• These resistant strains of TB do not respond to the standard six months treatment
with first-line anti-TB drugs and can take two years or more to treat with drugs
that are less potent and much more extensive.
• DOTS is helping to reduce the spread of MDR strains of TB.

PREVENTING TB

• Contact tracing and the subsequent testing of contacts for the bacterium are essential
parts of controlling TB. Contracts are screened for symptoms of TB infection, but the
diagnosis can take up to two weeks.
• What is contact tracing?
Contract tracing is finding everyone who has come in direct contact with a sick TB
patient. Contracts are watched for signs of illness for 21 days from the last day they
came in contact with the TB patient.
Contract tracing can stop the TB outbreak in the tracks.
Fig. 4 on page no. 246 (contract tracing )
• THE ONLY VACCINE AVAILABLE FOR TB IS THE BCG VACCINE, which is derived from
M. Bovis and protects upto 70-80% of people who receive it.

Fig. 1 on page no.247 (vaccination)

• The effectiveness of the vaccine decreases with age unless there is exposure to TB.

248
• Many countries with high numbers of people with TB use the BCG vaccine to protect
children from getting the disease.
• Countries such as the UK and US do not include BCG vaccination in their immunization
programmes.
• Instead, it may be given only to people who are at high risk of becoming infected
because, for example, they live with an adult who is being treated for the disease.
• Fig. 2 on page no. 247
• TB can be transmitted between cattle and humans.
• To prevent people catching TB in this way, cattle are routinely tested for TB and any
found to be infected are destroyed.
• TB bacteria are killed when milk is pasteurized.
• These control methods are very effective and have reduced the incidence of human
TB caused by M. Bovis considerably, so that it is virtually eliminated in countries
where these controls operate.
• In the UK, less than 1% of the 9000 new cases of TB each year are due to M. Bovis.

Measless
• Measles is caused by a virus which enters the body and multiplies inside cell in the
upper respiratory tract (nasal cavity and trachea).
• There are no symptoms for 8-14 days after the initial infection and then a rash
appears and a fever develops.
• Other symptoms are a runny nose, a cough, red and watery eyes (conjunctivitis) and
small white spots that may develop inside the cheeks.
• Fig.3 on page no. 247
• Treatment involves bed rest and taking medicines to lower the fever, there are no
specific medicines for measles.
• After about ten days the disease clears up and there are rarely any complications.
• If complications do occur, they tend to be serious, pneumonia, ear and sinus
infections, brain damage and convulsions may follow a measles infection.

249
• Some of these cases are fatal and among malnourished children living in overcrowded
conditions, measles is a serious disease and a major cause of death.
• About 10% of all infant deaths in developing countries are the result of measles
infections.
• Measles is also responsible for many cases of childhood blindness.
• Measles is one of the MOST CONTAGIOUS DISEASES.
• When people infected with measles sneeze or cough they release droplets containing
many millions of virus particles.
• If these are inhaled by uninfected people who have no immunity to these disease, it is
almost inevitable that they will become infected and develop symptoms.
• The disease rarely affects infants under eight months of age, as they have passive
immunity in the form of antibodies that have crossed the placenta from their mother.
• However, as these antibodies are gradually destroyed the children lose their
immunity.
• Measles used to be a common childhood disease in developed countries, but the
incidence of the disease fell steeply after the introduction of a vaccine in the early
1960s.
• Measles is a major disease in developing countries, particularly in cities where people
live in overcrowded, insanitary conditions and where there is a high birth rate.
• The measles virus is transmitted easily in these conditions and it infects mainly
malnourished infants suffering from vitamin A deficiency.
• There are estimated to be over 20 million cases of measles worldwide each year, most
of which are in Africa, South-East Asia, India, Pakistan, Bangladesh and some
countries of the middle east.
• The death rate from the disease has fallen from 630000 in 1990 to 158000 in 2011,
largely as a result of a mass vaccination programme.
• There are very few cases of measles in developed countries and most outbreaks are
caused by someone entering the country with the disease that they caught elsewhere.
• When outbreaks occur, as an Indiana in the USA in 2005 and in South Wales in the UK
in 2012/13, they can spread rapidly among those at risk.

250
• These two epidemics occurred mainly among children whose parents had not had
them vaccinated.
• Adults who were not vaccinated and who did not have measles as a child are at risk
of severe complications in epidemics like this.

Antibiotics
• An antibiotic is a drug that kills or stops the growth of bacteria, without harming the
cells of the infected organism.
• Antibiotics are derived from living organisms, although they are often made more
effective by various chemical processes.
• There are a wide range of antibiotics to treat bacterial infections.
• Other antimicrobial drugs such as isoniazid, used for the treatment of TB, are
synthetic (made in laboratories).
How antibiotics work
Fig.1 on page no.248 (how antibiotic works)
• These include:
• Synthesis of bacterial cell walls
• Activity of proteins in the cell surface membrane
• Enzyme action
• DNA synthesis
• Protein synthesis
• Fig.2 on page no. 248 (how penicillin works)
• Bacterial cells have walls made of peptidoglycans.
• These are long molecules containing peptides (chains of amino acids) and
sugars.
• In the bacterial cell wall, peptidoglycans are held together by corsslinks that
form between them.
• Penicillin prevents the synthesis of the cross-links between the peptidoglycan
polymers in the cell walls of bacteria by inhibiting the enzymes that build these
cross-links.

251
 • This means that penicillin is only active against bacteria while they are
growing.
• When a newly formed bacterial cell is growing, it secretes enzymes called
autolysins, which make little holes in its cell wall.
• These little holes allow the wall to stretch so that new peptidoglycan chains
can link together.
• Penicillin prevents the peptidoglycan chains from linking up, but the autolysins
keep making new holes.
• The cell wall therefore becomes progressively weaker.
• Bacteria live in watery environments and take up water by osmosis.
• When they are weakened, the cell walls cannot withstand the pressure
potential exerted on them by the cell contents and the cell burst.

Lysis
of E.Coli
By penicillin
Etinne Maisonneuve & Kenn Gerdes

Centre for Bacterial Cell Biology

University of Newcastle, UK
http://www.ncl.ac.uk/cbcb/

Antibiotics

Penicillin kills by interfering with the production of cell wall as these E.Coli grow
in size, the weakened cell wall ruptures

• This explains why penicillin does not affect human cells.

• Our cells do not have walls.

252
 • This also explains why penicillin and other antibiotics do not affect viruses, which
do not even have cells, let alone cell walls.
• For example, when a virus replicates, it uses the host cell’s mechanism for
transcription and translation and antibiotics do not bind to the proteins that host
cells use in these processes.
• Eukaryotic cells have proteins that are different from those in bacteria so they are
unaffected by such antibiotics.
• Other drugs, called antivirals, are used to control viral infections.
• There are fewer antivirals than there are antibiotics.
• Penicillin first became available for treating disease in the 1940s.
• It was hailed as a wonder drug that could be used to wipe out all the diseases
caused by bacteria.
• To begin with, this seemed to be true, but very quickly it became clear that this
was not going to happen even though other antibiotics, such as streptomycin, soon
became available.
• Some types of bacteria are not sensitive to particular antibiotics: for example,
penicillin is not effective against M. tuberculosis.
• Even amon the types of bacteria that were killed by penicillin, there were certain
strains that were not affected.
• These strains had become resistant to antibiotics.
• During the 70 years since the introduction of antibiotics, most pathogenic bacteria
have become resistant to one or more types of antibiotics.
Fig. 1 on page no. 250
Fig. 2 on page no. 250
Fig. 3 on page no.250
• Penicillin has no effect on M. tuberculosis because the thick cell wall of this
bacterium has a gene that codes for an enzyme that catalyzes the breakdown of
penicillin.

253
• Proteins in the membranes of other species of bacteria can inactivate antibiotics so
they have no effect; bacterial membranes also have proteins that pump out antibiotics
if they enter the cytoplasm.
• In some cases, the antibiotics simply cannot bind to the intended site of action.
• Bacteria that are sensitive to an antibiotic are described as being susceptible to that
antibiotic.
• They may become resistant if they gain a gene coding for a protein that protects them
from the antibiotic.
• Soil bacteria have many resistance mechanisms as they grow in an environment
where there are many molecules that interfere with their metabolism.
• These resistance mechanisms are very similar to those found in pathogenic bacteria.
• Before the introduction of antibiotics, enzymes known as beta-lactamases were not
common among pathogenic bacteria.
• The genes for these enzymes have spread into many different forms of bacteria and it
is believed that they have come from soil bacteria.
• Penicillin has a structure that can be broke down by β-lactamase (penicillinase)
enzymes.
• Pathogenic bacteria that have become resistant to penicillin have often done so
because they have acquired the genes that code for these enzymes.
• Antibiotic resistance can arise when an existing gene within the bacterial genome
changes spontaneously to give rise to a nucleotide sequence that codes for a slightly
different proteins that is not affected by the antibiotic.
• This change in DNA is a mutation.
• When someone takes penicillin to treat a bacterial infection, bacteria that are
susceptible to penicillin will die.
• In most cases, if the dose is followed correctly, this will be the entire population of the
disease-causing bacteria.
• However, if the dose is not followed, perhaps because people stop taking the penicillin
when they feel better as the symptoms disappear, then some susceptible bacteria
survive and if any mutations occur these might confer resistance.

254
• The next time there is an infection with this strain of bacteria, penicillin may not be
effective.
• Bacteria have only one copy of each gene, since they only have a single loop of double
stranded DNA.
• This means that a mutant gene will have an immediate effect on any bacterium
possessing it.
• These individuals have a tremendous advantage. Bacteria without this mutant gene
will be killed, while those bacteria resistant to penicillin survive and reproduce.
• Bacteria reproduce asexually by binary fission; the DNA in the bacterial chromosome
is replicated and the cell divides into two, with each daughter cell receiving a copy of
the chromosome.
• This happens very rapidly in ideal conditions, and even if there was initially only one
resistant bacterium, it might produce ten thousand million descendants within 24
hours. A large population of a penicillin-resistant strain of a bacterium would result.
• This method of spreading antibiotic resistance in a population of bacteria is called
vertical transmission.
• Fig. 1 (a-b) on page no.251
• Genes for antibiotic resistance often occur on plasmids, which are small loops or
double-stranded DNA.
• Plasmids are quite frequently transferred from one bacterium to another, even
between different species.
• This happens during CONJUGATION when a tube forms between two bacteria to
allow the movement of DNA.
• During conjugation, plasmids are transferred from a donor bacterium to a recipient.
• Transfer of part of the DNA from the bacterial chromosome also occurs in the same
way.
• This method of transmission is HORIZONTAL TRANSMISSION.
• Thus it is possible to resistance to a particular antibiotic to arise in one species of
bacterium and be passed on to another.
• Fig. 2 on page no.251

255
 • Fig. 3 on page no. 251
• The grey areas on the agar jelly in this petri dish are colonies of the bacterium
Escherichia Coli.
• The white discs are pieces of card impregnated with different antibiotics.
• Where there are clear areas around the discs the antibiotic has prevented the bacteria
from growing.
• However, you can see that this strain of E. Coli is resistant to the antibiotics on the
discs at the bottom left and has been able to grow right up to the discs.
• The more we misuse antibiotics, the greater the selection pressure we exert on
bacteria to evolve resistance to them.
• Antibiotic-resistant strains of bacteria are continually appearing.
• Antibiotic-resistant infections increase the risk of death, and are often associated
with long stays in hospital, and some time serious complications.
• There is a constant race to find new antibiotics as resistant strains keep arising.
• Where there is wide spread use of antibiotics, such as in hospitals or on farms,
resistance quickly spreads among different species of bacteria.
• Resistance may 1st appear in a non pathogenic bacterium, but then he passed to a
pathogenic species.
• Bacteria living where there is wide spread use of antibiotics may have plasmids
carrying resistance genes for several different antibiotics, giving multiple resistance.
• This presents major problems for doctors.
• For example, methicillin-resistant stephylococus auresus(MRSA) has become a
problem in hospitals around the world and in prisons in USA. It now also infecting
people in general population.

Figure.1 on page no. 252

• MRSA caused dangerous infections after surgery which were mostly controlled by
vancomycin- an antibiotic often used as a last resort for treating infections when
everything else has failed, so as to lessen the chances of development of more such
resistant organisms.

256
 • Then another bacterium common in hospitals, enterococcus faecalis develop
resistant to vancomicin and this resistance passed to staphylococcus auresus.
• Recently antibiotic called carbapens have been antibiotics of last resort for use on
bacteria with multiple resistance.
• In 2019, carbapen-resistant klebsiella pneumonia was found in Greece.
• By 2010, it was also found in Cyprus, Hungary and Italy, and in Greece the proportion
of infection of K. pneumonia that were carbapen-resistant has risen to over 25%.
• In 2010, Greece used more antibiotics per head of population than any other
European country.

CHOOSING EFFECTIVE ANTIBIOTICS

• Antibiotics should be chosen carefully.

• Testing antibiotics against the strain of the bacterium
isolated from people ensures that the most effective antibiotic
can be used in treatment.
• As fast as we develop new antibiotics bacteria seem to develop resistant to them.
• It follows from this that the is a constant search for new antibiotics especially once
that work in a completely different way from those currently in use.
• Fortunately, a bacterium resistant to a particular to a antibiotic may not be resistant
to that antibiotic with a slightly alter chemical structure.
• Chemist can make such semi synthetic antibiotic to extend the range available.
• However, many experts believe that we will not be able to keep up and that soon there
will be no antibiotic left to treat diseases.
• This is fast becoming the case with GONORRHEA, asexsual transmitted infection.

Gonnorhea is a sexually transmitted infection caused by the bacterium Neisseria

gonorrhea.

Men may have burning with urination, discharge from the penis or testicular pain.

Women may have burning with urination, vaginal discharge, vaginal bleeding between
periods or pelvic pain.

257
• Clearly we should try to reduce the number of circumstances in which bacteria
develop resistance to antibiotic some of the ways in which we can do this include:
• Using antibiotic only when appropriate and necessary, not prescribing them for viral
infection.
• Reducing the number of countries in which antibiotics are sold without doctors
prescriptions.
• Avoiding the use of so called wide spectrum antibiotic and using instead an antibiotic
specific to the infection(knows as narrow spectrum)
• Making sure that patient complete their course of medication.
• Making sure that patients do not keep on use antibiotics for self medication in the
future.
• Changes in the type of antibiotic prescribe for certain disease so that the same
antibiotic is not always prescribed for same disease.
• Avoiding using antibiotic in farming to prevent, rather than cure, infections

258