1

2

Carbohydrates:
Monosaccharides: sweet, water soluble, reducing sugars
Disaccharides: sweet, water soluble, most are reducing (not sucrose)
Sucrose: Glucose + Fructose
Lactose: Glucose + Galactose
Maltose: Glucose + Glucose
Polysaccharides: long chains of repeating subunits (monosaccharides) joined by condensation
reactions
Isomers: same molecular formula but a different structural formula
Triglycerides
Fatty acid chains can be saturated (no C=C) or
unsaturated (have C=C bonds). The more
unsaturated the fatty acid the lower the melting
point. Fats are insoluble ion water. A phospholipid
has a fatty acid replace with a phosphate group.
This means it has a hydrophilic region (phosphate
head) and hydrophobic regions fatty acid tail. It is
integral in the cell membrane.
General fatty acid
Glycerol
Formation of a triglyceride
Proteins
Polymers made up
of amino acids.
3

Globular Proteins
The vast majority of proteins are globular, i.e. they have a compact,
ball-shaped structure. This group includes enzymes, membrane
proteins, receptors and storage proteins. The diagram below shows a
typical globular enzyme molecule. It has been drawn to highlight the
different secondary structures.

Globular Proteins
Have complex tertiary and sometimes quaternary structures.
Folded into spherical (globular) shapes.
Usually soluble as hydrophobic side chains in centre of structure.
Roles in metabolic reactions.
E.g. enzymes, haemoglobin in blood.


Fibrous (or Filamentous) Proteins Fibrous proteins are long and thin,
like ropes. They tend to have structural roles, such as collagen (bone),
keratin (hair), tubulin (cytoskeleton) and actin (muscle). They are
always composed of many polypeptide chains. This diagram shows part
of a molecule of collagen, which is found in bone and cartilage.

Fibrous Proteins
Little or no tertiary structure.
Long parallel polypeptide chains.
Cross linkages at intervals forming long fibres or sheets.
Usually insoluble.
Many have structural roles.
E.g. keratin in hair and the outer layer of skin, collagen (a
connective tissue).


4

How the shape of the enzyme/protein molecules
is suited to its function
Each enzyme/protein has specific primary structure
(amino acid sequence)
this sequence determines where the H-bonds will
form during development of the secondary structure
Proteins have a unique tertiary structure (further
folding of the secondary structure) held by ionic and
hydrogen bonds and if amino acids containing
cysteine are present, disulphide bridges
Globular proteins have an active site with unique
structure;
shape of active site complementary to/ will only fit
that of substrate
Enzyme substrate complexes can form

Describe how enzymes break down substances
Lowering the activation energy
Substrate with a complementary shape to the active
enters the active site
An enzyme substrate complex is formed
Active site changes shape to mould around the
substrate (induced fit)
This weakens the bonds in the substrate (lowering
activation energy) by stretching and distorting them
The bonds are broken
Products leave the active site
Enzyme remains unchanged


Describe how an enzyme catalyses a condensation
reaction
Enzyme active site has a complementary shape to the
substrate
An enzyme substrate complex is formed
The reactive groupshydroxyl, hydroxyl/ amino, and
carboxylic / hydroxyl and carboxylic are brought close
together
Change in the shape of the active site (induced fit) lowers
the activation energy
Water is removed and a glycosidic, peptide or ester bond
forms
Products leave the active site
The enzyme remains unchanged

The effect of temperature on the rate of an
enzyme reaction
As temperature increases so does the rate of the
reaction as
Substrate and enzyme gain K.E and collide more
frequently
More enzyme substrate complexes form
Further increases in the temperature cause bonds
(ionic, disulphide, hydrogen) holding the tertiary
structure of the enzyme in place begin to break
The enzyme denatures,
The active site no longer complements the substrate
No enzyme substrate complexes can form

The effect of pH on the rate of enzyme activity
Changes in the pH affect the charges on the R
groups of the amino acids at the active site
Interactions between the substrate and enzyme are
disrupted
Enzyme substrate complexes are less likely to form
More extreme pH conditions can cause the bonds
(ionic, Hydrogen) holding the tertiary structure to
break
The enzyme denatures (active site is no longer
complementary to the substrate)
No enzyme substrate complexes can form

Explain how inhibitors affect enzyme activity
There are two types of inhibitor, competitive and
non-competitive
Competitive inhibitors have a similar shape to the
substrate
They can enter and bind with the active site
Prevents enzyme substrate complexes forming
The problem can be overcome by increasing the
substrate concentration
Non-competitive inhibitors
Have a different shape to the substrate
Bind at a point other than the active site
They cause a change in the shape of the active site
Prevent formation of enzyme substrate complexes

5

Explain how the small intestine is adapted to its function in the digestion and
absorption of the products of digestion.
Large surface area provided by villi and microvilli
Thin epithelium gives a short diffusion pathway
The dense capillary network for absorbing amino acids and sugars and the lacteal
for the absorption of digested fats; ensures a steep concentration gradient is
maintained
The many mitochondria in the epithelial cells supply ATP/ energy for active
transport
Carrier proteins (in membranes) provide a path for polar molecules to pass through
the membrane.
Enzymes built into the epithelial membrane make it more likely for enzyme
substrate complexes to form and ensure products for absorption are released close to
the carrier and channel proteins


Absorption of glucose in intestines
Glucose moves into the epithelial cell with sodium Via a symport carrier
protein;
Sodium is removed from the epithelial cell by active transport at the sodium-
potassium pump;
Maintaining a sodium concentration gradient between the lumen and
epithelial cell
Glucose moves into blood by facilitated diffusion
And is carried away in the Heaptic portal vein

Digestion of Carbohydrates and Disaccharides
Starch Digestion:

Amylase
Hydrolyses
Glycosidic bonds
Producing maltose

Maltase hydrolyses
maltose to glucose
Lactose intolerance
Cause: reduced lactase levels as we age
Symptoms: diarrhoea and gas and cramps

Explanation
Gas comes from bacteria breaking down the
sugar

Diarrhoea: sugar lowers the water potential of
the lumen compared to epithelial cells, water
moves into the lumen by osmosis
Food moves through the
digestive system by peristalsis.
Mouth = digestion of carbs
Stomach = digestion of protein
Duodenum = most digestion,
receives pancreatic juice from
pancreatic duct and bile form
gall bladder (produced in liver)
Ileum = most absorption
Colon = absorption of water and
minerals
6

7

Describe the structure of a cell membrane and phospholipids.
Described as fluid mosaic.
Fluid: molecules within the membrane able to move;
Mosaic: mixture of phospholipid and protein

Double layer of phospholipid molecules;
Phospholipid consists of glycerol;
To which are joined two fatty acids;
And a phosphate;
formed by condensation reaction
Phosphate head is hydrophilic/polar
Fatty acid tail is hydrophobic
the phospholipids are arranged as bilayer in membrane;
Heads on the outside and tails on the inside;

Intrinsic proteins molecules pass through entire bilayer
Some of the proteins have channels/pores;
Some have specific binding sites and are carrier proteins

Extrinsic proteins only in one layer
Those on the outer side often act as receptors for hormones
Molecules can move in membrane/dynamic/membrane contains
cholesterol;
Many of the proteins and phospholipids have carbohydrates
attached forming glycolipids and glycoproteins that make up the
Glycocalyx

How the membrane regulates the movement of substances into and out of cells.
Non-polar/lipid soluble molecules move through phospholipid bilayer;
Small molecules/water/gases move through phospholipid layer/bilayer;
Ions/water soluble substances move through channels in proteins;
Some proteins are gated;
Reference to diffusion;
Carriers identified as proteins;
Carriers associated with facilitated diffusion;
Carriers associated with active transport/transport with ATP/pumps;
Different cells have different proteins;
Correct reference to cytosis;

How plasma membrane is adapted for its
functions.
Phospholipid bilayer (as a barrier);
Forms a barrier to water soluble but allows
non-polar substances to pass so maintains
a different environment on each side
(compartmentalisation)
Bilayer is fluid: can bend to take up
different shapes for phagocytosis and form
vesicles
Channel proteins (intrinsic): let water
soluble/substances through (facilitated
diffusion)
Carrier proteins (intrinsic): allow facilitated
diffusion and active transport
Extrinsic proteins: act in cell recognition,
act as antigens or receptors;
Cholesterol: regulates fluidity / increases
stability;



Membrane and Movement
The membrane is a phospholipid bilayer
Where the hydrophobic tails point inwards and the hydrophilic heads face outwards
The membrane contains two types of proteins
Intrinsic proteins (allow transport of water soluble molecules): spanning the entire bilayer
Extrinsic proteins: found on one side of the membrane

Most molecules move across the membrane by diffusion down a concentration gradient
Small molecules (water/gases) and lipid soluble molecules diffuse between the phospholipids
Polar molecules require channel or carrier proteins to move them
Channels are water filled pores that can be open at all times or they can be gated (voltage/ligand
gated)
Carrier proteins have a specific binding site for the molecules/ions
This can be facilitated diffusion, a passive (no ATP required) process
Some molecules are actively transported across the membrane (against the concentration
gradient)
This requires ATP (released in respiration)
The ATP changes the shape of the protein to move the molecule across the membrane

How proteins are arranged in a plasma membrane and role in transport
1 Some proteins pass right through membrane;
2 Some proteins associated with one layer;
3 Involved in facilitated diffusion;
4 Involved in active transport;
5 Proteins act as carriers;
6 Carrier changes shape / position;
7 Proteins form channels / pores;
8 Protein allows passage of water soluble molecules /
charged particles / correct named example;

Describing the fluid-mosaic structure of a membrane
Phospholipids and proteins;
Phospholipid bilayer: Arrangement of phospholipid molecules Tails to tails;
Molecules can move in membrane;
Intrinsic proteins extend through bilayer: Channel and carrier proteins
Extrinsic proteins in outer layer only: Act as antigens, receptors
Glycoproteins and glycolipids form glycocalyx
Presence of cholesterol to help regulate fluidity

8

The phospholipids are arranged in a bilayer (i.e. a double layer), with their
polar, hydrophilic phosphate heads facing out towards water, and their
non-polar, hydrophobic fatty acid tails facing each other in the middle of
the bilayer. This hydrophobic layer acts as a barrier to most molecules,
effectively isolating the two sides of the membrane. Different kinds of
membranes can contain phospholipids with different fatty acids, affecting
the strength and flexibility of the membrane, and animal cell membranes
also contain cholesterol linking the fatty acids together and so stabilising
and strengthening the membrane.
The proteins usually span from one side of the phospholipid bilayer to
the other (integral proteins), but can also sit on one of the surfaces
(peripheral proteins). They can slide around the membrane very
quickly and collide with each other, but can never flip from one side to
the other. The proteins have hydrophilic amino acids in contact with
the water on the outside of membranes, and hydrophobic amino acids
in contact with the fatty chains inside the membrane.
9

10

11

Diffusion: net movement of molecules form a high concentration to a low concentration

Lipid soluble molecules can diffuse easily though the phospholipid bilayer along with small hydrophilic molecules like water, carbon dioxide and oxygen

Facilitated diffusion: a passive process, moving large, hydrophilic molecules down the concentration gradient. The molecules cannot pass though the
hydrophobic bilayer and must enter/exit the cell through channel proteins or carrier proteins that are specific to the molecules.
Active transport
Moves a molecule against the concentration gradient (low to high)
Requires a specific protein carrier
Energy/ATP is used to change the shape of the protein
Energy is released in respiration

Rate of movement of molecules in facilitated
diffusion is limited by the availability of
carrier/channel proteins in the membrane. As
concentration increase rate will eventually
level out as the channels or carriers are
working at their maximum rate/ fully
occupied.
12


































Mechanism of the heart beat
Cardiac muscle is myogenic
The SAN
Sends a wave of electrical activity (depolarisation) across the
atria
This triggers atrial systole
The impulse is relayed to the ventricles through the AVN
Passing down to the apex of the heart along the bundle of His
The impulse spreads along the ventricle walls via the purkyne
fibres
The ventricles contract from the bottom
The AVN delays ventricular systole to allow them to fill up
with blood
Some key facts to learn

Valves in the heart and blood vessels prevent back flow
Tricuspid valve is on the right side of the heart
Bicuspid valve is on the left side of the heart
These two valves are called the atrioventricular valves
These valves are prevented from inverting as they are attached to the papillary
muscle in the ventricle walls, by tendinous cords

Semilunar valves are located between the ventricles and the aorta and pulmonary
artery. Rare for arteries to have valves
Pulmonary artery carries deoxygenated blood to the lungs. Rare for arteries to carry
deoxygenated blood
Pulmonary vein carries oxygenated blood to the heart from the lung, rare for vein to
have oxygenated blood.


Double circulation: blood flows through
the heart twice for one circuit of the
body, needs re-pumped after losing
pressure in the lungs

Deoxygenated blood returns via the vena
cava to the RA (1).
Atrium contracts blood through tricuspid
into RV (2).
Ventricle contracts and tricuspid shuts so
blood enters the pulmonary artery (3)
Blood returns to the LA (4) via the
pulmonary vein.
The LA contracts and blood forced
through the bicuspid valve into the LV
(5).
The LV contracts and the bicuspid shuts
and oxygenated blood flows into the
aorta (6)
The left ventricle has a thicker, more muscular wall than the right ventricle as it has to pump blood around the whole
body, so must generate a higher pressure.

When ventricular pressure > atrial pressure (1) the
atrioventricular valves shut to prevent backflow, this is the
first sound in the heart beat (lub)
When the ventricular pressure < arterial pressure (3) the
semilunar valves shut, this is the second sound of the heart
beat (dup)
QRS = electrical activity in the ventricles, occurs just before
ventricle pressure increases
P = electrical activity in atria and PQ = time delay due to
AVN

Cardiac Output: is the amount of blood flowing through the heart each
minute. It is calculated as the product of the heart rate and the stroke
volume:
Cardiac output = heart rate x stroke volume

The heart rate can be calculated from the pressure graph by measuring
the time taken for one cardiac cycle and using the formula:
Heart rate (beats/minute) = 60 time for 1 cycle
The stroke volume is the volume of blood
pumped in each beat. Both the heart rate and the
stroke volume can be varied by the body. When
the body exercises the cardiac output can
increase dramatically so that
Oxygen and glucose can get to the muscles
faster
Carbon dioxide and lactate can be carried away
from the muscles faster
Heat can be carried away from the muscles
faster
13

14








Smoking
decreases conc. of antioxidants in blood: this increases the damage done to artery walls;
raises the number of platelets in the blood and makes them more sticky :more blood clots are likely to form;
causes constriction of coronary arteries: raises blood pressure and damage to the artery lining
carbon monoxide combines with haemoglobin so less available to transport oxygen
blood pressure increased: due to increased heart rate
Fat
blood cholesterol level increases;
LDLs transport cholesterol in the blood;
LDLs deposit cholesterol in arteries
atheroma formed
blood pressure increased, turbulence makes clotting more likely
Salt
Increased salt concentration in blood
decreases water potential of the blood
water moves into the blood
blood pressure increased



How atheroma causes an aneurysm
Fatty material within walls of arteries;
Vessels narrow;
Blood pressure rises;
Weakened blood vessels may burst;
Describe how atheroma may form and lead to a myocardial infarction
Cholesterol deposited in the artery wall
This atheroma narrows lumen of the artery
This creates turbulence and can damage to lining of artery
Turbulence increases risk the of blood clot (thrombus)
The blood clot may break off (embolus)
And lodge in coronary artery;
Reduced blood supply to heart muscle;
Reduced oxygen supply;
Reduced respiration
Leads to death of heart muscle
Atheroma:
A build-up of cholesterol
in the artery wall

15

There are thousands of different kinds of cell, but the biggest division is between
the cells of the prokaryote kingdom (the bacteria) and those of the other four
kingdoms (animals, plants, fungi and protoctista), which are all eukaryotic cells.

Prokaryotic cells are smaller and simpler than eukaryotic cells, and do not have a
nucleus.
Prokaryote = without a nucleus
Eukaryote = with a nucleus

To see cells we need microscopes, like a light microscope. To see the
ultrastructure of a cell (the organelles inside) we need electron microscopes.
20 m
20m
If given a scale bar as below then the formula to use is

Actual length of scale bar
Magnification =
Representative length of the scale bar

Ensure you work in the same units


Cm mm m nm
10 1000 1000
Cytoplasm
A B
Capsule Cell wall Ribosomes Genetic material
If no scale bar is given then the formula to use is

Image Size
Magnification =
Actual Size of image

Ensure you work in the same units and then convert to the units
they want at the end

Resolution: how close 2 points can be to each other and still be distinguished as 2 separate points.
Electron microscopes have a higher resolution than (light microscopes, as they use electrons that have a shorter wavelength than light
Shorter wavelengths (like electrons) allow better resolution than longer wavelengths (like light).

Explain the advantages and limitations of using a transmission electron microscope
Advantages
1 TEM uses (beam of) electrons;
2 These have short wavelength;
3 Allow high resolution/greater resolution/Allow more detail to
be seen/greater useful magnification;

Disadvantages
4 Electrons scattered (by molecules in air);
5 Vacuum established;
6 Cannot examine living cells;
7 Lots of preparation/procedures used in preparing specimens/ fixing/staining/sectioning;
8 May alter appearance/result in artefacts;
9 very thin specimens
10 black and white, images
How to use a microscope
to measure the size of an
object.

Measure with an eyepiece
graticule

Calibrate with the stage
mcirometer (an object of a
known size)

Repeat and calculate an
average

16

Magnification and Resolution
Magnification simply indicates how much bigger the image is that the
original object. It is usually given as a magnification factor, e.g. x100. By
using more lenses microscopes can magnify by a larger amount, but the
image may get more blurred, so this doesn't always mean that more detail
can be seen.

Resolution is the smallest separation at which two separate objects can be
distinguished (or resolved), and is therefore a distance (usually in nm). The
resolution of a microscope is ultimately limited by the wavelength of light
used (400-600nm for visible light). To improve the resolution a shorter
wavelength
of light is needed, and sometimes microscopes have blue filters for this
purpose (because blue has the shortest wavelength of visible light).
Electron Microscopes
This uses a beam of electrons, rather than electromagnetic radiation, to
"illuminate" the specimen. This may seem strange, but electrons behave like
waves and can easily be produced (using a hot wire), focused (using
electromagnets) and detected (using a phosphor screen or photographic
film). A beam of electrons has an effective wavelength of less than 1nm, so
can be used to resolve small sub-cellular ultrastructure. The development of
the electron microscope in the 1930s revolutionised biology, allowing
organelles such as mitochondria, ER and membranes to be seen in detail for
the first time.

There are two kinds of electron microscope.
Transmission electron microscopes (TEM) work much like a light
microscope, transmitting a beam of electrons through a thin specimen and
then focusing the electrons to form an image on a screen or on film. This is
the most common form of electron microscope and has the best resolution
(<1nm).
Scanning electron microscopes (SEM) scan a fine beam of electron onto a
specimen and collect the electrons scattered by the surface. This has poorer
resolution, but gives excellent 3-dimentional images of surfaces.
Light Microscopes
These are the oldest, simplest and most widely-used form of microscopy.
Specimens are illuminated with light, which is focused using glass lenses and
viewed using the eye or photographic film. Specimens can be living or dead,
but often need to be coloured with a coloured stain to make them visible.
Many different stains are available that stain specific parts of the cell such
as DNA, lipids, cytoskeleton, etc.
1. Using a Magnification Factor
Image length = 40 mm
The magnification = 1000

The actual length is 40/1000 = 0.04mm

Usually convert this to m = 40 m
Estimation of size
It is possible to estimate the size of a structure seen with a microscope by comparing the image with a known
linear scale. Two pieces of apparatus are commonly used:
a graticule (eyepiece micrometer)
a stage micrometer.
A stage micrometer is a slide with a fine scale of known dimension etched onto it. An graticule is a fine scale that
fits inside an eyepiece lens. This is shown in Fig 3.
17

18

19

20

Folded inner membrane (cristae) increases the
surface area for the enzyme reactions.
Site of oxidative phosphorylation.
Fluid region = matrix
Has own circular DNA and 70s ribosomes, to
synthesise enzymes for respiration and allow
replication independent of the cell
Site of photosynthesis, fluid region is the
stroma and the membranes running
through are the thylakoid which can be
stacked to form grana. Contains
chlorophyll. Has its own circular DNA
and 70s ribosomes to synthesise enzymes
for the reactions and replicate
Nucleolus = synthesis of ribosomes

Nucleus
Thin strands of chromatin that condense at
replication to chromosomes.
Controls the cell through coded information
DNA
Nuclear pores allow movement of material
in and out
The golgi body is a stack of flattened membrane sacs that are referred to as
cisternae)
This is involved in the modification and packaging of materials from both the rough
and smooth endoplasmic reticulum. Addition of carbohydrates to form glycolipids
and glycoprotein (like mucus). It will also produce lysosomes (digestive enzymes)
Endoplasmic reticulum: is a stack of interconnected flattened sheets called cisternae
ER is Continuous with the nuclear envelope and can be either smooth or rough (studded with
ribosomes). The smooth ER is involved with the production of sterols, phospholipids and
detoxification.
The rough ER is involved with protein production at the ribosomes. Usually for export out of the
cell
The ER provides a pathway for materials through the cell Dominates in cells producing proteins
for export) digestive enzymes
21

Nucleus. This is the largest organelle. It is surrounded
by a nuclear envelope, which is a double membrane
with nuclear pores large holes containing proteins
that control the exit of substances from the nucleus.
The interior is called the nucleoplasm, which is full of
chromatin the DNA/protein complex (see unit 2).
During cell division the chromatin becomes condensed
into discrete observable chromosomes. The nucleolus
is a dark region of chromatin, involved in making
ribosomes.
Mitochondrion: This is a sausage-shaped organelle
(8Vm long), and is where aerobic respiration takes
place in all eukaryotic cells (anaerobic respiration takes
place in the cytoplasm). Mitochondria release energy
(in the formATP) from carbohydrates, lipids and other
energy rich molecules. Cells that use a lot of energy
(like muscle cells) have many mitochondria.

Mitochondria are surrounded by a double membrane:
the outer membrane is simple and quite permeable,
while the inner membrane is highly folded into cristae,
which give it a large surface area. The space enclosed
by the inner membrane is called the mitochondrial
matrix, and contains small circular strands of DNA. The
inner membrane is studded with stalked particles,
which are the enzymes that make ATP.
Ribosomes: These are the smallest and most numerous
of the cell organelles, and are the sites of protein
synthesis. Ribosomes are either found free in the
cytoplasm, where they make proteins for the cell's own
use, or they are found attached to the rough
endoplasmic reticulum, where they make proteins for
export from the cell. All eukaryotic ribosomes are of the
larger, "80S", type.
Endoplasmic Reticulum (ER). This is a series of
interconnected membrane channels involved in
synthesising and transporting materials. Rough
Endoplasmic Reticulum (RER) is studded with numerous
ribosomes, which give it its rough appearance. The
ribosomes synthesise proteins, which are processed in the
RER (e.g. by enzymatically modifying the polypeptide chain,
or adding carbohydrates), before being exported from the
cell via the Golgi Body.
Smooth Endoplasmic Reticulum (SER) does not have
ribosomes and is used to process materials, mainly lipids,
needed by the cell.
Golgi Body (or Golgi Apparatus). Another series of flattened
stacks of membrane vesicles, formed from the endoplasmic
reticulum. Its job is to transport proteins from the RER to the
cell membrane for export. Parts of the RER containing proteins
fuse with one side of the Golgi body membranes, and are
modified (carbohydrate is added to form glycoproteins), while
at the other side small vesicles bud off and move towards the
cell membrane, where they fuse, releasing their contents by
exocytosis.
Lysosomes: These are small membrane-bound vesicles formed from
the RER containing a cocktail of hydrolytic enzymes. They are used
to break down unwanted chemicals, toxins, organelles or even
whole cells, so that the materials may be recycled. They can also
fuse with a feeding vacuole to digest its contents.
Chloroplast
Membranes arrangement and disc shape provides large
surface for light absorption;
layering of membrane allows a lot of pigment;
Permeable membrane allows diffusion of gases /carbon
dioxide;
membranes provide surface for attachment of electron /
hydrogen acceptors;
Contains chlorophyll for light absorption;
Contains different pigments to absorb different
wavelengths;
Stacking / arrangement of grana/thylakoids maximises
light catchment;
Stroma contains enzymes for photosynthesis;
Outer membrane keeps enzymes in chloroplast;
Starch grains / lipid droplets store products of
photosynthesis;
Ribosomes / DNA for enzyme/protein synthesis;
Shape of chloroplast gives large surface area for CO
2
,
absorption.



22

Parts of the prokaryotic cell and the function
A) Cell membrane: regulates entry/exit/selectively permeable;
B) Mesosome: respiration/cell division;
C) Cell wall: (mechanical) protection/prevents (osmotic) lysis;
D) Slime layer/capsule: protection (against e.g. antibiotics);
E) Flagellum: movement of cell;
F) DNA: molecule/bacterial chromosome, genetic information;

A
B
C
D
E
F
Vibrio Cholera
Rod shaped bacterium
Causes dehydration through diarrhoea (5lts/day)

Ingested through
Contaminated water
Food (shellfish, bottom dwellers where sewage leakage has
occurred) or preparation by infected person.
Affects body through a toxin it produces. Toxin only affects the
upper regions of the small intestine as this is the only region with
membrane receptors that complement the toxin.
Mechanism of infection
Many of the organisms are destroyed by the stomach acid
but those that survive pass into the small intestine, and
using flagella in a corkscrew motion to get through the
mucus layer in upper region of the small intestine
(duodenum) and anchor itself.

Bacteria produce an exotoxin (protein secreted by
microbes), that binds to specific receptors on the cell
surface membrane (receptors located only in the upper
region of the small intestine).

Toxin causes chlorine ion channels to open and chlorine
ions flood the intestinal lumen (other ions enter like
sodium and potassium)

Excess ions in the lumen, lowers the water potential (more
negative) so water leaves the cells by osmosis. Ions enter
the cells to replace those lost and along with the loss of
water the cells water potential becomes more negative than
the blood so water leaves blood leading to dehydration.

[An endotoxin is not secreted, but is a component of the
bacteria released when they are lysed]

Difference between an endotoxin and
an exotoxin.
Endotoxins produced from the
breakdown of bacteria (cell walls);
exotoxins secreted (from living cells)
Endotoxins are lipopolysaccharides;
exotoxins are protein;

Treating effects of cholera
Oral rehydration therapy (ORT)
replaces lost water and salts;

Mechanism
1. Contains glucose/starch/ sugar;
2. Sodium/salt;
3. makes use of Co-transport protein
4. Sodium and glucose taken up (from
lumen by co-transport protein);
5. Lowers water potential in cells/
increases water potential gradient;
6. Water taken up by osmosis

How the distribution of cell membranes in a prokaryotic cell differs from that in a cell from eukaryotic.
Absence of nuclear membrane, No membrane bounded organelles; Such as mitochondria/chloroplast/vacuole/lysosome; No membrane systems/endoplasmic reticulum/Golgi; there are
mesosomes in prokaryotes;

23

How V.cholerae causes Diarrhoea

1. The cholera bacterium adheres to the epithelium and secretes the cholera toxin CT. CT enters the
epithelial cells and activates a chloride ion channel in the cell membrane.
2. This causes chloride ions to diffuse out of the cells into the lumen.
3. This lowers the water potential in the lumen of the gut.
4. So water is lost from cells to the lumen by osmosis, producing diarrhoea and dehydration.
Treatment for Diarrhoea

The treatment for diarrhoea was revolutionised in the 1960s, with the development of oral rehydration therapy (ORT).
This simple and cheap treatment consists of drinking an oral rehydration solution (ORS) of glucose and salt (NaCl), and
sometimes other ions like potassium and bicarbonate. ORT makes use of the sodium-glucose co-transporter protein that
normally absorbs glucose into the ileum epithelial cells.

1. If both Na+ and glucose are present in the lumen, they bind to the sodium-glucose co-transporter protein. Transport
only works if both molecules are present, which is why salt alone is not an effective treatment. ORS contain equimolar
concentrations of glucose and salt.
2. The transporter protein carries the Na+ and glucose into the cell, down their concentration gradients.
3. This lowers the water potential inside the epithelial cells. So water diffuses from the lumen into the epithelial cells by
osmosis, rehydrating cells and reducing diarrhoea.
24


Cell fractionation: Separating different parts and organelles of a
cell, so that they can be studied in detail. The most common
method of fractionating cells is to use differential centrifugation.

Organelles are separated based on different densities

There are 3 key stages
1) Homogenisation
2) Filtration
3) Centrifugation

Homogenisation: Pestle mortar (using and as abrasive component)
In a blender
This breaks open cells. Done in a
Buffered solution: Preventing pH changes that may damage the organelle
by denaturing enzymes or proteins affecting function

Isotonic: prevent osmotic movement where swelling of organelle may
cause lysis or shrinkage of organelle both resulting in destruction of
natural function

Cold (5
0
C): This reduces enzyme activity and will prevent autolysis (self-
destruction) of the organelle.

2) Filtration: removes any debris (unbroken cells, cartilage, and sand). This
will prevent contamination of the pellets resulting from centrifugation


3) Centrifugation: Homogenate (solution of organelles) is centrifuged at
different speeds. Low speeds will cause more dense organelles to separate
and higher speeds will cause least dense organelles to sediment.

After spinning the pellet of organelle is at the bottom the remaining
solution is called the supernatant and poured (decanted) for
centrifugation.

Most dense organelle Nucleus
Mitochondria/Chloroplasts
ER, Golgi, Lysosomes
Least dense organelle Ribosomes

80s in eukaryotic cells will sediment before 70s in prokaryotic cells as they
are larger (more dense)

Describe how you would obtain a sample
of undamaged chloroplasts from leaves.

1. Chop up leaves in a.
2. Cold: reduces enzyme activity
3. Buffered solution: prevents pH
changes affecting enzymes
4. Isotonic: prevents osmosis and
possible lysis or shrinkage of organelles
5. Filter and centrifuge filtrate
6. Centrifuge supernatant
7. at higher speed;
8. Chloroplasts in (second) pellet;

Key to remember you must separate
organelles in order, most dense first
then least dense, if you try and jump to
a particular stage the sample will be
contaminated


25




The rate at which a substance can diffuse is given by Fick's law:
From Fick's law we can predict that, in order to support a fast rate of diffusion, exchange surfaces must have:
1) A large surface area 2) a small distance 3) a mechanism to maintain a high concentration gradient across the gas exchange surface.
Describe and explain the difference in the composition of
gases in inhaled and exhaled air.
1 inhaled air contains more oxygen than exhaled air;
2 inhaled air contains less carbon dioxide than exhaled air;
3 inhaled air contains less water (vapour);
4 relative amount/percentage of nitrogen also changes;
5 respiration results in lower blood O2 / higher blood CO2;
6 oxygen enters blood / carbon dioxide leaves blood in alveoli;
7 by diffusion;
8 water vapour diffuses from moist surface;


A thin surface and a diffusion gradient are both features of gas
exchange surfaces. Describe how these are achieved at the gas
exchange surfaces of a mammal;
Wall of alveoli / capillaries have single epithelial layer/
Alveoli and capillaries are close together;
the epithelium is flattened (squamous)
Ventilation maintains high O
2
/low CO
2
concentration(in alveoli);
blood flow/circulation maintains high CO
2
/ low O
2
concentration(in
blood);



Describe and explain how the structure of the mammalian breathing system and
red blood cells enables efficient uptake and transport of oxygen.
Alveoli provide a large surface area;
Walls of alveoli/capillary are thin (single layer of flattened/squamous epithelial
cells) to provide a short diffusion pathway;
Capillary are close to alveoli provides a short diffusion pathway;
narrow capillaries slows blood flow - more time for diffusion; and forces close
contact between red blood cells and capillary walls thus reduces diffusion path;
Many blood capillaries provide a large surface area and keeps oxygen in blood
around alveoli low, thus maintains a steep diffusion gradient
Intercostal muscles and diaphragm muscles enable ventilation (keeps oxygen in
alveoli high) to maintain a steep diffusion
wide trachea and branching of bronchi/bronchioles for efficient flow of air;
cartilage rings keep airways open;
Cell membrane permeable to gases;
RBC biconcave discs so large surface area;
They have no nucleus/mitochondria;
They contain Haemoglobin for transport of oxygen;

Explain how inspiration occurs.
Diaphragm contracts/moves down/ flattens;
External intercostal muscles contract rib cage
upwards and out
Increases volume (of thorax);
Decrease in pressure;
Air moves from high to lower pressure in lungs

Explain how Expiration occurs
A passive process, where the external
intercostals and the diaphragm relax.

However in forced expiration, the internal
intercostals muscles are involved (contracting
to pull the ribs down) and the abdominal
muscles are involved (contracting to force the
diaphragm up).

26

Change in
lung volume
/ dm
0.5
0
0 0.5 1.0 1.5 2.0 2.5 3.0 3.5
Time / s
3
Inspiration Expiration
Graph 1
Pulmonary Ventilation is the volume air ventilating the lungs each minute. It is calculated as the
product of the ventilation rate and the tidal volume.

Pulmonary ventilation = ventilation rate x tidal volume

The ventilation rate can be calculated from the pressure graph by measuring the time taken for one
ventilation cycle and using the formula:

Ventilation rate (breaths/minute) = 60 time for 1 cycle

So for diagram shown Ventilation rate = 60 3 = 20

The tidal volume is the normal volume of air breathed in each breath (also called the breathing depth).
It can be measured from the volume graph.
Both the ventilation rate and the tidal volume can be varied by the body. When the body exercises the
pulmonary ventilation can increase so that
Oxygen can diffuse from the air to the blood faster
Carbon dioxide can diffuse from the blood to the air faster
These changes allow aerobic respiration in muscle cells to continue for longer.
27

Describe the transmission and course of infection of pulmonary tuberculosis.
1 (Bacteria transmitted in) droplets / aerosol;
2 (Bacteria) engulfed / ingested by phagocytes / macrophages;
3 (Bacteria) encased in named structure e.g. wall / tubercle / granuloma / nodule;
4 (Bacteria) are dormant / not active / not replicating;
5 If immunosuppressed, bacteria activate / replicate / released;
6 Bacteria destroy alveoli / capillary / epithelial cells;
7 (Leads to) fibrosis / scar tissue / cavities /calcification;
8 (Damage) leads to less diffusion /less surface area / increases diffusion distance;
9 (Activation / damage allows bacteria) to enter blood / spreads (to other organs);


People with emphysema may feel weak and tired. Explain why.
Alveoli break down: Less surface area for gas exchange
Walls thicken (scar tissue): increases diffusion distance
Loss of elastin (elastic tissue) due to elastase from white blood cells involved: Alveoli cannot recoil so it
is more difficult to expel air Reduced diffusion gradient
Less oxygen enters blood and then tissues;
Less respiration / less energy released / less ATP produced;

You can recognise the difference between healthy lung tissue and that with emphysema because, with
emphysema
There are a smaller number of alveoli, larger air spaces per alveolus, there are thicker walls.

Risk increased with.
Infection e.g. (chronic) bronchitis; heredity;
Industrial pollution - must contain reference to inhalation of particles (dust)
Smoking

Asthma is caused by physical factors called allergens in the environment. These allergens include pollen, dust mites faeces and fur.
These allergens trigger an inflammatory response by the immune system.

White blood cells called mast cells release histamines, which cause the smooth circular muscles of the bronchioles to contract, narrowing the airways
(bronchoconstriction).

The epithelial cells also secrete more mucus, which further blocks the airways.

The constricted bronchioles stimulate wheezing and coughing as the lungs try to loosen the mucus. The constrictions reduce the tidal volume, so
alveolar air is only replaced slowly. The oxygen concentration gradient across the alveolar epithelium is reduced, so the rate of diffusion in the alveoli
is reduced by Ficks law. Less oxygen diffuses into the blood, so less oxygen is available for cellular respiration throughout the body.
Symptoms of Asthma
Difficulty breathing due to constriction of airways and mucus build up.
Wheezing due to narrow pathway and sir flowing through it
Tight feeling in the chest due to constriction of smooth muscle
Heavy coughing to remove obstructions

Chronic bronchitis
Tar from smoking leads to the enlarging of goblet cells and excess mucus production.
The tar will also destroy or reduce effectiveness of the cilia so mucus removal is less
effective. This results in heavy coughing to try and shift the obstruction, heavy
coughing can result in damage to the epithelial cells, this can cause scar tissue
development and also induce an inflammatory response where even more mucus is
produced furthering the obstruction.

The trapped mucus increases the susceptibility to chest infections (mucus traps
microbes in the air) which again can result in tissue damage (microbial activity) and
result in immune responses such as inflammation and mucus production and also the
attraction of phagocytic cells.

Pulmonary fibrosis: when scars form on the epithelia that are damaged, increasing the diffusion
pathway, loss of elasticity in lung tissue, which reduces the concentration gradient, narrowing of
vessels, reducing air flow and concentration gradient. Results in shortness of breath, dry cough,
tiredness (insufficient oxygen for respiration)
28

Disease is a general term meaning a disorder of the body. Diseases can be
caused by many different factors:
Infectious Diseases are caused by pathogenic organisms (usually microbes)
living in or on the body and so causing harm (e.g. cold, TB, AIDS).
Dietary Deficiency Diseases are caused by a lack of specific nutrients in the
diet, e.g. kwashiorkor (protein), scurvy (vitamin C), and rickets (vitamin D).
Environmental Diseases are caused by non-living factors in the
environment. They include skin cancer (caused by radiation), lung cancer
(caused by smoking), asthma (caused by dust), pulmonary fibrosis (caused by
dust or pollution), and Creutzfeldt-Jakob disease (caused by prions).
Social Diseases are caused by human activities and lifestyle. They include
alcoholism, emphysema, coronary heart disease, anorexia, drug addiction and
even accidents.
Ageing Diseases are caused by degeneration of body tissues and include
arthritis, arteriosclerosis and cataracts.
Genetic Diseases are caused by genes inherited from parents. These are
really characteristics that are unusual in the population and are life-
threatening (e.g. muscular dystrophy, cystic fibrosis, haemophilia).
In fact all diseases are affected by genetics, but these single gene disorders
are governed entirely by the action of a single allele and are not influenced by
any other factor.
For a pathogen to cause a disease these steps must take place:
1. The pathogen must be transmitted to the human host (through drinking water, eating food, breathing
aerosol droplets, animal bites, or direct contact)
2. The pathogen must gain entry inside the human body. The human body is protected by a tough layer
of endodermis (skin), but pathogens can enter via cuts in the skin (e.g. malaria); or the thinner
epithelium exchange interfaces, such as the digestive system (e.g. cholera) or lungs (e.g. tuberculosis).
3. The pathogen must evade the defences of the host. Humans have a range of defences, such as
stomach acid, lysozyme enzymes and the immune system, and these defences are usually very effective
at preventing disease. But it only takes a few pathogen cells resisting the defences to multiply and cause
a disease.
4. The pathogen must harm the host. Pathogens harm their hosts in two ways.
First, by reproducing inside host cells, using up cellular resources and preventing the cell from carrying
out its normal reactions. The microbes then usually burst out of the host cell, rupturing the cell
membrane and killing the cell in the process.
Second, by producing toxins chemicals that interfere with the body's reactions. These chemicals may
inhibit enzymes, bind to receptors, bind to DNA causing mutations, interfere with synapses and so on.
A persons lifestyle affects their chances of suffering from
diseases (except the single gene disorders).
Lifestyle factors can include diet, exercise, work environment,
sexual habits, smoking, drinking and drug-taking. Some of these
factors have obvious association with disease (like smoking),
while others are less obvious (like occupation), but all the factors
have an associated risk.

Different disease have specific risk factors, i.e. factors that
specifically increase the risk of getting that disease. A few
examples are:
Disease Risk Factors
Lung cancer: smoking and cleanliness of the environment.
Skin cancer: exposure to sunlight and colour of skin.
CHD: diet, age, genetics and exercise.
Diabetes: genetics diet and exercise.
AIDs: sexual habits, drug habits and genetics.

Some of these risk factors are beyond our control, e.g. genes and
age. But the others are lifestyle factors and so within our power
to change.
1) This graph
shows there is no
pattern between
income and
incidence of lung
cancer
2) This graph
shows a correlation
between smoking
and cancer. But it
does not prove
causation as other
factors can
influence results,
age, diet, and
genetics
3) To show causality
controlled experiments
are needed. Here arsenic
(component of cigarette
smoke) inhibits DNA
ligase which repairs
damaged DNA. Thus in
cells this could lead to
cancer, now we have a
mechanism to explain
graph 2, and we have
evidence for a causal
relationship between
smoking and cancer.
4) There is a correlation
between alcohol and
cancer. Lab studies have
not found a causal link
between the two, so
alcohol is not a risk factor.
The correlation is indirect,
where heavy drinkers,
tend to be heavy smokers
29

Defence mechanisms of the body reduce the chance of entry by a pathogen.
Skin: layer of dead cells acts as a barrier to microorganisms
impregnated with keratin
fatty acids in sebum inhibit growth of microorganisms;
commensal bacteria compete with other microorganisms;

Tears (also saliva ): contain lysozyme; capable of digesting bacterial cell walls;;
Mucus in respiratory system traps pathogens;
microorganisms are moved up the respiratory tract by cilia and are swallowed;
killed by acid in the stomach;

Stomach contains hydrochloric acid which destroys bacteria;

Blood clot prevents entry;







Describe phagocytosis.
1. Phagocyte attracted by chemicals
2. (Pathogen) engulfed;
3. Enclosed in vesicle (phagosome)
4. (Vesicle) fuses with lysosome; (phagolysosome)
5. Lysosome contains hydrolytic enzymes;
6. Pathogen digested/ molecules hydrolysed;
7. Phagocyte may display some of the foreign
antigen on its cell membrane becoming an
Antigen Presenting Cell (APC)

Antibodies
B lymphocytes make antibodies/immunoglobulins. They are proteins
that bind to specific antigens forming an antigen-antibody complex

They are 4 polypeptide chains consisting of 2 heavy and 2 light chains,
held by disulphide bridges. The constant region is the same in all
immunoglobulins (same amino acid sequence), but the variable regions
vary greatly due to different amino acid sequences, it is this region that
binds to the antigen.

protein molecules are suited to carry out the role of antibodies because
A Large variety of different molecules is possible
a variety of shapes can be created;
As the Tertiary shape will vary as the primary sequence varies;
thus it is likely that a shape complementary to a specific antigen; will be
created
The Immune System is the bodys defence system against disease. It is made up of
white blood cells (or leukocytes), which are found in the blood, lymph, tissue fluid and
body cavities (such as alveoli). There are dozens of different kinds of leukocytes, which
fall into four categories: there are two branches to the immune system
1) Non-specific: chemical and cellular (phagocytosis)
2) Specific: Humoral (B cells) and cell mediated (T cells)
Only vertebrates have a specific immune system. The key feature of the
specific immune system is that it can recognise foreign cells, those distinct
form its own cells. This is called self/non-self recognition. It can do this
because of antigens: Protein/glycoproteins/lipoproteins that stimulate the
immune response


How antibodies work
1. By binding to antigens on viruses and bacteria they prevent the
viruses or bacteria attaching to cells and so infecting them.
2. By binding to free toxin proteins they change the shape of the
active region so that these proteins
can no longer take part in the reactions that caused disease.
3. By linking cells together. Because each antibody molecule has
two antigen-binding sites (one on each arm of the Y), antibodies
can stick cells together into large clumps. This process, called
agglutination, immobilises viruses and cells, and precipitates
soluble toxins so that they can easily be destroyed by phagocytes
or cytotoxic T-cells. Large antigen-antibody complexes also
stimulate the
30

The role of B and T cells in the defence of the body against a virus infection.
B lymphocytes produce antibodies/involved in humoral response;
T lymphocytes involved in cell mediated immunity;
Macrophages present antigens;
(specific) B lymphocytes recognise/bind to antigen;
increase in numbers by mitosis;
produce plasma cells (which make antibodies);
antibodies bind to and clump/ agglutinate virus;
memory cells produced by 1
st
exposure/cloned on 2
nd
exposure;
T lymphocytes(helpers) produce lymphokines/chemicals;
which aid B lymphocyte cloning;
encourages phagocytes to engulf clumped virus;
killer T cells kill virus infected cells
Explain how the body responds both generally and specifically to pathogens
that enter the blood.
Non-specific response
1) Chemical response: Inflammatory response: histamines are released body
temperature increased Interferon production (details on sheet)
2) Action of phagocytes: detail explained on this sheet

Specific immune response
1) Humoral response involving B cells, T helper release cytokines to encourage
mitosis in B cells forming clones. B plasma cells produced, these secrete
antibodies that. Agglutinate pathogens making phagocytosis more likely and
they can neutralise toxins/pathogens

2) Cell mediated response involving T cells, cytotoxic cells destroy infected
body cells

Memory cells produced;
On further exposure to same microorganism; Antigen recognised;
Faster response; Greater production of antibodies;


Inflammation: release chemicals, like histamines and
kinins (cause inflammation with added effect of
attracting phagocytes) which stimulate:
Vasodilation to increase the flow of blood to the area,
so the area turns red.
Capillary leakage so that phagocytes and granulocytes
can enter the local tissue fluid. The area swells
Sensory neurone impulses, so the area is tender or
painful.
Blood clotting to seal a wound, so a scab is formed.

Interferon is released form viral infected cells,
stimulating nearby cells to produce chemicals to
prevent the entry of the virus.
Foreign antigens need to be presented on antigen-presenting cells (APC) to initiate the specific immune
response. The most important antigen-presenting cells are macrophages. Whenever they find a non-self
antigen the macrophage ingests the antigen and its cell by phagocytosis. Some of the antigens pass to the
surface of the macrophage, which thus becomes an antigen-presenting cell. This method of presenting
antigens amplifies the number of foreign antigens in the blood without increasing the number of pathogens.
The macrophage also secretes chemicals to stimulate the next stage of the immune response clonal
selection.

APC interacts with the Helper T-cells (T
H
cells), and when they binds, the T
H
cell begins to secrete chemicals
called cytokines which activate T and B lymphocytes to divide mitosis (clonal selection) and differentiate.

T-lymphocytes differentiate into cytotoxic T cells bind to complementary antigens and destroy the cells by
making pores in them which leads to cell lyses.

Activated B cells differentiate into plasma cells (rich in RER), which secrete antibodies

Some activated B and T cells differentiate into memory cells which remain in the body for years, they
provide the secondary immune response.
31

Give two ways in which
passive immunity differs from
active immunity.
Antibodies not produced by
body;
No memory cells;
Short-term / not lifelong;
Antibodies (or context
established) donated by mother /
across placenta / in milk;


Some pathogens antigens remain constant, others develop
new strains with different antigens (as a result of
mutations) thus the memory cells no longer recognises
(compliments) the antigen. These pathogens show
antigenic variability.
32

What is vaccination?
Injection of antigens/toxins/dead weakened pathogens
(Antigen from) attenuated microorganism/non-virulent microorganisms/dead
microorganisms/isolated from microorganism;
Stimulates the formation of memory cells;

Methods used to prepare vaccines.
Killed microorganism
Modified toxin
Attenuated/heat treated/UV treated microorganism
genetically engineered antigens
isolated antigen


Explain how a host is made less susceptible by the use of vaccination.
Nature of vaccine e.g. attenuated strain or isolated antigen or toxin
vaccine introduces antigen into host
stimulates lymphocytes;
memory cells produced;
if host subsequently meets active pathogen
production of same (B/T) lymphocytes;
antibodies and cytotoxic cell production can occur in large number s and rapidly
Pathogen destroyed before it can affect host;

Vaccines protect against disease by stimulating the production of memory cells.
Describe how memory cells protect the body from disease.
On further exposure to same microorganism;
Antigen recognised;
Faster response;
Greater production of antibodies;

Explain why it is necessary to vaccinate elderly against the flu every year.
Influenza virus mutates;
different strains / different - shaped antigen; shows antigenic variability
mutant forms will not be recognised by lymphocytes memory cells
immune system; accept elderly have weaker immune system

Suggest two reasons why parents may decide against vaccination for their children.
consider vaccines to be unsafe / have side effects / damage immune system;
consider natural immunity to be more effective; allow in (a) if not here
religious / ethical objections qualified e.g. objections to use of fetal /
animal tissue;
consider low risk of disease when high percentage of population already
vaccinated/Ref. to Head Effect




Immunisation programmes may use either attenuated or dead microorganisms. Suggest why
there might be problems for the patient when using these vaccines.
Process of killing organisms might not be 100% efficient;
live organisms might give rise to full-blown disease;
attenuated organisms are non-virulent;
but might mutate to virulent forms;
immunity can decline - booster injections required;
named side effects, eg allergies;
less effective due to changed antigens;


Give two ways in which passive immunity differs from active immunity.
Antibodies not produced by body;
No memory cells;
Short-term / not lifelong;
Antibodies (or context established) donated by mother /across placenta / in milk;


If enough people are vaccinated in a population (typically 85-95%), then even the few that are not, or
cannot be, vaccinated are protected by herd immunity, since there are not enough hosts for the pathogen
to survive and reproduce.


Success of vaccination programmes
- Able to provide for all the vulnerable groups and at the same time to
interrupt the transmission
- Few (if any) side effects
- Easy to produce and store

Why does it not eliminate disease?
Not all people will develop immunity
Attenuated antigen mutates (antigenic variability)
Not all people want to get the vaccine
Booster injections may be required

33

Monoclonal Antibodies
The unique tertiary structure of each antibody protein allows it to bind specifically and tightly
to one particular antigen. Scientists quickly realised that the remarkable specific binding
property of antibody proteins in vivo would make them very useful tools in medicine and
research in vitro. [In vivo means in life, i.e. in a living organism; and in vitro means in glass,
i.e. in a test tube.] Monoclonal antibodies are antibodies of one particular shape made by a
clone of a single B-lymphocyte.

Making Monoclonal Antibodies
Antibody proteins are far too complicated to be synthesised chemically in vitro: they have to be
made by living cells. In 1975 Kohler and Milstein developed a method to make monoclonal
antibody proteins using mice.

1. Inject a mouse with the antigen protein that you want antibodies for. The mouse will show a
primary immune response and make a clone army of B-lymphocytes with antibodies specific for
that antigen.

2. After a few days, extract B-lymphocyte cells from the rabbits blood. The blood contains a
mixture of thousands of different B-cells, each making their own specific antibodies, so we
need to isolate the B-cell we want. Dilute the blood cells into hundreds of wells in an
immunoassay plate, so that there is just one cell per well. The cells multiply in their wells and
secrete antibodies a different antibody in each well.

3. Test each well for production of the antibody required and row the B-cells from that well in
a culture flask, where they multiply by mitosis, making millions of identical cloned cells, each
secreting identical antibodies monoclonal antibodies.
Using Monoclonal Antibodies
Monoclonal antibodies have many uses, but they are all based on the same
principle. If monoclonal antibodies are mixed with a sample containing a
mixture of proteins, the antibodies will bind specifically and tightly to only one
protein in the sample.
The monoclonal antibodies can have another molecule chemically attached to the
constant region, which can make the antibody coloured, or fluorescent, or attach it to
a surface. This allows the target protein to be visualised.
Some uses of monoclonal antibodies include:

Antibodies can be used as a magic bullet to target drugs to one specific cell type in
the body. Monoclonal antibodies are made to an antigen only found on the target
cell, and the drug is bound to the constant region of the antibody. The antibody/drug
complex is then be injected into the patient and the antibody will ensure that the
agent is carried only to the target cells and nowhere else.

Antibodies can be made to target a toxic agent (e.g. a radioactive substance) to
cancer cells and nowhere else in the body.

Antibodies to the protein hormone hCG, produced in pregnancy, are bound to a test
strip and used to detect the presence of hCG in urine in a pregnancy test strip.

Antibodies are used to detect the presence of specific proteins in very low
concentrations in the ELISA assay.

Fluorescent antibodies are used to stain particular cell organelles in microscope
slides.

Antibodies can be used directly in passive immunity to help the body's normal
immune response to a serious infection
What is a monoclonal antibody
A hybrid cell from tumour (cancerous,
myeloma) and B-lymphocyte called a
hybridoma
it produces the same antibodies as it is
derived from one type of plasma cell;
this means the antibodies are specific
(complementary) to fits only one
antigen
34

35

36

37







































Difference between the glucose molecules is the
orientation of the OH group at C1.
Basic structure of starch
Storage polysaccharide
Insoluble (no effect on water potential and
thus osmotically inactive)
Not a pure substance but a mixture of

Amylose: a chain of alpha glucose held by 1,4
glycosidic bonds. It forms a helix, held by H
bonds within the chain

Amylopectin: a polymer of alpha glucose
with 1,4 glycosidic bonds and a small number
of 1,6 branches. This gives it an open
structure and the branches are quickly
hydrolysed


Cellulose structure and function

Is a polymer of beta glucose.
Condensation polymerisation occurs
The monomers are held by1,4 glycosidic bonds.
Alternate beta glucose molecules are inverted
The chain is long and straight
Adjacent chains can be held together by H-bonds

This forms micro-fibrils which are rigid and can link to
form cellulose fibres
Cellulose is a component of the cell wall
Its strength means it can resist osmotic pressure
Starch: structure for function
Role = storage
Features: Insoluble stays in cell
Features: Osmotically inactive cell does not absorb
water
Feature: good respiratory substrate provides many
glucose molecules on hydrolysis
Feature: Amylose is Coiled lots of glucose in a small
space
Feature: amylopectin branched is rapidly
hydrolysed to glucose because enzymes can begin to
operate on all brnaches
Basic structure of glycogen

Glycogen is similar to amylopectin.

It is polymer of (1-4) alpha glucose with 9%
(1-6) branches, though more than starch.
Because it is so highly branched, it can be
mobilised (broken down by glycogen
phosphorylase to produce glucose for energy)
very quickly, reflects the grater metabolic
demands of animal over plant

Animals storage polysaccharide

Found mainly in muscle and liver.


Starch/Glycogen vs Cellulose
Starch/Glycogen
1. (1,4 and) 1,6 bonds/contains
1,6 bonds /branching
2. All glucoses/ monomers same
way up
3. Helix/coiled/compact
4. Alpha glucose
5. No (micro/macro)
fibrils/fibres

Celluose
1. 1,4 bonds / no 1,6 bonds /
unbranched / straight;
2. Alternate glucoses/monomers
upside down;
3. Straight;
4. Beta glucose;
5. Micro/macro fibrils/fibres;

38


Q. Describe how the structure of a chloroplast is adapted to its function in photosynthesis.
Membrane is permeable to gases
Disc shape gives a large surface area for absorption of light and gas
Contains chlorophyll to absorb light
Contains a range of pigments to increase the range of wavelengths that can be absorbed
Stacking of the thylakoids (grana) maximises light absorption
Stroma contains enzymes for the reactions of photosynthesis
Stroma contains ribosomes (70s) and DNA for making enzymes needed in photosynthesis

Q. Photosynthesis generally takes place in a leaf.
Describe how the leaf is adapted to allow this process
to occur effectively.
Large surface area to collect solar energy;
transparent nature of cuticle to allow light penetration;
position of chlorophyll to trap light;
stomata to allow exchange of gases;
thin / max. surface area to volume ratio for diffusion of
gases;
spongy mesophyll / air spaces for carbon dioxide store;
Describe how the structure of xylem is related to
its function.
Vessels;
Have no end walls / hollow / no cytoplasm;
Allows unrestricted flow of water.
Lignification;
Provides support / strength / impermeability;
Pits allow lateral transport;
Tracheids with porous end walls.


Q. Palisade cells are the main site of
photosynthesis. Explain one way in which a
palisade cell is adapted for photosynthesis. Find a
detailed diagram of the palisade cell to include the
typical organelles of a eukaryotic cell.
Contains many chloroplasts thus lots of chlorophyll:
To trap or absorb light (energy);

Elongated cells with long axis perpendicular to the
surface:
Light has a longer pathway allowing maximum light
absorption / light penetration;

Chloroplasts move;
To trap or absorb light (energy)

Range of pigments;
Can absorb a range of wavelengths / colours / for
max light absorption;

Large S.A. or cell wall feature e.g. thin / permeable;
For (rapid) CO
2
absorption;
39







































Haemoglobin (Hb): Hb is a molecule that transports oxygen. It has a quaternary structure (association of 4 polypeptides there are two types of polypeptide chains, 2 alpha
and 2 beta chains). Each polypeptide chain is associated with a haem prosthetic group that contains a ferrous group(Fe++) and each can bind one oxygen molecule. So one
Hb molecule can carry 4 oxygen molecules forming oxyhaemoglobin
Hb + 4O2 HbO8 (deoxyhaemoglobin is a different colour to oxyhaemoglobin and so the % saturation can be measured with a colorimeter). Oxygen drives the process to
the right, and H+ drives it to the left.
The oxygen dissociation curve: shows the change in the affinity of Hb for oxygen at different PPO
2
.
Affinity = the tendency for Hb to bind oxygen, this changes with pH, temperature (similar effect to pH, associated with
respiration which raises temp) and PPO
2
.

Effect of PPO
2

At high PPO
2
the percentage saturation is very, around 95% and at low PPO
2
the percentage saturation is low.

So Hb coming from the lungs where PPO
2
is high is heavily saturated with oxygen. As it reaches the tissues where
respiration is occurring at PPO
2
is low it is only about 50% saturated.

Thus Hb coming from the lungs is carrying a lot of oxygen, and as it reaches the tissues it releases this oxygen, which is then
used for respiration

The S-shaped curve is explained by the behaviour of Hb. It shows cooperative bonding, its affinity for oxygen changes as
the amount of oxygen bound changes. The addition of the first oxygen is difficult, but once bound, it changes the shape of
the Hb molecules making it easier for the 2
nd
and 3
rd
to bind, it is harder for the fourth.
This behaviour is reflected in the curve. The curve is quite shallow up to around 2kPa, where only one oxygen molecule sis
bound, then the curve rises steeply as the 2
nd
and 3
rd
oxygen bind, here small changes in PPO
2
causes large changes in %
saturation.
The effect of pH
Hb is even better at releasing O2 in areas where it is required than
the neutral pH graph suggests.
The graph shows the effect of low pH in reducing the affinity of Hb
for oxygen. A more acidic environment reduces the % saturation at
all PPO
2
. More oxygen is unloaded at the same PPO
2
.this is exactly
what is needed because, where a lot of CO
2
is being produced, a lot
of respiration is occurring and thus the demand for oxygen is higher.

The shift in the curve is called the Bohr Effect. So in fact, in normal
respiring tissues the saturation is not 50% but 20-25%.

Why does it happen: carbon dioxide forms carbonic acid in the red
blood cell cytoplasm, then dissociates to H
+
and HCO
3
-
. Hb combines
with the H ions, so has a buffering effect

Describe how Hb normally loads oxygen in the lungs and
unloads oxygen in tissues.
Hb has a high affinity and loads O2 in high PPO2
Oxygen diffuses into the cell
Attaches to the iron containing haem group
Becomes saturated with O2
Oxyhaemoglobin forms
Hb unloads oxygen in low PPO2
This arises in actively respiring tissues which use O2
Carbon Dioxide shifts dissociation curve to right
Further reduces Hb affinity for O2
So more O2 unloaded and used in respiration

40





































Different Haemoglobins
Different animals possess different types of haemoglobin with different oxygen transporting properties. These properties are related to the animals way of life, so they are
an adaptation that helps the animal survive in its environment.
A human foetus makes a different kind of
haemoglobin from an adult. Foetal haemoglobin
has a higher affinity for oxygen at low partial
pressures, so its oxygen dissociation curve is
shifted up.
A developing foetus obtains its oxygen, not
through its lungs, but from its mothers blood in
the placenta. So this different haemoglobin
allows oxygen to diffuse from the mothers
blood to the foetus, and to be unloaded in the
foetal tissues. Foetal haemoglobin is gradually
replaced by adult haemoglobin during the first
year after birth.
Lugworms live in the mud in estuaries and
seashores. When the tide is out the lugworm
stays in a burrow filled with sea water. But the
oxygen concentration in this burrow can fall very
low as the lugworm respires, so the lugworm has
haemoglobin with a very high affinity for oxygen:
its oxygen dissociation curve is shifted up. This
allows the lugworm to obtain oxygen even when
the PO2 is as low as 2kPa.
Mice lose heat very quickly due to their large
surface area: volume ratio, so they have a high
metabolic rate to generate more heat. Their
tissues therefore have a constant demand for
oxygen for respiration. The oxygen dissociation
curve for mouse haemoglobin is shifted down
compared to humans, so plenty of oxygen is
unloaded to all tissues all the time.
Myoglobin is a pigment found in
muscles that are involved in sustained
contractions. . It has a higher affinity for
oxygen than Hb. It can act as an oxygen
store only releasing oxygen when PPo2
is very low, thus it delays anaerobic
respiration as long as possible
41

DNA: Deoxyribonucleic acid
A long linear molecule wound around histone proteins to form chromatin. This can be further coiled to form chromosomes prior to
replication. (
DNA is double-stranded, so there are two polynucleotide stands alongside each other. The strands are antiparallel, i.e. they run in
opposite directions.
The two strands are wound round each other to form a double helix
Sugar-phosphate backbone;
It is an organic compound made up of nitrogenous base, a sugar, and a phosphate group.
There are 4 bases (A, T, C and G)ssociated with DNA (a 5
th
type U is found in RNA (it replaces T)
specific base-pairing occurs between these bases (AT/ CG);
Two types of bases purines (A + G) or pyrimidines (C, T or U in RNA replaces T)
Purines have a double ring structure and pyrimidines a single ring structure.
bases are held by H bonds


Nucleotides
Have a phosphate group
A sugar component: Two key types, deoxyribose with O missing at C2 and ribose (found in RNA) with a hydroxyl group (OH) at C2
A nitrogenous base: In DNA there are 4 types, A, T, C and G. A and T are complimentary base pairs and C and G are complimentary base
pairs. In RNA the base T is replaced by U.
In prokaryotic cells: the DNA is smaller, circular and not associated with proteins
How the structure of DNA is related to its function
Sugar - phosphate backbone gives strength;
Long molecule stores large amount of information
Coiling gives compact shape;
Sequence of bases can be in any order and it can encode information
Information can be replicated due to complementary base pairing;
Double helix protects weak hydrogen bonds and prevents molecular damage
Chains held together by weak hydrogen bonds; but many hydrogen bonds together give molecule stability;
Hydrogen bonding allows chains to split/unzips easily for replication or transcription
Double stranded which means that each can act as a template in replication, replication is semi conservative


What are the interesting points about the genetic code?
The code is degenerate, i.e. there is often more than one codon for an amino acid. The degeneracy is on the third base of the codon, which is
therefore less important than the others, this minimises the effect of mutations
20 amino acids and 64 codons so some codon means "start" i.e. the start of the gene sequence. It is AUG, which also codes for methionine.
Thus all proteins start with methionine (although it may be removed later). AUG in the middle of a gene simply codes for methionine.
Three codons mean "stop" i.e. the end of the gene sequence. They do not code for amino acids.
The code is non-overlapping; each base is part of one codon
The code is universal: the same sequence of bases code for the same amino acids in most organisms

42

A lot of the DNA in eukaryotes does not code for polypeptides. The rest, called
non-coding DNA, does not form genes. There are two kinds of non-coding DNA:

Non-coding regions of DNA within a gene are called introns (for interruption
sequences), while the coding parts of DNA are called exons (for expressed
sequences). No one knows what these introns are for, but they need to be
removed before proteins are made.

Non-coding regions of DNA between genes are called satellites. Satellite DNA often
contain simple base sequences repeated many times (sometimes thousands of
times).
Genes: short sections of DNA. They a
specific order of bases that
determine the primary structure of a
polypeptide.
Alleles are alternate forms of a gene

Proteins are made from amino acids. Polymers formed by condensation reactions, held by
peptide bods.
Different proteins have different numbers and sequences of amino acids
The order of nucleotide bases in a gene that determines this sequence of amino acids
Each amino acid is coded for by a sequence of 3 bases (triplet/codon)

The genetic code is degenerate meaning there is more than one codon for most amino acids.
For example CCA, CCT, CCC and CCG all code for the amino acid glycine.
There are 20 amino acids and 64 codons

Non-coding DNA was original termed junk DNA, but in fact it probably serves many
different functions. Some may be structural, helping to coil the DNA molecule into
chromosomes; some may have a control function, regulating when genes are expressed;
some is involved in DNA replication; and some contains unused copies of genes.
Homologous
chromosomes: two
chromosomes of the same
size and shape, one
originating from each
parent. They contain the
same genes, but may have
different alleles.
43

Explain how a sequence of DNA is replicated
The process is described as semi-conservative replication, because both strands are used (as templates); and
(Daughter) DNA has one new strand and one original/parent strand;
Uncoiling of the helix occurs then
Unzipping of the DNA hydrogen bonds broken (DNA helicase)
nucleotides line up to their complementary/specific base pair ( A and T / C and G)
DNA polymerase catalyses the polymerisation of the nucleotides
new complementary strands form and identical DNA molecule produced
Replication is speeded up by occurring at many sites simultaneously, at regions called replication forks


Mutations: Mutations are changes in genes, which are passed on to daughter cells. DNA is a
very stable molecule, but bases can change when DNA is being replicated. Normally replication is
extremely accurate, and there are even error-checking procedures in place to ensure accuracy, but
very occasionally mistakes do occur (such as a T-C base pair). So a mutation is a base-pairing error
during DNA replication. Mutagens, UV light, X-rays, High energy radiation can cause
mutations also

A change in a gene could cause a change in the protein encoded by the gene, and so cause a change
in the cell function: Many of the proteins in cells are enzymes, and most changes in enzymes will stop
them working, so a reaction in a cell doesn't happen,. It's just possible (though unlikely) that a
mutation could make a modified enzyme that actually worked faster than the original enzyme. This
means cell's function could be improved.
How a mutation may cause an inactive enzyme

Mutation (addition, deletion or substitution)
Changes in the sequence of nucleotides/bases
Changed order of amino acids different tertiary structure;
Inactive enzyme if shape of active site is changed
Enzyme-substrate complex does not form

Since mutations change genes, they give rise to new alleles (i.e. different versions of genes). A cell
with the original, functional gene has one allele, while a cell with a mutated, non-functional version
of the same gene has a different allele..
44

This replication mechanism is sometimes called semi-conservative replication, because each new DNA molecule contains one new strand and one old strand.

Alternative theories suggested that a "photocopy" of the original DNA could be made, leaving the original DNA conserved (conservative replication), or the old DNA molecule could be dispersed
randomly in the two copies (dispersive replication).

The evidence for the semi-conservative method came from an elegant experiment performed in 1958 by Matthew Meselson and Franklin Stahl


45

If replication was
conservative then the
results for generation 1
would have given one
band in the original
position and another
band higher up made
form only the lighter
isotope.

The results show the
new DNA is
intermediate to the
control and the
original, so a hybrid is
indicated, however it
could be dispersive

The second generation
disproves dispersive
replication as we
would get a single
wide band as each
band would be a mix
of both isotopes. We
get two distinct bands
as shown

As generations
increase the proportion
of N14 increases
46

Variation is the differences that exist between species and within species. It can be a result of

1) Genetic factors:
Independent assortment of chromosomes in meiosis I and chromatids in meiosis II.
Crossing over of genetic information during meiosis I.
Random fertilisation
2) Environmental factors: exercise, light, oxygen, nutrients, pH depending on whether it is
plant or animals.
3) Combination of both genes and environment
4) Mutations causes changes in the DNA

Characteristics that are susceptible to environment are usually controlled by a number of gene,
polygenic, height and mass. These form a continuum

Characteristics that are not influenced by environment are usually controlled by a single gene
and give rise to discrete variation, like Blood group.
Meiosis halves the chromosomes number so that in fertilisation the diploid number will be restored. This is essential so the
chromosome number remains constant generation to generation. Meiosis introduces variation within a species and this is key to
the survival of the organisms

The basics of the process
DNA replication (during interphase) and 2 cell divisions that result in 4 unique haploid daughter cells

Meiosis I: homologous chromosomes pair up (bivalents),
Crossing over can occur: equivalent portions of chromatids are exchanged giving new combination of alleles. The point of cross
over is called the chiasma. There can be numerous chiasma in one bivalent
Independent assortment occurs, where the arrangement of the bivalents in metaphase is random.
The chromosomes are separated to the poles of the cell by contraction of spindle fibres

Meiosis II: the sister chromatids are separated.
The key points in each stage of cell division are

Interphase: 90% of the cell cycle. Cells get bigger, increase in
number of organelles, cytoplasm volume, cell mass, protein
synthesis Like histones) and of course DNA replication, increase in
AYP stores

Prophase: chromosomes condensed and become visible, nuclear
envelope breaks down, nucleolus breaks down

Metaphase: chromosomes arrange themselves at the cell equator
attaching to spindle fibres by their centromere

Anapahse: Separation of bivalents in meiosis I/sister chromatids in
meiosis II and mitosis, to the poles of the cell along spindle fibres

Telophase: Nuclear envelope reforms, spindles break down,
chromosomes uncoil, cytokinesis occurs, were cells split
Describe what happens to chromosomes in meiosis. (6)
1. Chromatin condenses and Chromosomes are visible
2. Chromosomes associate in homologous pairs
(bivalents)
3. Chiasma form and Crossing-over occurs
4. Chromosomes join to spindle fibres
5. By their centromere
6. At equator (middle of cell) independent assortment
creates genetic variation
7. Homologous chromosomes move to opposite poles
8. Chromatids are separated in 2
nd
division again this is
random assortment depending on the orientation of the
chromosomes
47

48



























Cell Cycle is subdivided into

1) Interphase: which is further categorised as
Growth 1 (G1): cell grows, protein synthesis occurs, organelles
replicated
Synthesis (S): DNA replication and association with histone
proteins
Growth 2 (G2): ATP stores increased

Overall in interphase
Increase in volume of cell, increase in mass
Increase in number of organelles
synthesis of protein
DNA replication
ATP stores increased

2) Mitotic phase, consisting off
Prophase chromosomes condense;
Metaphase chromosomes move to equator or centre of cell
attach to spindle;
Anaphase chromatids separate/centromeres divide;
Telophase Chromosomes uncoil
Followed by cytokinesis (cell division)

Importance of mitosis
1. Growth / increase in cell number;
2. Replace cells / repair tissue / organs
/body;
3. Genetically identical cells;
4. Asexual reproduction /cloning;

Describe the behaviour of chromosomes during
mitosis and how this results in two genetically
identical cells. (7)
1 chromosomes condense;
2 chromosomes appear with two identical
chromatids (due to replication);
3 chromosomes move to equator of the cell;
4 attach to individual spindle fibres by centromere;
5 spindle fibres contract and centromeres divide
6 (sister) chromatids move to opposite poles
7 each pole identical copies of each chromosome;
8 nuclear envelope forms around each group of
chromosomes at each pole;


Cell division occurs at 12 and 24 hours. DNA replication
occurs 6-9 and 18-21 hours

Calculating the time spent in a phase of the cell cycle.

(N
o
of cells in that phase total N
o
of cells) Time for 1 complete cycle
49


50


What is a carcinogen: an agent that can cause the development of cancer

Mutagens: UV light, X-rays, Gamma rays, Uranium are, increase the chances of mutations

A tumour is mass of identical cells (clones) formed by uncontrolled cell division.

A benign tumour grows slowly, remains encased in a capsule and does not spread far. Benign tumours
are usually harmless (e.g. warts), though they may cause harm by pressing on blood vessels or other
tissues. They can often be treated easily by surgery or radiation.

A malignant tumour grows quickly and spreads throughout the surrounding tissue, affecting its normal
function and so causing harm (e.g. lung cancer reduces elasticity of alveoli). These tumours are more
difficult to treat without damaging the whole tissue.

A metastasis is a tumour that has spread to the bloodstream or lymphatic system and so can spread to
other parts of the body, causing secondary tumours there. These are the most difficult to treat.


Cancer cells are characterised by
High rate of division
Ability to separate form adjacent cells
Ability to migrate to other tissues (metastasis)

Cancer develops due to an accumulation of mutations to
genes which disrupts normal cell processes

Cell growth and division is controlled by genes, like proto-
oncogenes that regulate cell growth and may exert their
effect in a variety of ways producing proteins like, growth
factors and regulators of replication.

A second set of genes called tumour suppressor genes
prevent the over expression of proto-oncogenes

If mutations occur in the genes that regulate cell division,
then cells can grow out of control, continuing to divide and
forming a tumour. Cancer is a tumour that invades
surrounding tissue.


Describe how altered DNA may lead to cancer.

Cancer develops due to an accumulation of mutations
A mutation is a change in the base sequence in the DNA
These mutation become more likely when exposed to
mutagens like UV, x-ray
Cell cycle is controlled by a number of genes
Mutations must arise in these different genes
Proto-oncogenes are involved in cell growth and if they
mutate to form oncogenes then cell division may begin to
occur at a higher rate and a tumour can develop
If tumour suppressor genes produce proteins that inhibit cell
division
A mutation here results in proteins that do not function
Cell division is not inhibited
Mitosis goes uncontrolled
A tumour develops


51

52



A community: all the organisms living in a habitat Species diversity: the variety of species in the community
Species diversity tell us: it tells us about the complexity, quality and stability of an ecosystem

Sampling techniques to measure species diversity
Quadrats: area divided into a grid, random numbers are generated 9avoid bias) that act as co-ordinates where the quadrats will be placed. A large number of quadrats must be used; this allows it
to be representative of the area and allows patterns to be established (random, uniform clustered distribution) also allows statistical analysis to be carried out. The percentage cover, abundance or
number of different species can be calculated

Transects: are used when there is an environmental gradient. A line or belt transect can be used. Belt transects give greater detail, using quadrats to measure abundance at each interval. Intervals
may be required and the size of the interval will be determined by the distance being sampled

Q. What is the difference between species richness and abundance?
Species richness is the number of different species in a sample Abundance is the number of each species present

To best measure diversity we should account for both richness and abundance. The Simpson index of diversity does this. The formula is.


N = total number of
organisms of all species

n = total number of
organisms of each species

Species Diversity and human influence on diversity
Agricultural practices (intensive farming) why things are done and the consequences
of these processes

Agriculture
1. Selective breeding: done to select for certain favourable characteristics
reduces genetic diversity.
2. Destruction of hedgerows: Makes large farms with large fields are cheaper
and more efficient to run by easing the moving machinery and harvesting.
Hedgerows provide habitats for at least 30 species of trees and shrubs, 65
species of nesting birds, 1500 species of insects and 600 species of
wildflowers. These in turn provide food for small mammals. Hedgerows also
act as wildlife corridors, allowing animals to move safely between woodlands.
3. Monoculture: increases the productivity by growing the best crops, which can
be sowed and harvested quickly using dedicated machinery. This increases
yield and reduces labour costs. It reduces genetic diversity and renders all
crops in a region susceptible to disease. Reduces animal species diversity,
because there are few niches.
4. Fertilisers: maintain soil fertility, but they can pollute surrounding
groundwater causing eutrophication and killing aquatic animals.
5. Pesticides: are sprayed on crops to prevent attack by insects and other
invertebrate animals, but many pesticides have a broad spectrum, killing a
wide range of animals and so reducing diversity.
6. Herbicides: kill competing plants (weeds) that might reduce crop yield.


The Two main reasons humans clear forests are:
To use the land for agriculture, housing, mining or reservoirs
To use the timber for fuel, charcoal, paper or building materials.

Forests have a high biodiversity because a mature forest has many different species of plants in
several layers; each adapted to their own conditions of light and nutrient availability. The
different plants have different animals feeding on them and living in them; and the different
primary consumers have different secondary consumers feeding on the. So forests contain
complex food webs with high diversity.

By contrast, a field of crops has very low diversity with very few plants (often just the crop and
a few weeds) and so few animals. Deforestation therefore reduces biodiversity.
As the diagram shows, forests have a deeper and more extensive root system, so binding
the soil together.
Without this root system, soils can be eroded, leading to desertification (fertile land
becomes desert). Forests also have a high productivity: i.e. there is a lot of plant material
produced per square meter of land, and a lot of photosynthesis takes place. So deforestation
reduces the rate at which carbon dioxide is removed from the atmosphere and so increases
the greenhouse effect and global warming.


53


Genetic diversity:
Caused by
1) Environment: factors affecting plants and animals need to be considered. Characteristics controlled by many genes (polygenic) are most susceptible to environmental influence).
Characteristics controlled by single genes are not influenced by the environment.

2) The genes we inherit: we inherit our genes from parents in the sperm and egg during sexual reproduction. These gametes are formed by meiosis; this can cause variation by.crossing over
and independent assortment. Sexual reproduction involves the random fusion of gametes. Mutations can also result in variation

A genetic bottleneck happens when a population is drastically reduced in size due to a
natural catastrophe. The few individuals left will only have a small range of alleles
between them, so if they reproduce and the population increases again there will be
reduced genetic diversity. Many of the original variety of alleles will have been lost in
individuals who didnt survive.

The founder effect occurs when a small number of individuals colonise a new habitat and start a
new, isolated population. Since the few individuals will only have a small range of alleles
between them, the founder effect is an example of a genetic bottleneck, and is sometimes called
a colonisation bottleneck. These modern populations will have low genetic diversity, reflecting
the small range of alleles in the small founding population. In extreme cases a founding
population can be as small as a single pregnant female animal or a single plant seed.

Selective breeding
Selective breeding, or artificial selection, means the controlled
breeding of animals or plants by humans so that only individuals
with certain characteristics are allowed to reproduce. Since these
characteristics are (at least partly) genetically controlled, this
means selecting certain alleles and rejecting others, so the
genetic diversity of these animals and plants is reduced.

Advantages of selective breeding:
Used to produce high yielding domestic plants and animals
Reliable and cheap source of food is established as higher yields can be obtained
The standard of living has been raised

Disadvantages/ethics of selective breeding
Do animals have value or are they solely for human usefulness
What features do we select for?
Should humans decide what is appealing in animals?
Animal welfare must be considered, for example, humans select animals that grow quickly (leads to high infection
risk and joint problems) and that are better suited to living in sheds than open fields which is their natural habitat
Reduction in genetic variation (genetic uniformity) makes animals vulnerable mass declines in numbers
Loss of alleles that may be beneficial to humans in the future
Lower resistance to disease
Decreased fertility
Development of physical problems in animals that would normally be selected against in the wild

54

So the rate of exchange of substances therefore depends on
the organism's surface area that is in contact with the
surroundings. As organisms get bigger their volume
increases at a greater rate than the surface area able to
supply it. So as organisms get bigger their surface area:
volume ratio gets smaller. To maximise their rate of gas
exchange multicellular organisms have developed particular
body shapes to increase their surface area: volume ratio.
The openings of the tracheae (spiracles) have valves
which can open and close. When open water loss is
possible and they are when closed gas exchange is not
possible. The opening of the valves is carbon dioxide
sensitive, so at rest they will remain closed until a
critical level of carbon dioxide is reached. When active
(in flight) they will open regularly as carbon dioxide is
being produced in respiration. The valves can open
independently. Tracheal system relies mainly on
diffusion so limits the size of the insects.



How the gas exchange system of an insect is efficient
The high number of tracheoles and the high degree of branching
results in a large surface area for gas exchange.
The ends of the tracheoles penetrate deep into the muscle and no
cell is far away from a source of oxygen
The ends of the tracheoles are fluid filled and this gives the insect
some control over the rate of gas exchange. In high activity
respiration increases, oxygen is not sufficient lactic acid will begin to
build up, lowering the water potential of the tissues, so fluid is drawn
out of the tracheoles and this increases the surface area for gas
exchange, so diffusion is faster.



Explain how the gills of a fish are adapted to form a
specialised exchange surface.
Structure of filaments with a large number of lamellae;
and secondary lamella provide a large surface area for
exchange
The lamellae have a thin surface, a single layer of
flattened epithelial cells provide a short distance
between water and blood / short diffusion pathway
Counter-current flow of water and blood across lamella
maintains a diffusion gradient along length of lamellae
and prevents oxygen concentrations reaching
equilibrium;
Ventilation mechanism in producing water flow over
gills and circulation of blood maintains a diffusion
gradient.

How a fish ventilates the gills
Mouth opens, whilst opercular valve
shuts;
The floor of mouth lowers
Volume increases and pressure
lowers so water
Mouth closes, opercular valve opens
floor raised results in increased
pressure due to decreased volume;
High pressure forces water over
gills;

The difference in oxygen concentration between he water and blood is
maintained by
Fish has ventilation system which replaces water, which is high in oxygen
The circulatory system brings in blood with low concentration of oxygen and
removes oxygenated blood Water flows in opposite direction to blood across
(gill) lamellae; so difference in concentration maintained and Diffusion occurs
over full length of lamellae.

55

Gas exchange occurs by Diffusion via stomata and air spaces. The
spongy mesophyll has large air spaces to facilitate faster diffusion, the
cell walls are thin to give a short diffusion pathway, there is a large
surface area to volume ratio, and the palisade cells are cylindrical in shape
so that even when close packed air spaces remain for gas diffusion.

Gas exchange system of mammals adaptations

Large surface area

Wall of alveoli and capillaries have single epithelial layer
epithelium flattened (squamous)
Alveoli and capillaries close together
all give a short diffusion pathway

Ventilation maintains high O
2
/low CO
2
concentration (in alveoli)
blood flow/circulation maintains high CO
2
/low O
2
concentration(in blood)
Steep diffusion gradient

Narrow capillaries slow blood flow - more time for diffusion;
and the close contact between cells and capillary walls reduces diffusion path;






56


















How tissue fluid is formed and returned to circulation
High hydrostatic pressure of blood high at arterial end of
capillary

Forces fluid out of the capillary

Plasma proteins are too large to leave the capillary and
remain

The water potential in the capillary becomes more
negative than the surrounding tissue fluid

Water moves back into the capillary at the venous end

By osmosis

Lymph system collects any excess tissue fluid;

Returns the fluid to the circulatory system link at the vena
cava/

The hydrostatic pressure decreases from the arterial end to the venule end due to
Frictional force (resistance) of capillary the capillary walls
loss of fluid (not all (filtered) fluid is returned, some drains to lymph)


Oedema (build-up of tissue fluid)
Lack of protein in the diet (less plasma proteins so the osmotic
potential of the blood is not as effective (does not become as negative
in relation to the tissue fluid)
Raised blood pressure will increase the hydrostatic pressure further
than usual so more tissue fluid generated than normal
Infection (histamine), causes capillaries to dilate so more permeable
than usual proteins may leak out and thus the water potential at the
venous end is not as negative so less water absorbed by osmosis
Fluid retention (poor kidney function) so raises the volume of blood
plasma and thus the hydrostatic pressure.
Parasites that develop inside the lymph system blocking it

There is a net loss of
water from a capillary at
the arterial end
because
At the arterial end the
hydrostatic pressure is
greater than pressure of
water potential

The lymphatic system has three different functions:
It drains excess tissue fluid
It absorbs fats from the small intestine, via the lacteals inside each villus.
It is part of the immune system. There are networks of lymph vessels at various places in the body (such as tonsils
and armpits) called lymph nodes where white blood cells develop. These become swollen if more white blood cells
are required to fight an infection.
57

58



Artery
Thickest wall, enabling it to carry blood at high pressure /
withstand pressure surges;
most elastic tissue, which smooth out flow/maintains
pressure; it can distend when ventricles contract and can
recoil
most muscle which maintains pressure;
muscle in wall to control blood flow, contracts,
vasoconstriction occurs altering blood flow to organs. The
proportion of muscle increases in the arterioles and elastics
tissue declines

Vein
Thin wall does not have to withstand high pressure; so they have less elastic
tissue and muscle tissue, but have a larger lumen to reduce friction as blood is
under lower pressure
Presence of veins to prevent back flow
Blood flow is a result of skeletal muscle contraction, squeezing it along vessels,
residual blood pressure from the heart, negative pressure form the thorax

The capillary is adapted for the exchange of substances
between blood and the surrounding tissue.
1. Permeable capillary wall/membrane;
2. Single cell thick/thin walls, reduces diffusion distance;
3. Flattened (endothelial) cells, reduces diffusion distance;
4. Fenestrations, allows large molecules through;
5. Small diameter/ narrow, gives a large surface area to
volume/ short diffusion distance;
6. Narrow lumen, reduces flow rate of cells through the
capillary giving more time for diffusion;
7. Red blood cells are in close contact with the capillary wall
give short diffusion distance



59

Property
Tissues
present in
wall
Mean diameter
of vessel
Mean
thickness of
wall
Artery
4.0 mm
1.0 mm
5.0 mm
0.5 mm
8.0 m
0.5 m

Capillary Vein
Blood vessel
Relative thickness (shown by length of bar)
Endothelium
Elastic tissue
Muscle
Aorta Small
arteries
Arterioles Capillaries Venules Veins
Elastic
fibres
Muscle
fibres
Permeability
As you move away from the heart the proportion of elastic
tissues in the vessels decrease and the proportion of muscle
increases. This reflects the decreasing pressure surges and need
for elasticity, but the increased need for maintaining pressure
and controlling the blood flow to different organs.
Permeability increases at the capillaries, the site of exchange,
they are one cell thick
Veins have a lot less muscle and elastic tissue than arteries,
thinner walls as a consequence. A wider lumen helps to reduce
restriction to flow
60

How water enters and moves across the plant roots to the xylem.
Water enters root hair cells by osmosis;
because active uptake of mineral ions has created a WP gradient
water moves through the cortex;
by osmosis down a WP gradient
water moves through two paths
1) Symplastic pathway through cell vacuoles and cytoplasm and membranes
2) Apoplastic pathway through cell walls only
At the endodermis all water enters the symplastic pathway due to the
waterproof layer of the casparian strip
This gives the plant control over the materials entering the xylem

Describe and explain how water moves via the apoplastic and symplastic pathways from the soil to
the xylem in a root.
Apoplastic Via cell walls
As far as endodermis / Casparian strip / layer of wax;
Caused by transpiration pull;
Cohesion / hydrogen-bonding between water molecules;
Symplastic Through cell surface membrane/ vacuoles membrane;
High to low;
Diffusion / osmosis;
Cell-to-cell via plasmodesmata / via strands of cytoplasm;
Secretion / active transport of ions into xylem by endodermis;
OR
Active uptake of ions from soil at epidemis;
s
in xylem / increases osmosis into xylem;


Explain how the structure of the endodermis affects the passage of water by
this apoplastic pathway.
Casparian bands; (accept ref to suberin)
which are impermeable/waterproof;
lower water potential in the cytoplasm of endodermis cell;
enters symplastic pathway / cytoplasm of cell;
by osmosis;

Evidence for root pressure
The reduction in root pressure caused by metabolic inhibitors: prevent the release of energy by the mitochondria, thus the active loading of minerals would decrease or stop meaning the
water potential gradient was not as great so the osmotic influx of water would cease or reduce causing a smaller hydrostatic pressure.
The effect of temperature on the root pressure: an increase in temperature will cause an increase in root pressure and vice versa, linked with the kinetic energy of molecules, but also due
to the temperature sensitivity of enzymes involved in cellular respiration and proteins involved in transport. At very high temperatures enzymes will denature, change in the tertiary
structure so the active site no longer complements the substrate.
A lack of oxygen will also reduce the root pressure; oxygen is required for the release of energy in aerobic respiration. A lack of oxygen results in less energy resulting in less active loading
of the ions into the xylem hence a reduction in root pressure.


61

Describe the roles of root pressure and
cohesion-tension in moving water through
the xylem.

Root pressure
Involves active transport of ions into xylem;
Water potential in xylem reduced
Water potential gradient established between
xylem and surrounding cells Water enters
xylem by osmosis
Volume of water in xylem increases;
Pressure in xylem increases (and forces water
upwards);

Cohesion tension
Solar energy source;
Evaporation of water from leaves through
stomata
Water potential gradient created across leaf /
mesophyll cells;
therefore water moves out of xylem (into
surrounding tissues) by osmosis this creates a
pull/tension on the water in xylem
Which is in a continuous column
Cohesion (or description) of water molecules
maintains column;
Due to H-bonding / polarity / charges of water
molecules ;
Column doesnt break because of adhesion
with xylem walls;
Lignified walls keep xylem (vessels) open;
capillarity due to narrow lumen of xylem
(vessels);




Evidence that supports the root pressure theory
Guttation: (only) upward pressure could force
liquid water out of leaves;
OR
Sap exuding from a cut, rooted stem: (only)
upward force could make this happen;

Evidence to support supports cohesion
The diameter of a tree is less during the day, when the tree is transpiring, than it is at night. Evaporation from leaves
during daytime; tension/negative pressure (on water) in xylem creates inward pull (on walls of xylem vessel);
xylem vessels become narrower; due to adhesion of water molecules (to walls of xylem vessels);
If air enters the xylem the transpiration stream can cease as cohesion is disrupted between the water molecules.
If the xylem breaks air is sucked into the vessel suggesting a negative pressure inside

The argument against is that rot pressure would force the xylem wider and thus increase the diameter of the tree
62


Water evaporates from the leaves but some is also used by the plant. Describe the
ways in which this water could be used by the plant.

Water is used in the light-dependent reactions of photosynthesis;
electrons from water enable ATP production / H
+
are used to reduce NADP /
produces O
2
;
(water can be used in) hydrolysis reactions within the plant;
to create turgor;
as a solvent for transport;
as a medium for chemical reactions;
component of cells / cytoplasm; 6





Sunken stomata: water evaporation into pit creates local humidity, increased humidity
reduces gradient for water evaporation; air movement across stomata is reduced boundary
layer of humid air builds up.
Close arrangement of stomata: diffusion shells of individual stomata overlap so interferes
with water diffusion and slows evaporation;
Restriction of stomata to lower side of leaf: rate of air movement below leaf less/ heating
effect of sun less; gradient for water evaporation reduced/ water molecules have less
kinetic energy;

Thick cuticle (on upper surface): waxy layer is waterproof; water unable to to evaporate
easily, diffusion distance has been increased

Presence of hairs: traps air which traps water and becomes saturated close to leaf surface;
increased humidity reduces gradient for water evaporation;

Reduced leaves/spines: less surface area for evaporation; more distance across leaf for
water to diffuse; fewer stomata for evaporation

Rolled leaves: stomata enclosed in localised humidity; increased humidity reduces gradient
for water evaporation; reduced exposure to air currents

Stomata are close together: diffusion shells overlap

Explain how the presence of hairs and rolled leaves reduce water loss in xerophytic
plants.
Trap moist air / increase humidity;
Reduce air flow (around leaf surface / stomata);
Lower WP / water vapour concentration gradient (between inside
and outside of leaf);
Shield stomata from high temperature / high light
intensity / wind; ignore sun
Reduce transpiration / evaporation / diffusion of water (vapour);

Explain how xerophytic adaptations reduce the rate of diffusion of water from
the leaf, make reference to Ficks law.
Reduced number of stomata; reduced surface area;
Thick waxy cuticle; increases diffusion distance;
Leaves reduced to spines; reduced surface area ;
(epidermal) hairs; reduce diffusion gradient;
Sunken stomata; reduced concentration gradient;
curled leaves; reduce concentration ; difference
Statement of Ficks law:
Rate of diffusion SA exchange surface conc difference
Thickness of exchange surface;
Low surface area, low concentration difference and high thickness/equivalent reduce
loss / candidate clearly relates features to equation to show how rate is reduced;



63







































Courtship behaviour is innate, in other words it is genetically programmed, so all members of the same
species show exactly the same courtship behaviour, while members of different species show different
behaviours.
The rituals relay on sign stimuli (specific external stimuli, sound, scent, colour, movements) that elicit
specific responses in potential mates. The males action can serve as a sign stimulus to the female who
responds with an action of her own, encouraging the male to carry out a further action. The stimulus
response chain.
It can be simple (releasing chemicals or complex involving displays
The behaviour is species specific; this prevents interbreeding and makes courtship more successful

Females of many species are only receptive to mating for a short period of time. Courtship is used by
males to determine whether the female is receptive, if she responds with the appropriate response the
courtship continues. If she is not receptive the female exhibits a different pattern of behaviour and he
ceases the courtship.
In courtship animals use signals to communicate.

Elaborate courtship rituals develop by natural selection
Male with best display more likely to mate, they are thus, more likely to pass on genes; the genes for
features of display passed on; more young from these males exist in population and are thus more likely to
survive and reproduce, the process repeated through many generations.



Why is courtship behaviour necessary?
Acts as (sign) stimulus for mating behaviour /courtship signal;
Assists species recognition;
Identify members of the opposite sex recognition;
Identify members of the same species; as only they can have fertile
offspring
Identify members of the species that are sexually mature;
Synchronises mating behaviour;
Reduces aggression and allows animals to approach each other
Forms a pair bond
Indicates fittest / healthy male;
This specificity of courtship rituals means they can be used in classification:
The courtship displays that are more similar are like to be species that are
close evolutionary relatives. The more closely related a species the more
similar the behaviour

There are 5 species represented in this table
B and C are most closely related because they have a similar sequence (just one element
missing for duck C). Whilst duck F is the most distant as the sequence of behaviour is least
like any of the others
64

Phylogenetics: Aims to classify species in to groups and reflect their evolutionary
history (phylogeny), it looks for the point of divergence from a common ancestor.
The process can be supported by various techniques

1) Protein and Base sequencing: here the same protein of gene is used. With proteins
the amino acid sequence is determined. The greater the differences in the amino acid
sequence then this would indicate a greater difference in the base/gene sequence
that coded for it. Suggesting a more distant evolutionary relationship, where more
time has passed to allow mutations to change the base sequence. Sequencing the
bases is a better approach, due to the degeneracy of the genetic code where the
same protein could be coded for by a different codon, so although the proteins are
the same the actual bases/codons could be entirely different.
2) DNA Hybridisation: using the DNA sequence for the same gene in both species
The DNA of the species to be compared is separated by heating, breaks H
bonds.
The DNA is mixed and cooled; this allows H-bonds to reform between specific
base pairs. The result will be a mixture of species A, B and the desired hybrid
DNA.
Hybrids are isolated and then heated to break the H-bonds. The temperature
required to break the H-bonds is recorded.
A high temperature suggests a high degree of H bonds; this could only have
happened if there were many complementary base pairs, indicating the DNA is
similar. A low temperature suggests few H-bonds and thus the base sequence
must have been different.

3) Immunological comparison.
The same proteins form different species are compared. The
process relies upon the specificity of antibodies for antigens
Serum albumin from A is injected into B.
B produces antibodies specific c to all the antigen sites on
the albumin from species A.
Serum is extracted from B; containing antibodies specific to
the antigens on the albumin from A.
Serum from species B is mixed with serum from the blood of
a third species C.
The antibodies respond to their corresponding antigens on
the albumin in the serum of species C.
The response is the formation of a precipitate.
The greater the number of similar antigens, the more
precipitate is formed and the more closely the species are
related. The fewer the number of similar antigens, the less
precipitate is formed and the more distantly the species are
related.
65

Classification/Taxonomy
Kingdom (5 possibilities: animal, protoctist, plant, fungi, prokaryote)
Phylum
Class
Order
Family
Genus
Species

*king, Philip, Came, Over, From, Germany, Swimming*

Binomial naming system= Genus and species

What is meant by a hierarchy?
Large groups split into smaller groups (which do not overlap);

How does a phylogenic system differ to a simple hierarchy? 3 max
(phylogenetic) based on evolutionary history;
shows ancestry of groups / points of divergence;
example, e.g. reptiles and birds separated after mammals / reptiles
and birds more closely related than mammals;
(hierarchical) based on shared characteristics (seen today);

Explain the principles which biologists use to classify organisms into
groups. (3)
large groups are divided into smaller groups; (not just hierarchical)
members of a group have features in common; based on
anatomy/fossils/embryology/DNA/specific aspect of cell biology or
homologous structures, reflecting evolutionary history; phylogeny. Process
starts with species grouped into genus then grouped into family, order, class,
phylum. As the groups get larger there is a more distant common ancestry.

Explain the principles biologists use to classify organisms into groups
compared to older models.
Consider phylogeny
Look at evolutionary lineage/history
Find the point of divergence from a common ancestor
Consider, genetic, biochemical, embryology, homology of anatomy
Organisms are arranged in a hierarchy where large taxa (groups) are
subdivided into smaller taxa
(K, P, C, O, F, G,S)
As groups get smaller the similarities between the species increase
Each species is given a binomial name using the genus and species
Older models of classification used observable features to group organisms

How does a phylogenic system differ to a simple hierarchy? 3 max
Hierarchical classification, large groups are divided into smaller groups
Process starts with species grouped into genus then grouped into family, order, class, phylum. As the groups get larger
there is a more distant common ancestory
Initially this was based on shared easily observable characteristics
(phylogenetic) based on evolutionary history;
shows ancestry of groups / points of divergence;
members of a group have features in common; based on anatomy/fossils/embryology/DNA/specific aspect of cell
biology or homologous structures, reflecting evolutionary history; phylogeny.
Describe the principles on which the system of classification of living organisms is based. (4)
hierarchy / groups within groups / KPCOFGS;
no overlap;
common structures / similar characteristics;
reflecting evolutionary history;
binominal nomenclature / example;
definition of a species;


Scientists analysis of blood proteins has indicated a lack of genetic diversity in populations of some organisms.
Describe the processes that lead to a reduction in the genetic diversity of populations of organisms.
1. Reduced variety/number of different alleles/DNA / reduced gene pool (in new population);
2. Founder effect;
3. A few individuals from a population become isolated/form colonies:
4. (Genetic) bottlenecks;
5. (Significant) fall in size of population
6. Selective breeding / artificial selection;
7. Using organisms with particular alleles/traits/phenotypes/characteristics;

Scientists studied two species of North American seahorse. They thought that these two species are closely related.
Describe how comparisons of biological molecules in these two species could be used to find out if they are closely
related.
(Compare) DNA;
Sequence of bases/nucleotides;

DNA hybridisation;
Heat and Separate DNA strands / break hydrogen bonds;
Mix DNA/strands (of different species);
Measure Temperature/heat required to separate (hybrid) strands indicates relationship;
Higher temp more closely related

Compare same/named protein;
Sequence of amino acids /primary structure;

Immunological evidence not a mark
Inject (seahorse) protein/serum into animal
(Obtain) antibodies/serum;
Add protein/serum/plasma from other (seahorse) species;
Amount of precipitate indicates relationship


66

Antibiotics are antimicrobial agents produced naturally by other microbes (usually fungi or bacteria).

Many chemicals kill microbes. But a therapeutically useful antimicrobial agent must be selectively toxic i.e. it must kill pathogenic microbes already growing in human tissue, without also killing
the host human cells. Antibiotics do this by inhibiting enzymes that are unique to prokaryotic cells, such those involved in synthesising the bacterial cell wall or 70S ribosomes. For example:
Penicillin (and related antibiotics ampicillin, amoxicillin and methicillin) inhibits an enzyme involved in the synthesis of peptidoglycan for bacterial cell wall. This weakens the cell wall, killing
bacterial cells by osmotic lysis.
Streptomycin, tetracycline and erythromycin inhibit enzymes in ribosomes. This stops protein synthesis so prevents cell division.
How antibiotics work
Prevent cell wall synthesis: this only works on growing bacteria. It stops cross links between
peptidoglycan monomers in the cell wall from forming, so it is weak. Water will often enter
bacteria by osmosis as the water potential inside the cell is more negative than the surrounding
medium. The pressure inside the cell increases which is usually resisted by the cell wall (pressure
potential) which is now less effective and the cell may rupture (osmotic lysis)

Prevent DNA replication: prevents the bacteria from reproducing/dividing so the population stops
increasing allowing time for an immune response. The joining of nucleotides in replication is
controlled by enzymes and the active site of the enzyme is blocked so the reactions cannot be
catalysed.
Prevent mRNA synthesis (transcription): No genetic code is transferred to the ribosomes, so no
protein synthesis of either structural or functional (enzymes/channel/carrier proteins). This
antibiotic may carry out its effect by cutting DNA in to sections so it cant be transcribed to mRNA.

Prevent translation (protein synthesis): stopping protein synthesis means that no structural or
functional proteins can be made. This may be done by binding to the ribosomes so tRNA can not
bind and peptide bonds cannot form.
This is a broad spectrum antibiotic as all bacteria produce proteins. It does not affect host, possibly
as it only attaches to 70s ribosomes and humans are eukaryotic and have 80s, it is also possible
that the mechanism by which it enters the cell requires a special carrier that is only found on
bacterial cells



Antibiotics can be
Bacteriocidal: kills the microbes
Bacteriostatic: Inhibits the growth of microbial
population allows time for immune response
Broad spectrum: antibiotics effective against a
range of microbial species
Narrow Spectrum: antibiotics that are effective
against a few species of microbes

Desired characteristics of antibiotics
Cheap to make
Easy to administer
Minimal/no side effects
Effective against targeted pathogenic
organism
Persists long enough and in suitable
concentrations within the host to be
effective

1) Given to patients with a viral infection, not because it will have an effect on the
virus as they..
Have no organelles
Are inside the cells
Are acellular (not made of cells)
Have a protein coat and not a cell wall
Do not have their own metabolism

But it will prevent a secondary infection developing from opportunistic bacteria which
may result from the weakening of the immune system caused by the viral infection.

2) Tissue culturing (prophylactic)

3) Used in cattle feed:
Stimulates growth, increase in biomass thus can be sold quicker so profit
increase
Keeps them healthy (prophylactic: preventative measure)

[Unwanted dosage may pass to people consuming the meat leading to an immune
response and results in the inability to use it to treat infections and can lead to the
spread of antibiotic resistance within the same species and other species of bacteria
as seen in the flow chart below]


67

Development of resistance
Natural mutation (change in the genetic code) results in resistance (ability to produce an enzyme like
penicillinase, change in the targeted receptor protein that uptakes the antibiotic into the bacterial cell so it no
longer complements the shape of the carrier protein)
This bacteria has an advantage over others
Use of the antibiotic selects for the resistant strain of the bacteria
Resistant strain survives and reproduces passing on the allele to next generation (vertical transmission)
Genes for resistance are usually found on the plasmid and this can be exchanged between bacteria
(conjugation) of the same species of different species (horizontal transmission)
Frequency of the resistant allele increases within bacterial populations.


Preventing resistance
Use a lower dose of the Antibiotic to prevent selection for the resistant forms
Use less antibiotics (particularly for trivial ailments) reduces selection for the resistant form
Vary the type of antibiotics used reduces selection for resistance
Use a high dose of the antibiotic for a short time killing all
Complete the course prevents re-emergence of dormant forms
Do not use other peoples antibiotics or antibiotics from other illnesses
Do not use them in animal feeds

Imagine a community of different bacterial species
living in your gut, and one particular cell has just
mutated to become resistant to penicillin. What
happens next? It will reproduce by binary fission
and pass on its resistance gene to all its offspring,
forming a new strain of bacteria in your gut. If
there is no antibiotic present in your gut (most
likely) this mutated strain may well die out due to
competition with all the other bacteria, and the
mutation will be lost again. However, if you are
taking penicillin, then penicillin will be present in
the bacteria's environment, and these mutated cells
are now at a selective advantage: the antibiotic kills
all the normal bacterial cells, leaving only the
mutant cells alive. These cells can then reproduce
rapidly without competition and will colonise the
whole environment. This
a good example of natural selection at work. The
mutant cells have been selected by the environment
and
so the frequency of the mutated gene in the
population has increased.
Bacteria have a trick that no other organisms can do: they can transfer genes between each other
by conjugation. This is the transfer of DNA between bacterial cells via a cytoplasmic bridge or
pilus. From time to time two bacterial cells can join together (conjugate), and DNA passes from
one (the donor) to the other (the recipient). The transferred DNA can be one or more plasmids,
or can be all or part of the whole bacterial chromosome (in which case the donor cell dies).
Conjugation is sometimes referred to as bacterial sex or mating, but it is quite distinct from
sexual reproduction, because the gene exchange is not equal, it can take place between different
species, and bacteria do not use conjugation for reproduction. It is better thought of as an
alternative to sex, where these asexual organisms gain some of the advantages of genetic
exchange.
Conjugation means a resistance gene can spread from the bacterium in which it arose to other,
perhaps more dangerous, species. It is also the cause of multiple resistance. It is highly unlikely
that a single strain will mutate twice to develop resistance to antibiotics, but it is perfectly likely
that it could receive genes for
resistance to different antibiotics by horizontal gene transfer. This has led to strains of bacteria
that are resistant to many (or even all) antibiotics.
68

















Explain how resistance to an antibiotic could
become widespread in a bacterial population
Genetic mutations naturally occurs resulting in
resistant allele
Frequent use of antibiotic creates selection
pressure/ antibiotic kills bacteria;
bacteria with mutation have (selective) advantage
over others / described;
survive to reproduce more than other types;
Pass on advantageous allele/ mutated allele in
greater numbers;
frequency of (advantageous) allele increases in
subsequent generations;
6. Frequency of resistant types increases in
subsequent generations;


Ways in which an antibiotic could act against
bacteria

Disruption of cell wall;
inhibit mRNA translation / protein synthesis;
inhibits nucleic acid synthesis / DNA replication;
interfere with functioning of bacterial membrane;






Describe ways in which antibiotics can act against bacteria. Explain
why this mode of action is effective against the bacteria.

Prevent DNA replication
Prevent m-RNA synthesis (transcription)
Prevent transfer of amino acids to ribosomes (translation)
Prevent cell wall synthesis


Preventing DNA replication:
bacterial cell will be unable to divide;
Prevent reproduction population of bacteria will not increase;


Preventing m-RNA synthesis
no m-RNA means code not passed to ribosomes for transcription;; no
protein synthesis means no new enzymes;

Preventing transfer of amino acid
No proteins made at ribosomes
No translation means no enzymes / no proteins structures;

Prevent cell wall synthesis
Water potential of bacterial cell lower than surrounding solution
Water enters by osmosis
Osmotic lysis occurs


69

We can compare members of different species to show interspecific variation, or we can look at members of the same species to look
at intraspecific variation. Variation arises due.
1) Genetic differences: independent assortment in meiosis, crossing over in meiosis, mutations, random fusion of gametes
2) Environmental influence

We onlt measure a smaple of the population, and it must be chosen carefully.
Randomly: avoid bias
It must be large to be representative and minimise anomalies

If the numbers are large enough and the results can be plotted as a graph (the characteristic is measured quantitatively) the data
usually follows a normal distribution curve and are described as continuous (usually characteristics that are polygenic: controlled by
more than one gene). This curve is described by
1) The mean:
2) The standard deviation

Characteristics controlled by a single gene (and that are not influenced by environmental factors) do not show intermediate values
and are described as discontinuous/discrete
Standard deviation gives an indication of the value range either side of
the mean. It shows the variability in the data.

2 sets of data could have the same mean or range suggesting the
populations are similar, but the variation around this mean could differ
considerably, and so standard deviation would show this.

So standard deviation is a more effective when comparing variation in
populations as the range and mean are affected by a single outlier, SD
shows variation about the mean, standard deviation can b eused in
statistical analysis to determine significance of differences
In the graph opposite we can see that both sets of
data have the same mean value, however
There is a more uniform population with a small
standard deviation
And a more varied population where the standard
deviation is large
Data set A: 46, 42, 44, 45, 43 mean = 44
Data set B: 52, 80, 22, 30, 36 mean = 44
Calculating the standard
deviation
Geneteic variation is important in evolution because
It creates a rnage of phenotypes, some of which are better
adapted to the environment
These organismssuvive and reproduce (natural selcteion)
Passing on their useful alleles
Alleic frequencies change over time.
70