Biology

Biology
Biology is the study of living things: the science of life. Although we cannot define life with a
single simple sentence, we can come up with a series of seven characteristics shared by
living systems:

● Cellular organization. All organisms consist of one or more cells. Often too tiny to
see, cells carry out the basic activities of living. Each cell is bounded by a membrane
that separates it from its surroundings.
● Ordered complexity. All living things are both complex and highly ordered. Your body
is composed of many different kinds of cells, each containing many complex
molecular structures. Many nonliving things may also be complex, but they do not
exhibit this degree of ordered complexity.
● Sensitivity. All organisms respond to stimuli. Plants grow toward a source of light,
and the pupils of your eyes dilate when you walk into a dark room.
● Growth, development, and reproduction. All organisms are capable of growing and
reproducing, and they all possess hereditary molecules that are passed to their
offspring, ensuring that the offspring are of the same species.
● Energy utilization. All organisms take in energy and use it to perform many kinds of
work. Every muscle in your body is powered with energy you obtain from your diet.
● Homeostasis. All organisms maintain relatively constant internal conditions that are
different from their environment, a process called homeostasis. For example, your
body temperature remains stable despite changes in outside temperatures.
● Evolutionary adaptation. All organisms interact with other organisms and the
nonliving environment in ways that influence their survival, and as a consequence,
organisms evolve adaptations to their environments.

Hierarchical organization of living systems

Life forms a hierarchy of organization from atoms to complex multicellular organisms. Atoms
are joined together to form molecules, which are assembled into more complex structures
such as organelles. These in turn form subsystems that provide different functions. Cells can
be organized into tissues, then into organs and organ systems such as the goose’s nervous
system pictured. This organization then extends beyond individual organisms to populations,
communities, ecosystems, and finally the biosphere.

Each level builds on the level below it:

1. The cellular level. At the cellular level, atoms, the fundamental elements of matter,
are joined together into clusters called molecules. Complex biological molecules are
assembled into tiny structures called organelles within membrane-bounded units we
call cells. The cell is the basic unit of life. Many independent organisms are composed
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 only of single cells. Bacteria are single cells, for example. All animals and plants, as
well as most fungi and algae, are multicellular, composed of more than one cell.
2. The organismal level. Cells in complex multicellular organisms exhibit three levels of
organization. The most basic level is that of tissues, which are groups of similar cells
that act as a functional unit. Tissues, in turn, are grouped into organs—body
structures composed of several different tissues that act as a structural and
functional unit. Your brain is an organ composed of nerve cells and a variety of
associated tissues that form protective coverings and contribute blood. At the third
level of organization, organs are grouped into organ systems. The nervous system, for
example, consists of sensory organs, the brain and spinal cord, and neurons that
convey signals.
3. The populational level. Individual organisms can be categorized into several
hierarchical levels within the living world. The most basic of these is the
population—a group of organisms of the same species living in the same place. All
populations of a particular kind of organism together form a species, its members
similar in appearance and able to interbreed. At a higher level of biological
organization, a biological community consists of all the populations of different
species living together in one place.
4. The ecosystem level. At the highest tier of biological organization, populations of
organisms interact with each other and their physical environment. Together
populations and their environment constitute an ecological system, or ecosystem.
For example, the biological community of a mountain meadow interacts with the
soil, water, and atmosphere of a mountain ecosystem in many important ways.
5. The biosphere. The entire planet can be thought of as an ecosystem that we call the
biosphere.

As you move up this hierarchy, the many interactions occurring at lower levels can
produce novel properties. These so-called emergent properties may not be predictable.

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Core Concepts in Biology
Structure determines function

A major unifying theme of biology is the relationship between structure and function. Said
simply, the proper functioning of molecules, of cells, and of tissues and organs depends on
their structure. When we know the function of a particular structure, we can infer the
function of similar structures found in different contexts, such as in different organisms.
When structure is altered, function is disrupted, with potential physiological consequences.

Living systems transform energy and matter

From single cells to the highest level of biological organization, the biosphere, living systems
have a constant need for energy. If we trace this all the way back, the original energy source
for the biosphere is the sun. Without this energy, living systems would not exhibit their
characteristic highly organized state. While this sounds simple, it means that the basic
nature of life is to constantly transform both energy and matter. We break down “food”
molecules for energy, then use this energy to build up other complex molecules. The energy
from the sun is trapped by photosynthetic organisms, which use this energy to reduce CO2
and produce organic compounds. The rest of us, who need a constant source of energy and
carbon, can oxidize these organic compounds back to CO2, releasing energy to drive the
processes of life.

Living systems depend on information transactions

The most obvious form of information in living systems is the genetic information carried in
every cell in the form of deoxyribonucleic acid (DNA). Each DNA molecule is formed from
two long chains of building blocks, called nucleotides, wound around each other. Four
different nucleotides are found in DNA, and the sequence in which they occur encodes the

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information to make and maintain a cell. The continuity of life from one generation to the
next— heredity—depends on the faithful copying of a cell’s DNA into daughter cells. The
entire set of DNA instructions that specifies a cell is called its genome. The sequence of the
human genome, 3 billion nucleotides long, was decoded in rough-draft form in 2001.
However, the importance of information goes beyond genomes and their inheritance. Cells
are highly complex nanomachines that receive, process, and respond to information. The
types in time and space, leading to changes over developmental time into different tissue
types—even though all cells in an organism carry the same genetic information. Living
systems are able to collect information about the environment, both internal and external,
and then respond to this information.

Carbon: The Framework of Biological Molecules

The framework of biological molecules consists predominantly of carbon atoms bonded to
other carbon atoms or to atoms of hydrogen, oxygen, nitrogen, sulfur, or phosphorus.
Because carbon atoms can form up to four covalent bonds, molecules containing carbon can
form straight chains, branches, or even rings, balls, tubes, and coils. Molecules consisting
only of carbon and hydrogen are called hydrocarbons. Because the oxidation of hydrocarbon
compounds results in a net release of energy, hydrocarbons make good fuels.

Functional groups account for differences in molecular properties

Carbon and hydrogen atoms both have very similar electronegativities. Electrons in C—C and
C—H bonds are therefore evenly distributed, with no significant differences in charge over
the molecular surface. For this reason, hydrocarbons are nonpolar. Most biological
molecules produced by cells, however, also contain other atoms. Because these other atoms
frequently have different electronegativities, molecules containing them exhibit regions of
partial positive or negative charge. They are polar. These molecules can be thought of as a
C—H core to which specific molecular groups, called functional groups, are attached. One

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such common functional group is —OH, called a hydroxyl group. Functional groups have
definite chemical properties that they retain no matter where they occur. Both the hydroxyl
and carbonyl (C==O) groups, for example, are polar because of the electronegativity of the
oxygen atoms. Other common functional groups are the acidic carboxyl (COOH), the
phosphate (PO4−), and the basic amino (NH2) groups. Many of these functional groups can
also participate in hydrogen bonding.

The primary functional chemical groups

These groups tend to act as units during chemical reactions and
give specific chemical properties to the molecules that possess
them. Amino groups, for example, make a molecule more basic,
and carboxyl groups make a molecule more acidic. These
functional groups are also not limited to the examples in the
“Found in” column but are widely distributed in biological
molecules.

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Isomers have the same molecular formulas but different structures
Organic molecules having the same molecular or empirical formula can exist in different
forms called isomers. If there are differences in the actual structure of their carbon skeleton,
we call them structural isomers. Glucose and fructose are structural isomers of C6H12O6.
Another form of isomers, called stereoisomers, have the same carbon skeleton but differ in
how the groups attached to this skeleton are arranged in space. Enzymes in biological
systems usually recognize only a single, specific stereoisomer. A subcategory of
stereoisomers, called enantiomers, are actually mirror images of each other. A molecule that
has mirror-image versions is called a chiral molecule. When carbon is bound to four different
molecules, this inherent asymmetry exists.

Figure 1 Polymer macromolecules. The four major biological macromolecules are shown. Carbohydrates,
nucleic acids, and proteins all form polymers and are shown with the monomers used to make them. Lipids do
not fit this simple monomer–polymer relationship. The triglyceride shown is constructed from glycerol and fatty
acids. All four types of macromolecules are also shown in their cellular context

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Biological macromolecules include carbohydrates, nucleic acids, proteins, and lipids
Remember that biological macromolecules are traditionally grouped into carbohydrates,
nucleic acids, proteins, and lipids. In many cases, these macromolecules are polymers. A
polymer is a long molecule built by linking together a large number of small, similar chemical
subunits called monomers. The nature of a polymer is determined by the monomers used to
build the polymer. For example, complex carbohydrates such as starch are polymers
composed of simple ring-shaped sugars. Nucleic acids (DNA and RNA) are polymers of
nucleotides, and proteins are polymers of amino acids. These long chains are built via
chemical reactions termed dehydration reactions and are broken down by hydrolysis
reactions. Lipids are macromolecules, but they really don’t follow the monomer–polymer
relationship. However, lipids are formed through dehydration reactions, which link the fatty
acids to glycerol.

The dehydration reaction

Despite the differences between monomers of these major polymers, the basic chemistry of
their synthesis is similar: To form a covalent bond between two monomers, an —OH group is
removed from one monomer, and a hydrogen atom (H) is removed from the other. This
reaction is the same for joining nucleotides when synthesizing DNA or joining glucose units
together to make starch. This reaction is also used to link fatty acids to glycerol in lipids. This
chemical reaction is called condensation or a dehydration reaction, because the removal of
—OH and —H is the same as the removal of a molecule of water (H2O). For every subunit
added to a macromolecule, one water molecule is removed. These and other biochemical
reactions require that the reacting substances are held close together and that the correct
chemical bonds are stressed and broken. This process of positioning and stressing, termed
catalysis, is carried out within cells by enzymes.

The hydrolysis reaction

Cells disassemble polymers into their constituent monomers by reversing the dehydration
reaction—a molecule of water is added instead of removed. In this reaction, called
hydrolysis, a hydrogen atom is attached to one subunit and a hydroxyl group to the other,
breaking the covalent bond joining the subunits.

Figure 2 Making and breaking macromolecules. a. Biological macromolecules are polymers formed by linking
monomers together through dehydration reactions. This process releases a water molecule for every bond
formed. b. Breaking the bond between subunits involves hydrolysis, which reverses the loss of a water molecule
by dehydration.

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Carbohydrates: Energy Storage and Structural Molecules
Monosaccharides are simple sugars
Carbohydrates are a loosely defined group of molecules that all contain carbon, hydrogen,
and oxygen in the molar ratio 1:2:1. Their empirical formula (which lists the number of
atoms in the molecule with subscripts) is (CH2O)n, where n is the number of carbon atoms.
Because they contain many carbon–hydrogen (C—H) bonds, which release energy when
oxidation occurs, carbohydrates are well suited for energy storage. Sugars are among the
most important energy-storage molecules, and they exist in several different forms. The
simplest of the carbohydrates are the monosaccharides (Greek mono, “single,” and Latin
saccharum, “sugar”). Simple sugars contain as few as three carbon atoms, but those that
play the central role in energy storage have six (figure 3.6). The empirical formula of
6-carbon sugars is:
C6H12O6 or (CH2O)6
Six-carbon sugars can exist in a straight-chain form, but dissolved in water (an aqueous
environment) they almost always form rings. The most important of the 6-carbon
monosaccharides for energy storage is glucose, which you first encountered in the examples
of chemical reactions in chapter 2. Glucose has seven energy-storing C—H bonds. Depending
on the orientation of the carbonyl group (C=O) when the ring is closed, glucose can exist in
two different forms: alpha (α) or beta (β).
Sugar isomers have structural differences Glucose is not the only sugar with the formula
C6H12O6. Both structural isomers and stereoisomers of this simple 6-carbon skeleton exist
in nature. Fructose is a structural isomer that differs in the position of the carbonyl carbon
(C=O); galactose is a stereoisomer that differs in the position of —OH and —H groups
relative to the ring. These differences often account for substantial functional differences
between the isomers. Your taste buds can discern them: Fructose tastes much sweeter than
glucose, despite the fact that both sugars have identical chemical composition. Enzymes that
act on different sugars can distinguish both the structural and stereoisomers of this basic
6-carbon skeleton. The different stereoisomers of glucose are also important in the polymers
that can be made using glucose as a monomer, as you will see later in this section.

Figure 3 Monosaccharides. Monosaccharides, or simple sugars, can contain as few as three carbon atoms and are often
used as building blocks to form larger molecules. The 5-carbon sugars ribose and deoxyribose are components of nucleic
acids). The carbons are conventionally numbered (in blue) from the more oxidized end.

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Figure 4 Structure of the glucose molecule. Glucose is a linear, 6-carbon molecule that forms a six-membered ring in
solution. Ring closure occurs such that two forms can result: α-glucose and β-glucose. These structures differ only in the
position of the —OH bound to carbon 1. The structure of the ring can be represented in many ways; shown here are the
most common, with the carbons conventionally numbered so that the forms can be compared easily. The heavy lines in the
ring structures represent portions of the molecule that are projecting out of the page toward you.

Disaccharides serve as transport molecules in plants and provide nutrition in

animals
Most organisms transport sugars within their bodies. In humans, the glucose that circulates
in the blood does so as a simple monosaccharide. In plants and many other organisms,
however, glucose is converted into a transport form before it is moved from place to place
within the organism. In such a form, it is less readily metabolized during transport. Transport
forms of sugars are commonly made by linking two monosaccharides together to form a
disaccharide (Greek di, “two”). Disaccharides serve as effective reservoirs of glucose
because the enzymes that normally use glucose in the organism cannot break the bond
linking the two monosaccharide subunits. Enzymes that can do so are typically present only
in the tissue that uses glucose. Transport forms differ depending on which monosaccharides
are linked to form the disaccharide. Glucose forms transport disaccharides with itself and
with many other monosaccharides, including fructose and galactose. When glucose forms a
disaccharide with the structural isomer fructose, the resulting disaccharide is sucrose, or
table sugar. Sucrose is the form most plants use to transport glucose and is the sugar that
most humans and other animals eat. Sugarcane and sugar beets are rich in sucrose. When
glucose is linked to the stereoisomer galactose, the resulting disaccharide is lactose, or milk
sugar. Many mammals supply energy to their young in the form of lactose.

Polysaccharides provide energy storage and structural components

Polysaccharides are longer polymers made up of monosaccharides that have been joined
through dehydration reactions. Starch, a storage polysaccharide, consists entirely of
α-glucose molecules linked in long chains. Cellulose, a structural polysaccharide, also
consists of glucose molecules linked in chains, but these molecules are β-glucose. Because
starch is built from α-glucose we call the linkages α linkages; cellulose has β linkages.

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Nucleic Acids: Information Molecules
The biochemical activity of a cell depends on production of a large number of proteins, each
with a specific sequence. The information necessary to produce the correct proteins is
passed through generations of organisms, even though the proteins themselves are not
inherited. Nucleic acids carry information inside cells, just as disks contain the information in
a computer or the GPS system in a car displays information needed by travelers. Two main
varieties of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).
Genetic information is stored in DNA, and short-lived copies of this are made in the form of
RNA, which is then used to direct the synthesis of proteins during the process of gene
expression. Unique among macromolecules, nucleic acids are able to serve as templates for
producing precise copies of themselves. This characteristic allows genetic information to be
preserved during cell division and during the reproduction of organisms. The role of RNA in
cells is much more complicated: RNA carries information; it is part of the organelle
responsible for protein synthesis, and recent work indicates it is also involved in the control
of gene expression. As a carrier of information, the form of RNA called messenger RNA
(mRNA) consists of transcribed single-stranded copies of portions of the DNA. These
transcripts serve as blueprints specifying the amino acid sequences of proteins.
Structure of a nucleotide. The nucleotide subunits of
DNA and RNA are made up of three elements: a 5-carbon
sugar (ribose or deoxyribose), an organic nitrogenous
base (adenine is shown here), and a phosphate group.
Notice that all the numbers on the sugar are given as
“primes” (1´, 2´, etc.) to distinguish them from the
numbering on the rings of the bases.

Nucleic acids are nucleotide polymers

Nucleic acids consist of long polymers of repeating subunits called nucleotides. Each
nucleotide consists of three components: a pentose, or 5-carbon, sugar; a phosphate (-PO4−)
group; and an organic nitrogen-containing, or nitrogenous, base. In DNA, the sugar is
deoxyribose and in RNA it is ribose. Polynucleotides are formed by joining the phosphate of
one nucleotide to a hydroxyl group on the sugar of another nucleotide in a dehydration
reaction. We call this a phosphodiester bond because the two sugars are linked by ester
bonds to a phosphate. A nucleic acid, then, is simply a chain of 5-carbon sugars linked
together by phosphodiester bonds with a nitrogenous base protruding from each sugar.
These chains of nucleotides, polynucleotides, have polarity, or different ends: a phosphate
on one end and an —OH from a sugar on the other end. We conventionally refer to these
ends as 5´ (“five-prime,” -PO4−) and 3´ (“three-prime,” —OH) taken from the carbon
numbering of the sugar. Nucleotides have five types of nitrogenous bases. Two of these are
large, double-ring molecules called purines that are each found in both DNA and RNA; the
two purines are adenine (A) and guanine (G). The other three bases are single-ring

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molecules called pyrimidines that include cytosine (C, in both DNA and RNA), thymine (T, in
DNA only), and uracil (U, in RNA only).

DNA stores genetic information

Organisms use sequences of nucleotides in DNA to encode the information specifying the
amino acid sequences of their proteins. This method of encoding information is very similar
to the way in which sequences of letters encode information in a sentence. A sentence
written in English consists of a combination of the 26 different letters of the alphabet in a
certain order; the code of a DNA molecule consists of different combinations of the four
types of nucleotides in specific sequences, such as CGCTTACG. DNA molecules in organisms
exist as two polynucleotide chains wrapped about each other in a long linear molecule in
eukaryotes, and a circular molecule in most prokaryotes. The two strands of a DNA polymer
wind around each other like the outside and inside rails of a spiral staircase. Such a spiral
shape is called a helix, and a helix composed of two chains is called a double helix. Each step
of DNA’s helical staircase is composed of a base-pair. The pair consists of a base in one chain
attracted by hydrogen bonds to a base opposite it on the other chain. The base-pairing rules
arise from the most stable hydrogen bonding configurations between the bases: Adenine
pairs with thymine (in DNA) or with uracil (in RNA), and cytosine pairs with guanine. The
bases that participate in base-pairing are said to be complementary to each other.

Figure 5 The structure of DNA. DNA consists of two polynucleotide chains running in opposite directions wrapped about a
single helical axis. Hydrogen bond formation (dashed lines) between the nitrogenous bases, called base-pairing, causes the
two chains of DNA to bind to each other and form a double helix.

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Cell Structure
Cell Theory
Cells are too small for you to be able to see. Although there are exceptions, a typical
eukaryotic cell is 10 to 100 micrometers (μm) in diameter, and most prokaryotic cells are
only 1 to 10 μm in diameter. Because cells are so small, they were not discovered until the
invention of the microscope in the 17th century. English natural philosopher Robert Hooke
was the first to observe cells in 1665, naming the shapes he saw in cork cellulae (Latin,
“small rooms”). They are known to us as cells. In 1838, German botanist Matthias Schleiden
stated that all plants “are aggregates of fully individualized, independent, separate beings,
namely the cells themselves.” In 1839, German physiologist Theodor Schwann reported that
all animal tissues also consist of individual cells. Thus, the cell theory was born.
The cell theory includes the following three principles:
1. All organisms are composed of one or more cells, and the life processes of
metabolism and heredity occur within these cells.
2. Cells are the smallest living things, the basic units of organization of all organisms.
3. Cells arise only by division of a previously existing cell.
The advantage of small cell size is readily apparent in terms of the surface area-to-volume
ratio. As a cell’s size increases, its volume increases much more rapidly than its surface area.
For a spherical cell, the surface area is proportional to the square of the radius, whereas the
volume is proportional to the cube of the radius. Thus, if the radii of two cells differ by a
factor of 10, the larger cell will have 102, or 100 times, the surface area, but 103, or 1000
times, the volume of the smaller cell. The cell surface provides the only opportunity for
interaction with the environment, because all substances enter and exit a cell via this
surface. The membrane surrounding the cell plays a key role in controlling cell function.
Because small cells have more surface area per unit of volume than large ones, control over
cell contents is more effective when cells are relatively small. Although most cells are small,
some quite large cells do exist. These cells have apparently overcome the surface
area-to-volume problem by one or more adaptive mechanisms. For example, some cells,
such as skeletal muscle cells, have more than one nucleus, allowing genetic information to
be spread around a large cell. Some other large cells, such as neurons, are long and skinny,
so that any given point within the cell is close to the plasma membrane. This permits
diffusion between the inside and outside of the cell to still be rapid.

MODERN STATUS OF THE CELL THEORY

1. All livings things are made up of one more cells
2. New cells arise from preexisting cells division.
3. The cells is the foundamental structure and fuctional unit of all living organisms.
4. Energy flow occurs within cells.
5. Cells contain ereditary information (DNA) wich is passed from cell to cell division.

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 Surface area-to-volume ratio
As a cell gets larger, its volume increases at a faster rate than its
surface area. If the cell radius increases by 10 times, the surface
area increases by 100 times, but the volume increases by 1000
times. A cell’s surface area must be large enough to meet the
metabolic needs of its volume.

Microscopes allow visualization of cells and components

The reason we can’t see such small objects
is the limited resolution of the human eye.
Resolution is the minimum distance two
points can be apart and still be
distinguished as two separate points. When
two objects are closer together than about
100 µm, the light reflected from each
strikes the same photoreceptor cell at the
rear of the eye. Only when the objects are
farther than 100 µm apart can the light
from each strike different cells, allowing
your eye to resolve them as two distinct
objects rather than one.

Types of microscopes
The first microscopists used glass lenses to
magnify small cells and cause them to
appear larger than the 100-µm limit
imposed by the human eye. The glass lens
increases focusing power. Modern light
microscopes, which operate with visible
light, use two magnifying lenses (and a
variety of correcting lenses) to achieve very
high magnification and clarity. Microscopes
that magnify in stages using several lenses
are called compound microscopes. They can
resolve structures that are separated by at
least 200 nanometres (nm). One way to
avoid overlap is by using a beam of
electrons rather than a beam of light.
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Electrons have a much shorter wavelength, and an electron microscope, employing electron
beams, has 1000 times the resolving power of a light microscope. can resolve objects only
0.2 nm apart (which is only twice the diameter of a hydrogen atom!). A second kind of
electron microscope, the scanning electron microscope, beams electrons onto the surface of
the specimen. The electrons reflected back from the surface, together with other electrons
that the specimen itself emits as a result of the bombardment, are amplified and
transmitted to a screen, where the image can be viewed and photographed. Scanning
electron microscopy yields striking three-dimensional images. This technique has improved
our understanding of many biological and physical phenomena.

Cellular organization
The general plan of cellular organization varies between different organisms, but despite
these modifications, all cells resemble one another in certain fundamental ways. Before we
begin a detailed examination of cell structure, let’s first summarize four major features all
cells have in common: (1) a nucleoid or nucleus where genetic material is located, (2)
cytoplasm, (3) ribosomes to synthesize proteins, and (4) a plasma membrane.
Centrally located genetic material
Every cell contains DNA, the hereditary molecule. In prokaryotes, the simplest organisms,
most of the genetic material lies in a single circular molecule of DNA. It typically resides near
the center of the cell in an area called the nucleoid. This area is not segregated, however,
from the rest of the cell’s interior by membranes. By contrast, eukaryotic cells, found in
more complex organisms, have DNA segregated into a nucleus, which is surrounded by a
double-membrane structure called the nuclear envelope. In both types of organisms, the
DNA contains the genes that code for the proteins synthesized by the cell.
The cytoplasm
A semifluid matrix called the cytoplasm fills the interior of the cell. The cytoplasm contains
all of the sugars, amino acids, and proteins the cell uses to carry out its everyday activities.
Although it is an aqueous medium, cytoplasm is more like Jell-O than water due to the high
concentration of proteins and other macromolecules. We call any discrete macromolecular
structure in the cytoplasm specialized for a particular function an organelle. The part of the
cytoplasm that contains organic molecules and ions in solution is called the cytosol to
distinguish it from the larger organelles suspended in this fluid.
The plasma membrane
The plasma membrane encloses a cell and separates its contents from its surroundings. The
plasma membrane is a phospholipid bilayer about 5 to 10 nm (5 to 10 billionths of a meter)
thick, with proteins embedded in it. Viewed in cross section with the electron microscope,
such membranes appear as two dark lines separated by a lighter area. This distinctive
appearance arises from the tail-to-tail packing of the phospholipid molecules that make-up
the membrane. The proteins of the plasma membrane are generally responsible for a cell’s
ability to interact with the environment. Membrane proteins give cells identity, and provide
for a variety of functions, including transport and communication with other cells and the
environment. This interaction between cell surface molecules is especially important in

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multicellular organisms, whose cells must be able to recognize one another as they form
tissues.

When cells were visualized with microscopes, two basic cellular architectures were
recognized: eukaryotic and prokaryotic. These terms refer to the presence or absence,
respectively, of a membrane-bounded nucleus that contains genetic material.

Prokaryotic Cells
Prokaryotes are the simplest organisms and are small. They consist of cytoplasm surrounded
by a plasma membrane and are encased within a rigid cell wall. They have no distinct
interior compartments. Prokaryotic cells do not have an internal membrane system or
numerous membrane-bounded organelles. The plasma membrane of a prokaryotic cell
carries out some of the functions organelles perform in eukaryotic cells. Because a
prokaryotic cell contains no membrane-bounded organelles, the DNA, enzymes, and other
cytoplasmic constituents have access to all parts of the cell. Reactions are not
compartmentalized as they are in eukaryotic cells, and the whole prokaryote operates as a
single unit.

Generalized cell organization of a prokaryote. The nucleoid is

visible as a dense central region segregated from the
cytoplasm. Some prokaryotes have hairlike growths (called pili
[singular, pilus]) on the outside of the cell. Flagella (singular,
flagellum) are long, threadlike structures protruding from the
surface of a cell that are used in locomotion. Prokaryotic
flagella are protein fibers that extend out from the cell.

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Figure 6 Structure of an animal cell. In this generalized diagram of an animal cell, the plasma membrane encases the cell,
which contains the cytoskeleton and various cell organelles and interior structures suspended in a semifluid matrix called
the cytoplasm. Some kinds of animal cells possess fingerlike projections called microvilli. Other types of eukaryotic cells—for
example, many protist cells—may possess flagella, which aid in movement, or cilia, which can have many different
functions.

Eukaryotic cells
Eukaryotic cells are far more complex than prokaryotic cells. The hallmark of the eukaryotic
cell is compartmentalization. This is achieved through a combination of an extensive
endomembrane system that weaves through the cell interior and by numerous organelles.
These organelles include membrane-bounded structures that form compartments within
which multiple biochemical processes can proceed simultaneously and independently. Plant
cells often have a large, membrane-bounded sac called a central vacuole, which stores
proteins, pigments, and waste materials. Both plant and animal cells contain
vesicles—smaller sacs that store and transport a variety of materials. Inside the nucleus, the
DNA is wound tightly around proteins and packaged into compact units called
chromosomes. All eukaryotic cells are supported by an internal protein scaffold, the
cytoskeleton.
The largest and most easily seen organelle within a eukaryotic cell is the nucleus. The
nucleus contains the genetic information that enables the synthesis of nearly all proteins of
a living eukaryotic cell. Most eukaryotic cells possess a single nucleus, although the cells of
fungi and some other groups may have from several to many nuclei.
The surface of the nucleus is bounded by two phospholipid bilayer membranes, which
together make up the nuclear envelope. The outer membrane of the nuclear envelope is
continuous with the cytoplasm’s interior membrane system, called the endoplasmic
reticulum.

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Cell Structures:
Golgi Complex
It is a morphologically and biochemically
differentiated part of the smooth endoplasmic
reticulum. It consists of flattened and stacked sacs
which in section appear as a series of parallel
membranes, dense vesicles and vacuoles. It’s involved
in secretion phenomena. It shows a definite polarity,
with a cis receiving side vesicles containing the
products of the endoplasmic reticulum and the trans
face releasing new vesicles.
The Golgi apparatus is a smooth, concave, membranous structure. It receives material for
processing in transport vesicles on the cis face and
sends the material packaged in transport or
secretory vesicles off the trans face. The substance
in a vesicle could be for export out of the cell or for
distribution to another region within the same cell.
Proteins and lipids manufactured on the rough and
smooth ER membranes are transported into the Golgi apparatus and modified as they pass
through it. The most common alteration is the addition or modification of short sugar
chains, forming glycoproteins and glycolipids. In many instances, enzymes in the Golgi
apparatus modify existing glycoproteins and glycolipids made in the ER by cleaving a sugar
from a chain or by modifying one or more of the sugars.

Ribosomes
Ribosomes are small granules measuring 15-20 nm and are made up of rRNA. They consist
of two subunits, major and minor, between which a strand of mRNA runs.
They are constantly associated with forming polyribosomes and preside over protein
synthesis for the renewal of intracellular protein material, of new free ribosomes, or to
synthesize proteins, which are released outside the cell.
So Ribosomes:
- Build proteins from amino acids in cytoplasm
- may be free-floating or may be attached to ER
- made of RNA.

17
Newly synthesized or modified glycoproteins and
glycolipids are accumulated at the end of the Golgi body in
folds of lamellar membranes called cisternae.
Periodically, the membranes of the cistern buds, generate
small membranous secretory vesicles that contain
glycoproteins and glycolipids.
These vesicles can then diffuse to other sub-cellular
locations, distributing the newly synthesized molecules to
their correct destination.
An additional function of the Golgi apparatus is the
synthesis of cell wall components.
Polysaccharides that do not include cellulose, but form the wall of vegetal cells, are
synthesized in the Golgi apparatus and sent to the plasma membrane where they can add to
cellulose, which is present on the outside of the cell.
Other polysaccharides created by plants are also synthesized in the Golgi apparatus.
Lisosomes
. Lysosomes are formed from vesicles budding off the Golgi. They contain hydrolytic enzymes
that digest particles or cells taken into the cell by phagocytosis, and break down old
organelles. They are part of the endomembranous system. They derive from the Golgi
apparatus and contain high levels of digestive
enzymes, which catalyze the rapid degradation of
proteins, nucleic acids, and carbohydrates. Digest
both materials from outside the cell, or help recycle
endocellular waste materials.
During the life of a eukaryotic cell, lysosomal
enzymes degrade old organelles and recycle the
molecules that compose them. This process allows the
formation of new Organelles.
The digestive enzymes in the lysosome are optimally active at
acid pH. Lysosomes are activated by fusing with a food vesicle
produced by phagocytosis or by fusing with an old or worn-out
organelle. The fusion event activates proton pumps in the
lysosomal membrane, lowering the internal pH. As the interior
pH falls, the digestive enzymes contained in the lysosome
become active. This leads to the
degradation of macromolecules in the
food vesicle or the destruction of the
old organelle.

18
Mitochondrion
makes the cell’s energy
the more energy the cell needs, the more mitochondria it
has. They are organelles surrounded by a double membrane
system that convert energy into forms useful for promoting
cellular reactions. Mitochondria divide like bacteria and can
change position, shape and number within the cells
according to the tissue energy. Their internal structure is
preserved to ensure the production of ATP.
Glycolysis occurs in the cytosol. The subsequent oxidation
of pyruvate occurs within the mitochondria. The citric acid
cycle or krebs cycle occurs in the mitochondrial matrix.
Oxidative phosphorylation occurs on mitochondrial ridges.
Matrix
It contains a highly concentrated mixture of hundreds of enzymes, including those necessary
for the oxidation of pyruvate and fatty acids and for the citric acid cycle. The matrix also
contains the mitochondrial genome, special mitochondrial ribosomes, tRNA, and various
enzymes needed for the expression of mitochondrial genes.
Inner membrane
It is folded in numerous ridges that greatly increase its total area. It contains proteins with
three types of functions:
1) those that carry out the oxidation reactions of the respiratory chain;
2) an enzyme complex called ATPsintase that produces ATP in the matrix;
3) specific transport proteins that regulate the passage of metabolites in and out of the
matrix.
Outer membrane
It contains a large protein (called porin) that forms a channel, so the outer membrane is
permeable to all molecules below 5000 daltons. Other proteins of this membrane include
enzymes involved in mitochondrial synthesis of lipids and enzymes that convert lipid
substrates into forms that are subsequently metabolized in the matrix.
Inter-membrane Space
It contains several enzymes that use ATP coming out of the matrix to phosphorylate other
nucleotides.
Mitochondria divide like bacteria and can change position, shape and number within the
cells according to the tissue energy.

19
Peroxisomes
Peroxisomes are spherical organelles that may contain a large crystal structure composed of
protein. Peroxisomes contain digestive and detoxifying enzymes that produce hydrogen
peroxide as a by-product. A peroxisome has been colored
green in the electron micrograph.
Microbodies of 0.5-1 nano meters of diameters. They are
delimited by a single membrane.
They are rich in enzymes such as peroxidase, catalase, D
amino acid oxidase, uricoxidase.
They participate in the formation of a-keto acids and play an
important role in gluconeogenesis starting from lipids and
other non-carbohydrate precursors.

Centrioles
Grane es that constitute the kinetic center of the cell and requlate both its movements
towards the environment (cilia, flagella) and the movements that occur within it (movement
of chromosomes).
The centriole is a hollow cylindrical or lle whose wall is made up of nine triplets of tubules
(microtubules). Together with an electron-dense material surrounding them, called
“pericentriolar material” (PCM), they constitute the
centrosome, the most important “microtubule
organizing center” (WTOC) of the cell.
During cell division the centrioles duplicate, but
remain united into a single centrosome. At the end of
cell division the two pairs separate, migrating to
opposite poles of the cell and giving rise to two
distinct centrosomes.

Citosol
It's the part of the cytoplasm that remains if we remove the plasma membrane and the
membranous organelles. It's comparable to an aqueous gel in which many small and large
molecules are immersed. It's a very dynamic compartment.

Cytoskeleton
The cytoplasm of all eukaryotic cells is crisscrossed by a network of protein fibers that
supports the shape of the cell and anchors organelles to fixed locations. This network, called
the cytoskeleton, is a dynamic system, constantly assembling and disassembling. Individual
fibers consist of polymers of identical protein subunits that attract one another and
spontaneously assemble into long chains. Fibers disassemble in the same way, as one
subunit after another breaks away from one end of the chain. Cytoskeleton it’s only typical
of eukaryotic cells.
The cytoskeleton is also closely associated with signaling and cell adhesion processes.

20
They are made of microtubules and are found throughout cytoplasm. This gives shape to cell
& moves.
There are three types of fibers that compose the cytoskeleton:
1. Microfilaments, with a diameter of 7 nm, are polymers of the protein actin: are
useful to maintain the structure of the cell and generate movement.

2. The intermediate filaments in each cell type are formed by polymers of different
proteins, but all similar in size and structure.

3. Microtubules are composed of the tubulin protein and have a diameter of 25 nm.

that do not work alone, but associated with accessory proteins, essential for the assembly of
cytoskeletal structures and their functioning.
The cytoskeleton is involved in:
- Spatial organization of the cytoplasm
- Intracellular movements of organelles
- Segregation of chromosomes during cytokinesis cell division
- Cell movement on the substate
- Muscle contraction
Actin filaments and microtubules often coordinati activities to carry out cellular processes.
For example, during cell replication, new chromosomes move to opposite sides of dividing
cells because they are attached to shortening microtubules.
Muscle cells also use actin filaments which run along the filaments of the motor protein
myosin to drive muscle contraction. The blinking of eyelashes, the flight of the wing and the
crawling of a baby depend on the movements of the skeleton of muscle cells.

21
Plant cell
The plant cell is a type of eukaryotic cell, with several peculiarities that differentiate it from
animal cells, fungal cells and other living kingdom cells:
The presence of the cell wall made up of cellulose (a polymer whose elementary unit is
glucose, proteins and, as a result of modifications, lignin, suberin... ) and its related
plasmodesmas, channels in the cell wall through which plant cells are in communication
with each other.
Plastids, and especially chloroplasts which, thanks to chlorophyll, enable plant cells to
produce sugar (glucose) and oxygen monomers from CO2 using solar energy, this process is
defined as chlorophyll photosynthesis.
The characterizing presence of numerous vacuoles (not organelles but endocellular cavities,
wrapped by a membrane, called tonoplast) that occupy a large part of the cell and whose
main function is to maintain the cell turgor. They are involved in the control of the passage
of molecules from the lymph to the cytosol, in the maintenance of the optimal pH of the
cytosol and perform reserve functions of various substances.
The absence of animal cell centrioles.

Figure 7 7 Structure of a plant cell. Most mature plant cells contain a large central vacuole, which occupies a major portion
of the internal volume of the cell, and organelles called chloroplasts, within which photosynthesis takes place. The cells of
plants, fungi, and some protists have cell walls, although the composition of the walls varies among the groups. Plant cells
have cytoplasmic connections to one another through openings in the cell wall called plasmodesmata. Flagella occur in
sperm of a few plant species, but are otherwise absent from plant and fungal cells. Centrioles are also usually absent.

22
Viruses
Due to their characteristics they are not considered
living organisms. Viruses have a simple acellular
structure and reproduce exclusively by exploiting
the metabolic mechanisms of a host cell. Since
they are not cellular and cannot reproduce
independently, they cannot be assigned to
traditional classification categories.
Viruses are specific and this specificity also extends
to the type of cells that the individual virus is capable of infecting. For example,
bacteriophages are viruses that infect bacteria. The image illustrates the replication of a
virus in an animal cell, which follows the following stages:
1. Hooking
2. Penetration
3. Biosynthesis
4. Maturation
5. Release

Biological Features
They are unable to reproduce independently and are equipped with their own genome and
protein structures essential to protect it and allow the attack and invasion of target cells.

Virus infection can be lytic, i.e. the infected cell produces new viruses and then the escape
of the virus determines its lysis or lysogenicity.
The genome of the virus can integrate into the genome of the host cell (provirus) and
replicate with it with various effects such as predisposition to tumor development.
Eukaryotic cells exhibit distinctive features that characterise their structure and function:

23
In terms of structural organization, these cells possess a defined nucleus separated from the
cytoplasm by a nuclear envelope containing intricate pores. Chromosomes within the
nucleus are complex structures formed by DNA and associated proteins, capable of
compacting during mitosis.
The cytoplasm houses various membranous organelles, such as the endoplasmic reticulum,
Golgi complex, lysosomes, endosomes, peroxisomes, and glyoxysomes. Specialized
organelles like mitochondria facilitate aerobic respiration, while chloroplasts enable
photosynthesis.
A complex cytoskeletal system, including microfilaments, intermediate filaments, and
microtubules, is present, along with associated motor proteins.
Eukaryotic cells also feature motility structures like complex flagella and cilia. They possess
the capability to ingest fluids and particulate matter through vesicle formation from the
plasma membrane, involving endocytosis and phagocytosis.
In the case of plant cells, a distinctive cell wall composed of cellulose is present. Cell division
involves a mitotic spindle with microtubules aiding in chromosome separation.
Genetically, eukaryotic cells are diploid, containing two copies of genes inherited from each
parent. The synthesis of RNA is orchestrated by three different enzymes known as RNA
polymerases.
Furthermore, the reproductive strategy involves sexual reproduction, necessitating
processes like meiosis and fertilization. These characteristics collectively define the
complexity and versatility of eukaryotic cells.

Lipids
Lipids are the chief concentrated storage form of energy forming about 3.50/0of the cell
content. Lipids are organic substances relatively insoluble in water but soluble in organic
solvents (alcohol, ether).
Functions :
1. They are the concentrated fuel reserve of the body.
2. Lipids are constituents of membrane structure and regulate the membrane
permeability.
3. They serve as sourceof fat soluble vitamins.
4. Lipids are important cellular metabolic regulators
5. Lipds protect the internal organs and serve as insulating materials
Lipids, characterized by their abundant nonpolar covalent bonds (C-H), are hydrophobic due
to their insolubility in water. Unlike polymers, they do not constitute long chains of repeating
units.
Fats and oils, collectively known as neutral fats or glycerides, serve as a means of energetic
storage. Phospholipids play a crucial structural role in biological membranes, contributing to
their formation and integrity.

24
Steroids, with regulatory and structural functions, play essential roles in digestion.

Simple Lipids
Simple lipids, classified into two main types, are esters formed by the combination of fatty
acids with alcohol.
1. Neutral or True Fats: These are esters of fatty acids with glycerol. True fats consist
mainly of carbon, hydrogen, and oxygen, with a lower proportion of oxygen. A fat
molecule comprises two essential components: Glicerol and Fatty Acids.
2. Waxes: Waxes are esters of fatty acids with alcohols other than glycerol, presenting a
different composition and functional role compared to true fats.
Glycerol:
Glycerol is a three-carbon molecule, each carbon bearing a hydroxyl (-OH)
group.
Fatty Acids:
Fatty acids, occurring in 1-3 molecules, can be of the same or different
long-chained varieties. Fatty acids are
unbranched chains of carbon atoms,
featuring a carboxyl (-COOH) group at
one end and hydrogen atoms bonded to
nearly all carbon atoms. Fatty acids can
be categorized as saturated or
unsaturated.

Saturated Fatty Acids:

In saturated fatty acids, all carbon bonds are saturated with hydrogen atoms. These
molecules are rigid and tightly pack together, leading to a solid state at room temperature.
Unsaturated Fatty Acids:
Unsaturated fatty acids have one or more double bonds in their hydrocarbon chain,
introducing kinks in the molecule. These kinks prevent the alignment of unsaturated fat
molecules with adjacent ones, resulting in a more fluid consistency at room temperature, as
observed in oils.

25
 Figure 8 (a) satured (b) unsatured fatty acids

TRIGLYCERIDES:
Triglycerides, found abundantly in both animal and vegetable fats, constitute a prevalent
type of glyceride. Comprising a molecule of glycerol (a three-carbon alcohol) combined with
three molecules of fatty acid, triglycerides serve as a primary form of energy storage in
organisms.

WAXES:
Waxes are lipids consisting of long-chain saturated fatty acids and high-molecular-weight
saturated alcohols, distinct from glycerol. Examples include beeswax, derived from the
abdominal glands of worker honey bees, and lanolin or wool fat, obtained from sheep's
wool. Sebum, cerumen (earwax), and various plant waxes also fall under this category, each
serving unique biological functions.

COMPLEX LIPIDS:
Complex lipids, derivatives of simple lipids, feature additional groups such as phosphate,
nitrogenous bases, or proteins. This category includes Phospholipids, Glycolipids, and
Lipoproteins.

Phospholipids:
Phospholipids consist of a glycerol or another alcohol molecule with a phosphate group at
one of its -OH groups, two fatty acid molecules at the other -OH groups, and a
nitrogen-containing base attached to the
phosphate group.
Amphipathic in nature, phospholipids have a
hydrophobic tail (fatty acids) and a hydrophilic
head (phosphate group). In the presence of
water, the hydrophilic ends point toward the
26
aqueous environment, while the hydrophobic tails point in the opposite direction,
contributing to their unique structural and functional properties.
Cell Membrane:
The cell membrane is composed of a double layer of phospholipids, forming a lipid bilayer.
Phospholipids are the major components of cellular membranes.
Multiple Sclerosis:
Multiple sclerosis is a chronic inflammatory
demyelinating disease with autoimmune origins,
affecting the central nervous system, including the
brain and spinal cord. The autoimmune response
leads to the destruction of myelin sheaths,
resulting in the blocking or slowing of nerve
impulses. Areas where myelin is damaged are
referred to as plaques, giving rise to the term
"sclerosis with plaques." Symptoms include
physical weakness, paralysis, and ocular disorders,
characterized by periods of exacerbation and remission.

LIPOPROTEINS:
Lipoproteins, crucial constituents of membranes, contain lipids and proteins. They are
present in milk and egg yolk and play a role in transporting lipids in the blood and lymph.
There are five types of lipoproteins: chylomicrons, VLDL, LDL, HDL, and Free fatty acid
albumin complex.
Lipoproteins - Introduction, Structure, and Function:
Lipoproteins are globular, micelle-like particles with a hydrophobic core of triacylglycerols
and cholesterol esters, surrounded by an amphipathic coat of protein, phospholipid, and
cholesterol. Apolipoproteins on the surface aid in solubilizing lipids and targeting
lipoproteins to specific tissues. The five types of lipoproteins (VLDL, LDL, IDL, HDL) have
distinct functions and properties, facilitating the transport of triacylglycerols, cholesterol,
and phospholipids throughout the body.
The major fuction of lypoproteins is to transport triacylglycerols, cholesterol and
phospholipids around the body.

DERIVED LIPIDS:
Derived lipids are obtained through the hydrolysis of simple and complex lipids. Examples
include steroids, terpenes, and prostaglandins.

Steroids:
Steroids, included in lipids due to their fat-like properties, are composed of four fused
carbon rings. They do not contain fatty acids but are vital components in lipids. Common
steroids are sterols that include cholesterol, vitamin D, testosterone, and adrenocortical
hormones.

27
Cholesterol:
Cholesterol, a key constituent of biological membranes,
regulates membrane fluidity. It reduces fluidity at moderately
elevated temperatures and prevents membrane solidification at
lower temperatures by disrupting the regular packing of
phospholipids.
Terpenes:
Terpenes, major components of essential oils produced by plants, contribute to the
fragrance of plant parts. Examples include phytol in vitamins A, E, and K, carotenoid pigment
as a precursor for vitamin A, and gibberellins, a plant hormone classified as a terpene.

Structure and function of biological membranes

The plasma membrane serves as the boundary of the cell, separating the internal
environment from the external one. While it does not entirely block exchanges, it plays
crucial roles in cellular processes.
This membrane plays a vital role in controlling the
access of solutes and solvents (a mechanical barrier),
allowing the cytoplasm to maintain distinct
characteristics compared to external liquids.
Moreover, it facilitates the entry and exit of
macromolecules and larger structures, contributing to
the dynamic nature of cellular processes.
In response to external signal molecules, the plasma
membrane triggers internal reactions, showcasing its
active role in cellular communication. It carries
specific molecules that aid in cell recognition,
adhesion to other cells, and intercellular
communication. The membrane's flexibility allows the cell to exhibit movement, expansion,
and changes in shape.
Membranes found in various cellular locations share a common composition of lipids and
proteins. Lipids form a double layer, constituting the basic structure and providing an
impermeable barrier to most water-soluble molecules. Proteins, unique to each membrane,
execute diverse functions essential for cellular activities.
Lipids in cell membranes possess an amphipathic
nature, with hydrophilic heads interacting with
aqueous solutions and hydrophobic tails avoiding
them. This amphipathic arrangement is critical for
the spontaneous formation of lipid bilayers in
aqueous environments.
Membrane lipids include phospholipids
(phosphoglycerides, sphingolipids), sterols (cholesterol, ergosterol, stigmasterol), and

28
glycolipids modified by adding carbohydrates.
Glycolipids play roles in protection, electrical effects,
insulation, and cell recognition.
The absence of covalent bonds between lipid molecules
allows for individual movement within each layer,
rendering membranes "fluid". This fluidity is attributed
to lateral and transversal diffusion of lipids.
Cellular processes involving membranes include
diffusion and flip-flop movements of lipid
molecules. Membranes also participate in signal
transduction, influencing cellular processes such
as apoptosis, caspase activation, and immune
responses. They express "eat-me" signals on
apoptotic cells, facilitating recognition and
removal. Additionally, membranes contribute to
peptide presentation, influencing immune
responses. Overall, the intricate structure and
dynamic functions of biological membranes are
indispensable for cellular integrity,
communication, and responsiveness to the
environment.

Membrane fluidity
Ease with which its lipid molecules migrate into the bilayer.
Membrane fluidity: ease with which its lipid molucules migrate into the bilayer. At a given
temperature it depends on:

29
 - Lipid composition;
- Nature of hydrocarbon tails;
The fundamental properties of hydrocarbon tails for membrane fluidity are: tail length and
number of double bonds. The shorter the chains, the greater the fluidity. Length between
16-20 carbon atoms. Generally of the 2 chains:
1. One is always saturated (no double bonds) 2;
2. The other always unsaturated (1 or more double bonds);
In animal cells, fluidity is influenced by the presence of cholesterol. Cholesterol has a polar
head group, a rigid planar steroid ring structure and a nonpolar hydrocarbon tail. It is
positioned between 2 phospholipids, characterizing a cholesterol-stiffened region. So low
cholesterol 🡪 more fluid region. The fluidity degree is characterized by the presence of the
variability in concentrations of cholesterol. In fact, in extreme high temperatures membrane
is more fluid and needs more cholesterol; viceversa with extreme low temperatures.
The MEMBRANE is asymmetrical
Another main feature of the plasma membrane is the asymmetry. The two lipid layers have
different compositions which makes the bilayer “asymmetric”.
This asymmetry is already determined at the level of biogenesis of the different membranes
and is maintained in all passages of membrane portions from one organelle to another. In
the extracellular space we have a lot of glycolipdis; phosphotidylserine is mainly in the
citosol 🡪 hence, the asymmetry.
The asymmetry is established during biogenesis in the ER and is also guaranteed thanks to
the presence of phospholipid translocators, a special class of transmembrane enzymes that
catalized the flip-flop.
Lipid asymmetry is functionally important:
Outer Membrane: glycolipids for signaling and recognition reactions.
Inner Membrane: phosphadylethanolamine and phosphatidylserine, for the transmission of
signals within the cell.
Membranes are dynamic structures and are constantly forming, transforming, melting and
breacking down.
Two different model of biological membranes: models.
Membrane proteins and the fluid mosaic model 2 different models of biological membranes.
In 1935 the sandwich model by Davison-Danielli: they though that the protein covered both
sides of the cellular membrane.
La membrana può essere assimilata ad un sandwich di fosfolipidi tra due trati proteici. Si è
poi visto che questo modello, accettato per più di 20 anni, non è corretto. Questo modello è
stato superato nel 1972 dall’attuale modello a mosaico fluido: la membrana ha un doppio

30
strato fosfolipidico e da un mosaico di proteine associate che attraversano questo bilayer e
sono in continuo cambiamento.
In this model the globular proteins are embedded in the bilayer. The fluid mosaic model: a
double layer fluid lipid in which proteins floats. In this model, called fluid mosaic, a mosaic of
proteins floats within or on top of fluid lipid bilayer. The membranes have different
compositions.
In this model, called fluid mosaic, a mosaic of proteins floats. In the bilayer they are the
protein embedeed, that have the fuctions of (sostentamento).
Chemical composition of membrane: protein, lipids and carbs, but in different size, depends
on the membrane type.
Today we divide membrane proteins in two categories based on how they are associated
with the membrane:
- Integral membrane proteins: are incorporated into the membrane;
- Pheriphelical proteins, are associated with the membrane surface;

1. Transmembrane Proteins;
2. Monolayer associated proteins belong to the integral proteins;
3. Lipid-linked to integral membrane proteins;
4. Protein- attached are peripheral proteins (associated with other integral membrane
proteins)
Transmembrane Proteins:
- Transporters: Membranes are very selective, allowing only certain solutes to enter or
leave the cell, through either channels or carriers composed of proteins. Specific
function: actively pump ions in and out of the cell
- Anchors: Surface proteins that interact with other cells are often anchored to the
cytoskeleton by linking proteins. For examples integrins link intracellular actin
filaments to extra cellular matrix protein
- Receptors: Membranes are exquisitely sensitive to chemical messages, which are
detected by receptor proteins on their surfaces.
- Enzymes: Cells carry out many chemical reactions on the interior surface of the
plasma membrane, using enzymes attached to the membrane.
Water transport
Water can freely diffuse through cell membranes by Osmosis: is the movement of water
molecules from solution with a high concentration of water molecules to a solution with a
lower concentration of water molucules, through a cell’s partially permeable membrane.
So osmosis is the flow of water across a semipermeable membrane due to differences in
solute concentrations. Differences in concentrations of non permeant solutes determine
differences in osmotic pressure, this difference in osmotic pressure.

31
Hypotonic solution: the compartment with high concentration of molecules and a lower
concentration of water.
The quantity of water that enters the cell depends on the difference in the concentration of
solutes in the cell and in the extracelluloar fluid. This difference is mesaured as osmotic
pressure, defined as the force necessary to stop the osmotic flow.
- Hypertonic Solution: If two solutions have unequal osmotic concentrations, is the
solution with the higher concentration (Greek hyper, “more than”).
- Hypotonic Solution: The solution with the lower concentration (Greek hypo, “less
than”).
- Isotonic Solution: When two solutions have the same osmotic concentration (Greek
iso, “equal”).
The terms hyperosmotic, hypoosmotic, and isosmotic are also used to describe these
conditions.
Cellula in soluzione isotonica: se una cellula si trova in una soluzione con una concentrazione
di soluti pari a quella intracellulare. Equilibrio dinamico.
Cellula in soluzione ipertonica: shrinkage.
Cellula in soluzione ipotonica: swelling of the cell.

Aquaporins: Water channels

In some cases water transport may be facilitated by transmembrane water channels.
Osmosis is a slow process but in some conditions water transport needs to be as fast as
possible for example a livello renale. * I meccanismi di osmosi sono molto lenti, dunque c’è
bisogno di accellerare questi processi. Ci aiutano dunque dei canali particolari: Aquaporine.
Water flow in living cells is facilitated by aquaporins, which are specialized channels for
water. In some cases osmosis is not sufficient: so aquaporins are needed. Aquaporins sono
formate da Alfa eliche all’esterno con all’interno gli amminoacidi polari che formano un
canale di passaggio.

Figure 9 Sono composte da alphaeliche, proteine a spirale, che facilitano il passaggio dell’acqua.

With the exception of solutes that cross the membrane by simple diffusion, most solutes can
cross the plasma membrane only through the use of channels or transporters.
Type of transport:

32
 - Simple diffusion: passage of solutes according to a concentration gradient without
the aid of channels or transporters.
- Passive transport: transport of solutes mediated by channels or transporters along a
concentration gradient: ex. Ion channels: Always opened or with regulated opening.
The channels form aqueous transmembrane pores that allow the passive movement
of small water-soluble molecules into or out the cell.
- Active transport: transport of solutes mediated exclusively by transporters against
the concentration gradient.
Ion channels are not simple membrane pores. They have 2 important properties:
Selectivity depends on: channel diameter, channel shape, distribution of the amino acids
that cover its walls.
Ion channels are not always open, but they open following a specific stimulus. Compared to
transporters, channels do not gave to undergo a change in conformation every time an ion
passes.
Active transport
This process requires the expenditure of energy, typically from ATP, and is therefore called
active transport.

ATP-powered pumps: Hydrolyse ATP to drive counter gradient transport.

Coupled Transporters: link the upgradient transport of one solute to the upgradient
transport of another solute.
Photo Powered pumps: they exploit the energy coming. Ex. The sodium–potassium pump.

Figure 10 The sodium–potassium pump is a protein carrier that transports sodium (Na+) and potassium (K+) across the
plasma membrane. For every three Na+ transported out of the cell, two K+ are transported into it. Energy for the process is
provided by ATP hydrolysis. The pump changes conformation between high affinity for Na+ and high affinity for K+ based on
phosphorylation state.

Uniporter: trasportatori uniporto, che assistono un passaggio di un tipo di molecola, in una

sola direzione.

33
Symporter: 2 tipi di molecole, nella stessa direzione. 2 movimenti.
Antiporter: “”, in direzioni opposte. (sodium–potassium pump).
example: P-type pump, sodium/potassium pump —> sodio è più presente all’esterno della
cellula e viene pompato verso l’esterno (quindi contro gradiente; il potassio è più presente
all’interno e viene pompato verso l’interno.
The gradient driving the entry of Na allows sugar molecules
to be transported against their concentration gradient. A
membrane protein transports Na+ into the cell, down its
concentration gradient, at the same time it transports a
glucose molecule into the cell. The gradient driving the Na+
entry allows sugar molecules to be transported against their
concentration gradient. The Na+ gradient is maintained by
the Na+/K+ pump. ADP = adenosine diphosphate; ATP =
adenosine triphosphate; Pi = inorganic phosphate.

34
Hemochrome
It is one of the most popular blood tests, as its results help keep track of a person’s overall
health.

Connective tissue

Blood
Blood, a specialized connective tissue within the
cardiovascular system, is a viscous fluid with a
slightly alkaline pH of 7.4 and a distinctive red
color. It circulates throughout the vascular district,
with slight variations between males and females.
The primary functions of blood encompass a range
of vital roles:
- Respiratory Gas Transport: Facilitating the
movement of oxygen to tissues and carbon
dioxide to the lungs.
- Nutrient Transport and Distribution: Conveying essential nutrients to various parts of
the body.

35
 - Waste and Toxin Transport: Moving waste and toxic substances from peripheral
tissues to excretion sites.
- Hormone Transport: Serving as a carrier for hormones, ensuring their effective
distribution.
- pH and Electrolyte Regulation: Maintaining acid-base balance, buffering against lactic
acid produced by muscles.
- Immune Defense: Acting as a key component in the body's defense against
pathogens, contributing to immunity.
- Coagulation for Vessel Protection: Preventing fluid loss through damaged vessels via
the process of coagulation.
- Thermoregulation: Absorbing and distributing heat to contribute to the body's
temperature regulation.
In essence, blood plays a multifaceted
role in maintaining physiological
balance and supporting various
essential functions within the body.
It consists of: Plasma (Liquid portion)
and Cellular elements: red blood cells,
White blood cells and Platelets
(Corpuscular or figured portion).

Figure 11 Plasma Composition

36
 Figure 12 Cellular elements Composition

Hemopoiesis:
Hemopoiesis is the process of forming blood cells that appear in the circulation during the
third week of embryonic development and colonize organs. Is the process that leads to the
formation of mature blood cells. The sites of hematopoiesis are different in the fetus than in
the adult.

Hemocateresis: is the process that leads to the destruction of blood cells

37
Blood Cells

Formed elements
- Erythrocytes (Red blood cells): 99% of cells and transport oxygen.
- Leukocytes (White blood cells) - Protect against infection, they can be:
a) Granular (Polymorphonucleates), basophils, eosinophils, neutrophils.
b) Agranular Leukocytes - Lymphocytes and Monocytes.

- Platelets: Cell fragments, Coagulation.

Leucocytes
Leukocytes, commonly known as white blood cells, play a pivotal role in the body's defense
against foreign substances and contribute to the maintenance of overall health. They can be
classified into two main categories:
Granulocytes are cells that feature specific granules in the cytoplasm, including:
- Neutrophils: Small phagocytic cells.
- Basophils: Release histamine, intensifying the inflammatory response.
- Eosinophils: Attenuate the inflammatory response.
Agranulocytes (Agranulars) are Lacking cytoplasmic granules, this category includes:
- Lymphocytes: Primarily involved in immunity.
- Monocytes: Transform into macrophages.

White blood cells typically range between 6,000 and 10,000/mm³, and their count can
indicate conditions like leukocytosis or leukocytopenia. Importantly, they don't execute their
functions within the circulation but leverage it as a means of transportation. Upon reaching
their destination, they exit through vessel endothelium and engage in activities within
38
connective tissues. While they maintain a round form in vessels, they assume various shapes
in connective tissues.
Leukocytes serve as key defenders, acting against foreign substances, including
microorganisms, and participating in the removal of dead cells and cellular debris. Their
primary function is executed through diapedesis, which refers to the passage of leukocytes
through vessel walls.
In essence, leukocytes are dynamic cellular defenders that mobilize through the
bloodstream to safeguard the body from external threats, demonstrating their versatility in
immune response and tissue maintenance.

39
Immune system
Immunity: Defence capacity of the body to combat diseases- or to counter infection.
Immune System: Molecules, cells, tissues and organs which provide non- specific and
specific protection against, Microorganisms, Microbial toxins, Tumour.
Major fuctions:
● Protection against disease-causing invaders.
● Removal of dead/damaged tissue and cells.
● Recognition & removal of abnormal cells (tumour, infected cells etc).

The two components of immunity

The key to the function of the immune system is the ability to distinguish self from nonself
cells, and the two branches of immunity do this in very different ways
Schematically, defence against microbes can be mediated by Early Reactions of Natural (or
Innate) Immunity or Late Reactions of the Acquired (Specific/Adaptative) Immunity.
Innate Immunity: The Immediate Shield
Innate immunity acts as the frontline defender with immediate and generalized nonspecific
responses. This early reaction does not show an increased resistance after infection. The key
players in innate immunity include:
● Macrophages
● Granulocytes: Neutrophils, Basophils, Eosinophils
● Natural Killers
● Soluble Mediators: Complement proteins, acute phase proteins, cytokines, and
chemokines.
Adaptive Immunity: Tailored and Specific
Adaptive immunity, in contrast, orchestrates a late and specific response, enhancing
resistance after infection. The principal components involved are:
● Lymphocytes: B lymphocytes for Humoral Immunity and T lymphocytes for
Cell-Mediated Immunity
● Soluble Mediators: Antibodies, cytokines, and chemokines.

40
Natural or Innate or Nonspecific Immunity
Natural immunity is mediated by pre-existing molecules and cells in the body, does not
increase in the presence of the pathogen, and is nonspecific; in practice it acts as the body's
first line of defence but at the same time also serves as a trigger and auxiliary "work force"
for the subsequent specific immune response coordinated by T-helper lymphocytes N.

41
Nonspecific immunity
Nonspecific immunity, constituting innate defenses, encompasses a range of dynamic
mechanisms that act as the body's initial line of defense against potential threats. These
defenses include:
● Physical barriers: the skin, mucous membranes, cilia movement.
● Phagocytic cells: macrophages and eosinophils.
● Host microbial flora: it has antagonistic activity.
● Chemicals: complement, lysozyme, natural antibodies (Ig anti-A....) interferons.
● Cellular products: such as mucus that traps microorganisms.
● Environmental conditions: such as pH.
Important components of innate immunity: They are Factors that limit entry of
microorganisms into the body.
These factors collectively limit the entry of microorganisms into the body. Key components
include:
● Keratin Layer of Intact Skin
● Lysozyme in Tears and Secretions
● Respiratory Cilia
● Low pH in Stomach and Vagina
● Fatty Acids in Skin
● Surface Phagocytes
Mode of Action:
These components act as mechanical barriers, degrade bacterial cell walls, elevate mucus
containing trapped organisms, retard microbial growth, and facilitate the ingestion and
destruction of microbes.
Cells of Innate Immunity:
Engaged in the identification, ingestion, and elimination of microorganisms:
● Neutrophils: Produce lysozyme, collagenase, elastase, and microbicidal substances.
Half-life: 6 hours.
● Monocytes: Transform into macrophages, contributing to the elimination of
pathogens.
● Cells of innate immunity: Identification-ingestion and elimination of microorganisms

42
Identification-ingestion and elimination of microorganisms

Phagocytosis and intracellular killing

43
Acquired Immunity
Acquired immunity, on the other hand, is induced by the pathogen and is activated upon its
entry into the body. It is specific and the organism retains a memory of it, that is, it is
protected over time.
Myeloid Origin
From a common myeloid progenitor originates all cells of natural immunity: granulocytes
(NEUTROPHILS, BASOPHILS, EOSINOPHILS) and MONOCYTES/MACROPHAGES. Natural
immunity cells provide the first immune response.
Lymphoid Origin: From a common lymphoid progenitor, lymphocytes originate instead.

Limphocytes
Are composed the 20-40% of all circulating white blood cells. Appearance: Eccentric, dense
nucleus occupying about 90% of the cell. Cytoplasm sparse, pale blue in colour with a few
bluish granules 8-10 µm. Lymphocytes are a type of small (6-10 µm), roundish leukocytes
with sparse cytoplasm and few granules. Upon activation, the lymphocyte undergoes a
series of morphological changes: the cytoplasm becomes larger, the nucleus moves to the
side, and organelles and the amount of cytoplasmic RNA increase.

Lymphocytes are a morphologically homogeneous population, but functionally they are

divided into two types, T and B lymphocytes, to which are added "non-T non-B"
lymphocytes such as natural killer NK cells.
Functions:
They do not perform activities in the circulation, but in the
connective tissue. They migrate into the lymph nodes and
spleen, where they form clones of identical cells. After
stimulation by antigen, they proliferate and differentiate into
two populations: Memory cells, which do not participate in
the immune response, but remain in the clone and are ready
to respond to that antigen. Effector cells, immunocompetent
lymphocytes that can be classified as B and T lymphocytes.

44
B lymphocytes:
- Are formed and become immunocompetent in the bone marrow.
- Responsible for humoral immune response;
- Can differentiate into plasma cells and produce antibodies.
B lymphocytes differentiate in the bone marrow. They perform two actions: they present
antigen and activated produce antibodies by differentiating into plasma cells.
T lymphocytes:
- They migrate from the bone marrow to the thymus where they mature.
- They constitute the cells of specific immunity, or the body's ability to respond to a
specific pathogenic stimulus.

Natural Killer (NK) Cell:

- Innate Immune lymphocyte.
- 5-20% of blood and spleen lymphocyte population (CD56+CD3-).
- Kill target cells (e.g. leukemic blasts).
- Regulated by specific NK receptors.
- Produce immunoregulatory cytokines (e.g. IFN-γγ that activate macrophages to kill
phagocytosed microbes.
- They regulate immune responses.
Antibodies
Also called immunoglobulins, they are a group of glycoproteins found in blood and tissue
fluids. They are the antigen specific products of B lymphocytes in their terminal
differentiation or PLASMA CELL stage.

45
Ways in which Antibodies limit the infectious process: Neutralization - Opsonization -
Complement activation - ADCC (antibody dependent cell mediated cytotoxicity).

46
 Figure 13 ADCC

Cytokines
Cytokines are a broad and loose category of small proteins important in cell signaling. They
act through cell surface receptors and are especially important in the immune system;
cytokines modulate the balance between humoral and cell-based immune responses, and
they regulate the maturation, growth, and responsiveness of particular cell populations.
Classification:
Interleukins, Chemokines, Members of the Tumor Necrosis Factor family, Interferons, Colony
Stimulating Factors, Growth Factors.

Each
cytokine
binds to a
unique cell
membrane
receptor and triggers specific intracellular pathways.

47
Anatomy of the immune system
In order to optimize the cellular interactions required for the recognition and activation
steps of RI, the various cellular elements are concentrated in anatomically defined tissues or
organs, where Antigens are also transported and concentrated. The organs of the immune
system are divided into primary and secondary:
- Primary ones are the site of origin of immune system cells (Bone marrow and
thymus).
- Secondary ones are lymph nodes, spleen, cutaneous SI and mucosal- associated SI
(Peyer's plaques, tonsils, adenoids and palatine lymphatic tissue).
Lymphatic organs
Central (primary): produce lymphocytes:
- Bone marrow: immunocompetence for B and NK
lymphocytes.
- Thymus: immunocompetence for T lymphocytes.
Peripheral (secondary): activation and proliferation of specific
clones of lymphocytes:
- Lymph nodes.
- Spleen.
The lymphatic organ system is a complex of anatomical
structures in which lymphocytes multiply, differentiate and
from which they migrate to perform specific defensive
functions.

48
Platelets
Cellular remnants resulting from the rupture of
megakaryocytes in the marrow. Cytoplasmic fragments
surrounded by membrane2-4 µm Ø, discoidal shape,
clear peripheral region called Hyalomere, central region
called Granulomere. 250-400,000/mm3
(thrombocytopenia vs thrombocytosis). Main reserve in
the spleen. Present many organelles but lack nucleus.
Essential for coagulation.
Fuctions
They represent a key component in blood coagulation: in fact, they thicken and aggregate
(i.e., weld together) in places where vessel lesions occur, thus triggering the process of
coagulation or hemostasis.
Studi di aggregazione piastrinica
Obiettivo del test è valutare la funzione delle piastrine circolanti.

1) Sampling
2) The sample is centrifuged at a relatively low speed to sediment the larger white and
red blood cells from platelets. Platelets remain in plasma.
3) Free floating platelets .
4) Platelet activators added to test tubes 🡪 functioning platelets clump together and
fall to the bottom - platelets that work poorly, no.
5) Results expressed as a Intensity fluctuations of the light transmitted by the sample
exposed to a light source (measured spectrophotometrically in a platelets
aggregometer).

49
 Figure 14 Esempio di profilo di aggregometria

Method for counting WBC, RBC and PLT

- MANUAL: carried out under a microscope in dedicated counting chambers, on
treated and diluted blood samples manually.
LIMITATIONS: -poor precision, due to the small number of cells counted and manual
sample preparation techniques. -Time -Operator training and experience.
- AUTOMATED: performed by blood cell counters on samples of whole blood treated
and diluted automatically with Impedance Principle.
ADVANTAGES: -High precision and accuracy: high number of cells counted and
automation -Analysis speed (up to 150 blood counts/hour.
Conteggio delle cellule ematiche con conta globuli automatizzato per la conta differenziale
dei leucociti

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Coulter Impedance Principle
Coulter Principle: It is based on the fact that cells are
poor conductors of electricity while the diluent is
excellent conductor.
Operation:
1) Dilution of the blood in solution
buffered isotonic electrolysis.
2) Sample aspiration through diameter
orifices specific.
3) Maintaining constant current between two electrodes (one external and one
internal with respect to the counting chambers).
Coulter method: Each cell that passes through the
electric field opening, replaces a volume of diluent
equal to its size. Each cell causes a change in
current, generating a measurable electrical impulse.
The passage of the cell through the electric field
produces a pulse whose amplitude is directly
proportional to its volume. The number of recorded
pulses allows counting of cellular elements. The
analysis of the pulse amplitude allows the
differentiation of the cells that produced the pulses.

Red Blood Cell (RBC) and Platelet (PLT)

Counting
The counting and volumetric sizing of RBCs and platelets are performed in the same reading
chamber, employing the Impedance Principle. Dimensional analysis of impulses, with
specific volumetric thresholds, allows for the differentiation of RBCs from platelets.
RBC Analysis
Impulses ranging from 36 to 360 fL are classified as RBCs and are distributed across a
dimensional histogram divided into 256 volumetric channels (1.3 fL/channel). From the RBC
histogram, measurements of Mean Corpuscular Volume (MCV) and Red Cell Distribution
Width (RDW) are obtained.
Platelet Analysis
Data from the triple count are collected, and an initial histogram is generated with
thresholds ranging from 2 fL to 20 fL.
- Lower limit of 2 fL: Removes interference from electronic noise.
- Upper limit of 20 fL: Ensures exclusion of microcytes and RBC fragments from the
count.

51
 Figure 15 istogramma PLT-Normale

This approach ensures precise and accurate quantification and sizing of both RBCs and
platelets, enhancing the diagnostic capabilities of the analysis. The application of volumetric
principles and specific thresholds contributes to the reliability and efficiency of the
Impedance Principle in hematological analysis.

Leucocytes Formula Analysis through VCS Technology

It is a simultaneous, multiparametric analysis of native leukocytes in a single detection
channel where the five leukocyte populations are all directly determined. The system takes
three measurements for each of the cells passing through an optical quartz flow cell, with
hydrodynamic focusing.
The measurements carried out are:
1. A low frequency current measures the change in impedance caused by the passage
of cells, determining their total volume.
2. A high-frequency current measures cell conductivity, providing information on the
internal chemical and physical constituents of the cell (size and density of the
nucleus, nucleus/cytoplasm ratio).
3. The scatter of light emitted by a neon-helium laser provides information about the
shape, membrane and granularity of the cell.

Volumetric Analysis: Volume

Structural Analysis: Internal Cellular Complexity 🡪 Conductivity:
The level of cellular complexity directly influences the resistance a cell presents to the
passage of current, determining its opacity. Among the internal structural components, the
nucleus exhibits the lowest resistance, making it an efficient current conductor. Cells with a
high N/C (nucleus-to-cytoplasm) ratio, such as lymphocytes, demonstrate lower opacity
compared to cells with a low N/C ratio, like granulocytes.
Laser Light Scattering: Analysis of Internal Granulation and Cell Surface
The study of laser light scattering provides insights into internal granulation and the surface
of cells.

52
 Figure 16 Cell Population separation system using AccuGate

Morphological Analysis of Red Blood Cells (RBC):

Parameters such as Anisocytosis, Microcytosis, Macrocytosis, Hypochromia, Poikilocytosis,
and the presence of Double RBC Populations, Micro RBC/Fragments, and RBC Agglutination
are assessed.
Analysis of RBC Parameters (MCV, MCH, MCHC, HCT, RDW) and Histogram Evaluation:
Analyzers use these parameters to provide valuable flags for morphological abnormalities.
These flags aid the laboratory in the microscopic evaluation of peripheral blood smears.
In summary, these analytical techniques encompass volumetric assessments, conductivity
studies based on cellular complexity, laser light scattering for granulation and surface
analysis, and comprehensive morphological evaluations of red blood cells, all contributing to
a detailed understanding of cellular characteristics and potential abnormalities.

53
Immunofluorescence
Detection of antigens:
The antibody reacts specifically; natural recombinant antibodies are used as bait, which,
thanks to their binding specificity, recognize the antigen and bind to the cell.

KIND OF CD MARKERS:
Various CD markers categorize immune cells, such as lineage markers (CD3, CD4, CD8),
activation markers (CD69, CD25, CD62L), memory markers (CD45RO, CD27), tissue homing
markers (α4/β7), chemokine receptor markers (CCR7, CCR5, CXCR4, CCR6), and intracellular
markers (FoxP3, cytokines, proliferation markers, antigen-specific markers).

How to uniquely identify a certain type of cell?

The process of indirect immunofluorescence:
The incubation involves a secondary antibody: a fluorochrome linked to the antibody. The
fluorochrome is an excitable molecule that, when stimulated, emits a specific fluorescence.
Therefore, if I detect a fluorescence signal, it means there is positivity for a certain antigen
(marker). Thus, the fluorochrome binds to the marker.
First, the cells to be detected are treated with an unlabelled antibody to form a complex
with a specific antigen, and then fluorescently labelled antibodies of anti-antibodies were
used to detect the presence of specific antigens in the cells, which had fluorescence
enhancement effect.
An antigen is a molecule that provokes a specific immune response. The most effective
antigens are large, complex molecules such as proteins. The greater their “foreignness,” or
put another way, their phylogenetic distance from the host, the greater will be the immune
response they elicit. Antigens may be components of a microorganism or a virus, but they
may also be proteins or glycoproteins on the surface of transfused red blood cells or on
transplanted tissue.

54
Multiparametric Flow Cytometry: Cytoflorimetria a flusso
It is a technique that allows us to interrogate the markers of these cells. It is based on two
phenomena:

Behavior of light on cells

A part of the light is deviated, and another part manages to be transmitted. The two
portions of light: scattered and transmitted. Based on the physical characteristics of the cell
(including internal characteristics), the transmitted light can vary.
Principles of immunofluorescence and flow cytometry analysis:

Input: Exclusive antibody labeling is used. Antibody (reactant A) fluorescein (green

fluorescence) and phycoerythrin (red fluorescence). The fluorescence intensity is captured in
a two-dimensional signal.
Output: Two-dimensional Cartesian diagram, signal overlap may occur: depends on the
position in the quadrants (signals related to a specific antibody), and there may be an
overlay if it is located between the two signals.

The cytometer can study characteristics specific to the examined cells.

55
Components of the cytometer:

1. fluidics: from tube to irrigation point good and bad flow conditions.
2. optics: illuminating cells measuring fluorescence
3. electronics: from photons to electrons electron quantification.

Fluidics:
It serves to bring the cells to the interrogation point, where the cells are intercepted. A
sample is aspirated, diluted in a sheath fluid, the flow is formed, and it is brought to the
interrogation point, where laser beams act.

The problem is: which signal belongs to that particular cell? Solution: sequential alignment
of cells. In a capillary flow, forming a single row of cells one after another. In this way, I sort
the cells and the fluorescence signals.

56
The impact of the flow rate will have repercussions on how specific the analysis is. If the flow
is fast, it does not allow for analysis, and there may be more coincident events.
Flow cell: Hydrodynamic focusing
- High-pressure injection: a high number of cells analyzed.
- Low-pressure injection: a low number of cells analyzed, but a narrow column, for
optimal interaction with the laser.

Optics: Lasers, Filters, Detectors:

Laser light intercepts the flow cell, capturing the emitted fluorescence and directing it
towards the detectors, converting the photons of light into an electrical current. Lasers can
be blue, red, etc. Parametric analysis with up to 50 markers is possible using multiple lasers
with different fluorochromes that are excited depending on the wavelength.
LASER: Light Amplification by Stimulated Emission of Radiation: High Intensity, Polarized,
Coherent.

Types of light:

Laser light scatter (FSC E SSC) e Fluorescence

57
Laser light scatter
FSC (Forward Scatter): is directly proportional to the size of the cell.

SSC (Side Scatter): is a signal at a 90-degree angle, directly

proportional to the complexity of the cell.

Fluorescence
Emission of light by a compound that has absorbed a photon of light.

Each fluorochrome has a different excitation spectrum. The most commonly used source is
the Argon laser, with power ranging from 15 mW to 5W. It is expensive and usable at a few
wavelengths, namely 488 nm (blue), 345 nm, and 514 nm. The Argon laser source has high
intensity, allowing the focusing of radiation on a very small section (cellular dimensions) and
the excitation of numerous fluorophores.

58
A series of filters are used to facilitate the optical path of fluorescence to the detectors.

Detectors convert the luminous signal into a difference in potential, and the photodetector
converts the signal into the graph with the dots.

Spillover effect: The spillover effect, which is the overlap of a fluorochrome into a second
channel due to the physics of fluorescence, is a fundamental process to recognize and
correct for the contamination of fluorescence.
Compensation (Compensazione del campione:): Compensation, in the context of sample
analysis, refers to the mathematical correction of spillover. This involves a mathematical
procedure where the contaminating fluorescence in the emission spectrum of one
fluorochrome is subtracted from an adjacent fluorochrome.

59
It is crucial to automate compensation using software-based methods, ensuring accurate
calculations with proper controls. This automated approach enhances precision and
reliability in the compensation process.

Electronics:
It causes the conversion into digital signal. From photoelectric current to dots on a plot.

Every cell passing through a LASER creates a signal pulse in every detector. Area is the
default measurement on most instrument. Height and Width are useful for i.e., doublets
exclusion. The threshold (or trigger) controls what pulses are considered events in the
system. The signal pulse is converted into a discrete digital value via an Analog Digital
Converter (ADC).

All the values are stored in a FCS file, along with many other information.
Data visualization:
One optical parameter Histogram Plot.

Two optical parameters: Dot plot, density plot.

60
Gating strategies refer to post-acquisition analysis approaches. Initially, physical
characteristics (FSC) are selected, followed by the investigation of CD3 and CD45 expression.
Further subsets (gating) are then defined, progressing from general to specific features of
interest.
The number of colors available determines the depth of subcategories that can be queried.
However, as the number of colors increases, the analyses become more complex, and
overlapping occurrences become more frequent..

61
The Cell Cycle
The key roles of cell division
The organism’s ability to reproduce best distinguishes living things form non-living matter.
The continuity of life is based upon the reproduction of cells, known as Cell Division. The cell
division process is an integral part of the cell cycle.

Stages of the cell cycle:

INTERPHASE:
Interphase, consisting of G1, S, and G2, collectively represents the phase between cell
divisions in the cell cycle.
G1 Phase: Primarily focused on the growth and metabolic activities of the cell, often referred
to as gap phase 1.
S Phase: Dedicated to the synthesis and replication of DNA, commonly known as the
synthesis phase.
G2 Phase: Involves further growth and preparation, sometimes called gap phase 2.
MITOSIS:
Mitosis is the phase of the cell cycle in which the spindle apparatus assembles, binds to the
chromosomes, and moves the sister chromatids apart. Mitosis is the essential step in the
separation of the two daughter genomes. It is traditionally subdivided into five stages:
prophase, prometaphase, metaphase, anaphase, and telophase.
P – Chromosomes are condensed; (prophase)
M – Chromosomes align at cell centre; (metaphase)
A – The duplicated DNA segregates; (anaphase)
T – Chromosomes are decondensed. (telophase)
CYTOKINESIS: Cytokinesis is the process where a cell divides into two daughter cells, each
possessing identical genomes.
RESTING PHASE (G0): During the resting phase, known as G0, cells may exit interphase and
enter a non-dividing quiescent state.

62
The cell cycle consists of Interphase and Mitotic Phase (M Phase). During Interphase the cell
grows, and nuclear DNA is duplicated. During the Mitotic phase, duplicated chromosomes
undergo segregation and are distributed into daughter nuclei. Subsequently, the cytoplasm
undergoes division, resulting in the formation of two daughter cells.
It's important to note that not all cells proceed through the mitotic phase. Some cells have
the capability to exit the cell cycle and enter a quiescent state known as the G0 phase.
The duration of the cell cycle varies depending on cell type, in fact cells often pause in G1
before DNA replication and enter a resting state called the G0 phase; cells may remain in this
phase for days to years before resuming cell division. At any given time, most of the cells in
an animal’s body are in G0 phase. Some, such as muscle and nerve cells, remain there
permanently; others, such as liver cells, can resume G1 phase in response to factors released
during injury.
The centromere is a point of constriction on the chromosome containing repeated DNA
sequences that bind specific proteins. These proteins make up a disklike structure called the
kinetochore. This disk functions as an attachment site for microtubules necessary to
separate the chromosomes during cell division.

Chromosomes vary in size, staining properties, the location of a visible constricted region
called the centromere, and the relative length of the two arms on either side of the
centromere. The particular array of chromosomes an individual organism possesses is called
its karyotype.
Meyosis: When defining the number of different chromosomes in a species, geneticists
count the haploid (n) number of chromosomes.

Interphase
During interphase, the cell undergoes a series of distinct phases:
G1 Phase: The cell undergoes recovery from the previous division, doubling its organelles,
growing in size, and accumulating raw materials essential for DNA synthesis.
S Phase (Checkpoint): DNA replication occurs during this phase, accompanied by the
synthesis of proteins associated with DNA.

63
G2 Phase: Positioned between DNA replication and the onset of mitosis, the cell actively
synthesizes proteins necessary for the upcoming division process.

Mitosis phase
The mitotic phase incluses two essential processes:
Mitosis (karyokinesis): This involves the division of the nucleus, where chromosomes are
distributed to form two daughter nuclei.
Cytokinesis: This is the division of the cytoplasm, resulting in the creation of two genetically
identical daughter cells.

Prophase
Chromatin condenses and forms visible chromosomes. Centrosomes separate, moving to
opposite ends of the nucleus. The centrosomes start to form a framework (mitotic spindle)
made of microtubules that is used to separate the two sister chromatids. Nucleolus
disappears and nuclear envelope disintegrates.

Metaphase
Centrioles are at opposite poles and aligned. Kinetochores of sister chromatids are attached
to microtubules and chromosomes align on an axis called the metaphase plate.

Anaphase
Each centromere splits making two chromatids free, and each chromatid moves toward a
pole. Cell begins to elongate due to microtubules not associated to the kinetochore. Each
pole now has a complete set of chromosomes.

Telophase
Formation of nuclear membrane and nucleolus. Short and thick chromosomes begin to
elongate to form long and thin chromatin (less coiled) and the formation of the cleavage
furrow (a shallow groove in the cell near the old metaphase plate).

Cytokinesis
Division of the cytoplasm.

64
Results of Mitosis:
1. Two daughter cells;
2. Each cell has the same chromosome number as parent cell (2n);
3. Genetically identical to each other and to parent cell.

65
Figure 17. Photos depict mitosis and cytokinesis in a plant, the African blood lily (Haemanthus katharinae), with
chromosomes stained blue and microtubules-stained red.

MEIOSIS
Meiosis causes the formation of gametes (eggs and sperm) and it’s called reduction-division.
Preceded by interphase which includes chromosomes replication. Meiosis in a diploid
organism consists of two rounds of division, called meiosis I and meiosis II, with each round
containing prophase, metaphase, anaphase, and telophase stages. Original cell is diploid (2n)
🡪 Daughter cells are haploid (n).
The gametes (eggs and sperm) each contained two
chromosomes, but all of the non reproductive cells, or
somatic cells, of embryos and mature individuals each
contained four. Each gametes containing half the
complement of chromosomes found in other cells, fuse to
produce a single cell called a zygote. The zygote, like all of
the cells ultimately derived from it, contains two copies of
each chromosome. The fusion of gametes to form a new
cell is called fertilization.
Two haploid (1n) gametes are brought together through
fertilization to form a diploid (2n) zygote.

66
So, gamete formation must involve some mechanism that reduces the number of
chromosomes to half the number found in other cells. If it did not, the chromosome number
would double with each fertilization, and after only a few generations, the number of
chromosomes in each cell would become impossibly large. The number of chromosomes
does not explode in this way because of a special reduction division, meiosis. Meiosis occurs
during gamete formation, producing cells with half the normal number of chromosomes.
(In un organismo haploide, ogni cromosoma è presente in una singola copia anziché in due,
come in un organismo diploide.)

67
MEIOSIS I
Consist of Prophase I, Metaphase I, Anaphase I and Telophase I.
Phrophase I is further subdivided in:
● Leptotene
● Zygotene
● Pachytene
● Diplotene
● Diakinesis
PROPHASE I

A physical exchange of chromosome pieces.

During early prophase I of meiosis, homologous chromosomes find each other and become
closely associated, a process called pairing, or synapsis.
The Crossing over, or chromosomal crossover, means the exchange of genetic material
between Homologous chromosomes. This produces Recombinant Chromosomes, therefore
the chromatids held together by the centromere are no longer identical. The result is the
mixture of parental characteristics in the offspring. It is essential for normal segregation of
chromosomes during meiosis.

METAPHASE I
Homologous pairs of chromosomes align along the equator of the cell.

68
MEIOSIS II
Phropase II
Following a brief interphase, with no S phase, meiosis II begins. During prophase II, a new
spindle apparatus forms in each cell, and the nuclear envelope breaks down.
Methaphase II
In metaphase II, chromosomes align along equator of cell (the metaphase plate in each cell).
Anaphase II
Sister chromatids are pulled to opposite poles of the cells, as in mitosis.
Telophase II
In telophase II, the nuclear membranes re-form around four die rent clusters of
chromosomes. After cytokinesis, four haploid cells result. No two cells are alike due to the
random alignment of homologous pairs at metaphase I and crossing over during prophase I.

69
Random orientation of chromosomes on the metaphase plate: The number of possible
chromosome orientations equals 2 raised to the power of the number of chromosome pairs.
In this hypothetical cell with three chromosome pairs, eight (23) possible orientations exist.
Each orientation produces gametes with different combinations of parental chromosomes.

Results of meiosis:
Four haploid cells with one copy of each chromosome.

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Mitosis VS Meiosis

INTRACELLULAR CONTROL OF THE CELL CYCLE

The cell cycle is controlled by regulator molecules that can:
- Promote the cell cycle process (Positive regulators);
- Stop it from progressing (Negative regulators).

Positive regulators
Were identified proteins that were produced in synchrony with the cell cycle, so called
Cyclins: regulatory subunits of the protein kinases that control cell cycle.
Cyclin-dependent kinase (Cdk) is a catalytic subunit of the protein Kinase (enzyme) that is
only active when is associated with cyclin.

Negative Regulators:
They are tumour suppressor genes. These genes code for signalling protein in a inhibitory
pathway. If a tumour suppressor gene mutates the result can be an active cell division.
Retinoblastoma protein (Rb):
Prevents cells from moving to S phase by binding to transcription factor. When Rb is
phosphorylated, it cannot bind so the cell can move to S phase.

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p53:
Prevents the division of cells with damaged DNA by inhibiting Rb pathway.
Halts cell division, if damaged DNA is detected, using different mechanisms:
1. Stimulates repair enzymes to fix DNA;
2. Forces the cell into G0 resting stage;
3. Keeps the cell in G1 arrest;
4. Causes apoptosis of damaged cells.
In cancer cells, p53 activity is shut down, causing a failure of cell division control.
Cyclin proteins are produced in synchrony with the
cell cycle. These proteins complex with
cyclin-dependent kinases to drive the cell cycle.
Three checkpoints exist in the cell cycle: the G1/S
checkpoint, the G2/M checkpoint, and the spindle
checkpoint.
The cell cycle can be halted at these checkpoints if
the process is not accurate. Anything that damages
DNA, such as environmental factors like the
chemicals in cigarette smoke, or UV radiation in
sunlight, can lead to mutations. Random mutations
also arise with each round of DNA replication. If a
gene is mutated that encodes one of the proteins
involved in controlling cell division, this can lead to
uncontrolled growth.

Sum-up: Mechanism of cycle regulation

The initiation of the cell cycle occurs with the receipt of a signal (e.g., growth factor ligand)
by a cell in G0 or G1. The signal induces the synthesis of G1 and G1/S phase cyclin-CDKs that
activate the transcription of genes encoding DNA synthesis enzymes and S phase cyclin
CDKs.
S phase cyclin-CDKs are initially held in check by inhibitors until G1/S phase cyclin-CDKs
phosphorylates the inhibitors. The polyubiquitination is triggered by SCF ubiquitin ligase and
there is the degradation through the proteasome. The released S phase cyclin-CDs then
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phosphorylate regulatory proteins bound to chromosomal replication origins, promoting
initiation of DNA synthesis. The synthesis of mitotic cyclin-CDKs increases in S and G2
phases. The activities of these complexes initially are blocked by phosphorylation of CDK
subunits, and then are activated later by dephosphorylation.
Once activated, miotic cyclin-CDKs phosphorylate a large number of proteins that control
chromosome condensation, retraction pf the nuclear envelop, formation of the mitotic
spindle, and alignment of chromosome at the metaphase plate. Subsequently, the anaphase
promoting complex (APC/C), another ubiquitin ligase, polyquinanes a protein called securing
which helps hold the sister chromatids of metaphase chromosomes together.
Due to the loss of mitotic cyclin-CDK kinase activity proteins responsible for chromosomal
condensation, etc. are dephosphorylated. Chromosomes then decondense and nuclear
membranes are re-synthesized.
Cell next move forward into telophase where cytokinesis occurs, completing the cell cycle. In
the ensuing G1 phase, replication origin regulators are synthesized, and prereplication
complexes assemble at origins. This prepares cells for another round of DNA synthesis in the
next S phase.
Due to degradation of regulatory proteins at the G1/S, metaphase/anaphase and
anaphase/telophase boundaries, the passage of cells through the cell cycle is irreversible.
The G1/S transition (“START”) is a major checkpoint after which passage through the cycle
becomes independent of mitogens (e.g., growth factors).

In Italiano:
Il ciclo cellulare è il processo attraverso il quale le cellule si preparano, si dividono e si
replicano. Questo processo è controllato in modo rigoroso per garantire che le cellule si
moltiplichino in modo corretto e che eventuali errori vengano corretti. Ecco come funziona:

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1. Inizio del Ciclo Cellulare: Il ciclo cellulare inizia quando una cellula riceve un segnale,
ad esempio da un fattore di crescita, che le dice di iniziare a dividere. Questa fase è
chiamata G1 o fase di Gap 1.
2. Sintesi del DNA: Il segnale induce la sintesi di proteine chiamate cicline-CDK, che
attivano la trascrizione dei geni necessari per la sintesi del DNA. Questo passaggio
avviene nella fase G1 e G1/S.
3. Fase S: Durante la fase S (S sta per "sintesi"), il DNA della cellula viene replicato,
producendo una copia esatta di ciascun cromosoma.
4. Controllo delle Proteine Regolatorie: Le proteine regolatorie impediscono che il ciclo
cellulare prosegua troppo rapidamente. In questa fase, le proteine CDK (Chinasi
dipendente dalla ciclina) della fase S vengono inizialmente inibite da inibitori.
Tuttavia, le proteine CDK di fase G1/S fosforilano gli inibitori, permettendo alle
proteine CDK di fase S di essere attivate.
5. Degradazione delle Proteine: Le proteine inibitrici delle fasi precedenti vengono
quindi marcate per la degradazione attraverso il sistema proteasoma, innescato dalla
ligasi ubiquitina SCF.
6. Fase M e Anafase: Successivamente, il ciclo prosegue nella fase M (M sta per
"mitosi"), dove le proteine CDK mitotiche fosforilano altre proteine che controllano la
divisione cellulare. Questo include la condensazione cromosomica, la formazione del
fuso mitotico e l’allineamento dei cromosomi alla metafase.
7. Securina e Anafase Promoting Complex (APC/C): Durante l’anafase, l’APC/C (un’altra
ligasi ubiquitina) poliubiquitina una proteina chiamata securina. Questo permette
alle cromatidi sorelle di separarsi e migrare verso i poli opposti della cellula.
8. Telofase: Alla fine dell’anafase, le proteine che controllano la condensazione
cromosomica vengono defosforilate, i cromosomi si decondensano e le membrane
nucleari vengono ricostruite.
9. Citosinesi e Fine del Ciclo: La cellula procede quindi nella citodieresi, dove avviene la
divisione cellulare. Alla fine di questo processo, la cellula ha completato il ciclo
cellulare.
10. Checkpoint G1/S: Il checkpoint G1/S è un punto critico nel ciclo cellulare in cui la
cellula decide se deve o meno iniziare un nuovo ciclo. Dopo questo checkpoint, la
cellula diventa indipendente dai segnali esterni e prosegue nel ciclo.

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Gruppi sangugni
Probability Calculation for Association with a Specific Pathology: Understanding
Odds Ratio
The probability of being associated with a particular
pathology is expressed as a ratio, known as the odds
ratio. When equal to 1, the odds ratio is null. If it
surpasses 1, it indicates an unfavorable factor
associated with the condition. A positive association
suggests an increased likelihood, while a ratio below 1
implies a negative association, providing a degree of
protection.

Erythrocytes (Red Blood Cells): Oxygen Transport

and Hemoglobin Structure
Erythrocytes, or red blood cells, play a crucial role in
transporting oxygen to tissues. Their distinctive
biconcave disc shape facilitates the accommodation of
hemoglobin—a protein comprising two alpha and two
beta chains, each binding to two iron molecules (heme
group), imparting the characteristic red color to blood.
Hemoglobin is well-known for its oxygen transport function
(oxyhemoglobin), but it can also bind to other gases, forming
carboxyhemoglobin (CO2) and transporting nitric oxide (NO). Variants
include HbA1 (comprising 96% of total alpha and beta helices), HbA2
(2% of total), and the remaining HbF. They transport oxygen from lungs
to tissues and carbon dioxide from tissues to lungs.
Erythrocytes are vital not only for oxygen transport but also for their plasma membrane
composition, consisting of 50% proteins, 40% lipids, and 10% carbohydrates. Proteins
beneath the plasma membrane provide structural support, crucial for maintaining flexibility
and integrity within blood vessels. The carbohydrate composition at the extracellular surface
determines blood group affiliation and acts as antigens—molecules capable of eliciting an
immune response. This antigenic nature poses
challenges in the context of transfusions.
Understanding the intricacies of erythrocyte
structure and function is paramount, as it not only
influences oxygen transport but also holds
implications for immune responses and blood
compatibility in transfusion scenarios.
Erythrocytes are rich in a tetrameric protein called hemoglobin from p.m. 68 kDa. The
protein consists of four equal chains two by two, (usually two α chains and two β chains).
Each chain is linked to an iron-containing heme group (such that it gives the red color).

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Co-dominant System: Blood Type Antibodies and ABO Groups
In a co-dominant system, individuals produce antibodies against what is different—leading
to agglutination when encountering diverse erythrocytes. The ABO blood groups,
determined by genetic factors, involve protein-sugar complexes acting as antigens on red
blood cells. The immune system, tolerant to its own antigens, produces antibodies against
differing antigens, causing agglutination and lysis of foreign red blood cells. IgM antibodies,
initially responding to bacterial carbohydrates in our normal flora, also recognize
monosaccharide differences on red blood cells.
Genotype and Phenotype: Decoding Blood Types
The genotype represents the genetic constitution, only partially expressed in the living body.
The phenotype encompasses all manifested characteristics in an organism. The assignment
of blood groups is driven by the genotype, revealing the genetic makeup that is only partially
expressed in the body.

Genes and Mendelian Principles: Unraveling Blood Type Acquisition

Genes, hereditary units controlling traits, follow Mendelian principles in their transmission.
Each gene has different alleles, and during meiosis, alleles segregate independently,
determining the formation of gametes. The acquisition of an individual's blood type is a
result of the complex interplay of these genetic factors.
Antigens vs. Immunogenicity: Recognizing and Responding
An antigen is any protein substance recognized by the immune system, capable of evoking
an immune response for elimination. It can originate externally (bacteria, viruses) or
internally, developing within the body. Understanding the distinction between antigens and
substances that evoke an immune response is essential in comprehending immune
reactions.

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Mendel's Principles and Heredity
Mendel's principles of heredity elucidate the transmission of traits. Genes exist in pairs, with
one copy from each parent. Each gene may have different alleles, and during meiosis, alleles
segregate independently. This independence in the segregation of alleles is crucial in
understanding the inheritance of blood types and the manifestation of traits.

Terminology:
● Locus: the position of a gene on a chromosome eg. sex determining gene, SRY SRY is
located on chromosome Y at position p11;
● Alleles: alternative forms of a gene eg. in the beta globin gene, the codon (triplet of
nucleotides) encoding for the sixth aminoacid:
Normal allele A .……………...GAG..... (glutamic acid);
Sickle allele S .……................GTG..... (valine) mutation;
Cristallin allele C .…………….AAG..... (lysine) mutation.
Polimorfismo: mutazioni che non si associano a una patologia ma ci caratterizzano come
individui diversi dagli altri.
● Homosygote: carries 2 identical alleles at the same locus (bAbA🡪 suplice copia uguale
in questo caso dell’emoglobina OR bSbS).Individui che trasporta per quell dertermiato
locus alleli identici.
● Heterozygote: carries 2 different alleles at the same locus (bAbS);
● Compound heterozygote: carries 2 mutant alleles (2 different mutations) at the same
locus (bSbC);
● Carrier (healthy): asintomatic heterozygote; (portatore, 1 mutatnt allele, gene
recessivo);
● Proband (Propositus): the affected individual through which a family is directed to
analysis;
● Genotype: genetic constitution of an individual, either as a whole or referred to a
specifc gene, eg. Sickle cell anemia, bSbS.
● Phenotype: observable feature of an individual, or a cell.
Mendelian inheritance of tracts
Dominant: each trait or character that is expressed in the heterozygote, that is, in case of
illness, where only one copy of the defective gene is sufficient to express the affected
phenotype; (se il 50 percento basta per manifestare la malattia);
Recessive: each trait or character that is expressed in the homozygote, that is, in case of
illness, where both copies of the defective gene are to be present in order to express the
affected phenotype; (se il 100 percento basta per manifestare la malattia);
Codominant: : in cases where heterozygous state expresses a distinct phenotype respect to
the two homozygous states, eg. blood groups, blood cell enzymes etc

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Codominance
ABO blood groups:
3 alleles at one genes locus: i, IA e IB
Each individual possesses 2 copies These alleles determine the expression of an antigen,
recognised by the immune system. Nei gruppi sanguigni vale la codominanza: il caso di AB.

Codominanza e allelia multipla

Genotypes, Phenotypes, and Compatibility in Blood Groups
Inherited along with blood types are specific antibodies, known as agglutinins, which
significantly influence blood transfusions by causing the agglutination of red blood cells. The
ABO blood group system involves three alleles: IA and IB, which are codominant and
dominant over i, which is recessive. This results in six genotypes: three homozygous and
three heterozygous combinations. The expression of these genotypes gives rise to four
distinct phenotypes or blood groups: A, B, AB, and O.
Understanding the genetic basis of blood types, including the interaction of codominant and
recessive alleles, is crucial in comprehending the compatibility of blood transfusions and the
potential for agglutination reactions. The four blood groups, each associated with specific
antigens and antibodies, play a key role in determining the compatibility of donor and
recipient blood during transfusions.
Punnet square:
The Punnett square, devised by British geneticist Reginald Punnett, serves as a valuable tool
in biology to predict the likelihood of different phenotypes resulting from the crossing of
various genotypes. In this diagram:
● - The probability of transmitting an allele is 50%.
● - The probability of each possible combination is 25%.
These 2x2 matrices aid in calculating and determining the potential phenotypes resulting
from the cross of different genotypes. By placing the parental genotypic aspects on the rows
and columns, it becomes possible to deduce all potential phenotypes.

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Hereditary Transmission of the Blood Group
The AB0 blood group system follows Mendelian principles of heredity:
● - Each individual's genome contains two alleles.
● - There's a 50% chance for each allele to be transmitted to offspring.
But what if we only know the phenotype of the blood type without knowledge of the
genotype?
Understanding blood group phenotypes alone may not
provide a complete picture of the underlying genotypes.
However, Punnett squares and genetic analyses allow for
inference and determination of potential genotypes,
shedding light on the hereditary transmission of blood
groups even when genotypic information is not directly
available.

Example:

Medical Oncology
Cancer-Tumors-Neoplasms:definition
They constitute numerous pathological
conditions characterized by the
appearance in the context of a tissue or
organ of a mass of cells that has gradually
undergone a progressive series of damage
in its genome. Disease of "genes" causing:
Loss or reduction of properties typical of
the cell of origin or acquisition of new
properties absent in the latter.
Tumor cells are characterized by
continuous and unrestricted proliferation and survival, extending indefinitely in both time

79
and space. Their ability to spread to distant locations, known as metastasis, raises questions
about the factors contributing to the multiplicative autonomy of
these cells.
The uncontrolled growth of tumor cells hinges on the disruption of
the delicate balance observed in normal tissues, where the rates of
cell growth and cell death are harmonized. This imbalance may stem
from unregulated cell growth or the inhibition of apoptosis, the
mechanism responsible for the programmed cell self-destruction.
Mitogenic stimuli, such as growth factors and adhesion to the extracellular matrix, play a
role in promoting cell growth. Conversely, anti-mitogenic stimuli, including cell-to-cell
contact, absence of growth factors, lack of adhesion to the extracellular matrix, and
apoptosis, act as checks on cell proliferation. Tumor cells exhibit a remarkable ability to
proliferate independently of external stimuli and demonstrate increased survival by evading
anti-mitogenic signals.

The development of neoplasms is influenced by various factors, with cancer often

considered a disease associated with aging. This association is rationalized by the
accumulation of sequential mutations in cellular DNA and a reduced efficiency in the control
systems that maintain cellular homeostasis.

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Carcinogenesis is a multistep process:
● Initiation: during which, one or more mutations transform a normal cell into a latent
neoplastic cell.
● Promotion: as a result of 'accumulation of further mutations and/or az. of
proliferative stimuli, there is the appearance of a clinically evident tumor consisting
of the progeny of the transformed cell that has begun to multiply.
● Progression: further appearance of mutations responsible for the metastatic
phenotype and reproductive overgrowth.
Molecular mechanisms of carcinogenesis: Oncogenes, oncosuppressors, and DNA repair
genes.
→ Oncogenes: mutated versions of genes that normally and positively preside over the
mechanisms of cell proliferation, survival, and differentiation (proto-oncogenes) that as a
result of the activating mutations affecting them, code for proteins abnormal in number
and/or function that promote an uncontrolled increase in cell number with a
growth-accelerating function. Not all genes can mutate into oncogenes but only
"proto-oncogenes" can form an oncogene.

What functions do proto-oncogenes normally

perform? Heterogeneous class of genes that
code for STIMULATING signals of the cell cycle,
include: Growth factors, Membrane receptors
for growth factors, Proteins involved in signal
transduction, Transcription factors capable of
binding to DNA, Cyclins, cyclin-dependent
kinases and their inhibitors and activators (= cell
cycle regulators).
Transforming Proto-oncogenes into Oncogenes:
Mutation Mechanisms
The conversion of proto-oncogenes into oncogenes involves mutations categorized as
'acquisition of function' or gain-of-function mutations, typically exhibiting dominant
traits even in heterozygosity. Various types of mutations contribute to this
transformation:

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 1. Point Mutations:
- Alter the properties of the encoded protein.
2. Gene Amplification:
- Involves the replication of genes, resulting in an increased number of copies,
sometimes in the hundreds.
3. Chromosomal Translocations:
- Movement of chromosomal segments that relocate the proto-oncogene into
transcriptionally active regions.
4. Chromosomal Translocations Creating Chimeric Genes:
- Fusion of two different genes through chromosomal translocations, forming a hybrid
or chimeric gene.
Point mutations in coding sequences, such as those seen in the RET gene in multiple MEN2
neoplastic endocrine syndromes, represent one category of mutations turning
proto-oncogenes into oncogenes. Additionally, mutations in regulatory sequences and gene
amplification phenomena, exemplified by the c-myc proto-oncogene in Burkitt's lymphoma,
also contribute to this transformation.

→ Oncosuppressors or antioncogenes: genes that encode for proteins with a restraining

function on cell growth mechanisms and that as a result of the inactivating mutations they
have undergone, encode for products that are inefficient
at carrying out these arresting functions.
In the intricate landscape of cancer development, a lone
mutated oncogene is typically insufficient to induce
cancer. This is attributed to the presence of tumor
suppressor genes, a vital class that effectively puts the
brakes on unregulated growth.
Tumor suppressor genes, also known as oncosuppressors
or antioncogenes, encode proteins crucial for restraining
cell growth mechanisms. In the unfortunate event of
inactivating mutations, these genes produce products that fail to perform their arresting
functions effectively.

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Diverse Functions of Oncosuppressors
The functions of oncosuppressors are remarkably varied, including the production of factors
inhibiting the cell cycle, pro-apoptotic factors, and factors involved in maintaining genome
stability. Examples encompass genes directly regulating the cell cycle (e.g., RB1 and TP53),
those involved in cell-cell contact growth inhibition (e.g., NF2), and genes crucial for DNA
damage repair and genomic integrity (e.g., BRCA1, BRCA2, MLH1, MLH2).
Among the major players in tumor suppression is TP53. TP53 produces the p53 protein that
performs numerous functions. p53 has been termed the 'guardian of the genome,' is
strongly involved in the G1-S checkpoint, blocks cells with damaged DNA by allowing repair
of the damage or by inducing the cell to go into apoptosis.

TP53 mutations are implicated as one of the most frequent genetic changes in tumor
transformation. Germline mutations in TP53 lead to Li-Fraumeni syndrome, an autosomal
dominant disorder associated with a predisposition to diverse cancers.

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Li-Fraumeni syndrome, arising from germline mutations in TP53, presents a
rare autosomal dominant disorder. Afflicting young individuals, it predisposes
them to a spectrum of cancers, including breast, lymphoma, osteosarcoma,
brain tumors, and sarcomas. The prevalence of Li-Fraumeni syndrome is
estimated at 1-9 cases per 100,000 individuals.

→ DNA Repair System genes: genes that code for proteins whose normal function is to
correct errors to duplicating DNA prior to cell division, and which as a result of the
inactivating mutations undergone, are responsible for the abnormal accumulation of
mutations and their transfer to cell progeny.

Benign and Malignant Tumors

Although they differ equally from normal cells in their replicative autonomy, tumors are, on
the basis of morphological features, proliferative activity, and especially biological behavior
toward neighboring tissues and the whole organism, divided into two major groups: that of
benign tumors and that of malignant tumors.

Benign Tumors:
- Not categorized as true cancers.
- Can be surgically removed.
- Lack recurrence.
- Do not metastasize to distant sites.
- Rarely pose a life-threatening risk.

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Benign tumors exhibit an expansive growth pattern and are typically encapsulated by fibrous
connective tissue, creating a clear demarcation from surrounding tissues (e.g., glandular
epithelium adenomas). While they compress neighboring tissues during expansion, benign
tumors do not invade the basement membrane and, when excised, seldom recur. Their
growth remains localized, confined to the site of origin, without infiltrative-type expansion
or metastatic spread.
Malignant Tumors:
- Genuine cancers.
- Characterized by cellular abnormalities:
- Uncontrolled cell division.
- Local invasion.
- Invasion of blood vessels and lymphatic system.
- Capable of metastasis.
Malignant tumors infiltrate and replace
neighboring tissues, breaching the basal
membrane, invading the dermis, and
reaching vascular and lymphatic beds.
Malignant cells suspended in blood or
lymph form neoplastic emboli, circulating
to distant sites and potentially leading to
metastatic formations. Malignant tumors
induce cachexia, a severe metabolic
imbalance resulting in substantial weight
loss, asthenia, and apathy.
Cachexia Associated with Malignant
Tumors:
Malignant tumors cause cachexia, a progressive and rapid decay of the
body that goes into massive weight loss, asthenia, and apathy. It is a
metabolic imbalance that results in depletion of fat and muscle tissue
reserves. Anorexia, chronic inflammation, insulin resistance, and
increased protein catabolism of muscle tissue are conditions
frequently associated with, and believed to be equally responsible for,
the phenotypic picture of cachexia. The overall metabolic picture is
thus characterized by:
- increased glucose, protein and vitamin requirements: inability to introduce energy
substrates with food as a result of anorexia, nausea and vomiting;
- reliance on gluconeogenesis with depletion of protein and lipid reserves and
consequent weight loss;
- difficult utilization of newly formed glucose due to hypoinsulinemia and/or
peripheral insulin resistance;

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 - oxidative damage induced by O2 free radicals on DNA, membrane lipoproteins, and
enzymes and coenzymes central in regulating major cellular metabolic pathways.
Naming Neoplastic Entities: Criteria for Nomenclature and Classification of tumors
1. Histogenetic Criterion:
Takes into consideration the recognition of
the particular tissue from which the tumor
originated, that is, its histogenesis and, more
specifically, the progenitor cell type of the
neoplastic cell population.
2. Prognostic Criterion:
Focuses on predicting the biological behavior,
distinguishing between benign and malignant
tumors.

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