CELLULAR MEMBRANE

Membrane
About Cell Membranes
1. All cells have a cell membrane
2.Functions:
a. Controls what enters and exits the cell to maintain an internal balance called homeostasis
b. Provides protection and support for the cell
3. Structure of cell membrane
Lipid Bilayer -2 layers of phospholipids
a. Phosphate head is polar (water loving-hydrophillic)
b. Fatty acid tails non-polar (water fearing- hydrophobic)
c. Proteins embedded in membrane

4. Cell membranes have pores (holes) in it

a. Selectively permeable: Allows some molecules in and keeps other molecules out
b. The structure helps it be selective!

CHEMICAL COMPOSITION
1. Lipids -four major classes of lipids are commonly present in the plasma membrane: phospholipids, spingolipids, glycolipids and sterols.
2. Proteins-plasma membrane contains about 50% protein. Plasma membranes contains structural proteins, transport proteins and enzymes.
Some of them acts as receptors.
3. Carbohydrates – Attached to proteins or lipids on the extracellular side of the membrane (forming glycoproteins and glycolipids)

Cell Transport
• Cells need to:
– Take in things they need
– Get rid of the things they don’t need
– Communicate with one another
• Two main types
– Passive Transport
– Active Transport

Passive Transport
• The movement of materials across the cell membrane without using energy
• Caused by concentration gradient
• Works in both directions
• Three types:
– Diffusion
– Facilitated diffusion
– Osmosis

Diffusion
• The process by which molecules spread from areas of high concentration to areas of low concentration (Moving across the concentration
gradient)
• No energy is required
• When the molecules are even throughout a space - it is called EQUILIBRIUM

Diffusion across a membrane

• Permeable – membrane through which molecule scan pass
• Move from high concentrations to low until equilibrium is reached
• Small, uncharged particles (oxygen, carbon dioxide, most lipids, water)
• No energy required

Facilitated Diffusion
• Molecules that cannot directly diffuse across the mem brane pass through special protein channels
• NO ADDITIONAL ENERGY NEEDED
• Ions & large molecules (Cl- and glucose)
• Protein channels (“carriers”)
• Specific for each ion/molecule

Osmosis
• Osmosis is the diffusion of water through a selectively permeable membrane down its concentration gradient
• From an area of high-water concentration to an area of lower water concentration

Osmotic Solutions
• Isotonic- concentrations of solute and solvent are equal
• Hypertonic- higher concentration of solutes
• Hypotonic- lower concentration ofsolutes

Osmotic Pressure
The pressure required to prevent the passage of water through a semi-permeable membrane from a region of low concentration of solutes to
one of higher concentration, by osmosis. Cell walls create this pressure this prevents plant cells from breaking.

Osmolarity vs. Osmolality

● The term osmolarity refers to the number of particles of solute per liter of solution; milliosmoles osmotic pull per kilogram of water
(mOsm/kg)
 ● osmolality refers to the number of particles of solute per kilogram of solvent. Osmolarity is the osmotic pull per liter of solution
(mOsm/L).

III. Osmosis & osmotic pressure

✓ When two solutions of different concentrations are separated by a semi permeable membrane (impermeable to solute and permeable to
water) water mols. diffuse from solution having less conc. To the sol. having higher concentration.

Osmotic pressure (again)

Active Transport
• Movement against a concentration gradient
• Requires Energy (ATP)
• Allows for stockpiling
• One direction
• Types:
– Primary active Transport
– Secondary Active Transport

Primary Active Transport

• They use the energy directly from the hydrolysis of ATP
– Sodium Potassium Pump
– Calcium Pump
– Hydrogen Potassium Pump
– Hydrogen/Proton Pump

Sodium Potassium Pump

•Present in all eukaryotic cells
•Functions
1. Maintains sodium potassium concentrarion difference across the cell membrane
2. Maintains the volume of the cell
3. Causes negative electrical charge inside the cell- electrogenic pump
4. Essential for oxygen utilization by the kidneys

Calcium Pump
● found in the membrane of the sarcoplasmic reticulum.
● Calcium pumps play a crucial role in proper cell signalling by keeping the intracellular calcium concentration roughly 10,000 times
lower than the extracellular concentration.[1] Failure to do so is one cause of muscle cramps.
● The release of a large amount of free Ca2+ can
-trigger a fertilized egg to develop,
-skeletal muscle cells to contract

Hydrogen Potassium Pump

• Hydrogen Potassium Adenosine Triphosphatase (H+-K+ATPase)
○ are gastric proton pumps that function to maintain an acidic environment within the stomach.
○ enzyme primarily responsible for the acidification of the stomach contents and the activation of the digestive enzyme pepsin

Proton Pump
• Hydrogen Adenotriphosphatase (H+ATPase)
○ Present in lysosme and endoplasmic Reticulum
○ Pumps proton from cytosol into these organelles
• The most important active transporter in the plasma membrane
○ It hydrolyzes ATO and uses the released energy to pump hydrogen ions (H+) out of the cell
○ Creates a membrane potential or voltage because the proton pump moves posiive charge (H+) outside the cells, making the inside
of the cell negative in charge relative to the outside

Secondary Active Transport

• Energy is utilized in the transport of one substance helps in the movement of other substance
• Co-transport/Symport
• Counter transport/Aniport

Co-Transport/Symport
• The transport of Na+ via its concentration gradient is coupled to the transport of other substances in t he same direction
• Make use of carrier protein
• E.g. Sodium glucose Co-transport

Counter Transport/ Antiport

• The transport of Na+ via its concentration gradient is coupled to the transport of other substances in the opposite direction
• Sodium-Hydrogen counter transport in the proximal tubule of the kidneys
• Sodium-Calcium exchanger in the cardiac cells

Secondary Active Transport

Peculiarities of active transport
1) Carrier mediated transport
2) Rapid rate of transport
3) Transport takes place against electrochemical gradient
4) Expenditure of energy by transport protein which incorporates ATPase activity
5) Carrier protein shows specificity, saturation competitive inhibition, blocking
6) Substances transported – Na+, K+, H+, Cl -, I -, Glucose, Amino acids

Vesicular Transport
• is the predominant mechanism for exchange of proteins and lipids between membrane-bound organelles in eukaryotic cells.
○ Endocytosis
○ Pinocytosis
○ Phagocytosis
 ○ Exocytosis

Endocytosis
● The process by which cells absorb material (molecules such as proteins) from outside the cell by engulfing it with their cell
membrane
● Two Types
○ Pinocytosis
○ Phagocytosis

Extracellular material to be tackled by lysosomes is brought into the cell by endocytosis.

3 types
Receptor mediated endocytosis- Requires ATPase, Ca, microfilaments
Phagocytosis – specialized cells
Pinocytosis - All cells

TYPES OF ENDOCYTOSIS
Phagocytosis Pinocytosis
• Means “cell eating” • Means” cell drinking”
• An extension of cytoplasm surrounds a particle• Tiny pockets form along the cell membrane fill
and package it into a food vacuole with liquid and form vacuoles within the cell

• requires an enormous amount of energy • Requires enormous amounts of energy

• movement of solids into the cell • Movement of liquids into the cell

B. Receptor mediated endocytosis – highly selective process to import imp. Specific large molecules. Requires energy & Ca++.

EXOCYTOSIS
• The process where cells release large amount of materials
• Vacuole surrounding material fuses with
the cell membrane, forcing the contents out of the cell
• Also requires energy
A cellular process where cells eject waste products or chemical transmitters (such as hormones) from the interior of the cell.
• Exocytosis is similar in function to Endocytosis but works in the opposite direction.

Passive transport Active transport

• No expenditure of energy molecules • Expenditure of energy mol. ( ATP )

• Takes place along conc., electrical, & • Can take place against conc. Gradient
pressure gradient
• Carrier may or may not be required • Carrier is always required

• Rate is proportional to conc. difference • Rate is proportional to availability of carrier

& energy. (Vmax)

Simple Diffusion Facilitated Diffusion

• Passive transport • Passive transport
• For small molecules • For large molecules
• No carrier required • Carrier mediated
• Rate of transport is directly proportional to • Initially rate is proportional to conc. gradient
onc. Gradient. ill Vmax (saturation of carriers)
• Examples: Examples: glucose, amino acids
Lipid soluble – O2, CO2, alcohol
Lipid insoluble –urea, Na+, K+

Osmosis in the Body

• Gargling With Saltwater Provide Relief from Sore Throat
• Digested Food Absorbed in Small and Large Intestines
• Pathogenic Bacteria Interfere with Intestinal Cells

BIOCHEMISTRY OF WATER AND ELECTROLYTES

WATER MOLECULE
- H20 is made up of three atoms (one oxygen and two hydrogen)
- A hydrogen bond between water molecules forms side with oxygen has a negative charge; the hydrogen side-positive one

THE WATER MOLECULE

🞆 There is a greater concentration of electrons around the nucleus of the oxygen than around the hydrogen.
🞆 Therefore, the hydrogen end is slightly positive and the oxygen end is slightly negative (Dipolar)

WATER
🞆most abundant substance
🞆vital molecule found in living organism
🞆colorless, odorless, and tasteless
🞆possesses unique chemical properties that’s suit biological systems
🞆water is a polar covalent molecule.
🞆molecule of water has two hydrogen atoms covalently bonded to a single oxygen atom.
🞆 Functions
●
● acts as heat-exchange agent
● carrier in the bloodstream, distributing oxygen, minerals, vitamins, and hormones.
● biochemical reactions take place in an aqueous environment.
● excellent solvent for polar substances-DNA, proteins, and carbohydrates
● Regulates rate at which air temperature changes.

PHYSICAL PROPERTIES OF WATER

STATES OF WATER
🞆Water is the only substance found on Earth in all three states (phases):
1. Liquid
2. Solid (Ice)
3. Gas (Steam or Vapor)

Evaporation - When water molecules move faster, they tend to break their hydrogen bonds.
Condensation - When gas or vapor molecules slow down, they clump or join together

As water becomes cooler, it becomes less dense:

1 gram per cubic cm at 39.2o
0.95865 gram per cubic cm at 212o

UNIQUE PHYSICAL PROPERTIES OF WATER

1. Water has a high specific heat capacity.
2. Water in a pure state has a neutral pH.
3. Water conducts heat more easily than any liquid except mercury.
4. Water molecules exist in liquid form over an important range of temperature from 0 - 100° Celsius.
5. Water is a universal solvent. 🞆6. Water has a high surface tension

WATER AS A SOLVENT
Since water can dissolve more things than any other natural substance, it is known as the “ Universal Solvent”
Properties of Solvents include:
🞆 Interacts with other polar compounds
🞆 Is repelled by non-polar compounds
🞆 Small size allows it to saturate areas
🞆 Can convey other substances in solutions
Water is especially good at dissolving salts
- Salts form from the combination of particles with opposite electrical charges (or ions)
- EX. Na++ Cl-= NaCl

SURFACE TENSION
- Next to mercury, water has the highest surface tension of all liquids.
- This is a result of the tendency of water molecules to attract to one another, or cohere, at the surface of any
accumulation of water
- Surface tension allows insects to walk on water.
- Interaction between hydrogen bonding and the earth’s gravitational pull
🞆 Cohesion- the attraction between water molecules to each other through Hydrogen bonds (H)
🞆 Adhesion- the attraction of water molecules to another substance.

CHEMICAL PROPERTIES OF WATER

THE WATER MOLECULE
🞆Water is formed when two hydrogen atoms bond to one oxygen atom.
🞆 Not symmetrical
🞆 Electrons spend more time near the (O) and less time near the (H)
🞆Water molecule is polar
Water molecules are held together by a hydrogen bond

The Water Molecule

- Covalent bonds between H+ and 02-
- Polarity shows hydrogen Bond
- Hydrogen bonds constantly forms and break, giving water its strcture
COVALENT BOND
- mutual sharing of one or more pairs of electrons between two atoms.
HYDROGEN BOND
- interaction involving a hydrogen atom located between a pair of other atoms having a high affinity for electrons
POLARITY
- A molecule in which opposite ends have opposite electrical charges is called a POLAR molecule
- Water is a polar molecule

THE WATER MOLECULE

When water molecules attract to one another, the hydrogen end of one molecule is attracted to the oxygen end of another.
Chemical Reactivity:
- water can participate in chemical reactions: Example involvement of water molecules in dehydration synthesis and
hydrolysis

When minerals dissolve in water they separate into positive and negative ions.
Charged low-molecular molecules present in the plasma and the cytosol : Potassium (K), Chloride (Cl), Sodium (Na), Manganese (Mn),
Magnesium (Mg), Calcium (Mg)

Electrolytes are minerals in the blood, tissues, and elsewhere throughout the body. Each electrolyte plays a
specific role in the body

The body’s nerve reactions and muscle function are dependent upon the proper exchange of these electrolyte ions outside and
inside cells.
They control important physiologic functions such as calcium, chloride, magnesium, phosphate, potassium and sodium.
The body’s nerve reactions and muscle function are dependent upon the proper exchange of these electrolyte ions outside and
inside cells.
They control important physiologic functions such as calcium, chloride, magnesium, phosphate, potassium and sodium.
These substances are present in the blood, bodily fluids, and urine. They’re also ingested with food, drinks, and supplements.
An electrolyte disorder occurs when the levels of electrolytes in the body are either too high or too low.
Electrolytes need to be maintained in an even balance for body to function properly. Otherwise, vital body systems can be
affected.

ELECTROLYTES
Positively charged-ion - cation (sodium, potassium, calcium, and magnesium)
Negatively charged ion - anion (chloride, phosphates, and bicarbonate.)
Ions are invisible charged particles in the air – either molecules or atoms, which bear an electric charge.

BENEFITS OF NEGATIVE IONS

🞆 They neutralize free radicals.
🞆 They revitalize cell metabolism.
🞆 They enhance immune function.
🞆 They purify the blood.
🞆 They balance the autonomic nervous system, promoting deep sleep and healthy digestion.
🞆 clear the air of airborne allergens such as pollen, mold spores, bacteria and viruses.
🞆 also clear the air of dust, pet dander and cigarette smoke.

SODIUM (NA)
🞆It Helps control fluids in the body, impacting blood pressure.
🞆This is necessary for muscle and nerve function.
🞆Regulate osmotic pressure and the body’s water content, transmit nerve signals, contract muscles, etc.

CHLORIDE (CL)
🞆 Regulate osmotic pressure and the body’s water content, enable the secretion of stomach acid, etc.
🞆 It helps balance electrolytes
🞆 As it balances acidity and alkalinity, which helps maintain a healthy pH.
🞆 This is essential for digestion.

POTASSIUM
🞆It regulates the heart and blood pressure 🞆As it helps balance electrolytes
🞆Then, aids in transmitting nerve impulses 🞆Moreover, contributes to bone health 🞆This is necessary for muscle contraction

MAGNESIUM
🞆 This is important for the production of DNA and RNA
🞆 As it contributes to nerve and muscle function
🞆Moreover, it helps maintain heart rhythm 🞆 Therefore, it regulate blood glucose levels 🞆 It enhances the immune system

CALCIUM
🞆 It is an essential component of bones and teeth 🞆 This is important for the movement of nerve impulses and muscle
movement
🞆 As it contributes in blood clotting

PHOSPHATE & BICARBONATE

🞆 Phosphate
This strengthens bones and teeth
It helps cells produce the energy needed for tissue growth and repair
🞆 Bicarbonate
It helps the body maintain a healthy pH
It regulates heart function

Type IV Solution Uses Nursing Considerations

Type IV solution Uses Nursing considerations

Isotonic 0.9% Normal Saline (0.9% NaCl) Fluid resuscitation for hemorrhaging, Monitor closely for hypervolemia,
severe vomiting, diarrhea, GI suctioning especially with heart failure or renal
losses, wound drainage, mild failure.
hyponatremia, or blood transfusions.
 Lactated Ringer’s Solution (LR) Fluid resuscitation, GI tract fluid losses, Should not be used if serum pH is greater
burns, traumas, or metabolic acidosis. than 7.5 because it will worsen alkalosis.
Often used during surgery. May elevate potassium levels if used with
renal failure.

5% Dextrose in Water
(D5W) *starts as isotonic and then
changes to hypotonic when dextrose is
metabolized
Provides free water to help renal
excretion of
solutes, hypernatremia, and some
dextrose supplementation.
Should not be used for fluid resuscitation
because after dextrose is metabolized, it
becomes hypotonic and leaves the
intravascular space, causing brain
swelling. Used to dilute plasma
electrolyte concentrations.

Hypotonic 0.45% Sodium Chloride (0.45% NaCl) Used to treat intracellular dehydration
and hypernatremia and to provide fluid
for renal excretion of solutes.
Monitor closely for hypovolemia,
hypotension, or confusion due to fluid
shifting into the intracellular space,
which can be life-threatening. Avoid use
in patients with liver disease, trauma,
and burns to prevent hypovolemia from
worsening. Monitor closely for cerebral
edema.

5% Dextrose in Water (D5W)
Provides free water to promote renal
excretion of solutes and treat
hypernatremia, as well as some
dextrose supplementation.
Monitor closely for hypovolemia,
hypotension, or confusion due to fluid
shifting out of the intravascular space,
which can be life-threatening. Avoid use
in patients with liver disease, trauma,
and burns to prevent hypovolemia from
worsening. Monitor closely for cerebral
edema.

5% Dextrose and Lactated Ringer’s Used to treat severe hyponatremia and

(D5LR)D10
Hypertonic 3% Sodium Chloride (3% NaCl)
5% Dextrose and 0.45% Sodium
Chloride (D50.45% NaCl)
cerebral edema. Monitor closely for hypervolemia,
hypernatremia, and associated
respiratory distress. Do not use it with
patients experiencing heart failure, renal
failure, or conditions caused by cellular
dehydration because it will worsen these
conditions

BUFFER SYSTEM
Introduction to Buffers
• A buffer is a solution that can resist pH change upon the addition of an acidic or basic components. It is able to neutralize small amounts of
added acid or base, thus maintaining the pH of the solution relatively stable.
• This is important for processes and/or reactions which require specific and stable pH ranges.
• Buffer solutions have a working pH range and capacity which dictate how much acid/base can be neutralized before pH changes, and the
amount by which it will change.
• Buffer systems play important roles in nature and in laboratory settings. In nature, they offer protection to living organisms, while in labs
they're used to create an environment with a stable pH.

What is a buffer composed of?

• To effectively maintain a pH range, a buffer must consist of a weak conjugate acid-base pair, meaning either
a. a weak acid and its conjugate base, or
b. a weak base and its conjugate acid.
• The use of one or the other will simply depend upon the desired pH when preparing the buffer.

ACID BUFFER AND BASE BUFFER

● Acidic solutions contain high concentrations of hydrogen ions (H+) and have pH values less than seven.
● It is generally made from a weak acid and one of its salts (often called conjugate*). Commonly used acidic buffer solutions are a
mixture of ethanoic acid and sodium ethanoate in solution, which have a pH of 4.76 when mixed in equal molar concentrations.
● Basic solutions contain high concentrations of hydroxide ions (OH-) and have pH values greater than seven.
● A very commonly used example of an basic buffer solution is a mixture of ammonia and ammonium chloride solution.
● Neutral solutions contain equal concentrations of hydrogen and hydroxide ions and have a pH of 7.
● Our blood is a buffer system that keeps pH between 7.35 and 7.45. It is important that the pH does not stray too far from this range;
blood that is too acidic or basic can damage bodily tissues. Seawater is also a buffer; the average pH of ocean water is 8.4. The
buffering ability of ocean water protects sea life from the corrosive effects of acids and bases.

Buffer Systems in the Body

• Carbonic acid bicarbonate buffer system
● The carbonic acid-bicarbonate buffer system plays an extremely important role in maintaining pH homeostasis of the blood.
● How? It is regulated in the blood by sodium When sodium bicarbonate (NaHCO3), comes into contact with a strong acid, such as
HCl, carbonic acid (H2CO3), which is a weak acid, and NaCl are formed. When carbonic acid comes into contact with a strong base,
such as NaOH, bicarbonate and water are formed.
● Carbonic acid-bicarbonate buffer system converts strong bases to a weak base (bicarbonate ion), and strong acids to a weak acid
(carbonic acid).
• Phosphate buffer system
● Phosphates are found in the blood in two forms: sodium dihydrogen phosphate (Na2H2PO4−), which is a weak acid, and sodium
monohydrogen phosphate (Na2HPO42- ), which is a weak base.
● How? When Na2HPO42- comes into contact with a strong acid, such as HCl, the base picks up a second hydrogen ion to form the
weak acid Na2H2PO4 − and sodium chloride, NaCl. When Na2HPO42− (the weak acid) comes into contact with a strong base,
such as sodium hydroxide (NaOH), the weak acid reverts back to the weak base and produces water.
• Protein buffer system.
● Nearly all proteins can function as buffers. Proteins are made up of amino acids, which contain positively charged amino groups
and negatively charged carboxyl groups.
● How? The charged regions of these molecules can bind hydrogen and hydroxyl ions, and thus function as buffers. Buffering by
proteins accounts for two-thirds of the buffering power of the blood and most of the buffering within cells.

Biochemistry for nurses

Acid-Base Regulation
● The regulation of acid-base balance is concerning the proper balance between chemical acids and bases in the extracellular fluids
(which includes the blood plasma). Acid-Base regulation is also called Body Ph.
● The body is very sensitive to its extracellular pH level otherwise outside the acceptable range of pH, proteins are denatured and
digested, enzymes lose their ability to function, and death may occur.
● The body’s acid-base balance is normally tightly regulated by buffering agents, the respiratory system, and renal systems, keeping the
blood pH between 7.35 and 7.45.
● Nurses require a good knowledge of the normal body mechanisms which regulate the Acid-Base balance because many ill patients
have damaged respiratory, renal and/or metabolic functions.

● In normal conditions, blood pH is between 7.35-7.45

● To maintain constant the blood pH, two organs play an essential role:
1. LUNGS: breathing in removing carbon dioxide (CO2) has an effect on the blood Ph.
2. KIDNEYS: eliminating protons (H+) and retaining bicarbonate (HCO3) has an effect on the blood Ph.
● Kidneys and Lungs contribute to the maintenance of acid-base balance of the body but for various reasons, this balance can be broken
(ACIDOSIS and ALKALOSIS)

Respiratory Regulation of Acid-Base Balance

○ The respiratory system contributes to the balance of acids and bases in the body by regulating the blood levels of carbonic acid.
○ CO2 in the blood readily reacts with water to form carbonic acid, and the levels of CO2 and carbonic acid in the blood are in
equilibrium.
○ When the CO2 level in the blood rises (as it does when you hold your breath), the excess CO2 reacts with water to form
additional carbonic acid, lowering blood pH. Increasing the rate and/or depth of respiration (whichyou might feel the “urge” to
do after holding your breath) allows you toexhale more CO2
Con’t
○ The loss of CO2 from the body reduces blood levels of carbonic acid and thereby adjusts the pH upward, toward normal levels.
○ Excessive deep and rapid breathing (as in hyperventilation) rids the blood of CO2 and reduces the level of carbonic acid, making
the blood too alkaline. This brief alkalosis can be remedied by rebreathing air that has been exhaled into a paper bag.
Rebreathing exhaled air will rapidly bring blood pH down toward normal.
○ The chemical reactions that regulate the levels of CO2 and carbonic acid occur in the lungs when blood travels through the lung’s
pulmonary capillaries. Minor adjustments in breathing are usually sufficient to adjust the pH of the blood by changing how much
CO2 is exhaled.
○ This situation is common if you are exercising strenuously over a period of time. To keep up the necessary energy production, you
would produce excess CO2 (and lactic acid if exercising beyond your aerobic threshold). In order to balance the increased acid
production, the respiration rate goes up to remove the CO2. This helps to keep you from developing acidosis.
○ The body regulates the respiratory rate by the use of chemoreceptors, which primarily use CO2 as a signal. Peripheral blood
sensors are found in the walls of the aorta and carotid arteries.
○ These sensors signal the brain to provide immediate adjustments to the respiratory rate if CO2 levels rise or fall.
○ Changes in the pH of CSF affect the respiratory center in the medulla oblongata, which can directly modulate breathing rate to
bring the pH back into the normal range.
○ Hypercapnia, or abnormally elevated blood levels of CO2, occurs in any situation that impairs respiratory functions, including
pneumonia and congestive heart failure. Reduced breathing (hypoventilation) due to drugs such as morphine, barbiturates, or
ethanol (or even just holding one’s breath) can also result in hypercapnia.
○ Hypocapnia, or abnormally low blood levels of CO2, occurs with any cause of hyperventilation that drives off the CO2, such as
salicylate toxicity, elevated room temperatures, fever, or hysteria.

Renal Regulation of Acid-Base Balance

○ The renal regulation of the body’s acid-base balance addresses the metabolic component of the buffering system. Whereas the
respiratory system (together with breathing centers in the brain) controls the blood levels of carbonic acid by controlling the
exhalation of CO2, the renal system controls the blood levels of bicarbonate.
○ A decrease of blood bicarbonate can result from the inhibition of carbonic anhydrase by certain diuretics or from excessive
bicarbonate loss due to diarrhea. Blood bicarbonate levels are also typically lower in people who have Addison’s disease (chronic
adrenal insufficiency), in which aldosterone levels are reduced, and in people who have renal damage, such as chronic nephritis.
○ Bicarbonate ions, HCO3– , found in the filtrate, are essential to the bicarbonate buffer system.
○ The hydrogen ions also compete with potassium to exchange with sodium in the renal tubules. If more potassium is present than
normal, potassium, rather than the hydrogen ions, will be exchanged, and increased potassium enters the filtrate. When this
occurs, fewer hydrogen ions in the filtrate participate in the conversion of bicarbonate into CO2 and less bicarbonate is
conserved. If there is less potassium, more hydrogen ions enter the filtrate to be exchanged with sodium and more bicarbonate is
conserved.

MAJOR BIOCHEMICAL PATHWAY

Introduction
• Metabolism is the term used to describe
- the interconversion of chemical compounds in the body
- The pathways taken by individual molecules,
- Their interrelationships, and the mechanisms that regulate the flow of metabolites through the pathways
• It falls mainly in 3 categories: catabolism, anabolism and amphibolic pathways
• Anabolic pathways
- Involved in the synthesis of larger and more complex compounds from smaller precursors
- Ex: Synthesis of protein from amino acids and the synthesis of reserves of triacylglycerol and glycogen.
- Anabolic pathways are endothermic.
• Catabolic pathways
- Involved in the breakdown of larger molecules, commonly involving oxidative reactions;
- They are exothermic, producing reducing equivalents, and, mainly via the respiratory chain
• Amphibolic pathways
- Occur at the “crossroads” of metabolism, acting as links between the anabolic and
catabolic pathways
- Ex: Citric acid cycle
Metabolism
• A 70-kg adult human being requires about 1920-2900 kcal from metabolic fuels each day, depending on
physical activity.
• This energy requirement is met from
– Carbohydrates (40%-60%)
– Lipids (mainly triacylglycerol, 30%-40%)
– Protein (10%-15%), as well as alcohol.
• There is a constant requirement for metabolic fuels throughout the day
• Most people consume their daily intake of metabolic fuels in two or three meals, so there is a need to form reserves
• Reserves of
– Carbohydrate: glycogen in liver and muscle
– Lipid: triacylglycerol in adipose tissue
– Labile protein
• If the intake of metabolic fuels is consistently greater than energy expenditure.
– Surplus is stored, largely as triacylglycerol in adipose tissue, – Leading to the development of obesity
• If the intake of metabolic fuels is consistently lower than energy expenditure
– Reserves of fat and carbohydrate, and amino acids are used for energy yielding metabolism
– This leads to emaciation, wasting, and, eventually, death
• All the products of digestion are metabolized to acetyl-CoA - oxidized by the citric acid cycle

Carbohydrate Metabolism
• Carbohydrates function
- Major energy source for sustainment of life (storage & generation)
- Cell wall of bacteria & aid in molecular recognition and communication.
- Such as blood group sugars on cell surface
- • Major disease is Diabetes Mellitus

Glucose Metabolism
▪ During a fast, the blood glucose level is kept constant by mobilizing the glycogen stores in the liver.
▪ During long fasts, gluconeogenesis is required to maintain blood glucose levels because glycogen stores are used up in
about 24-48 hours.
▪ An individual with a fasting blood glucose level >100 mg/dL is referred to as hyperglycemic.
▪ An individual with a fasting blood glucose level <50 mg/dL is referred to as hypoglycemic. update

Major biochemical pathways

• Glycolysis
– is a cytoplasmic pathway which breaks down glucose into two three-carbon compounds and generates energy.
– Phosphorylation pf CHO-hexokinase
– Use of ATP
– Glycolysis is used by all cells in the body for energy generation.
– The final product of glycolysis is pyruvate in aerobic settings and lactate in anaerobic conditions.
• Glycogenesis
– Glycogen synthesis
– important in liver and muscle
– is an anabolic process that requires energy.
– Glycogen- primary carbohydrate stored in the liver and muscle cells of animals, from glucose.
– takes place when blood glucose levels are sufficiently high
– is the process of storing excess glucose for use by the body at a later time.

• Tricarboxylic Acid
– also called Krebs cycle and citric acid cycle
– the second stage of cellular respiration
– Carried out in the intracellular structure-mitochondria
– plays a central role in the breakdown, or catabolism, of organic fuel molecules-glucose; other sugars, fatty acids and some
amino acids
Before these rather large molecules can enter the TCA cycle they must be degraded into a two-carbon compound called acetyl
coenzyme A (acetyl CoA). Once fed into the TCA cycle, acetyl CoA is converted into carbon dioxide and energy.

• Pentose Phosphate Pathways

– Also known as hexose monophosphate shunt
 – fundamental component of cellular metabolism.
– parallel to the glycolysis pathway and takes place in the cytoplasm
– used to produce ribose-5-phosphate and nicotinamide adenine dinucleotide phosphate (NADPH)
➢ribose-5-phosphate- precursor for the synthesis of nucleotides and nucleic acids.
➢NADPH- is an essential electron donor in all organisms, and provides the reducing power for anabolic reactions and redox balance

• Pentose Phosphate Pathways

– plays a critical role in regulating cancer cell growth by supplying cells with not only ribose-5-phosphate but also NADPH for
detoxification of intracellular reactive oxygen species, reductive biosynthesis, and ribose biogenesis.

• Glycogenolysis
– process by which glycogen is broken down into glucose to provide immediate energy and to maintain blood
glucose levels during fasting
– occurs primarily in the liver and is stimulated by the hormones glucagon and epinephrine (adrenaline).

• Gluconeogenesis
– occurs in the liver and kidneys.
– supplies the needs for plasma glucose between meals.
– stimulated by the diabetogenic hormones (glucagon, growth hormone, epinephrine, and cortisol).
– helps to maintain the glucose level in the blood so that the brain and muscle can extract sufficient glucose from it to meet
their metabolic demands.

Metabolic pathways
Glycolysis ------- Glucose Pyruvate Lactate
Glycogenesis ------- Glucose glycogen
Tricarboxylic acid cycle ------- Glucose pyruvate Co2+H20 +ATP
Pentose Phosphate Pathways ----- Glucose ribose+CO2+NADPH
Glycogenolysis ------------ Glycogen glucose
Gluconeogenesis --------- Non carbohydrates source, protein, fats glucose

Hormonal control and major site of glucose regulation

- Islets of Langerhans in the pancreas produce
1. Alpha cells secrete glucagon
2. Beta cells secretes insulin
3. Delta cells-somatostatin

Normal Blood Glucose

• Fasting blood glucose- 20-105 mg/dL
• May rise to 130-160 mg/dL about 1 hr after a glucose load or after a high carbohydrate meal (postprandial).
• After 2 hr, it drops back to normal fasting range
Hormones Affecting Blood Glucose Levels
• Insulin
• Glucagon
• Epinephrine
• Growth hormone
• Adreno-corticotropic hormone ACTH • cortisol
• Somatostatin
• T3 &T4

Insulin (hormone)
• Secreted by pancreas B cells of islets of Langerhans.
• Primary function is to decrease blood glucose & movement of glucose from blood into cells.
• Actions:
– Facilitate the entry of glucose into hepatocyte, adipocytes,red cells and monocytes by making the cell membrane
permeable to glucose.
– Increases uptake of glucose by the liver promotes glycogenesis and lipogenesis.
– inhibits hepatic output of glucose into circulation
– increases synthesis of protein in the liver, muscle and fat cells.
– decreases gluconeogenesis.
So insulin reduces plasma glucose; So insulin works to return the glucose level back to normal by lowering it and facilitates its
entry into the cells.
Glucagon (hormone)
• Secreted by α cells of islets, secretion is regulated by plasma glucose concentration. Insulin inhibits glucagon
release.
• Primary function:
– increases blood glucose conc and FFA, mobilization of energy stores.
• Actions:
– stimulates breakdown of liver glycogen into glucose (Glycogenolysis)
– increase liver gluconeogenesis (from non carb) - promotes hepatic lipolysis.

Epinephrine (hormone)
• Secreted by adrenal medulla
• Primary function: increases blood glucose and mobilizes energy stores.
• Actions:
– Increases glycogenolysis
– Stimulates glucagon secretion - Inhibits insulin secretion
 – Increases TG breakdown in adipose tissue (lipolysis).
– Physical or emotional stress causes increased secretion of epinephrine and an immediate increase in blood glucose
levels.

Growth Hormone
• Polypeptide secreted by the anterior pituitary • • Primary action: increase in blood glucose and mobilize energy stores.
• Actions:
– increases gluconeogenesis
– antagonizes insulin
– inhibits lipogenesis from carbohydrates

Adreno-corticotropic hormone ACTH

• Secreted by anterior pituitary
• Primary function: increases blood glucose & and mobilizes energy stores.
• Actions: - antagonizes insulin

Cortisol
• Secreted by adrenal cortex
• Primary function: increases blood glucose & and mobilizes energy stores.
• Actions:
– Promotes protein catabolism
– Promotes deamination of amino acids
– Promotes gluconeogenesis
– Inhibits glucose metabolism in the peripheral tissues – antagonist of insulin.

Thyroid hormones T4 & T3

• Secreted by the thyroid gland
• Primary function: increases blood glucose & and mobilizes energy stores.
• Actions:
– stimulates glycogenolysis
– increases intestinal absorption of glucose – accelerates the degradation of insulin.

Somatostatin
• Peptide hormone synthesized by delta by the alpha cells of the pancreatic islets of Langerhans.
• Inhibits both insulin, glucagon and growth hormone release , resulting in an increase in plasma glucose level

Hormones
• So Glucagon Epinephrine Growth hormone ACTH Cortisol T3 & T4 Increases blood glucose & and mobilizes energy stores.
• So which hormone increases in Hypoglycaemia???

ABNORMALITIES IN CARBOHYDRATES METABOLISM

Lactose Intolerance
• Lactose or milk sugar found in the milk of mammals - 4-6% in cow's milk and 5-8% in human milk.
– It is also a by-product in the manufacture of cheese.
• Lactose intolerance is the inability to digest significant amounts of lactose, the predominant sugar of milk
• This inability results from a shortage of the enzyme lactase, which is normally produced by the cells that line the small
intestine.
• Lactase breaks down the lactose, milk sugar, into glucose and galactose that can then be absorbed into the bloodstream.
• Common symptoms (severe to mild) include: o – abdominal pain
– abdominal bloating
– Gas
– diarrhea
– nausea
– uncomfortable 30 minutes to 2 hours after consuming milk and milk products
• Diagnostic Test
– Hydrogen Breath Test.
– Stool AcidityTest.

Hypoglycemia
• fasting blood glucose < 70 mg/d
• the body respond by secreting glucagon and epinephrine
• Causes of hypoglycemia
• starvation
• exaggerated insulin release -hyperinsulinemia
•Over administration of insulin or oral hypoglycemic agents
• Sever hepatic dysfunction
• Fasting blood levels < 50 mg/dL uncommon.
• Rapid drop in blood glucose (< 50 -55 mg/dL) will cause release of epinephrine leading to: – in early stage: anxiety,
dizziness, chills, tachycardia and increase perspiration.
• Blood levels < 20 mg/dL (later stages) lead to: – impaired nerve function & nerve damage, slurred speech, loss of motor
coordination, coma, lethargy, confusion, seizures

Hyperglycemia
• Remember: normal fasting blood glucose 70 to 105 mg/dL
• Hyperglycemia - fasting blood glucose > 105 mg/dL.
• Diabetes mellitus - a disorder in glucose metabolism producing hyperglycemia. • • Remember:
 hypoglycemia - fasting blood glucose < 70 mg/dL

Diabetes Mellitus
• commonly known as diabetes
• metabolic disease that causes high blood sugar.
• Untreated high blood sugar from diabetes can damage your nerves, eyes, kidneys, and other organs.
• There are a few different types of diabetes
– Type 1 diabetes is an autoimmune disease. The immune system attacks and destroys cells in the pancreas, where
insulin is made.
– Type 2 diabetes occurs when your body becomes resistant to insulin, and sugar builds up in your blood.
– Prediabetes occurs when your blood sugar is higher than normal, but it’s not high enough for a
diagnosis of type 2 diabetes.
– Gestational diabetes is high blood sugar during pregnancy. Insulin-blocking hormones produced by the placenta
cause this type of diabetes.

• General symptoms
– increased hunger
– increased thirst
– weight loss
– Frequent urination
– Blurry vision
– Extreme fatigue
– Sores that don’t heal

men with diabetes may have a decreased sex drive, erectile dysfunction (ED), and poor muscle strength.
Women with diabetes can also have symptoms such as urinary tract infections, yeast infections, and dry, itchy skin.

• Causes
– Decrease or absence of insulin secretion – Insulin resistance
– Abnormality in the control of insulin secretion
– Insulin action defect on target cell

Glycogen Storage Disease (GSD)

• GSD are group of inherited inborn errors of metabolism due to deficiency or dysfunction of the of enzymes
• Confined just to liver and muscles
• But some cause more generalized pathology and affect tissue such as kidney, heart and bowel
• The classification of GSD is based on the enzyme deficiency and the affected tissue
• Type 1 –Von Gierke’s Disease
– Affected enZyme-glucose 6 phosphatase
– Affected Tissue-liver and kidney
– Large quantities of glycogen are formed and stored in hepatocytes, renal and intestinal mucosa
cells. The liver and kidneys become enlarged
• Type II-Pompe’sDisease
– Deficiency of enzyme alpha 1, 4 glucosidase (acid maltase) lead to accumulation of glycogen in many tissues
– In infantile form accumulation of glycogen in cardiac muscles lead to cardiac failure
– liver-hepatomegaly and elevation of hepatic enzymes
– Muscle and peripheral nerves-hypotonia and weakness
– Blood vessels-intracranial aneurysms

• Type III-Cori Disease

– Enzyme-glycogen debranching enzyme
– Affected Tissue-liver and muscle
– Can lead to liver cirrhosis and hepatocellular carcinoma

▪ Type IV-Andersen’s Disease; Amylopectinosis

▪ Enzyme-glycogen debranching enzyme
▪ Affected Tissue-Many including liver
▪ Treatment: liver transplant
▪ Mostly death by age 4 due to cirrhosis and portal hypertension
• Type V Mcardle disease
– Myophosphorylase deficiency
– Affect muscle
– Early life-muscle strength and reflexes may be normal – Later adult life-persistent proximal weakness and
muscle wasting
• Type VI- Hers Disease
– Liver phosphorylase enzyme deficiency
– Affect liver, rare in cardiac form
– Hepatomegaly, hypoglycemia, growth retardation, hyperlipidemia
• Type VII Tarui disease
– Phosphofructokinase deficiency
– Muscle is affected
– Exercise intolerance, muscle cramping
• Type VIII-Fanconi-Bickel syndrome
– Glucose transporter GLUT 2
– Similar feature to Von Gierke’s disease
• Type 0 Lewis disease
 – Hepatic Glycogen synthase
– Affects lives
– Fatigue and muscle cramps after exertion
– Mild growth retardation in some case
• Type VI- Hers Disease
– Liver phosphorylase enzyme deficiency
– Affect liver, rare in cardiac form
– Hepatomegaly, hypoglycemia, growth retardation, hyperlipidemia

Major Biochemical
BLOOD GLUCOSE TEST
• measures the glucose levels in your blood.
• Glucose is a type of sugar. It is the body's main source of energy.
• Hyperglycemia-high blood glucose levels
• Hypoglycemia-Low blood glucose levels
• provides useful information for diabetes management.
• help monitor the effect of diabetes medications on blood sugar levels,
• Identify blood sugar levels that are high or low,
• track your progress in reaching your overall treatment goals,
• learn how diet and exercise affect blood sugar levels, and
• understand how other factors, such as illness or stress, affect blood sugar levels
• When To Get Tested?
– there are risk factors for diabetes or when you are 45 years of age or older, as recommended by the American
Diabetes Association
– experience symptoms suggesting high or low blood glucose
– have diabetes, may be instructed by the healthcare practitioner to check the glucose level up to several times a
day.
• Test Preparation
- Screening and diagnosis: in general, it is recommended that
– fast (nothing to eat or drink except water) for at least 8 hours (usually overnight) before having a blood glucose
test.
– may be done at a random time
– post-meal glucose

BLOOD GLUCOSE TEST

• Hemoglobin A1c
– is also called A1c or glycated hemoglobin test , is hemoglobin with glucose attached.
– evaluates the average amount of glucose in the blood over the last 2 to 3 months by measuring the percentage of
glycated hemoglobin in the blood.
– used to screen for and diagnose diabetes or risk of developing diabetes.
– monitor treatment for individuals diagnosed with diabetes
– NORMAL VALUE: A1c level is between 4% and 5.6%.
• an A1c of less than 7% indicates good glucose control and a lower risk of diabetic complications for the majority of
people with diabetes.
• Capillary Blood Glucose
– Synonym: finger stick blood glucose
– circulating blood glucose as measured by glucometer analysis of a fingerstick sample.
– Regular measurements of CBG allow diabetic patients to make frequent adjustments in their caloric intake, exercise levels,
and use of antidiabetic medications, esp. insulin.
Normal Value: 70-110 mg/dl
• Fasting Blood Sugar (FBS)
– give vital clues about how a person’s body is managing blood sugar.
– Blood sugar tends to peak about an hour after eating and declines after that.
• Oral Glucose Tolerance Test (OGTT)
– Glucose Tolerance Test for Gestational Diabetes; Glucose Tolerance Test, Pregnancy
– To screen for and diagnose diabetes that develops during pregnancy, also known as gestational diabetes
– Most women are screened for gestational diabetes between 24 and 28 weeks of pregnancy.
• Oral Glucose Tolerance Test (OGTT)
– One-step 2-hour oral glucose tolerance test: you will need to fast (usually overnight) before an initial blood sample is
drawn. You will then be given a 75-gram dose of glucose to drink and your blood will be drawn at 1 hour and 2 hours after
the dose.
– NORMAL VALUE- equal to or less than 140 mg/dL (7.8 mmol/L)
• Diabetes Autoantibodies
– Islet Autoantibodies in Diabetes; Islet Autoantibodies in Diabetes
– To identify people at increased risk for developing type 1 diabetes or requiring insulin treatment; to aid in the classification
of diabetes.
– When to get tested
• Any time that you have diabetes and your healthcare practitioner cannot clearly determine if you have
– glutamic acid decarboxylase, or GAD, antibodies.
– Type 1 diabetes that develops later in life is called latent autoimmune diabetes in adults (LADA).
–
GAD antibodies belong to a group of diabetes-associated antibodies that instruct the immune system to destroy the
insulin-producing pancreatic cells. When insulin production stops, diabetes develops.
– Normal Value-under 5 units/ml
• Albumin-to-Creatinine Ratio
– Albumin is a protein found in the blood.
 – Creatinine is a waste product that comes from the normal wear and tear on muscles of the body.
– To screen for and detect early kidney disease in people with diabetes or other risk factors, such as high blood pressure
(hypertension)
– A random sample of urine, a timed urine sample (such as 4 hours or overnight), or a complete 24-hour urine sample is
collected in a clean container.
– Albumin-to-creatinine ratio (ACR) is the first method of preference to detect elevated protein.
• if the results continue to show albumin in urine, it may mean you have early-stage kidney disease. • If your test
results show high levels of albumin, it may mean you have kidney failure.
• Normal Value-
– less than 30 - normal
– 30-300-microalbuminuris
– Above 300-macroalbuminuria

• Comprehensive Metabolic Panel (CMP)

• measures the sugar (glucose) level, electrolyte and fluid balance, kidney function, and liver function
– How your kidneys and liver are working
– Your blood sugar levels
– Your electrolyte levels
– How much protein is in your blood
– The balance of acid and base in your blood
– How your metabolism is working
– Liver profile
• ALP (alkaline phosphatase): 44-147 international units per liter (IU/L)
• ALT (alanine aminotransferase): 7-40 IU/L
• AST (aspartate aminotransferase): 10-34 IU/L
• Bilirubin: 0.3-1.9 milligrams per deciliter (mg/dL)
– Kidney profile
– BUN (blood urea nitrogen): 6-20 mg/dL
– Creatinine: 0.6-1.3 mg/dL

• Serum electrolytes

– Sodium: 136-145 milliequivalents per liter (meq/L)

– Potassium: 3.5-5.1 meq/L
– Chloride : 96-106 meq/L
– CO2 (carbon dioxide): 23-29 meq/L
• Protein

– Albumin: 3.4-5.4 grams per deciliter (g/dL)

– Total protein: 6.0-8.3 g/dL
– Glucose
• The normal range is 70-99 mg/dL.
– Calcium
• Total blood calcium: 8.5 to 10.3 milligrams per deciliter (mg/dL) · Ionized calcium: 4.4 to 5.4 mg/dl.

• CMP calcium test

– This is important for healthy muscles, nerves, and hormones.
– If calcium is abnormal, you might have a hormone imbalance or problems with your kidneys, bones, or pancreas.
– The normal range is 8.6-10.2 mg/dL.

• Basic Metabolic Panel (BMP)

– basic metabolic panel (BMP) is a group of 8 tests that measures several substances in your blood.
– provides information on your blood sugar (glucose) level, the balance of electrolytes and fluids, and the health of your
kidneys.
• Glycated Serum Protein (GSP)
– Fructosamine
– To help monitor your blood glucose (sugar) levels over time if you have diabetes, especially if it is not possible to monitor your
diabetes using the hemoglobin A1c test;
– to help determine the effectiveness of changes to your diabetic treatment plan that might include changes in diet, exercise or
medications, especially if they were made recently
– Normal values vary in relation to the serum albumin concentration and are 200-285 µmol/L, when the serum albumin
concentration level is 5 g/dL. [4] Reduction in serum albumin lowers the serum

Low blood glucose → pancreas –glucagon→ liver

High blood glucose → pancreas –insulin→ body’s cells

LIPID PANEL TEST

LEARNING OBJECTIVES

1. At the end of the exercise, the students should be able to:

2. Know the chemical properties of lipids
3. Differentiate the 5 different lipoproteins: chylomicrons, very low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL),
low-density lipoproteins (LDL), and high density lipoproteins (HDL).
 4. Know the different lipid panel test, normal value and significance.
5. Understand the common lipid disorders.

LIPIDS

− Lipids are an essential component of the cell membrane

− The structure is typically made of:
• glycerol backbone
• 2 fatty acid tails (hydrophobic)
• phosphate group (hydrophilic)
− In the cell membrane, phospholipids are arranged in a bilayer manner, providing cell protection and serving as a barrier to certain
molecules.
− The hydrophilic part faces outward and the hydrophobic part faces inward. This arrangement helps monitor which molecules can enter and
exit the cell. For example, nonpolar molecules and small polar molecules, such as oxygen and water, can easily diffuse in and out of the
cell. Large, polar molecules, for example, glucose, cannot pass freely so they need the help of transport proteins.

LIPOPROTEINS

− substances made of protein and fat that carry cholesterol through your bloodstream.
− main goal of lipoprotein is to help transport lipids (hydrophobic) in water.
− The structure of lipoprotein consists of:
• triglycerides
• Cholesterol
• Phospholipids
• Apolipoproteins-mainly function as carrier proteins but also serve as cofactors for enzymes that metabolize lipoproteins and help
in lipid component exchange among lipoproteins

EXAMPLES OF LIPOPROTEINS

• Chylomicrons
• very-low-density lipoproteins (VLDL)
• intermediate-density lipoproteins (IDL)
• Low-density lipoproteins (LDL)
• high-density lipoproteins (HDL).
• Each one is used in a different phase of lipid transport.

CHEMICAL PROPERTIES OF LIPIDS

1. Hydrolysis of triglycerols
- Triglycerols like any other esters react with water to form their carboxylic acid and alcohol– a process known as hydrolysis. 2.
Saponification:
- Triacylglycerols may be hydrolyzed by several procedures, the most common of which utilizes alkali or enzymes called lipases. Alkaline
hydrolysis is termed saponification because one of the products of the hydrolysis is a soap, generally sodium or potassium salts of fatty
acids.
3. Hydrogenation
- The carbon-carbon double bonds in unsaturated fatty acids can be hydrogenated by reacting with hydrogen to produce saturated fatty
acids.
4. Halogenation
- Unsaturated fatty acids, whether they are free or combined as esters in fats and oils, react with halogens by addition at the double
bond(s). The reaction results in the decolorization of the halogen solution.
5. Rancidity:
- The term rancid is applied to any fat or oil that develops a disagreeable odor. Hydrolysis and oxidation reactions are responsible for
causing rancidity. Oxidative rancidity occurs in triacylglycerols containing unsaturated fatty acids.

CHYLOMICRONS

- large triglyceride-rich lipoproteins produced in enterocytes from dietary lipids—namely, fatty acids, and cholesterol - also
known as ultra low-density lipoproteins (ULDL)
- are lipoprotein particles that consist of
• triglycerides (85–92%)
• phospholipids (6–12%)
• cholesterol (1–3%)
• proteins (1–2%)
- FUNCTION:
o transport lipids absorbed from the intestine to adipose, cardiac, and skeletal muscle tissue, where their triglyceride components
are hydrolyzed by the activity of the lipoprotein lipase, allowing the released free fatty acids to be absorbed by the tissues.

VERY-LOW DENSITY LIPOPROTEINS (VLDL)

− consist of mainly triglycerides, some cholesterol molecules, and less protein

− less dense than most lipoproteins because of its high lipid composition.
− made in the liver and is responsible for delivering triglycerides to cells in the body, which is needed for cellular processes. − As triglycerides
 get delivered to cells, VLDL is made up less of fat and more of protein, leaving cholesterol on the molecule. As this process occurs, VLDL will
eventually become an LDL molecule.

INTERMEDIATE-DENSITY LIPOPROTEIN (IDL’S)

− belong to the lipoprotein particle family and are formed from the degradation of very low-density lipoproteins as well as high-density
lipoproteins.
− one of the five major groups of lipoproteins (chylomicrons, VLDL, IDL, LDL, HDL) that enable fats and cholesterol to move within the
water-based solution of the bloodstream
− transports a variety of triglyceride fats and cholesterol and, like LDL, can also promote the growth of atheroma − An atheroma, or
atheromatous plaque ("plaque"), is an abnormal and reversible accumulation of material in the inner layer of an artery wall

LOW-DENSITY LIPOPROTEINS (LDL’S)

− sometimes called “bad” cholesterol”, makes up most of your body's cholesterol. High levels of LDL cholesterol raise your risk for heart
disease and stroke.
− The main transporters of cholesterol in the blood carry cholesterol from the liver to body cells, including those in the arteries, where it can
contribute to plaque

HIGH DENSITY LIPOPROTEINS (HDL’S)

− Also known as “good” cholesterol

− It absorbs cholesterol and carries it back to the liver.
− The liver then flushes it from the body.
− High levels of HDL cholesterol can lower your risk for heart disease and stroke
Lipid Profile PURPOSE NORMAL VALUE PREPARATION SIGNIFICANCE

Total Cholesterol a measure of LDL • less than 200 • Patient is required • To estimate your
cholesterol, HDL milligrams per to fast, consuming risk of heart attacks
cholesterol, and deciliter (mg/dL) are no food or liquids and other forms of
other lipid considered desirable other than heart disease and
components for adults water, for 9 to 12 diseases of the blood
• between 200 and hours before the vessels.
239 mg/dL is test
considered
borderline high

• 240 mg/dL and

above is considered
high.

High- density • measures the level • HDL levels that are • fasting for up to 12 • assess your
lipoprotein of good cholesterol below 40 mg/dL for hours before the test. cardiovascular
cholesterol (HDL-C) in your blood men and 50 mg/dL • shouldn’t have an health, including
for women indicate HDL test when your risk for
an increased risk of you’re sick. heart disease
heart disease. Cholesterol levels • may also be
are ordered as a
temporarily lower follow-up test if you
during acute illness, have high results on
immediately your
following a heart cholesterol-screening
attack, and during test.
stressful
events like surgery
or an accident. It’s
recommended that
you wait at least
six weeks
after any illness
before you have
your cholesterol

Low-density •measures the • LDL cholesterol • Patient is required •can provide

lipoprotein amount of LDL levels should be to fast, consuming information about
cholesterol (LDL-C) cholesterol in your less than 100 no food or liquids cardiovascular
blood mg/dL other than health, they may be
• Levels of 100 to 129 water, for 8 to 12 measured as part of
mg/dL are acceptable hours before the screening,
for people with no test diagnosis, or
health issues but may monitoring
be of more concern
for those with heart
disease or heart
disease risk factors
• 130 to 159 mg/dL is
borderline high
• 160 to 189 mg/dL is
high.
• A reading of 190
 mg/dL or higher is
considered very
high.

Triglycerides • measures the • 10 to 150 mg/dL • Patient is required • The results of this
amount of (the lower the to fast, consuming test help your
triglycerides in your number, the no food or liquids doctor
blood better) other than determine your risk
water, for 9 to 12 of developing heart
hours before the disease
test and only water
is permitted

Very low-density • measures the • normal VLDL • not to eat or drink • The amount of
lipoprotein amount of very cholesterol is typically for 9- 12 hours VLDL cholesterol in
cholesterol low-density between 2 and 30 before testing blood can help
(VLDL-C) lipoprotein (VLDL) in mg/dl doctors evaluate a
your blood • level higher than 30 patient’s risk of
mg/dL may put you heart
at an increased risk disease and stroke.
for
cardiovascular disease.

Non-HDL-C • is a way of • non-HDL • not to eat or drink • gives a better

measuring how cholesterol level for 9- 12 hours assessment of the
much of the bad goal should be 30 before testing risk for heart disease
kinds of cholesterol mg/dL higher than than
you have in your your LDL cholesterol measuring only LDL
blood level goal
• subtracting your
HDL cholesterol
from your
total cholesterol.

Cardiac Risk Assessment

− This is a group of tests and health factors that have been proven to indicate your chance of having a cardiovascular event such as a heart
attack or stroke
− the most important indicators for cardiac risk are your personal health history. These include:
• Age
• Family history
• Weight
• Cigarette smoking
• Blood pressure
• Diet
• Exercise, physical activity
• Diabetes
• Pre-existing heart disease, or already having had a heart attack

Non-invasive Tests

• electrocardiogram (ECG, EKG)

• a stress test, also called ECG stress test
• metabolic stress test

Invasive tests

• angiography/arteriography
• cardiac catheterization

Hyperlipidemia

− also known as dyslipidemia or high cholesterol, means you have too many lipids (fats) in your blood
− this condition can lead to serious problems, including heart attacks, stroke, and peripheral artery disease

Atherosclerosis

− a disease that occurs when plaque builds up inside arteries

− The arteries get hard and narrow, which can restrict blood flow and lead to blood clots, heart attack or stroke

Coronary Heart Disease

− the term that describes what happens when your heart's blood supply is blocked or interrupted by a build-up of fatty substances in the
coronary arteries
Carotid Artery Disease

− occurs when fatty deposits (plaques) clog the blood vessels that deliver blood to your brain and head (carotid arteries) − The
blockage increases your risk of stroke, a medical emergency that occurs when the blood supply to the brain is interrupted or seriously
reduced.

Peripheral Arterial Disease

− the narrowing or blockage of the vessels that carry blood from the heart to the legs
− It is primarily caused by the buildup of fatty plaque in the arteries, which is called atherosclerosis

HYPERTENSION

− also known as high or raised blood pressure, is a condition in which the blood vessels have persistently raised pressure. −
Common factors that can lead to high blood pressure include diet high in salt, fat, and/or cholesterol.

HEALTH TEACHINGS TO PREVENT LIPID DISORDERS

1. Eating a heart-healthy diet

− the best way to lower your cholesterol is reduce your intake of saturated fat and trans fat
− Reducing these fats means limiting your intake of red meat and dairy products made with whole milk. Choose skim milk, low-fat or fat free
dairy products instead. It also means limiting fried food and cooking with healthy oils, such as vegetable oil. 2. Becoming more physically
active
− At least 150 minutes of moderate intensity aerobic exercise a week is enough to lower both cholesterol and high blood pressure 3.
Quitting smoking
− Smoking and vaping lowers HDL cholesterol
4. Losing weight
− Being overweight or obese tends to raise bad cholesterol

PROTEIN STRUCTURES

− Proteins are of great nutritional value and are directly involved in the chemical processes essential for life. −
Jöns Jacob Berzelius-coined the term protein,
− Proteins are species-specific; that is, the proteins of one species differ from those of another species.
− They are also organ-specific; for instance, within a single organism, muscle proteins differ from those of the brain and liver. − A protein
molecule is very large compared with molecules of sugar or salt and consists of many amino acids joined together to form long chains, much
as beads are arranged on a string.
− Proteins are biological polymers composed of amino acids. Amino acids, linked together by peptide bonds, form a polypeptide chain. −
There are about 20 different amino acids that occur naturally in proteins.
− Proteins have complex shapes that include various folds, loops, and curves.
− Chemical bonding between portions of the polypeptide chain aid in holding the protein together and giving it its shape. −
There are two general classes of protein molecules: globular proteins and fibrous proteins.
• Globular proteins are generally compact, soluble, and spherical in shape.
• Fibrous proteins are typically elongated and insoluble. – Globular and fibrous proteins may exhibit one or more of four types of
protein structure.

PROTEIN SHAPES

• Globular Protein
• Elongated Proteins

Structure of Protein Introduction:

• Proteins are an important class of biological macromolecules which are the polymers of amino acids.
• Biochemists have distinguished several levels of structural organization of proteins.
• They are:
− Primary structure
− Secondary structure
− Tertiary structure
− Quaternary structure

PRIMARY STRUCTURE

• The primary structure of protein refers to the sequence of amino acids present in the polypeptide chain.
• Amino acids are covalently linked by peptide bonds.
• Each component amino acid in a polypeptide is called a “residue” or “moiety”
 • By convention, the 10 structure of a protein starts from the amino- terminal (N) end and ends in the carboxyl-terminal (C) end. • The primary
structure is the specific sequence of amino acids i.e. the order that they are bonded together. • The exact order that amino acids are bonded
together is determined by the information stored in genes.
• (contd)
− Through processes called transcription and translation,
− DNA provides all the necessary information for cells to produce the exact primary structure for thousands of different proteins. −
The primary structure determines the secondary and tertiary structures of proteins.

SECONDARY STRUCTURE

• The secondary structure of a protein is formed by hydrogen bonds between atoms along the backbone of the polypeptide chain. •
Remembering each amino acid has a carboxyl group and an amine group
• The slight negative charge on the oxygen of the carboxyl group forms a weak bond with the slight positive charge of a hydrogen atom on the
amine group of another amino acid
• Hydrogen bonds are weak but many of them create enough strength to influence the shape of a polypeptide chain.

TERTIARY STRUCTURE

• Is the final shape that the polypeptide chain takes and is determined by the R groups
• The attraction and repulsion between different R groups bends and folds the polypeptide to create the final 3D shape of a protein.

QUATERNERY STRUCTURE

• Not all proteins have a quaternary structure.

• A quaternary structure only results when multiple polypeptide chains combine together to form a large complex protein. • In
such cases, each polypeptide is referred to as a ‘subunit’.
• Hemoglobin is an example of a protein with quaternary structure. In most animals, hemoglobin is made from four globular subunits.

Quaternary structure of Hemoglobin

• Hemoglobin (HbA1) has 4 polypeptides chains (tetramer) associated by non-covalent bonds:

• 2 alpha chains + 2 beta chains
• It possesses quaternery structure (oligomeric). In this, R group contaccts are present between similar side chains and there is very little
contact between dissimilar

COMMON DISORDERS OF PROTEIN METABOLISM

Daily requirements

• Protein is essential to good health. The very origin of the word — from the Greek proteos, meaning “first” — reflects protein’s top-shelf status in
human nutrition.
• The DRI (Dietary Reference Intake) is 0.8- 1 grams of protein per kilogram of body weight, or 0.36 grams per pound. • This
amounts to:
o 56 grams per day for the average sedentary man.
o 46 grams per day for the average sedentary woman.
• However, requirement can increase in various physiological situations like
o Last half of pregnancy
o During lactation
o Infancy
o Childhood
o Adolescence

Metabolic Disturbance of Protein

★ Protein Energy Malnutrition (PEM)

• PEM is a spectrum of diseases whose essential feature is a deficiency of protein at one end as in Kwashiorkor and; • total
inanition (starvation of infant due to severe and prolonged restriction of all food at other end as in Marasmus.
• In the middle of spectrum, there is Marasmic Kwashiorkor in which there are clinical features of both disorders. •
Causes of PEM
o Dietary deficiency
o Serious illness
o Infections in babies
o Low socio-economic status
o Problems in mother leading to inadequate production of milk
o Dietary factors like inadequate breast feeding, ignorance of weaning

★ Kwashiorkor
• It's a maladaptive response to starvation due to lack of physiological adaptation to unbalanced deficiency where body utilizes proteins and
 conserves fat
• Occurrence – Seen predominantly between 1- 5 years of age
• Causes – Due to insufficient intake of proteins, as the diet of weaning child mainly consists of carbohydrates •
Signs and symptoms
o Weight is about 60-80 % of normal as loss of true weight is masked by increased fluid retention
o Generalized edema (due to decreased colloidal osmotic pressure) of pitting variety
o Sparing of subcutaneous fat and muscle mass as body utilizes its proteins to compensate the deficient state o
Swollen abdomen
o Moon face
o Characteristic skin lesions with alternating layers of hyperpigmentation, desquamation hypopigmentation; giving a flaky paint
appearance
o Hair changes like loss of colour or alternate bands of pale and darker colour, straightening and loss of firm attachment to scalp o
Diarrhoea (due to impaired synthesis of digestive enzymes and loss of electrolytes in stools)
o Eyelashes give a ‘Broomstick’ appearance
o Dehydration (due to diarrhoea and vomitting)
o Prone to infections
o Psychomotor changes (due to cerebral atrophy)
• Treatment
o Provide adequate nutrition by dietary supplements
o Restore normal body composition
o Dietary support in the form of 3-4 gm protein and 200 cal/kg body weight
o Cure the conditions that cause the deficiency
• Prevention
o Breastfeeding a baby for at least 6 months to prevent PEM
o Improved food distribution systems, public health programs and agricultural development should be promoted

★ Marasmus
• It's an adaptive response to starvation
• Seen mostly in the first year of life
• Causes – Due to deficiency of calories
• Clinical symptoms –
o Growth retardation due to low calorie intake
o Loss of muscle mass as muscle proteins are mobilised as use for fuel
o to provide body with a.a as a source of energy to compensate malnutrition.
o Due to muscle wasting rib cage is prominent
o Subcutaneous fat is lost due to lipolysis Lean body, so head appears too large
o Body weight falls to 60 % of normal
o Child looks older than his age
o Weakened body is under stress -More chances of infections
o Dry and baggy skin
o Sparse hair and wrinkled appearance (Old man face)
o No edema
o Bones are prominent due to absence of fat around them
o Hypotension and slow pulse
• Treatment
o Dietary support by giving food as proteins and supplements
o Bring the child out of starvation

Points of differentiation between kwashiorkor and marasmus

Kwashiorkor Marasmus

Malnutrition that occurs due to insufficient intake of Malnutrition occurs due to starvation (I.e deficiency of proteins,
proteins Large belly, diarrhoea, pigmented skin, hair carbs. And fats in diet.
changes Muscle wasting, skin foldings, prominent rib cage, and
Weaned from mother’s milk to a diet low in protein shrunken abdomen
Edema-present (pitting type) Children under 1 yr
60-80% of normal body wt. Failed breastfeeding (inadequate calorie intake)
Decreased plasma albumin Absent, rather wasting
Treated by High protein Less than 60% of normal
Normal/slightly decreased plasma albumin
Treated by a well balanced diet

★ Gout
• It is an inflammatory disease caused by deposition of monosodium urate monohydrate crystal around the synovial joints •
Epidemiology
− Levels are higher in men increase with age and are associated with body weight
• Pathophysiology
− it can due to diminished renal excretion
− increased intake of red meat
− Over production of uric acid
• Clinical features
− Severe pain
− Extreme tenderness
 − Marked swelling with overlying red shiny skin
− Crystal may deposit in joints and soft tissues to provuse irregular firm nodules called TOPHI
• TREATMENT
− Oral NSAIDS for pain relief
− Local icepacks
− Oral colchicine-decreasing swelling and lessening the buildup of uric acid crystals that cause pain in the affected joint(s). −
Joint aspiration with intraarticular steroid injections ▪ Oral corticosteroids-anti inflammatory
− Urate lowering therapy
− Allopurinol drug reduces conversion of hypoxanthine and xanthine to uric acid

In born errors of amino acids metabolism

★ PHENYLKETONURIA
• deficiency of hepatic enzyme PHENYLALANINE HYDROXYLASE result in accumulation of phenylalanine
• Phenylalanine is an essential aromatic amino acid in humans (provided by food), Phenylalanine plays a key role in the biosynthesis of
other amino acids and is important in the structure and function of many proteins and enzymes.
• Oral Manifestations
o Prominent cheek and jaw bones
o Widely spaced teeth
o Poor development of tooth enamel
o Patient are prone to tooth wear (erosion)
• Clinical manifestation
o Mental retardation due to its accumulation
o Hypopigmentation of hair and skin
• Treatment
o Diet low in phenylalanine amino acid-bread, pasta, biscuits and flour
o Phenylalanine –free formula for infants
o Meat, fish, eggs, milk, cheese, nuts are excluded
o Vegetables and fruits are allowed in measurable amount
o Diet with enough tyrosine-avocados bananas
★ Albinism
• It is due to lack of synthesis of pigment Melanin due to defect in tyrosinase-most responsible enzyme for Melanin synthesis •
Melanin from Greek: melas, "black, dark") is a broad term for a group of natural pigments found in most organisms. • CLINICAL
FEATURES
o Photophobia
o Susceptibility to Parkinson’s disease (as no DOPA formed
o Hypopigmentation in the form of
o Vitiligo-loss of pigmentation around the mouth, nose, eyes and nipples
o Leukoderma-loss of pigmentation begins with hands
• It’s a metabolic disorder of branched chain amino acids.
• The urine smells like maple syrup or burnt sugar
• SYMPTOMS
o Lethargy (lack of energy and enthusiasm)
o Maple syrup odor of urine
o Coma-A coma is a deep state of prolonged unconsciousness
o Mental retardation
• TREATMENT
o To feed a diet having a low branched of amino acids (leucine,isoleucine and valine)

ENZYMES

• enzyme is defined as a macromolecule that catalyzes a biochemical reaction.

• Substrates- the starting molecules in chemical reaction,
• Most enzymes are named by combining the name of the substrate with the -ase suffix (e.g., protease, urease). •
Most enzymes are globular proteins that are much larger than the substrate with which they interact.
• active site- contains one or more binding sites that orient the substrate in the correct configuration
• catalytic site-part of the molecule that lowers activation energy.
• Cofactor-a non-protein chemical compound or metallic ion that is required for an enzyme's activity
• could be a metal ion or an organic molecule, such as a vitamin.

Mechanism of Enzyme Action

• Lock and Key Theory

− first postulated in 1894 by Emil Fischer
− The lock is the enzyme, and the key is the substrate.
− Only the correctly sized key (substrate) fits into the keyhole (active site) of the lock (enzyme).
• Induced Fit Theory
− assumes that the substrate plays a role in determining the final shape of the enzyme and that the enzyme is partially flexible.
Ddifferent factors affect enzymatic activity.
Temperature End-products substrate

Radiation Inhibitors pH

Activators Enzyme water

TEMPERATURE

− The enzymatic activity will be optimum at normal temperature.

− The maximum activity is observed between 30-45 Celsius for most of the enzymes.
− Beyond 45 Celsius the enzyme activity is reduced drastically
− Beyond 60-60 Celsius the enzymes were denatured

WATER
− Enzyme activity is suppressed in the absence of water.
− Hydration of the cells is necessary for the enzyme activity
− Water provides a medium for the enzymatic reaction to take place
− In many cases, water is one of the reactants.

Hydrogen Ion Concentration (pH)

• Enzymes are active only over a limited range of pH.

• Most of the enzymes are specific to a particular pH.
• Ex:
▪ Trypsin is active in alkaline medium
▪ Diastase is active in the natural medium
▪ Pepsin show optimum activity acidic pH.

Concentration of ENZYMES

− A small amount of enzyme is enough for a large a amount of substrate.

− Increase in the concentration of enzyme will increase the rate of reaction (if there is enough substrates available in the medium. −
Increased number of enzyme molecules will have more active sites.

Concentration of SUBSTRATES

− Increase in the substrate concentration increases the activity of the enzyme until all the active sites of the enzyme molecule are saturated
with the substrate.
− Higher concentration of substrate can nullify the effects of competitive inhibitors in the medium.

Inhibitors

− Inhibitors in the reaction can inhibit enzymatic activity.

− Inhibitors are of different types
• Competitive inhibitors
• Noncompetitive inhibitors
• Uncompetitive inhibitors

Accumulation of end products

− The end product accumulation retards the enzymatic activity.

EFFECT OF ACTIVATORS

− Some enzymes require additional molecules for its optimum activity (generally known as prosthetic groups or co-factors). − The prosthetic
groups may be inorganic metal ions such as Mg2+, Cu2+, Ca2+, Na+, K+, etc, or complex organic vitamin derivatives such as NAD, FAD, etc.

EFFECT OF LIGHT RADIATION

− Some enzymes are sensitive to light

− Some enzymes require the presence of light for its catalytic activity
− Example: Photolyase enzyme involved in the photoreactivation DNA repair require light for its enzymatic activity. −
Usually, enzymatic activity is reduced under the influence of harmful radiations such as X-rays, UV rays, B and y rays.
ENZYMES
− Enzymes are biological catalysts.
− Diagnostic enzymes refers to the enzymes that are used directly or as components of the assay system for the determination of number of
substances
− Changes in the concentrations of various biomolecules are indications of abnormal metabolic activities, infections, infectious and non
infectious diseases and inflammatory conditions

LACTATE DEHYDROGENASE

− Important enzyme found throughout the body and involved in glucose metabolism
− LDH1 is found predominantly in heart muscle and in RBCs.
− LDH5 found in liver and skeletal muscle is the least stable and runs the shortest on electrophoresis
− LDH3 is found in a variety of tissues such as spleen,lung,endochrine glands and lymph nodes
− Normal value-180-360U/L

ASPARTATE TRANSAMINASE

− found in most tissues throughout the body, but especially in skeletal muscle, cardiac muscle, liver and kidney −
formally known as glutamate oxaloacetate transaminase (GOT)
− Useful in the diagnosis of myocardial infarction. ✓ Elevated AST levels is indicative of damage to the myocardium. −
Normal range-male 35<U/L, female 31<U/L

ALANINE TRANSAMINASE

− Formally known as glutamate pyruvate transaminase (GPT)

− Found in high concentrations in liver cells and in much smaller concentrations elsewhere
− markedly raised plasma activity indicates a severe liver disease, usually viral hepatitis or toxic liver necrosis −
Normal values-male<45U/L,female <34U/L

ALKALINE PHOSPHATASE

− High levels are found in liver,bone,placenta and intestine

− Used as a marker of cholestatic liver disease
− Cholestasis/ Cholestatic liver disease describes impairment in bile formation or flow which can manifest clinically with fatigue, pruritus, and
jaundice.

Sex Age Normal Range

Male/female 4-15 54-369

Male 20-50≥60 53-128

56-119

Female 20-50≥60 42-98

53-141

CREATININE KINASE

− Also known as creatine phosphatase (CPK)

− Mainly found in heart and skeletal muscle and in brain
− Thus 3 isoenzymes are found:
• CKBB the main form of brain,
• CKMB in heart muscle and diaphragm and
• CKMM-both heart and muscle
− Increased plasma activities of CK(MM+MB) results in severe damage to heart cells.
− Normal range=male-46-171U/L,female-34-145U/L28

ACID PHOSPHATASE
− Found in large amounts in prostate glands and its assay in plasma has been used in the diagnosis of prostatic carcinoma −
Also found in liver, red cells, platelets and bone.
− Normal range-0.1-0.4U/L

CHOLINESTERASE

− Presence is assumed when the muscle relaxant scoline is administered.

− Patients with low plasma activities of cholinesterase experience severe breathing difficulties for several hours after treatment with
scoline, so a preliminary assay may be carried out to see if it safe to administer the drug.
 − Normal range=male-40-78U/L,female-33-76U/L

A-GLUTAMYL TRANSFERASE

− Found in biliary ducts of the liver, in the kidney and pancreas with the largest amounts being in kidneY ✓ Also found in hepatocytes
where its enzyme activity can be induced by a number of drugs and in particular alcohol,
− a useful marker of alcohol induced liver disease and in particular liver cirrhosis.
− Normal range=male-<55U/L,female-<38U/L

AMYLASE

− Found in high concentrations in pancreas and salivary glands where it is secreted to digest complex carbohydrates − Useful in those patients
with acute abdominal pain, to differentiate between patients with acute pancreatitis and those with appendicitis − Patients with acute
pancreatitis will have high levels of amylase in their blood
− Normal range=28-100U/L

INBORN ERRORS OF METABOLISM

− In such conditions, the enzyme is not fully active, resulting in a pathological condition called an inherited disorder. −
The activity is measured in blood and sometimes in preparation of cells such as fibroblast
Disorder Enzyme affected

Phenylketonuria Phenylalanine hydroxylase

Alkaptonuria Homogentisic acid oxidase

galactosaemia galactokinase

Von gierke’s disease Glucose-6-phosphatase

Tay-sachs disease B-N-Acetyl glucosaminidase A

Hurler’s syndrome A-L-iduronidase

ENZYMES AS REAGENTS IN CLINICAL CHEMISTRY

− D-glucose in blood and other physiological fluids is commonly analyzed by means of procedures involving glucose oxidase. −
The reaction catalyzed by glucose oxidase is utilized for the screening of urine specimens.
− Used in the diagnosis of diabetes Mellitus
− Blood cholesterol may be determined by a reaction catalyzed by cholesterol oxidase,

ENZYMES IN FORENSIC MEDICINE

− For detecting body fluids and as a marker of genetic individuality

− Test for seminal acid phosphatase activity
− Used in DNA finger printing
− Forensic markers-adenosine deaminase, adenylate kinase, carbonic anhydrase, esterase D, glucose 6phosphatedehydrogenase, peptidase A,
glyoxalase, phosphoglucomutase

MINERALS
What Are Minerals?

• Inorganic elements essential to the nutrition of humans

• Fourteen minerals are essential to body function
− Play several key roles in overall health and well being
• Help chemical reactions take place in cells
• Help muscles contract
• Keep the heart beating
• Two groups: Major minerals – Trace minerals

Two Groups: Major and Trace Minerals

• Major Minerals (macrominerals)

− Need more than 100 mg/day.
− Min of 5 grams in the body.
− These Include:
o Calcium
 o Phosphorus
o Potassium
o Sulfur
o Sodium
o Chloride
− The major minerals are the 6 dietary minerals your body needs in the largest amounts.

Calcium

• Total content of calcium in the body is more than 1200 mg.

• 99% of total content is deposit in bones and teeth,
• 1% in blood and body fluids
• Intracellular calcium:
o cytosol
o mitochondria
o other microsomes
o regulated by "pumps"
• The serum level of calcium is closely regulated with a normal total calcium of 2 -2.75 mmol/L (9-10.5 mg/dL) and a normal ionized calcium of
1.1-1.4 mmol/L (4.5-5.6 mg/dL).
• Food sources: Milk, cheese, sardines, salmon, some dark green leafy vegetables
• Functions:
o Development of bones and teeth
o Transmission of nerve impulses
o Blood clotting
o Normal heart action
o Normal muscle activity
• Deficiency/Toxicity:
o Osteoporosis - means “porous bones” - characterized by low bone density or mass (reduced amount of bone tissue) and fragile
bones
o Osteomalacia - bones lose calcium and become softer and may deform
o Rickets - softening of bones in children potentially leading to fractures and deformity
o Tetany - involuntary contraction of muscles
o Retarded growth
o Poor tooth and bone formation
o hypocalcemia tetany
o increased neuromuscular excitability
o neurological disoders
o Result of vit. D deficiency
o Hypoparathyroidism
o renal insufficiency.
• Recommended daily amount:
• Children to age 11 – 1200 mg/day
• From age 11 to 24 – 800 mg/day
• From age 24 – 500 mg/day I
• Symptoms are: rickets (children), osteomatacia (adults)
• Toxicity: – hypercalcemia (normally does not to occur) Hyperparathyroidism, vitamin D intoxication, cancer.

Phophorus

• Phosphorous is a widely distributed in the body.

• Body distribution: About 80% of phosphorous is found in bones & teeth in combination with calcium. About 15% of phosphorous is present in soft
tissues. In the soft tissues most phosphate is organic, as a component of phospholipids, phosphoproteins, nucleic acids & nucleoproteins • About
1% is found in ECF. In the ECF phosphorous is present in the inorganic form, about 5g in brain & 2g in blood is found. Phosphorous is present in the
form of organic & inorganic
• Plasma phosphorous is 2.5 to 4.5 mg/dl in adults. In children’s it is about 5.0 to 6.0 mg/dl
• Food sources: Milk, cheese, lean meat, poultry, fish, whole-grain cereals, legumes, nuts
• Functions:
o Development of bones and teeth
o Maintains normal pH of the blood
o Constituent of all body cells
o CHO, CHON and fat metabolism
• Deficiency/Toxicity:
o Poor tooth and bone formation
o Weakness
o Anorexia
o General malaise
• DEFICIENCY:
o Rickets in children, osteomalacia in adults.
o Abnormalities in erythrocytes, leucocytes, platelets, liver.
o Depletion of phosphate occurs as a result of diminished absorption from intestine or excessive wasting through kidney. o
Hyperphosphatemia is associated with renal diseases.
o Hypophosphataemia -serum inorganic phosphate concentration <2.5 mg/dl .
o Hypothyroidism: Low PTH decreases phosphate excretion by the kidney and leads to high serum concentration. • Symptoms:
Increased serum phosphate levels causes decrease in serum calcium concentration; therefore tetany & seizures may be the presenting
symptoms.

Sodium

• Na+ is the major cation of extracellular fluid.

• Plasma concentration - 135 -145 meq/L
• ICT concentration - 3-10 mmol/L.
• Maintaining of total body fluid homeostasis and water balance.
• Decrease in blood pressure and decreases in sodium concentration result in the production of renin → aldosteron production → decreases the
excretion of sodium in the urine
• Food sources: table salt, beef, eggs, poultry, milk, cheese, Soy sauce, moderate amounts in breads and vegetables, large amounts in processed
foods
• Functions:
o Maintenance of fluid balance
o Transmission of nerve impulses
o Acid-base balance
o Muscle contraction
• Deficiency: nausea exhaustion muscle cramps
• Toxicity: hypertension edema
• Disease States:
o Hyponatremia
− Decreased sodium levels
− Diarrhea, vomiting, Chronic renal failure, Adisons disease (adreno cortical insufficiency), mild headache, moderate and severe, low
blood pressure & circulatory failure
− Overhydration, administration, of salt free fluids to patient
o Hypernatremia
− Increasing sodium levels
− Crushing’s syndrome
− Prolonged administration of steroid hormones (cortisone, ACTH, sex hormones)
− Severe dehydration (only water) as in case of diabetes insipidus -> increased blood volume -> hypertension

Potassium

• K+ is the principal cation of the intracellular fluid.

• Plasma concentration – 3.5 - 5meqlL.
• ICF concentration - 110 -160 mmol/L.
• Key role of K+ in skeletal and smooth muscle contraction
• The main dietary source is the cellular material we consume as foodstuffs.
• Sources: Oranges, bananas, dried fruits, vegetables, legumes, milk, cereals, meat
• Functions:
o Contraction of muscles
o Maintenance of fluid balance
o Transmission of nerve impulses
o Regular heart rhythm
o Cell metabolism
• Deficiency: hypokalemia muscle weakness confusion abnormal heartbeat
• Toxicity: hyperkalemia potentially life-threatening irregular heartbeats
• Disease states:
o Hypokalemia
− Serum K+ less than 3 meq/L
− Crushing syndrome
− Renal tubular acidosis
− Metabolic alkalosis, diarrhea & vomiting,
− In diabetic coma treatment, with insulin & glucose diuretics
− Muscle weakness, tachycardia, cardiac arrest
o Hyperkalemia
− Renal failure
− Addisons disease
− Severe dehydration
− Intravenous administration of fluids with excessive potassium salts
− Depression of CNS
− Bradycardia

Chlorine

• Intake, output and metabolism of sodium and chloride run in parallel.

• The homeostasis of sodium, potassium, and chloride are interrelated.
• Chloride is important in the formation of hydrochloric acid in gastric juice.
• Chloride ions are also involved in chloride shift.
• Chloride concentration in plasma is 96-106mEq/L.
• In CSF, it is about 125mEq/L.
• Chloride concentration in CSF is higher than any body fluids. Since CSF protein content is low. •
Chloride is increased to maintain Donnan membrane equilibrium.
• Food sources: Table salt, eggs, seafood, milk
• Function:
o Gastric acidity
o Regulates acid-base balance in the body
o Maintains fluid and electrolyte balance
o Formation of hydrochloric acid
• Deficiency/Toxicity:
o Imbalance in gastric acidity Imbalance in blood pH
o Nausea Exhaustion
• Deficiency:
o Hyperchloremia
− Is seen in
− Dehydration
− Cushing’ syndrome. Mineralocorticoids cause increased reabsorption from kidney tubules.
− Severe diarrhoea leads to loss of bicarbonate and compensatory retention of chloride.
− Renal tubular acidosis.
o Hypochloremia
− Excessive vomiting. HCl is lost, so plasma chloride is lowered. There will be compensatory increase in plasma bicarbonate. This is
called hypochloremic alkalosis.
− Excessive sweating.
− In Addison’s disease, aldosterone is diminished, renal tubular reabsorption of chloride is decreased, and more chloride is
excreted.
• Chloride channels
o The CFTR (Cystic Fibrosis Transmembrane Conductance Receptor) chloride conducting channel is involved in cystic fibrosis. o
In Cystic Fibrosis, a point mutation in the CFTR gene results in defective chloride transport.
o So water moves out from lungs and pancreas.

Magnesium

• Nearly 99% of the total body magnesium is located in bone or the intracellular space.
• Second plentiful cation of the extracellular fluids.
• Mg2+ is a cofactor of all enzymes involved in phosphate transfer reactions utilizing ATP and other nucleotide triphosphates as substrate. •
Required for the structural integrity of numerous intracellular proteins and nucleic acids.
• A substrate or cofactor for important enzymes such as adenosine triphosphatase, guanosine triphosphatase, phospholipase C, adenylate
cyclase, and guanylate cyclase.
• A required cofactor for the activity of over 300 other enzymes.
• A regulator of ion channels; an important intracellular signaling molecule.
• A modulator of oxidative phosphorylation.
• Food source: Green, leafy vegetables Whole grains, avocados, nuts, milk, legumes, bananas, nuts, seafood, chocolate, cocoa •
Function:
o Nerve transmission
o Synthesis of ATP
o Activation of metabolic enzymes
o Muscle activity
o Constituent of bones, muscles, and RBCs
• Deficiency and toxicity: Normally unknown Mental, emotional and muscle disorders
• Hypomagnesemia cause:
o changes in skeletal and cardiac muscle
o changes in neuromuscular function,
o hyperirritability, psychotic behaviour
o tetany
• Hypermagnesemia cause:
o muscle weakness
o hypotension
o ECG changes
o sedation and confusion
• Hypermagnesemia is usual due to renal insuficiency.

Trace Minerals

Iron Fluoride Molybdenum Boron Zinc Copper Arsenic Cobalt Iodine Manganese Nickel
Selenium Chromium Silicon

Iron
• Ferrous (Fe++ ) – Reduced
• Ferric (Fe+++ ) – Oxidized
• Chief functions in the body
o Part of the protein hemoglobin, which carries oxygen in the blood
o Part of the protein myoglobin in muscles, which makes oxygen available for muscle contraction
o Necessary for the utilization of energy as part of the cells’ metabolic machinery
 • Iron Sources – two forms of iron in food
1. Heme iron – meats, poultry, fish; readily absorbed
2. Nonheme iron – plants foods; less absorbed
a. Absorption enhanced by Vitamin C
b. Absorption diminished by phytates, oxalates, and tannic acid
• Food Sources: Muscle meat Poultry Shellfish Liver Legumes Dried fruits Whole grain or enriches breads and cereals Dark green and leafy
vegetables Molasses
• Functions:
o Transports oxygen and CO2
o Hemoglobin formation
o Component of cellular enzymes essential for energy production
• Deficiency: iron deficiency anemia
o Iron deficiency is the most common nutrient deficiency worldwide
o Can occur if inadequate intake or blood loss
o Females are prone due to menstruation, lower iron intake & pregnancy
o Preschoolers also at risk due to high milk diets
• Deficiency symptoms
o Microcytic, hypochromic anemia (low Hgb & Hct): weakness, fatigue, headaches
o Impaired work performance and cognitive function
o Impaired immunity
o Pale skin, nailbeds, mucous membranes, and palm creases
o Concave nails
o Inability to regulate body temperature
o Pica (craving for ice, clay, paste, and other non-food substances)
• Toxicity
• “iron overload”; more common in men
o Hemochromatosis – genetic defect
o Hemosiderosis – chronic ingestion of too much iron; more common in alcoholics
o Iron poisoning – acute toxicity from overdose of iron supplements; can be deadly in small children
• Toxicity symptoms
o GI distress
o Iron overload: infections, fatigue, joint pain, skin pigmentation, organ damage

Zinc

• Essential nutrient
• Better absorption from animal source
• Deficiencies cause growth retardation and poor sexual development
• Chief functions in the body
o Part of many enzymes, esp. enzymes for transfer of carbon dioxide
o Associated with the hormone insulin
o Involved in making genetic material and proteins
o Important to wound healing, taste perception, reproduction (the making of sperm), vision (transports and activates Vit. A) &
immune function
• Food Sources: Seafood, esp. oysters Liver Eggs Milk Wheat bran legumes
• Functions: Formation of collagen Wound healing Taste acuity Essential for growth Immune reactions
• Deficiency/Toxicity: Dwarfism Anemia Loss of appetite Skin changes Impaired wound healing Decreased taste acuity • Deficiency symptoms – In
children, growth retardation and delayed sexual maturation – Impaired immune function – Hair loss – Eye and skin lesions – Loss of appetite and
abnormal taste – Depressed immune function and poor wound healing - Night blindness • Zinc Absorption and Metabolism
o Zinc Transport
− Transported by the protein albumin
− Binds to transferrin
− Excessive iron and copper can lead to a zinc deficiency and excessive zinc can lead to an iron and copper deficiency.

Selenium

• An integral component of glutathion peroxidase (intracellular oxidant),

• A scavenger of peroxides,
• An essential element for immune function (selenoproteins)
• Selenoproteins catalyse oxido-reduction reactions, protective function from oxidative stress (macrophage or neutrophil-generated free-radical
species, UV in sunlight)
• The food content of Se is highly dependent upon the soil type in which the foodstuff are grown
• Readily absorbed
• Excreted through the urine and feces
• Co-factor for glutathione peroxidase
• Protects the heart and other cells from oxidative damage
• Works together with vitamin E
• Cancer prevention
• Deficiency of Selenium • Muscle pain • Muscle wasting • Cardiomyopathy

Iodine
• Found in an ion form, iodide
• Used for thyroid hormone synthesis
• Regulates metabolic rate, growth, development
• Thyroid gland enlarges (goiter) with low intake of iodide
• Cretinism is the stunting of fetal growth and mental development as a result of low iodine diet
• Chief functions in the body
o A component of two thyroid hormones that help to regulate growth, development, and metabolic rate
o 2001 RDA for adults: 150 µg/day
o Upper level: 1100 µg/day
• Significant sources –Iodized salt, seafood, bread, dairy products, plants grown in iodine- rich soil and animals fed those plants (content of
iodine in soils affects food content)
• Deficiency disease
o Simple goiter-enlarged thyroid; affects 200 million people worldwide
o Creatinism- severe mental and physical retardation in infants
• Deficiency symptoms – Underactive thyroid gland, goiter
• Toxicity symptoms – Underactive thyroid gland, elevated TSH, goiter (also causes enlarged thyroid gland)

Copper

• Aids in iron metabolism

• Absorption dependent on body’s needs
• Absorption decreased with high intakes of vitamin C, phytic acid, fiber, zinc, iron, certain amino acids
• Functions of Copper
o Increases iron absorption
o Formation of connective tissue
o In superoxide dismutase
o Immune system, blood clot, brain development, cholesterol metabolism
• Food Sources of Copper • Organ meats • Seafood • Cocoa • Mushroom, legumes, seeds, nuts, whole-grain • RDA is 900 ug/day for adults •
Daily Value is set at 2000 ug • Average intake is about or slightly below the RDA
• Deficiency of Copper • Anemia • Decrease WBC • Bone loss • Inadequate growth • Pre-term infants at risk • cardiovascular disease

Fluoride

• Role in prevention of cavities

• Resists acid and dental caries
• Fluoride inhibits bacterial growth that may cause cavities
• Fluoridated water (1ppm)
• “Food” Sources of Fluoride: Fluoridate water (~0.2 mg/cup) • Tea • Seafood, seaweed • Toothpaste • Adequate intake is 3.1 -3.8 mg/day for
adults • Typical fluoridated water contains 0.2 mg/cup
• Toxicity of Fluoride • Fluorosis • Mottling of the teeth in children • Limit toothpaste to pea size for children • In high amounts can weaken
teeth in children • Upper Level is 10 mg/day

Manganese

• Chief functions in the body

− Cofactor for several enzymes
− Component of bones and glands
• 2001 AI – Men: 2.3 mg/day – Women: 1.8 mg/day
• Upper level for adults: 11 mg/day
• Significant sources – Nuts, whole grains, leafy vegetables, tea
• Deficiency symptoms – Rare; requirements are low
• Toxicity symptoms – Nervous system disorders – Has occurred in miners inhaling large quantities of dust causes brain damage

NUCLEIC ACID

• Biopolymers/large biomolecules.
• Essential for life.
− Includes DNA and RNA
− Made from monomers.
• Known As nucleotides.
• Named because,
− First found in the nucleus of cells,
− Later on, discovered outside the nucleus.

History of Nucleic acid

− In 1869, Friedrich Miescher - Friedrich Miescher who discovered nucleic acids in 1871
− 1944, Avery, MacLeod and McCarty
• Expressed it as DNA & found as the genetic material.
 • By using bacterial DNA to transform the genetic material of other bacteria.

Composition of Nucleic Acids

• Linears polymers of nucleotides.

• Each nucleotide consists of 3 components:
− Purine or pyrimidine nucleobase (nitrogenous base)
− Pentose sugar
− Phosphate group

TYPES of NUCLEIC ACID

DNA and RNA

• Despite their different jobs, DNA and RNA have almost the same basic structure. • Each
one is made up of nucleotides.
− These tiny subunits are made of a sugar, nitrogen base, and phosphate group.

Structure of DNA

• DNA is a macromolecule made up of two antiparallel strands of bonded deoxyribonucleotides. • A

deoxyribonucleotide consists of
− a phosphate group,
− 5-carbon deoxyribose sugar, and
− one of DNA's four nitrogenous bases: thymine, cytosine, guanine, or adenine. • RNA
is a single-stranded molecule that consists of bonded ribonucleotides. • Similar to the
nucleotides found in DNA, ribonucleotides consist of a
− phosphate group,
− a 5-carbon ribose sugar and
− one of RNA's four nitrogenous bases: uracil, cytosine, guanine, or adenine.

What Are the Differences Between DNA and RNA?

Function of DNA (deoxyribonucleic acid)

• Permanent storage place for genetic information.

• Controls the synthesis of RNA (ribonucleic acid)
• The sequence of nitrogenous bases in DNA
• determines the protein development in new cells.
• The function of the double helix formation of DNA is to ensure that no disorders occur.
• This is because the second identical strand of DNA that runs anti- parallel to the first is a backup in case of lost or destroyed genetic
information.
• Examples: Down’s syndrome or sickle cell Anemia.

Functions of RNA (ribonucleic acid)

• Synthesized by DNA for the transportation of genetic information to the protein building apparatus in the cell. •
Synthesis of new proteins using the genetic information it has transported.
• mRNA (messenger ribonucleic acid) is used to transfer genetic information through plasma membranes
• RNA also directs the nucleic acid (specially DNA) carry out a vital role in the human body.
• In particular, nucleic acids play an essential role in:
A. Mitosis, Meiosis
B. Providing energy/cellular respiration.

TYPES of RNA

• mRNA (messenger) used as template to make proteins; carries instructions for polypeptide synthesis from nucleus to ribosomes in the
cytoplasm
• rRNA (ribosomal) makes up ribosomes; forms an important part of both subunits of the ribosome
• tRNA (transfer) matches amino acids to mRNA to help make proteins; carries amino acids to the ribosome and matches them to the coded
mRNA message.

Pharmaceutical Importance of Nucleic Acid

• Used in cancer, cardiovascular and autoimmune diseases.

• The pharmaceutical perspectives of Nucleic acid based therapy presents a comprehensive account of gene therapy. • Internationally
acclaimed scientists discuss the potential use of lipid, peptides and polymers for the vivo delivery of nucleic acids. • Control gene
regulation, transcription, translation and replication.

Gene Therapy

• It is a technique for the correction of a genetic deficiency in a cell by the addition of new DNA to the cell. This definition has been expanded to
include treatments of acquired diseases by the addition of new DNA.
• There are four approaches:
− A normal gene inserted to compensate for a nonfunctional gene.
− An abnormal gene traded for a normal gene
− An abnormal gene repaired through selective reverse mutation
− Change the regulation of gene pairs
• Gene Therapy Strategies
− Gene Augmentation Therapy (GAT)
− Gene inhibition therapy.
− Gene replacement therapy o Gene Correction (Chimeraplasty)
− Targeted killing of specific cells o Prodrug therapy
o Somatic gene therapy involves introducing a “good “gene into targeted cells with the end results of treating the patient-not the future
children
o Germline gene therapy involves modifying the genes in egg or sperm cells, which will then pass any genetic changes to future
generations as well

Somatic gene therapy

1. Ex vivo – cells removed from the body, incubated with vector and gene-engineered cells returned to body. 2. In
situ – vector is placed directly into the affected tissues
3. In vivo – vector injected directly into the blood stream
Application of gene therapy

• Cancers
• Inherited disorders
• Infectious diseases (viral or bacterial)
• Immune system disorders
• Vaccination
• Diabetes
• AIDS Cancer
• Parkinson’s Disease

Cancer Approaches

• Delivery of genes encoding toxic molecules to cancer cells to kill them.

• Delivery of genes encoding chemokines to cancer cells to activate the immune response to recognize and kill them. •
Antibody therapies: DNA vaccines with genes that encode antibodies to cancer specific proteins in tumor cells. • Insertion
of normal tumor suppressor genes into cells.
• Antisense therapy: DNA that blocks synthesis of proteins encoded by deleterious genes.

Problems with gene therapy

• Short Lived.
• Hard to rapidly integrate therapeutic DNA into genome
• Would have to have multiple rounds of therapy.
• Immune Response.
• new things introduced leads to immune response.
• increased response when a repeat offender enters.
• Viral Vectors.
• patient could have toxic, immune, inflammatory response.
• also may cause disease once inside.
• Multi gene Disorders.
• Heart disease, high blood pressure, Alzheimer’s, arthritis and diabetes are hard to treat because you need to introduce more than one gene.

Requirement for Gene therapy

• Understanding of the disease process.

• Structure/function of gene to be introduced.
• Efficient delivery of gene.
• Control of gene expression.
• Prevention/control of immune responses.
• Animal model and assessment of function.
• Clinical trial.

Vocabulary Review:

• Monomer: Any molecule that can react with other molecules of the same or different makeup to form a chain, or polymer

• Polymer: A chain of monomers made of a specific type of compound

• Macromolecule: A polymer that has a large (macro) molecular mass. There are four main types in organisms: carbohydrates, proteins, lipids, and
nucleic acids
• Nucleotides: The monomers of nucleic acids. Made up of a nitrogen group, a phosphorus group, and a 5-carbon ring
• Nucleic Acid: Complex polymer that stores information in a cell in the form of a code. DNA and RNA

IMMUNOCHEMISTRY

is the study of the chemistry of the immune system.

This involves the study of the properties, functions, interactions and production of the chemical components
(antibodies/immunoglobulins, toxin, epitopes of proteins like CD4, antitoxins, cytokines/chemokines, antigens) of the
immune system, immune responses and determination of immune materials by immunochemicals.

immunochemistry is the study of the identities and functions of the components of the immune system.

Immunochemistry is also used to describe the application of immune system components, in particular antibodies, to
chemically labelled antigen molecules for visualization.

One of the earliest examples of immunochemistry is the Wasserman test to detect syphilis.

Svante Arrhenius was also one of the pioneers in the field; he published Immunochemistry in 1907 which described
the application of the methods of physical chemistry to the study of the theory of toxins and antitoxins.
Various methods in immunochemistry have been developed and refined, and used in scientific study, from virology to
molecular evolution.

Immunochemical techniques include:

a.enzyme-linked immunosorbent assay

b. immunoblotting (e.g., Western blot assay)

c. precipitation and agglutination reactions

d. immunoelectrophoresis

e. immunophonotyping

f. immunochromatographic assay

g. cyclometry.

Immunochemistry is also studied from the aspect of using antibodies to label epitopes of interest.

Immunochemistry (IC)

The use of an antibody and subsequent visualization of a chemical reaction that produces a color change in order to
detect a target. ( cells)

Immunohistochemistry (IHC)

The use of an antibody and subsequent visualization of a chemical reaction that produces a color change in order to
detect a target in tissue.

Immunofluorescence (IF)

The use of an antibody and subsequent visualization using a fluorophore in order to detect a target.

Immunohistofluorescence (IHF)

The use of an antibody and subsequent visualization using a fluorophore in order to detect a target in tissue.

Immunocytofluorescence (ICF)

The use of an antibody and subsequent visualization using a fluorophore in order to detect a target in cells.

VARIOUS IMMUNOCHEMISTRY TECHNIQUES:

Immunochemical techniques - identification of a substance (especially a protein) by its action as an antigen.

An antibody, also known as an immunoglobulin, is a large Y-shaped protein used by the immune system to identify and
neutralize foreign objects such as bacteria and viruses.

The antibody recognizes a unique part of the foreign target, termed an antigen.

Each tip of the "Y" of an antibody contains a paratope (a structure analogous to a lock) that is specific for one
particular epitope (similarly analogous to a key) on an antigen, allowing these two structures to bind together with
precision Antigen and Antibody

Enzyme-linked immunosorbent assay (ELISA)

is one sub-type of solid-phase enzyme immunoassay. The term enzyme immunoassay includes all assay based on the
measurement of enzyme labeled antigen or antibody.

Two basic types :

homogeneous : EMIT (enzyme multiplied immunoassay technique).

It is used only for drug identification.

Heterogeneous : ELISA (Enzyme-linked immunosorbent assay)

it is used to detected the antigen or antibody .

ELISA is named because the technique involves the use of an immunosorbent- an absorbing material specific for one of
the components of the reaction, the antigen or antibody.

Agglutination (from latin ‘agglutino’ – to glue, to attach)

is an immunochemical technique in which a specific antibody reacts with the corpuscular antigen

Agglutination reaction is based on the formation of bridges between bivalent(IgG) or multivalent (IgM) antibodies and
antigenic particles with multiple epitopes.

Bivalency or multivalency of the used antibody and multiple antigenic determinants on the surface of particles are
necessary for the creation of cross-linking and the formation of a high-molecular-weight lattice that is observable
macroscopically.
Definition of terms:

Hemagglutination is a variant of agglutination technique in which red blood cells are used as the antigen bearing
particles.

Agglutination reactions are performed on slides, in test tubes or microtiter plates. They are more sensitive in
comparison with immunoprecipitation methods. The agglutination methods produce qualitative or semiquantitative
results.

Immunoprecipitation

Technique of precipitating protein antigens out of a solution using a specific antibody which is immobilized to a solid
support.

Widely used method for protein isolation from complex samples such as cell lysates, serum and tissue homogenates.

Applications:

1. to measure the molecular weight of the given protein.

2. to determine post translational modifications.

3. to analyze the expression level of a protein of interest in the study of protein -protein as well as protein- nucleic acid
interactions.

Immunoelectrophoresis

is a qualitative method that combines protein electrophoresis with immunodiffusion.

It is performed in two steps.

The first one involves the separation of antigens according to their charges/size in an electrical field.

The second step, a suitable antiserum (polyspecific or monospecific) is applied to grooves running parallel to the
electrophoresis migration zone.

Immunofixation

Immunofixation is a method used for the detection and isotyping of monoclonal immunoglobulins in serum, urine and
cerebrospinal fluid.

Two stages of Immunofixation.

a. serum proteins are separated by electrophoresis.

b. the monoclonal immunoglobulins are identified by means of immunoprecipitation with specific antibodies.

Precipitation methods in solution:

The precipitate, which shapes in an agarose gel, can also form in a solution : immunoturbidimetry and
immunonephelometry.

Turbidimetry and nephelometry: When a diluted antigen solution is combined with a solution of the corresponding
antibody, the formation of small aggregates (immunoprecipitates) results in turbidity (cloudy appearance) of the
solution.

Two approaches can be used to quantify this turbidity:

Turbidimetry is based on the measurement of intensity of light transmission; i.e., light that passes through the cuvette
is measured. Such measurement can be performed on a conventional spectrophotometer.

Nephelometry measures directly the intensity of scattered light. Nephelometry requires a special apparatus – the
nephelometer, which uses laser as the light source .

Immunoassay

Immunoassay is a biochemical method that identifies and quantifies (protein, lipid, nucleic acid etc.) in solution
(serum, urine etc.) using antibody-antigen reactions.

There are many different formats and variations of an immunoassay, but the key point is still specific antibody- antigen
recognition.

Competitive binding/ Competitive enzyme immunoassay

Competition binding assays are commonly used to measure the binding affinity of a ligand with its receptor. In such an
assay, the binding of a ligand labeled with a fluorescent or radioactive tag is typically measured at a single
concentration in the presence of varying concentrations of an unlabeled, competing ligand.
Non-competitive enzyme immunoassay (sandwich methods)

This kind of enzyme immunoassay can be adopted for measurement of either antigens or antibodies. It is a
heterogeneous immunoassay using a solid phase coated with antibody or antigen, which must always be in excess over
the analyte being measured.

It has been used extensively for detection of serum antibodies to viruses and parasites in serum, autoantibodies, and
IgE antibodies specific for a particular allergen.

C-Reactive Protein:

is a protein produced in the liver and found in the blood.

Plasma protein that appears in the blood stream during inflammation.

It is also classified as acute phase protein.

synthesized by the liver in response to the release of macrophages ( white blood cells that digest foreign substances,
cellular debris and cancer cells.

C-Reactive Protein can test:

a. cancer of lymph nodes

b. hearty diseases

c. rheumatoid arthritis

d. osteomyelitis

e. coronary heart disease risks

Spectrophotometric Techniques

is a standard and inexpensive technique to measure light absorption or the amount of chemicals in a solution.

It uses a light beam which passes through the sample, and each compound in the solution absorbs or transmits light
over a certain wavelength. The instrument used is called a spectrophotometer.

Immunofluorescence assay

is a technique allowing the visualization of a specific protein or antigen in tissue sections by binding a specific antibody
chemically conjugated with a fluorescent dye such as fluorescein isothiocyanate (FITC).

The specific antibodies are labeled with a compound (FITC) that makes them glow an apple-green color when observed
microscopically under ultraviolet light.

Fluorescence is the property of certain molecules or fluorophores to absorb light at one wave length and emit light at
longer wave length (emission wavelength) when it is illuminated by light of a different wavelength (excitation
wavelength)

Immunoelectronmicroscopy

Immuno-electron microscopy is used to localize molecules at the ultrastructural level by labeling them with specific
antibodies. The antibodies are visualized by electron-opaque markers (colloidal gold particles) attached to them.

Immunoelectron microscopy is one of the best methods for detecting and localizing proteins in cells and tissues. This
procedure can be used on practically every unicellular and multicellular organism, and often provides unexpected
insights into the structure-function associations.

Immunostaining

is any use of an antibody-based method to detect a specific protein in a sample.

The term "immunostaining" was originally used to refer to the immunohistochemical staining of tissue sections, as first
described by Albert Coons in 1941.

Immunostaining now encompasses a broad range of techniques used in histology, cell biology, and molecular biology
that use antibody-based staining methods.

Immunodetection

Immunodetection (immunological detection) is used to identify specific proteins blotted to membranes

is an invaluable technique for tagging cellular targets, usually via a two-step protocol comprising target labeling per se
with a primary antibody followed by a signal amplification with a secondary antibody conjugated to a fluorescent
reporter.
Immunoblotting

(western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies.

is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample
of tissue homogenate or extract.

IMMUNOGLOBULIN

Immunoglobulins also known as antibodies, are glycoprotein molecules produced by plasma cells (white blood cells).
They act as a critical part of the immune response by specifically recognizing and binding to particular antigens, such as
bacteria or viruses, and aiding in their destruction.

The various immunoglobulin classes and subclasses (isotypes) differ in their biological features, structure, target
specificity and distribution. Hence, the assessment of the immunoglobulin isotype can provide useful insight into
complex humoral immune response.

Assessment and knowledge of immunoglobulin structure and classes is also important for selection and preparation of
antibodies as tools for immunoassays and other detection applications.

The various antibodies produced by plasma cells are classified by isotype, each of which differs in function and antigen
responses primarily due to structure variability.

Five major antibody classes have been identified in placental mammals:

a. IgA

b. IgD

c. IgE,

d. IgG

e. IgM.

This classification is based on differences in amino acid sequence in the constant region (Fc) of the antibody heavy
chains.

IgG and IgA are further grouped into subclasses (e.g., in human IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) based on
additional small differences in the amino acid heavy chain sequences.

Based on differences in the amino acid sequence in the constant region of the light chain, immunoglobulins can be
further sub-classified by determination of the type of light chain (kappa light chain or lambda light chain).

A light chain has two successive domains: one constant domain and one variable domain. The ratio of these two light
chains differs greatly among species, but the light chains are always either both kappa or both lambda, never one of
each.

Determination of individual subclasses is relevant in assessing primary immunodeficiencies or immune responses,

especially if the total IgG or IgA concentration is not altered or elevated.

STRUCTURAL CHARACTERISTICS:

Heavy and Light Chains

All immunoglobulins have a four chain structure as their basic unit.

They are composed of two identical light chains (23kD) and two identical heavy chains (50- 70kD).

Disulfide bonds

Inter-chain disulfide bonds:

The heavy and light chains and the two heavy chains are held together by inter-chain disulfide bonds and by
non-covalent interactions.

The number of inter-chain disulfide bonds varies among different immunoglobulin molecules.

Intra-chain disulfide binds:

Within each of the polypeptide chains there are also intra-chain disulfide bonds.

Variable (V) and Constant (C) Regions

The variable regions are responsible for antigen binding.

the constant regions are responsible for various biologic functions eg, complement activation and binding to cell
surface receptors.

Hinge Region
This is the region at which the arms of the antibody molecule form a Y.

It is called the hinge region because there is some flexibility in the molecule at this point.

Domains

Three dimensional images of the immunoglobulin molecule show that it is not a straight molecule rather, it is folded
into globular regions each of which contains an intra-chain disulfide bond . These regions are called domains.

a. 1. Light Chain Domains - VL and CL 2.

b. Heavy Chain Domains - VH, CH1,CH2CH3 (or CH4)

Oligosaccharides

Carbohydrates are attached to the CH2 domain in most immunoglobulins.

However, in some cases carbohydrates may also be attached at other locations.

TYPES OF IMMUNOGLOBULIN AND ITS FUNCTIONS:

1. Immunoglobulin G (Ig G)

Most abundant class in serum.

Constitutes 80% total immunoglobulin.

Present in blood, plasma and tissue fluids.

Contains less carbohydrate than other immunoglobulins.

It has a half life of 23 days: the longest of all of the immunoglobulin isotypes

Crosses placenta and provide natural immunity to fetus and neonate at birth.

Acts against bacteria and viruses by opsonizing.

Neutralize toxin.

Activate complement by classical pathway.

Catabolism of IgG is unique in that it varies with its serum concentration.

Functions of Immunoglobulin G (Ig G)

1. IgG1, IgG3, IgG4 – cross placenta and protect foetus.

2. IgG3 activates complement.

3. IgG1 and IgG3 binds to Fc receptor on phagocytic cells, monocytes and macrophages and mediate opsinization.

2. Immunoglobulin A (Ig A)

Constitutes 10-15 % of total immunoglobulins.

Present in milk, saliva, tears, mucous of respiratory tract, digestive tract and genitourinary tract.

In serum exist as monomer.

In external secretions exist as dimer called secretory Immunoglobulin.

Has ‘J’ chain and secretory piece.

Half life: 6-8 days

Functions of Immunoglobulin A (Ig A)

1. Provides local immunity.

2. Secretory Ig A binds to surface antigens of microorganism and prevent its attachment and invasion of the mucosal
surfaces of respiratory and digestive tract- immune elimination.

3. Secretory IgA provides important line of defense against salmonella, Vibrio cholerae, N. gonorrhoeae, influenza virus
and poliovirus.

4. Secretory IgA present in breast milk protects newborn during first months of life.

5. Activates complement by the alternative pathway .

6. Promotes phagocytosis and intracellular killing of microorganisms.

3. Immunoglobulin M (Ig M)

Accounts for 5-10% of total serum proteins.

Polymer of five monomeric units (pentamer)

Held together by disulfide bonds and ‘J’ chain

Half life: 5 days

Most of IgM (80%) present intravascularly.

Present in low concentration in intercellular tissue fluids

Cannot cross placenta.

Presence of IgM antibody in serum of newborn indicate congenital infection.

Earliest immunoglobulin to be synthesized by foetus (20 weeks).

First immunoglobulin to be produced in primary response to antigen.

Relatively short-lived hence it’s demonstration in the serum indicates recent infection.

Monomeric IgM appears on the surface of unstimulated B lymphocytes and act as receptors for antigens.

Functions of Immunoglobulin M (Ig M)

1. it agglutinates bacteria

2. Activates complement by classical pathway.

3. Causes opsonization and immune heamolysis.

4. Believed to be responsible for protection against blood invasion by microorganisms.

4. Immunoglobulin E (Ig E)

Structure is similar to Ig G.

Has 4 constant region domains.

Mostly present extra cellularly.

Does not cross placenta

Produced in the lining of respiratory and intestinal tract. known as reagin antibody.

Does not activate complement nor agglutinate antigens.

Functions of Immunoglobulin E (Ig E)

> Responsible for symptoms of anaphylactic shock, hay fever and asthma.

> Play a role in immunity against helminthic parasites

> The physiological role of IgE appears to be protection against pathogens by mast cell degranulation and release of
inflammatory mediators.

Mediates P.K. reaction (PRAUSNITZ & KUSTNER)

The presence of a serum component responsible for allergic reaction was first demonstrated by Prausnitz and Kustner
in 1921.

Kustner was suffering from atopic hypersensitivity to certain species of fish. Kustner’s serum was injected
intracutaneously in Prausnitz. After 24 hrs small quantity of cooked fish antigen was injected at the same site. A wheal
and flare reaction occurred within minutes.

5. Immunoglobulin D (Ig D)

Structure is similar to IgG .

Constitutes 0.2% of total immunoglobulins

Half life: 3 days

IgD together with IgM is major membrane bound immunoglobulin on unstimulated B lymphocytes-acts as recognition
receptors for antigens

Role of different immunoglobulin classes:

1. IgG: Protects the body fluids

2. IgA: Protects the body surfaces

3. IgM: Protects the blood stream

4. IgE: Mediates type I hypersensitivity

5. IgD: Role not known

AUTOIMMUNE DISORDERS

AN AUTOIMMUNE DISORDER OCCURS WHEN THE BODY'S IMMUNE SYSTEM ATTACKS AND DESTROYS HEALTHY BODY
TISSUE BY MISTAKE. THERE ARE MORE THAN 80 TYPES OF AUTOIMMUNE DISORDERS.

THE BLOOD CELLS IN THE BODY'S IMMUNE SYSTEM HELP PROTECT AGAINST HARMFUL SUBSTANCES. EXAMPLES
INCLUDE BACTERIA, VIRUSES, TOXINS, CANCER CELLS, AND BLOOD AND TISSUE FROM OUTSIDE THE BODY. THESE
SUBSTANCES CONTAIN ANTIGENS. THE IMMUNE SYSTEM PRODUCES ANTIBODIES AGAINST THESE ANTIGENS THAT
ENABLE IT TO DESTROY THESE HARMFUL SUBSTANCES.

WHEN ONE HAS AN AUTOIMMUNE DISORDER, THE IMMUNE SYSTEM DOES NOT DISTINGUISH BETWEEN HEALTHY
TISSUE AND POTENTIALLY HARMFUL ANTIGENS. AS A RESULT, THE BODY SETS OFF A REACTION THAT DESTROYS
NORMAL TISSUES.

THE EXACT CAUSE OF AUTOIMMUNE DISORDERS IS UNKNOWN. ONE THEORY IS THAT SOME MICROORGANISMS
(SUCH AS BACTERIA OR VIRUSES) OR DRUGS MAY TRIGGER CHANGES THAT CONFUSE THE IMMUNE SYSTEM. THIS
MAY HAPPEN MORE OFTEN IN PEOPLE WHO HAVE GENES THAT MAKE THEM MORE PRONE TO AUTOIMMUNE
DISORDERS.

AN AUTOIMMUNE DISORDER MAY RESULT IN: THE DESTRUCTION OF BODY TISSUE, ABNORMAL GROWTH OF AN
ORGAN, CHANGES IN ORGAN FUNCTION. AN AUTOIMMUNE DISORDER MAY AFFECT ONE OR MORE ORGAN OR
TISSUE TYPES. AREAS OFTEN AFFECTED BY AUTOIMMUNE DISORDERS INCLUDE: BLOOD VESSELS, CONNECTIVE
TISSUES, ENDOCRINE GLANDS SUCH AS THE THYROID OR PANCREAS, JOINTS, MUSCLES, RED BLOOD CELLS AND SKIN

ACCORDING TO A 2014 STUDY, WOMEN GET AUTOIMMUNE DISEASES AT A RATE OF ABOUT 2 TO 1 COMPARED TO
MEN — 6.4 PERCENT OF WOMEN VS. 2.7 PERCENT OF MEN. OFTEN THE DISEASE STARTS DURING A WOMAN’S
CHILDBEARING YEARS (AGES 15 TO 44). SOME AUTOIMMUNE DISEASES ARE MORE COMMON IN CERTAIN ETHNIC
GROUPS. FOR EXAMPLE, LUPUS AFFECTS MORE AFRICAN-AMERICAN AND HISPANIC PEOPLE THAN CAUCASIANS.

CERTAIN AUTOIMMUNE DISEASES, LIKE MULTIPLE SCLEROSIS AND LUPUS, RUN IN FAMILIES. NOT EVERY FAMILY
MEMBER WILL NECESSARILY HAVE THE SAME DISEASE, BUT THEY INHERIT A SUSCEPTIBILITY TO AN AUTOIMMUNE
CONDITION.BECAUSE THE INCIDENCE OF AUTOIMMUNE DISEASES IS RISING, RESEARCHERS SUSPECT
ENVIRONMENTAL FACTORS LIKE INFECTIONS AND EXPOSURE TO CHEMICALS.

A “WESTERN DIET” IS ANOTHER SUSPECTED RISK FACTOR FOR DEVELOPING AN AUTOIMMUNE DISEASE. EATING
HIGH-FAT, HIGH-SUGAR, AND HIGHLY PROCESSED FOODS IS THOUGHT TO BE LINKED TO INFLAMMATION, WHICH
MIGHT SET OFF AN IMMUNE RESPONSE. HOWEVER, THIS HASN’T BEEN PROVEN.

A 2015 STUDY FOCUSED ON ANOTHER THEORY CALLED THE HYGIENE HYPOTHESIS. BECAUSE OF VACCINES AND
ANTISEPTICS, CHILDREN TODAY AREN’T EXPOSED TO AS MANY GERMS AS THEY WERE IN THE PAST. THE LACK OF
EXPOSURE COULD MAKE THEIR IMMUNE SYSTEM PRONE TO OVERREACT TO HARMLESS SUBSTANCES.

CLASSIFICATION OF AUTOIMMUNE DISORDERS.

HAEMOLYTIC AUTOIMMUNE DISEASES

A CLINICAL DISORDER DUE TO DESTRUCTIONS OF BLOOD COMPONENTS. AUTO AB ARE FORMED AGAINST ONE’S
OWN RBCS, PLATELETS OR LEUCOCYTES.

E.G. HAEMOLYTIC ANAEMIA, LEUCOPENIA, THROMBOCYTOPENIA , ETC.

LOCALIZED AUTOIMMUNE DISEASES OR ORGAN SPECIFIC AUTOIMMUNE DSE > A PARTICULAR ORGAN IS AFFECTED
DUE TO AUTO ABS.

FOR EXAMPLE:

1. THYROIDITIS (ATTACKS THE THYROID)

2. MULTIPLE SCLEROSIS (ATTACKS MYELIN COATING OF NERVE AXON

3. MYASTHENIA GRAVIS (ATTACKS NERVE-MUSCLE JUNCTION)

4. JUVENILE DIABETES OR TYPE I DM (ATTACKS INSULIN-PRODUCING CELLS)

SYSTEMIC AUTOIMMUNE DSE OR NON-ORGAN SPECIFIC AUTOIMMUNE DSE > IMMUNE COMPLEXES ACCUMULATE IN
MANY TISSUES AND CAUSE INFLAMMATION AND DAMAGE. AFFECTS MANY ORGANS OR THE WHOLE BODY .

FOR EXAMPLE:

1. SYSTEMIC LUPUS ERYTHEMATOSUS (ANTI-NUCLEAR AB.): HARMS KIDNEYS, HEART, BRAIN, LUNGS, SKIN…

2. RHEUMATOID ARTHRITIS (ANTI-IGG ANTIBODIES): JOINTS, HEARTS, LUNGS, NERVOUS SYSTEM…

3. RHEUMATIC FEVER : CROSS-REACTION BETWEEN ANTIBODIES TO STREPTOCOCCUS AND AUTO-ANTIBODIES.

COMMON AUTO IMMUNE DISEASES

THROMBOCYTOPENIA:

CHARACTERIZED BY LOW PLATELET COUNT DUE TO THE PRODUCTION OF ANTIPLATELET AB. (IGG TYPE)

MECHANISM: AN INTERACTION OF AB WITH BOUND DRUG OR NEW AG. CAUSES INTRAVASCULAR AGGLUTINATION OF
PLATELETS & CAN BE ELIMINATED BY PHAGOCYTIC CELLS.

GRAVES’ DISEASE (ANTI-THYROID STIMULATING HORMONE; ANTI-TSH)

IN GRAVES’ DISEASE, THE ANTIBODIES DO NOT DESTROY THE THYROID BUT ACT AS IF THEY ARE TSH (I.E., THEY BIND
AND ACTIVATE THE TSH RECEPTOR)

ADDISON’S DISEASE

IT IS DUE TO ADRENOCORTICAL DAMAGE & HENCE INSUFFICIENT SECRETION OF ADRENAL HORMONES. TISSUE
DAMAGE IS CAUSED BY AUTO AB AGAINST ZONA GLOMERULOSA CELLS OF ADRENAL CORTEX.

MYASTHENIA GRAVIS

IT IS CAUSED BY AUTO ANTIBODY AGAINST MUSCLE ANTIGEN & ACETYLCHOLINE RECEPTOR ANTIGEN. CHARACTERIZED
BY ↑ MUSCULAR WEAKNESS THAT MAKE ONE FATIGUE. SPEAKING, EATING & WALKING BECOME TIRED SOME.
EVENTUALLY DEATH FROM RESPIRATORY FAILURE.

ACETYLCHOLINE CANNOT BE PRODUCED, THUS NERVE IMPULSE CANNOT BE TRANSMITTED FROM THE NERVES TO THE
MUSCLES. HENCE THE NEUROMUSCULAR JUNCTION IS SEVERELY AFFECTED.

SYSTEMIC LUPUS ERYTHEMATOSUS (SLE)

IT IS A SKIN DISEASE DUE TO THE PRODUCTION OF ANTINUCLEAR FACTOR (ANF) OR ANTINUCLEAR AUTO AB.

ANF REACTS WITH THE BREAKDOWN PRODUCTS OF NUCLEI IN THE NORMAL WEAR & TEAR OF CELLS & FORM
IMMUNE COMPLEXES WHICH CAUSE THE TISSUE DAMAGE. IN THESE PATIENTS, LE CELL (A MATURE NEUTROPHIL)
APPEARS IN BLOOD & BONE MARROW.

CHARACTERISTICS OF SLE

1. MALAR RASH (SLE)

2. APPEARANCE OF BLOOD RED SPOTS OVER THE BRIDGE OF NOSE & CHEEKS. THE LESIONS TAKE THE SHAPE OF A
BUTTERFLY.

3. CONNECTIVE TISSUES OF THE SKIN, KIDNEY, HEART. SPEEL & BLOOD VESSELS ARE SEVERELY DAMAGED RESULTING IN
JOINT PAIN, FEVER & ANAEMIA.

4. GLOMERULONEPHRITIS DUE TO DEPOSITION OF IMMUNE COMPLEX IN THE GLOMERULUS REGION. IT IS A SYSTEMIC

DISEASE AFFECTING THE WHOLE BODY.

RHEUMATOID ARTHRITIS

IT IS A CHRONIC SYSTEMIC DISEASE OF THE JOINTS. CAUSED BY THE AUTO ANTIBODY OF IGM TYPE, CALLED AS
RHEMATOID FACTORS.

CHARACTERISTICS:

1. SYNOVIAL FLUID OF THESE PATIENTS CONTAIN INCREASED NO. OF T-CELLS & MACROPHAGES.

2. MARKED BY INFLAMMATORY CHANGES IN THE SYNOVIAL MEMBRANE & BY ATROPHY OF BONES.

3. IN LATER STAGE, DEFORMITY & ANKYLOSIS DEVELOPS.

HASHIMOTO’S THYROIDITIS

ATROPHY OF THYROID GLAND, WHICH RESULTS IN HYPOTHYROIDISM & DESTRUCTION OF THYROID FUN.

CHARACTERIZED BY GOITRE, ENLARGED THYROID GLAND, DEFICIENCY OF TH (THYROXIN) AND CAUSED BY AUTO AB OF
IGG & IGM TYPE AGAINST THE CONSTITUENTS OF THYROID GLAND (THYROID EPITHELIAL CELLS, COLLOID & NUCLEAR
COMPONENTS)

IT IS A T-CELL ASSOCIATED AUTO IMMUNE DISEASE.

Diagnosis of Auto immune disease

Diagnosed by clinical symptoms.

Confirmed by detecting the auto Ab in the serum of the patients.

Autoantibodies are demonstrated by immunoflurescent Ab test, haemagglutination, Complement fixation,

immunodiffusion, Radio immuno assay, etc.

Treatment

Some autoimmune diseases are treated with medications that alleviate specific symptoms.

Haemolytic anaemia: Treated with Vit B 12

Throtoxicosis: Treated with antithyroid drugs.

Myasthenia Gravis: Treated with Cholinesterase inhibitors

Rhemotoid Arthritis: Anti-inflammatory drugs

Lupes Erythematosus: Treated with immunosuppressive or antimitiotic drugs such as Corticosteoid, Cyclophosphamide
and azothioprine.
BIOCHEMISTRY OF HORMONES

INTRODUCTION:

Cells and tissues communicate with each other by releasing or responding to secreted substances that aim in altering
the specific function of a cell or tissue.

These messengers are termed hormones. This communication is required for the development and organization of
tissues, the control of growth, division and death and coordinating the diverse cellular activities.

This message system is termed the endocrine system.

The term hormone (hormacin/hormao-to excite) was first used by William M. Bayliss and his brother-in law Ernest H.
Starlng. In 1904, they showed that a chemical substances (secretin) from the intestine could stimulate the action f
pancreatic secretion. These substances were then called as “chemical messengers”.

Wenrt and Thimann (1937) defines a hormone “as a substance which produced in any part of an organism, is
transferred to another part and there influence a specific physiological process.

The tissue or organs where they are produces are called effectors and those where they exert their influence as
targets.

2 Types of Hormones based on their site of action:

Local hormones

➢ have specific local effects, whence their nomenclature.

➢ These may be exemplifies by acetylcholine, secretin, cholecystokinin, etc.

General hormones

➢ are secreted by specific endocrine glands and are transported in the blood that cause physiology actions at points
remote their place of origin.

➢ A few of the general hormones affects almost all cells pf the body, e.g., growth hormones (GH) and thyroid
hormones; whereas other general hormones however, affect specific tissues far more than other tissues. e.g.,
adrencorticotropin ( a hormone secreted from adenohypophysis and stimulating the adrenal cortex) and ovarian
hormones (affecting the uterine endometrium.

Classification of Hormones according to structures:

Peptide/Protein hormones:

➢ The structure of peptide hormones is that of a polypeptide chain (chain of amino acids).

➢ The peptide hormones include molecules that are short polypeptide chains, such as antidiuretic hormone and
oxytocin produced in the brain and released into the blood in the posterior pituitary gland. This class also includes
small proteins like growth hormones produced by the pituitary, and large glycoproteins such as follicle-stimulating
hormone produced by the pituitary.

➢ Secreted peptides like insulin are stored within vesicles in the cells that synthesize them. They are then released in
response to stimuli such as high blood glucose levels in the case of insulin.

➢ These hormones cannot pass through plasma membranes of cells; therefore, their receptors are found on the
surface of the target cells.

Steroid hormones or Lipid Hormone

➢ Most lipid hormones are derived from cholesterol and thus ara structurally similar to it. ➢ The primary class of
lipid hormones in humans is the steroid hormones. Chemically, these hormones are usually ketones or alcohols; their
chemical names will end in “-ol” for alcohols or“-one” for ketones.

➢ Examples of steroid hormones include estradiol, which is an estrogen, or female sex hormone, and testosterone,
which is an androgen, or male sex hormone.

Amino acid hormones

➢ The amino acid-derived hormones are relatively small molecules that are derived from the amino acids tyrosine
and tryptophan.

➢ If a hormone is amino acid-derived, its chemical name will end in “-ine”. Examples of amino acid-derived hormones
include epinephrine and norepinephrine, which are synthesized in the medulla of the adrenal glands, and thyroxine,
which is produced by the thyroid gland.

➢ The pineal gland in the brain makes and secretes melatonin which regulates sleep cycles.

Origin

Hormones

Functions

Hypothalamus
Stimulators inhibitors

Pituitary hormone regulation

Pituitary, anterior

ACTH, FSH, Growth Hormone, Prolactin, LH, TSH

Adrenal control, Gonad regulation, Growth regulation, milk production, Thyroid control

Pituitary, posterior

ADH, oxytocin

Water conservation, uterus contraction & milk ejection

Thyroid

Thyroxine

Metabolic rate control

Parathyroid

PTH, Calcitonin

Calcium regulation

Gut

Gut hormones

Food digestion

Pancreas

Insulin, Glucagon

Nutrient metabolism

Adrenals

Cortisol, Aldosterone, Epinephrine

Body preservation, salt preservation, and stress response

Ovaries

Estradiol, Progesterone

Female characteristics

Testes

Testosterone

Male characteristics

INSULIN

Classification: Protein Hormone

Gland: Pancreas

Activated by: The secretion of insulin may also be stimulated by certain amino acids, fatty acids, keto acids (products of
fatty acid oxidation), and several hormones secreted by the gastrointestinal tract. The secretion of insulin is inhibited
by somatostatin and by activation of the sympathetic nervous system.

Function/s: Hormone that regulates the level of sugar (glucose) in the blood. Insulin acts primarily to stimulate glucose
uptake by three tissues—adipose (fat), muscle, and liver— that are important in the metabolism and storage of
nutrients.

Target organ/s: liver, the skeletal muscle, and the adipose tissue.

Antagonist (if any): It is an enzyme system found in many body-tissues and is called insulinase..

Produced by the beta cells of the islets of Langerhans.

GLUCAGON

Classification: Protein Hormone

Gland: Pancreas

Activated by: Stimulated by the ingestion of protein.

Function/s: Glucagon plays a critical role in maintaining blood glucose concentrations during fasting and exercise. It
raises the concentration of glucose in the blood by promoting glycogenolysis, which is the breakdown of glycogen (the
form in which glucose is stored in the liver), and by stimulating gluconeogenesis, which is the production of glucose
from amino acids and glycerol in the liver.

Target organ/s: Liver

Antagonist (if any): Insulin

Produced by the alpha cells, found in the islets of Langerhans

ANTIDIURETIC HORMONE

Classification: Peptide Hormone

Gland: It's a hormone made by the hypothalamus in the brain and stored in the posterior pituitary gland.

Activated by: Secretion of antidiuretic hormone is also stimulated by decreases in blood pressure and volume,
conditions sensed by stretch receptors in the heart and large arteries

Function/s: ADH constantly regulates and balances the amount of water in your blood. Target organ/s: Kidneys

Antagonist (if any): Tolvaptan is a vasopressin receptor antagonist.

Adrenocorticotropic Hormone (ACTH)

Classification: Protein

Gland: Anterior pituitary gland

Activated by: Corticotrophin-releasing hormone.

Function/s: Adrenocorticotropic hormone (ACTH) plays a large role in how your body responds to stress. Stress
activates ACTH production and increases cortisol levels.

Target organ/s: Adrenal Cortex

Antagonist (if any): None

Other info: ACTH is released into the bloodstream and travels around the body. Production of ACTH is regulated by
corticotrophin-releasing hormone (CRH) from the hypothalamus and cortisol from the adrenal gland

Parathyroid Hormone (PTH)

Classification: Peptide

Gland: Parathyroid Glands

Activated by: Low calcium levels in the blood stimulate parathyroid hormone secretion.

Function/s: Parathyroid hormone regulates calcium levels in the blood, largely by increasing the levels when they are
too low. It stimulates the production of active vitamin D in the kidneys.

Target organ/s: kidneys, skeletal system, and intestine

Antagonist (if any): Current antagonists for the parathyroid hormone are N-terminally truncated or N-terminally
modified analogs of PTH (1-34) or PTHrP (1-34) and are thought to bind predominantly to the N-terminal extracellular
(N) domain of the receptor. Other info: A primary problem in the parathyroid glands, producing too much parathyroid
hormone causes raised calcium levels in the blood (hypercalcaemia) and this is referred to as primary

Calcitonin

Classification: Peptide Hormone

Gland: Thyroid Gland

Activated by: Calcitonin secretion is stimulated by increases in the serum calcium concentration and calcitonin
protects against the development of hypercalcemia. Calcitonin is also stimulated by gastrointestinal hormones such as
gastrin.

Function/s: Calcitonin is involved in helping to regulate levels of calcium and phosphate in the blood, opposing the
action of parathyroid hormone. This means that it acts to reduce calcium levels in the blood.

Target organ/s: Bone

Antagonist (if any): Calcitonin gene-related peptide (CGRP) receptor antagonists are a class of drug candidates that act
as antagonists of the calcitonin gene-related peptide receptor (CGRPR)

OTHER INFO: Calcitonin reduces calcium levels in the blood by two main mechanisms:

1. It inhibits the activity of osteoclasts, which are the cells responsible for breaking down bone. When bone is broken
down, the calcium contained in the bone is released into the bloodstream. Therefore, the inhibition of the osteoclasts
by calcitonin directly reduces the amount of calcium released into the blood.

2. It can also decrease the resorption of calcium in the kidneys, again leading to lower blood calcium levels.
NOREPINEPHRINE/EPINEPHRINE

Classification: Amine Hormone

Gland: Adrenal glands

Activated by: activated by a stressful event.

Function/s: Raise blood glucose level; increase rate of metabolism; constrict certain blood vessels.

Target organ/s: All tissues

Antagonist (if any): Tricyclic antidepressants, beta blockers, antipsychotics

OTHER INFO: Epinephrine and norepinephrine are very similar neurotransmitters and hormones. While epinephrine
has slightly more of an effect on your heart, norepinephrine has more of an effect on your blood vessels. Both play a
role in your body's natural fight-or-flight response to stress and have important medical uses as well.

Aldosterone (Mineralocorticoids)

Classification: Steroid Hormone

Gland: Adrenal Glands

Activated by: The release of aldosterone from the adrenal glands is regulated via the renin-angiotensin II-aldosterone
system.

Function/s: Aldosterone affects the body's ability to regulate blood pressure. The hormone causes the bloodstream to
re-absorb water with the sodium to increase blood volume. Indirectly, the hormone also helps maintain the blood's
pH and electrolyte levels.

Target organ/s: Kidney

Antagonist (if any): Eplerenone (Inspra), Spirinolactone (Aldactone)

OTHER INFO: Aldosterone is closely linked to two other hormones: renin and angiotensin, which create the
renin-angiotensin-aldosterone system. This system is activated when the body experiences a decrease in blood flow to
the kidneys, such as after a drop in blood pressure, or a significant drop in blood volume after a hemorrhage or
serious injury. Renin is responsible for the production of angiotensin, which then causes the release of aldosterone.

Cortisol

Classification: Steroid Hormone

Gland: Adrenal Gland

Activated by: Stimulated by the anterior lobe of the pituitary gland with ACTH.

Function/s: Cortisol can help control blood sugar levels, regulate metabolism, help reduce inflammation, and assist
with memory formulation. It has a controlling effect on salt and water balance and helps control blood pressure. In
women, cortisol also supports the developing fetus during pregnancy.

Target organ/s: liver, muscle, adipose tissue, and the pancreas.

Antagonist (if any): None

OTHER INFO: High cortisol levels can also contribute to changes in a woman's libido and menstrual cycle, even without
the presence of Cushing disease. Anxiety and depression may also be linked to high cortisol levels. Low cortisol levels
can cause a condition known as primary adrenal insufficiency or Addison disease.

Human Growth Hormone (hGH)

Classification: Peptide Hormone

Gland: Anterior Pituitary Gland

Activated by: Growth hormone levels are increased by sleep, stress, exercise, and low glucose levels in the blood. They
also increase around the time of puberty.

Function/s: Growth hormone acts on many parts of the body to promote growth in children. In adults, it does not
cause growth but it helps to maintain normal body structure and metabolism, including helping to keep blood glucose
levels within set levels.

Target organ/s: Thymus Gland

Antagonist (if any): Pegvisomant

OTHER INFO: excessive growth hormone for a long period of time produces a condition known as acromegaly, in which
patients have swelling of the hands and feet and altered facial features. Too little growth hormone (deficiency) results
in poor growth in children. In adults, it causes a reduced sense of wellbeing, increased fat, increased risk of heart
disease and weak heart, muscles, and bones.
Prolactin

Classification: Protein Hormone

Gland: Pituitary Gland

Activated by: Stimulation of the nipples and mammary gland, as occurs during nursing, leads to prolactin release.

Function/s: Stimulates milk production.

Target organ/s: Mammary Gland

Antagonist (if any): None

Other Info: One of the main regulators of the production of prolactin from the pituitary gland is the hormone called
dopamine, which is produced by the hypothalamus. Dopamine restrains prolactin production, so the more dopamine
there is, the less prolactin is released.

ESTROGEN

Classification: Steroid Hormone

Gland: Ovaries

Activated by: The stimulation for secretion of estrogen comes from the Luteinizing hormone (LH) from anterior
pituitary gland.

Function/s: Estrogen is responsible for female physical features and reproduction. Estrogen helps bring about the
physical changes that turn a girl into a woman. Estrogen helps control the menstrual cycle and is important for
childbearing.

Target organ/s: uterus, ovary, breast, bone marrow and brain

Antagonist (if any): Antiestrogens, also known as estrogen antagonists or estrogen blockers.

OTHER INFO: Estrogen also has other functions include: keeps cholesterol in control, protects bone health for both
women and men, and affects your brain (including mood), bones, heart, skin, and other tissues.

ANDROGEN

Classification: Steroid Hormone

Gland: Testes, ovaries, adrenal glands

Activated by: Insulin resistance leads to an increase in insulin secretion (hyperinsulinemia), which is thought to
stimulate ovarian androgen production.

Function/s: Androgens don’t just impact our sexual health; they also play a role in our metabolism, our insulin
sensitivity, and possibly our body composition. Androgens may also impact bone density and cardiovascular health in
cis-gender women

Target organ/s: Heart

Antagonist (if any): Antiandrogens, also known as androgen antagonists or testosterone blockers

OTHER INFO: The most well-known is probably testosterone

Luteinizing Hormone (LH)

Classification: Protein Hormone

Gland: Anterior Pituitary Gland

Activated by: The hypothalamus secretes GnRH in a pulsatile fashion, which triggers FSH and LH release from the
anterior pituitary.

Function/s: In men, luteinizing hormone stimulates Leydig cells in the testes to produce testosterone, which acts
locally to support sperm production. In women, luteinizing hormone carries out different roles in the two halves of the
menstrual cycle. The fine tuning of luteinizing hormone release is vital to maintaining fertility.

Target organ/s: Testes and Ovaries

Antagonist (if any): luteinizing hormone-releasing hormone antagonist OTHER INFO: Luteinizing hormone, like follicle
stimulating hormone, is a gonadotrophic hormone produced and released by cells in the anterior pituitary gland. It is
crucial in regulating the function of the testes in men and ovaries in women.

OXYTOXIN:

Classification: Peptide Hormone

Gland: Hypothalamus

Activated by: Induced by activation of sensory nerves in the skin, which are activated by touch, warmth, and stroking
in connection with skin-to-skin contact with the baby and, also by massage-like hand movements performed by the
baby.

– Function/s: Oxytocin is a hormone that acts on organs in the body (including the breast and uterus) and as a
chemical messenger in the brain, controlling key aspects of the reproductive system, including childbirth and lactation,
and aspects of human behavior. Oxytocin stimulates the uterine muscles to contract, and also increases production of
prostaglandins, which increase the contractions further. In the brain, oxytocin acts as a chemical messenger and has
been shown to be important in human behaviors including sexual arousal, recognition, trust, anxiety, and
mother–infant bonding.

– Target organ/s: Uterine smooth muscle Mammary gland

– Antagonist (if any): tocolytics

– Other Info:Oxytocin has been called the 'love hormone' or 'cuddle chemical'. Oxytocin is controlled by a positive
feedback mechanism where release of the hormone causes an action that stimulates more of its own release. A lack
of oxytocin in a nursing mother would prevent the milk-ejection reflex and prevent breastfeeding

Follicle-Stimulating Hormone (FSH)

Classification: Protein Hormone

Gland: Pituitary Gland

Activated by: The hypothalamus secretes GnRH in a pulsatile fashion, which triggers FSH and LH release from the
anterior pituitary.

Function/s: It regulates the functions of both the ovaries and testes. Follicle stimulating hormone is one of the
hormones essential to pubertal development and the function of women’s ovaries and men’s testes. In women, this
hormone stimulates the growth of ovarian follicles in the ovary before the release of an egg from one follicle at
ovulation. It also increases oestradiol production. In men, follicle stimulating hormone acts on the Sertoli cells of the
testes to stimulate sperm production (spermatogenesis).

Target organ/s: Ovaries and Testes

Antagonist (if any): A substance that blocks the pituitary gland from

making hormones called follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Other info: Follicle
stimulating hormone regulates testosterone levels and when these rises, they are sensed by nerve cells in the
hypothalamus so that gonadotrophin-releasing hormone secretion and consequently follicle stimulating hormone is
decreased. Lack or insufficiency of it can cause infertility or subfertility both in men and women.

Progesterone

Classification: Steroid Hormone

Gland: Ovaries

Activated by: Luteinizing hormone and follicle-stimulating hormone, which are produced by the pituitary gland,
promote ovulation, and stimulate the ovaries to produce estrogen and progesterone.

Function/s: It plays important roles in the menstrual cycle and in maintaining the early stages of pregnancy. It may also
be involved in the growth of certain cancers.

Target organ/s: Uterus

Antagonist (if any): Antiprogestin

Other InfO: Progesterone prepares the tissue lining of the uterus to allow the fertilized egg to implant and helps to
maintain the endometrium throughout pregnancy. During the early stages of pregnancy, progesterone is still produced
by the corpus luteum and is essential for supporting the pregnancy and establishing the placenta. If progesterone is
absent or levels are too low, irregular, and heavy menstrual bleeding can occur.

Testosterone

– Classification: Steroid Hormone

– Gland: Testes, Ovaries, and Adrenal Glands

– Activated by: LH stimulates the release of testosterone from the Leydig cells in the testes.

– Function/s: It plays a key role in reproduction and the maintenance of bone and muscle strength. Testosterone
initiates the development of the male internal and external reproductive organs during fetal development and is
essential for the production of sperm in adult life.

– Target organ/s: Reproductive Organs

– Antagonist (if any): Androgen antagonists or testosterone blockers

– OTHER ONFO: This hormone also signals the body to make new blood cells, ensures that muscles and bones stay
strong during and after puberty and enhances libido both in men and women. Testosterone is linked to many of the
changes seen in boys during puberty (including an increase in height, body and pubic hair growth, enlargement of the
penis, testes and prostate gland, and changes in sexual and aggressive behavior).

Thyroxine
Classification: Amine Hormone

Gland: Thyroid gland

Activated by: The hypothalamus secretes thyrotropin-releasing hormone which, in turn, stimulates the pituitary gland
to produce thyroid stimulating hormone. This hormone stimulates the production of the thyroid hormone, thyroxine.

Function/s: It plays vital roles in digestion, heart and muscle function, brain development and maintenance of bones.

Target organ/s: Most tissues, heart, muscles, brain, bones

Antagonist (if any): Antithyroid agent

OTHER INFO: The production and release of thyroid hormone thyroxine is controlled by a feedback loop system that
involves the hypothalamus in the brain and the pituitary and thyroid glands.

Triiodothyronine

Classification: Amine Hormone

Gland: Thyroid Gland

Activated by: The hypothalamus secretes thyrotropin-releasing hormone which, in turn, stimulates the pituitary gland
to produce thyroid stimulating hormone. This hormone stimulates the production of the thyroid hormone
triiodothyronine, by the thyroid gland.

Function/s: Triiodothyronine is a thyroid hormone that plays vital roles in the body's metabolic rate, heart and
digestive functions, muscle control, brain development and function, and the maintenance of bones.

Target organ/s: Most tissues, heart, digestive organs, muscles, bones

Antagonist (if any): Bisphenol A

OTHER INFO: The production and release of thyroid hormone triiodothyronine, is controlled by a feedback loop
involving the hypothalamus, pituitary gland and thyroid gland

Thyroid-Stimulating Hormone (TSH)

Classification: Protein Hormone

Gland: Pituitary Gland

Activated by: The hypothalamus, in the base of the brain, produces thyrotropin-releasing hormone (TRH). TRH
stimulates the anterior pituitary gland to produce TSH.

Function/s: Its role is to regulate the production of hormones by the thyroid gland. It controls production of the
thyroid hormones, thyroxine and triiodothyronine, by the thyroid gland by binding to receptors located on cells in the
thyroid gland.

Target organ/s: Thyroid Gland

Antagonist (if any): None

OTHER INFO: If a person has too little thyroid stimulating hormone, it is most likely that their thyroid gland is making
too much thyroid hormone, that is, they have an overactive thyroid or hyperthyroidism, which is suppressing the
thyroid stimulating hormone.

Cortisone (Glucocorticoids)

Classification: Steroid Hormone

Gland: Adrenal Gland

Activated by: ACTH binds to its receptor on the adrenals and induces glucocorticoid production and release.

Function/s: It plays a complex role in regulating body functions and is essential for survival. Cortisone is a
corticosteroid hormone (glucocorticoid). It decreases your body's natural defensive response and reduces symptoms
such as swelling and allergic-type reactions. Cortisone is active primarily as an agent for the rapid conversion of
proteins to carbohydrates (a glucocorticoid) and to some extent regulates the salt metabolism of the body (a
mineralocorticoid).

Target organ/s: All tissue

Antagonist (if any): None

OTHER INFO: Cortisol is also needed for the fight or flight response, which is a healthy, natural response to perceived
threats. The amount of cortisol produced is highly regulated by your body to ensure the balance is correct.

Adrenaline (or epinephrine)

Classification: Amine Hormone

Gland: Adrenal Gland

Activated by: Adrenaline is released mainly through the activation of nerves connected to the adrenal glands.

Function/s: Its major action, together with noradrenaline, is to prepare the body for 'fight or flight'. Key actions of
adrenaline include increasing the heart rate, increasing blood pressure, expanding the air passages of the lungs,
enlarging the pupil in the eye (see photo), redistributing blood to the muscles and altering the body’s metabolism, so
as to maximize blood glucose levels (primarily for the brain).

Target organ/s: Heart, all tissues

Antagonist (if any): Adrenergic antagonists (adrenoblockers)

OTHER INFO: Most people are exposed to stressful situations on occasion and so most of us are familiar with the
typical symptoms of adrenaline release, such as: rapid heartbeat, high blood pressure, anxiety, weight loss, excessive
sweating, and palpitations. However, this is a normal response of the body which is intended to help us respond to a
stressful situation; once the acute stress is over, the symptoms quickly disappear as adrenaline hyper-secretion stops

Noradrenalin (Norepinephrine)

Classification: Amine Hormone

Gland: Adrenal Gland

Activated by: Sympathetic activation of the adrenal glands causes the part called the adrenal medulla to release
norepinephrine (as well as epinephrine) into the bloodstream.

Function/s: Norepinephrine increases heart rate and blood pumping from the heart. It also increases blood pressure
and helps break down fat and increase blood sugar levels to provide more energy to the body. In the brain,
norepinephrine plays a role in the sleep-wake cycle, helping you to wake up, in increasing attention and focusing on
performing a task, and in memory storage. It is also important for emotions.

Target organ/s: All tissues, Heart

Antagonist (if any): Antagonist binding to the α2 noradrenalinereceptor

induces noradrenaline release

OTHER INFO: Problems with norepinephrine levels are associated with depression, anxiety, post traumatic stress
disorder and substance abuse. Bursts of norepinephrine can lead to euphoria (very happy) feelings but are also linked
to panic attacks, elevated blood pressure, and hyperactivity.

Negative and Positive Feedbacks:

• The hormone levels in the blood are regulated by a highly specialized homeostatic mechanism called feedback.

• Information regarding the hormone level or its effect is fed back to the gland that the hormone secreted from.

• These terms positive and negative are not meant to denote a good or bad response, but rather the type of response
the system has to the presence of the effector.

• Due to positive and negative feedback, our body will be in homeostasis

Positive feedback mechanisms:

➢ It amplifies changes rather than reversing them.

➢ The release of oxytocin from the posterior pituitary gland during labor is an example of positive feedback
mechanism. Oxytocin stimulates the muscle contractions that push the baby through the birth canal. The release of
oxytocin result in stronger or augmented contractions during labor. The contractions intensify and increase until the
baby is outside the birth canal. When the stimulus to the pressure receptors ends, oxytocin production stops and
labor contractions cease.

> Positive feedback mechanisms control self-perpetuating events that can be out of control and do not require
continuous adjustment. In positive feedback mechanisms, the original stimulus is promoted rather than negated.

> Positive feedback increases the deviation from an ideal normal value.

• Negative Feedback

> the response will reverse or cause the opposite effect of the original stimulus.

> Negative feedback can be explained with the process of insulin production and release. After a meal the blood sugar
level will be elevated due to the absorption of sugars from the digestive tract.

This triggers the release of insulin from pancreas. Insulin converts sugar into cells and hence the blood sugar level
drops. This lowblood sugar level will result in the cessation of insulin release.

CLINICAL CHEMISTRY

• Clinical chemistry refers to the biochemical analysis of body fluids. It uses chemical reactions to determine the levels
of various chemical compounds in bodily fluids.

• Several simple chemical tests are used to detect and quantify different compounds in blood and urine, the most
commonly tested specimens in clinical chemistry.

• Techniques such as spectrophotometry, immunoassays, and electrophoresis are also used in clinical chemistry to
measure the concentration of substances such as glucose, lipids, enzymes, electrolytes, hormones, proteins, and
other metabolic products present in human blood and urine.
• When an individual test alone is not sufficient to assess a medical condition, a combination of several tests may be
used. The pattern of results from the combination of tests may provide better insight into the status of the patient
than any single test result. Such tests, done on the same sample, are often ordered as a group called a panel or
profile.

• Blood is the most common biologic fluid collected for clinical laboratory testing. It is usually drawn from a vein (in
the arm) directly into an evacuated tube. Typically a tube will hold about 5 mL of blood – enough to perform many
clinical chemistry tests, since automated analyzers require only small amounts (usually from 2 to 100 μL) for a single
test.

• Occasionally, when collection of blood from a vein is difficult, a sample of capillary blood may be collected by
pricking the skin and collecting several drops of blood from the puncture site. An example is the use of heelstick blood
for testing of newborns.

• Phlebotomy – the act of drawing a blood sample from a blood vessel. For clinical chemistry testing, blood is usually
drawn from a vein, typically a vein in the arm or back of the hand. Collecting blood from a vein is called venipuncture.
The medical professional drawing the blood sample is called a phlebotomist.

• Other biologic fluids (matrices) often used for testing include urine, saliva, cerebrospinal fluid (CSF), amniotic fluid,
synovial fluid, pleural fluid, peritoneal fluid and pericardial fluid.

• These fluids often contain the same biologic analytes of interest – such as glucose and protein – but differ greatly
from each other in physical and chemical properties.

• These differences in fluid characteristics are termed matrix differences. Test methods that are designed for
determination of an analyte in blood plasma may not be suitable for determination of that same analyte in other
fluids (other matrices).

• When using a test method for analysis of a fluid other than blood plasma or serum, it is important to validate that
the method is acceptable for the type of fluid sample being used.

• Knowing the different normal lab values is an important step in making an informed clinical decision as a nurse.
Diagnostic and laboratory tests are tools that provide invaluable insights and information about the patient. Lab tests
are used to help confirm a diagnosis, monitor an illness, and the patient’s response to treatment.

Phases of Diagnostic Testing

• Diagnostic testing involves three phases: pretest, intratest, and post-test. Nurses have responsibilities for each phase
of diagnostic testing.

Pretest - the main focus is on preparing the client for the diagnostic procedure.

Nursing responsibility:

a. Assessment of the patient to assist in determining precautions.

b. Preparation of the equipment and supplies needed.

c. Preparation of a consent form, if required.

d. Providing information and answering client questions about the procedure.

Intratest - the main focus is specimen collection and performing or assisting with certain diagnostic procedures,

Nursing responsibility:

a. Use of standard precautions or sterile technique if necessary.

b. Providing emotional support to the patient and monitoring the patient’s response during the procedure.

c. Ensuring the correct labeling, storage, and transportation of the specimen.

Post test - During the last part of diagnostic testing, the nursing care revolves around observations and follow-up
activities for the patient. For example, if a contrast media was injected during a CT scan, the nurse should encourage
the patient to increase fluid intake to promote excretion of the dye.

Nursing responsibility:

a. Compare the previous and current test results.

b. Reporting of the results to the appropriate members of the healthcare team.

Erythrocyte Studies Normal Lab Values

Red Blood Cells (RBC)

Red blood cells or erythrocytes transport oxygen from the lungs to the bodily tissues. RBCs are produced in the red
bone marrow, can survive in the peripheral blood for 120 days, and are removed from the blood through the bone
marrow, liver, and spleen.

Normal values for red blood cell count:

Male adult: 4.5 – 6.2 million/mm3

Female adult: 4.5 – 5.0 million/mm3

Indications of RBC count:

Helps in diagnosing anemia and blood dyscrasia.

Hemoglobin (Hgb)

Hemoglobin is the protein component of red blood cells that serves as a vehicle for oxygen and carbon dioxide
transport. It is composed of a pigment (heme) which carries iron, and a protein (globin). The hemoglobin test is a
measure of the total amount of hemoglobin in the blood.

Normal values chart for hemoglobin count:

Male adult: 14 – 16.5 g/dL

Female adult: 12 – 16 g/dL

Indications of Hemoglobin count:

a. Hemoglobin count is indicated to help measure the severity of anemia (low hemoglobin) or polycythemia (high
hemoglobin).

b. Monitor the effectiveness of a therapeutic regimen.

Hematocrit (Hct)

Hematocrit or packed cell volume (Hct, PCV, or crit) represents the percentage of the total blood volume that is made
up of the red blood cell (RBC).

Normal values for hematocrit count:

Male adult: 42 – 52%

Female adult: 35 – 47%

Red Blood Cell Indices

Red blood cell indicates (RBC Indices) determine the characteristics of an RBC. It is useful in diagnosing pernicious and
iron deficiency anemias and other liver diseases.

Mean corpuscular volume (MCV): The average size of the individual RBC.

Mean corpuscular hemoglobin (MCH): The amount of Hgb present in one cell.

Mean corpuscular hemoglobin concentration (MCHC): The proportion of each cell occupied by the Hgb.

Normal Lab Values for RBC Indices are:

Mean corpuscular volume (MCV):

Male: 78 – 100 μm3;

Female: 78 – 102 μm3

Mean corpuscular hemoglobin (MCH): 25 – 35pg

Mean corpuscular hemoglobin concentration (MCHC): 31 – 37%

Serum Iron (Fe)

Iron is essential for the production of blood helps transport oxygen from the lungs to the tissues and carbon dioxide
from the tissues to the lungs.

Normal lab values for serum iron:

Male adult: 65 – 175 mcg/dL

Female adult: 50 – 170 mcg/dL

Indication of serum iron:

Helps in diagnosing anemia and hemolytic disorder.

White Blood Cells and Differential

White Blood Cells (WBC)

White blood cells act as the body’s first line of defense against foreign bodies, tissues, and other substances. WBC
count assesses the total number of WBC in a cubic millimeter of blood. White blood cell differential provides specific
information on white blood cell types:

• Neutrophils are the most common type of WBC and serve as the primary defense against infection.

• Lymphocytes play a big role in response to inflammation or infection. • Monocytes are cells that respond to
infection, inflammation, and foreign bodies by killing and digesting the foreign organism (phagocytosis).

• Eosinophils respond during an allergic reaction and parasitic infections. • Basophils are involved during an allergic
reaction, inflammation, and autoimmune diseases.

• Bands are immature WBCs that are first released from the bone marrow into the blood.

Normal lab values for white blood cell count and WBC differential: WBC Count: 4,500 to 11,000 cells/mm³
Neutrophils: 55 – 70% or 1,800 to 7,800 cells/mm³

Lymphocytes: 20 – 40% or 1,000 to 4,800 cells/mm³

Monocytes: 2 – 8% or 0.0 to 800 cells/mm³

Eosinophils: 1 – 4% or 0.0 to 450 cells/mm³

Basophils: 0–2% or 0.0 to 200 cells/mm³

Bands: 0–2 % or 0.0 to 700 cells/mm³

Coagulation Studies

Platelets (PLT)

Platelets are produced in the bone marrow and play a role in hemostasis. Platelets function in hemostatic plug
formation, clot retraction, and coagulation factor activation.

Normal values for platelet count: 150,000 to 400,000 cells/mm³

Activated Partial Thromboplastin Time (APTT)

Activated partial thromboplastin time (APTT) evaluates the function of the contact activation pathway and coagulation
sequence by measuring the amount of time it requires for recalcified citrated plasma to clot after partial
thromboplastin is added to it. The test screens for deficiencies and inhibitors of all factors, except factors VII and XIII.

Normal lab value for activated partial thromboplastin time: 20 to 60 seconds, depending on the type of activator used.

Indication for APTT:

a. Monitors the effectiveness of heparin therapy.

b. Detect coagulation disorders in clotting factors such as hemophilia A (factor VIII) and hemophilia B (factor IX).

c. Determine individuals who may be prone to bleeding during invasive procedures.

Prothrombin Time and International Normalized Ratio (PT/INR)

Prothrombin is a vitamin K-dependent glycoprotein produced by the liver that is essential for fibrin clot formation.
Each laboratory establishes a normal or control value based on the method used to perform the PT test. The PT
measures the amount of time it takes in seconds for clot formation, the international normalized ratio (INR) is
calculated from a PT result to monitor the effectiveness of warfarin.

Indication for PT and INR

a. Monitor response to warfarin sodium (Coumadin) therapy.

b. Screen for dysfunction of the extrinsic clotting system resulting from vitamin K deficiency disseminated intravascular
coagulation or liver disease.

Normal Lab Value for Prothrombin Time (PT)

Normal: 11 – 13 seconds

Critical value: >20 seconds for persons who do not use anticoagulants.

The INR standardizes the PT ratio and is calculated in the laboratory setting by raising the observed PT ratio to the
power of the international sensitivity index specific to the thromboplastin reagent used.

Nursing Care for Prothrombin Time

• If a PT is prescribed, the baseline specimen should be drawn before anticoagulation therapy is started; note the time
of collection on the laboratory form.

• Provide direct pressure to the venipuncture site for 3 to 5 minutes. • Concurrent warfarin therapy with heparin
therapy can lengthen the PT for up to 5 hours after dosing.

• Diets high in green leafy vegetables can increase the absorption of vitamin K, which shortens the PT.

• Orally administered anticoagulation therapy usually maintains the PT at 1.5 to 2 times the laboratory control value.

• Initiate bleeding precautions, if the PT value is longer than 30 seconds in a client receiving warfarin therapy.

Bleeding Time

Bleeding time assess the overall hemostatic function (platelet response to injury and vasoconstrictive ability).

Indication for Bleeding Time

Useful in detecting disorders of platelet function.

• Normal Values for Bleeding Time

Duke method: 1 to 3 minutes

Ivy method: 3 to 6 minutes

D-Dimer Test

is a blood test that measures clot formation and lysis that results from the degradation of fibrin.

Indication of D-Dimer Test

a. Helps to diagnose the presence of thrombus in conditions such as deep vein thrombosis, pulmonary embolism, or
stroke.

b. Used to diagnose disseminated intravascular coagulation (DIC). c. Monitor the effectiveness of treatment.

Normal Lab Value for D-Dimer: < 500 ng/mL

Serum Electrolytes

Electrolytes are minerals that are involved in some of the important functions in our body. Serum electrolytes are
routinely ordered for a patient admitted to a hospital as a screening test for electrolyte and acid-base imbalances.

Serum Potassium (K+)

• Potassium is the most abundant intracellular cation that serves important functions such as regulate acid-base
equilibrium, control cellular water balance, and transmit electrical impulses in skeletal and cardiac muscles.

• Normal Values for Serum Potassium

• Potassium: 3.5 – 5.0 mEq/L

• Indications for Serum Potassium

• Evaluates cardiac function, renal function, gastrointestinal function, and the need for IV replacement therapy.

Serum Sodium (Na+)

• Sodium is a major cation of extracellular fluid that maintains osmotic pressure and acid-base balance, and assists in
the transmission of nerve impulses. Sodium is absorbed from the small intestine and excreted in the urine in amounts
dependent on dietary intake.

• Normal Lab Values for Serum Sodium

• 135-145 mEq/L

• Indications for Serum Sodium

• Determine whole-body stores of sodium, because the ion is predominantly extracellular

• Monitor the effectiveness of drug, especially diuretics, on serum sodium levels.

Serum Chloride (Cl-)

• Chloride is a hydrochloric acid salt that is the most abundant body anion in the extracellular fluid. Functions to
counterbalance cations, such as sodium, and acts as a buffer during oxygen and carbon dioxide exchange in red blood
cells (RBCs). Aids in digestion and maintaining osmotic pressure and water balance.

• Normal Lab Value for Serum Chloride (Cl-)

• 95 – 105 mEq/L

Serum Bicarbonate

• Part of the bicarbonate-carbonic acid buffering system and mainly responsible for regulating the pH of body fluids.

• Normal Lab Value for Serum Bicarbonate

• 22 to 29 mEq/L

• Nursing consideration for Serum Bicarbonate

• Ingestion of acidic or alkaline solutions may cause increased or decreased results, respectively.

Calcium (Ca+)

• Calcium (Ca+) is a cation absorbed into the bloodstream from dietary sources and functions in bone formation,
nerve impulse transmission, and contraction of myocardial and skeletal muscles. Calcium aids in blood clotting by
converting prothrombin to thrombin.

• Normal Lab Value for Calcium

• Total: 4.5 – 5.5 mEq/L (8.5 to 10.5 mg/dL)

• Ionized: 2.5 mEq/L (4.0 – 5.0 mg/dL) 56% of total calcium

Phosphorus (P)

• Phosphorus (Phosphate) is important in bone formation, energy storage and release, urinary acid-base buffering,
and carbohydrate metabolism. Phosphorus is absorbed from food and is excreted by the kidneys. High concentrations
of phosphorus are stored in bone and skeletal muscle.

• Normal Lab Value for Phosphorus

• 1.8 – 2.6 mEq/L (2.7 to 4.5 mg/dL)

Magnesium (Mg)

• Magnesium is used as an index to determine metabolic activity and renal function. Magnesium is needed in the
blood-clotting mechanisms, regulates neuromuscular activity, acts as a cofactor that modifies the activity of many
enzymes, and has an effect on the metabolism of calcium.

• Normal Magnesium Lab Value

• 1.6 to 2.6 mg/dL

Renal Function Studies Normal Lab Values

Serum Creatinine (Cr)

• Creatinine is a specific indicator of renal function. Increased levels of creatinine indicate a slowing of the glomerular
filtration rate.

• Normal Lab Value for Serum Creatinine

• 0.6 to 1.3 mg/dL

Blood Urea Nitrogen (BUN)

• Urea nitrogen is the nitrogen portion of urea, a substance formed in the liver through an enzymatic protein
breakdown process. Urea is normally freely filtered through the renal glomeruli, with a small amount reabsorbed in
the tubules and the remainder excreted in the urine. Elevated levels indicate a slowing of the glomerular filtration
rate.

• Normal Lab Value for Blood Urea Nitrogen

• 8 to 25 mg/dL

Glucose Studies Normal Lab Values

Fasting Blood Glucose

• Fasting blood glucose or fasting blood sugar (FBS) levels are used to help diagnose diabetes mellitus and
hypoglycemia. Glucose is a monosaccharide found in fruits and is formed from the digestion of carbohydrates and the
conversion of glycogen by the liver. Glucose is the main source of cellular energy for the body and is essential for brain
and erythrocyte function.

• Normal Lab Value for Glucose

Glucose, fasting: 70 – 110 mg/dL

Glucose, monitoring: 60 – 110 mg/dL

Glucose, 2-hr postprandial: < 140mg/dL

Glucose Tolerance Test (GTT)

• The glucose tolerance test (GTT) aids in the diagnosis of diabetes mellitus. If the glucose levels peak at higher than
normal at 1 and 2 hours after injection or ingestion of glucose and are slower than normal to return to fasting levels,
then diabetes mellitus is confirmed.

• Normal Lab Values for Glucose Tolerance Test (GTT)

• 70 – 110 mg/dL (baseline fasting)

• 110 – 170 mg/dL (30 minute fasting)

• 120 – 170 mg/dL (60 minute fasting)

• 100 – 140 mg/dL (90 minute fasting)

• 70 – 120 mg/dL (120 minute fasting)

Arterial Blood Gas (ABG) Normal Lab Values

• Arterial Blood Gases (ABGs) are measured in a laboratory test to determine the extent of compensation by the
buffer system. It measures the acidity (pH) and the levels of oxygen and carbon dioxide in arterial blood. Blood for an
ABG test is taken from an artery whereas most other blood tests are done on a sample of blood taken from a vein.
• Normal Lab Values for Arterial Blood Gases

• pH: 7.35 – 7.45

• HCO3: 22 – 26 mEq/L

• PCO2: 35 – 45 mmHg

• PaO2: 80 – 100 mmHg

Liver Function Tests Normal Lab Values

Alanine Aminotransferase (ALT)

Alanine Aminotransferase (ALT) test is used to identify hepatocellular injury and inflammation of the liver and to
monitor improvement or worsening of the disease. ALT was formerly known as serum pyretic transaminase (SGPT).

• Normal Lab Value for Alanine Aminotransferase (ALT) • Male: 10 to 55 units/L

• Female: 7 to 30 units/L

Aspartate Aminotransferase (AST)

Aspartate Aminotransferase (AST) test is used to evaluate a client with a suspected hepatocellular disease, injury, or
inflammation (may also be used along with cardiac markers to evaluate coronary artery occlusive disease). AST was
formerly known as serum glutamic oxaloacetic transaminase (SGOT).

• Normal Lab Value for Aspartate Aminotransferase (AST)

• Male: 10 – 40 units/L

• Female: 9 – 25 units/L

Bilirubin

Bilirubin is produced by the liver, spleen, and bone marrow and is also a by-product of hemoglobin breakdown. Total
bilirubin levels can be broken into direct bilirubin, which is excreted primarily via the intestinal tract, and indirect
bilirubin, which circulates primarily in the bloodstream. Total bilirubin levels increase with any type of jaundice; direct
and indirect bilirubin levels help differentiate the cause of jaundice.

• Normal Lab Values for Bilirubin

• Total bilirubin: 0.3 – 1.0 mg/dL

• Direct bilirubin (conjugated): 0.0 to 0.2 mg/dL

• Indirect bilirubin (unconjugated): 0.1 to 1 mg/dL

• Critical level: > 12 mg/dL

Albumin

Albumin is the main plasma protein of blood that maintains oncotic pressure and transports bilirubin, fatty acids,
medications, hormones, and other substances that are insoluble in water. Albumin is increased in conditions such as
dehydration, diarrhea, and metastatic carcinoma; decreased in conditions such as acute infection, ascites, and
alcoholism. Presence of detectable albumin, or protein, in the urine is indicative of abnormal renal function.

• Normal Lab Value for Albumin

• 3.4 to 5 g/dL

Ammonia

Ammonia is a by-product of protein catabolism; most of it is created by bacteria acting on proteins present in the gut.
Ammonia is metabolized by the liver and excreted by the kidneys as urea. Elevated levels resulting from hepatic
dysfunction may lead to encephalopathy. Venous ammonia levels are not a reliable indicator of hepatic coma.

• Normal Lab Value for Ammonia

• Adults: 35 – 65 mcg/dL

• Nursing Considerations

Instruct the client to fast, except for water, and to refrain from smoking for 8 to 10 hours before the test; smoking
increases ammonia levels. > Place the specimen on ice and transport to the laboratory immediately.

Amylase

Amylase is an enzyme, produced by the pancreas and salivary glands, aids in the digestion of complex carbohydrates
and is excreted by the kidneys. In acute pancreatitis, the amylase level may exceed five times the normal value; the
level starts rising 6 hours after the onset of pain, peaks at about 24 hours, and returns to normal in 2 to 3 days after
the onset of pain. In chronic pancreatitis, the rise in serum amylase usually does not normally exceed three times the
normal value.

• Normal Lab Values for Amylase

• 25 to 151 units/L

Lipase
 Lipase is a pancreatic enzyme converts fats and triglycerides into fatty acids and glycerol. Elevated lipase levels occur
in pancreatic disorders; elevations may not occur until 24 to 36 hours after the onset of illness and may remain
elevated for up to 14 days.

• Normal Lab Values for Lipase

• 10 to 140 units/L

Serum Protein

Serum protein reflects the total amount of albumin and globulins in the plasma. Protein regulates osmotic pressure
and is necessary for the formation of many hormones, enzymes, and antibodies; it is a major source of building
material for blood, skin, hair, nails, and internal organs. Increased in conditions such as Addison’s disease,
autoimmune collagen disorders, chronic infection, and Crohn’s disease. Decreased in conditions such as burns,
cirrhosis, edema, and severe hepatic disease.

• Normal Lab Value for Serum Protein:

• 6 to 8 g/dL

Lipoprotein Profile

Lipid assessment or lipid profile includes total cholesterol, high density lipoprotein (HDL), low-density lipoprotein (LDL),
and triglycerides.

Cholesterol is present in all body tissues and is a major component of LDL, brain, and nerve cells, cell membranes, and
some gallbladder stones.

Triglycerides constitute a major part of very-low-density lipoproteins and a small part of LDLs. Increased cholesterol
levels, LDL levels, and triglyceride levels place the client at risk for coronary artery disease. HDL helps protect against
the risk of coronary artery disease.

> Normal Lab Values for Lipid Profile Cholesterol: Less than 200 mg/dL

riglycerides: Less than 150 mg/dL HDLs: 30 to 70 mg/dL

LDLs: Less than 130 mg/dL

Cardiac Markers and Serum Enzymes

1. Creatine Kinase (CK)

Creatine kinase (CK) is an enzyme found in muscle and brain tissue that reflects tissue catabolism resulting from cell
trauma. The CK level begins to rise within 6 hours of muscle damage, peaks at 18 hours, and returns to normal in 2 to
3 days. The test for CK is performed to detect myocardial or skeletal muscle damage or central nervous system
damage. Isoenzymes include CK-MB (cardiac), CK-BB (brain), and CK-MM (muscles):

• CK-MM is found mainly in skeletal muscle.

• CK-MB is found mainly in cardiac muscle

• CK-BB is found mainly in brain tissue

Normal Lab Values for Creatinine Kinase and Isoenzymes

•Creatine kinase (CK)

• Male: 38 – 174 U/L

• Women: 26 – 140 U/L

• Creatinine kinase isoenzymes

• CK-MM: 95% – 100% of total

• CK-MB: 0% – 5% of total

• CK-BB: 0%

2. Myoglobin

Myoglobin, an oxygen-binding protein that is found in striated (cardiac and skeletal) muscle, releases oxygen at very
low tensions. Any injury to skeletal muscle will cause a release of myoglobin into the blood. Myoglobin rise in 2-4
hours after an MI making it an early marker for determining cardiac damage.

• Normal Lab Values for Myoglobin

• Myoglobin: 5–70 ng/mL

BIOCHEMISTRY OF AGING

• Aging is an extremely complex, multifactorial process, and numerous aging theories have been proposed—such
as—stochastic, developmental, genome–based, and free radical.

• The most important of these are the genomic and free radical theories—the genes influence aging and longevity. A
comparison of the percentage of life expectancy and maximum life span is graphically presented.
• The immune and neuroendocrine systems clearly deteriorate with age.

• Magnesium, zinc, and selenium are some essential elements for critical biochemical processes.

• The heat shock protein (HSP) system is a ubiquitous cell defense mechanism involved in the response to
acutely-induced cell damage by numerous processes.

• Certain age–associated disorders discussed are atherosclerosis, neoplasia, cataracts and macular degeneration,
neurodegenerative disorders, and neuroendocrine disorders.

• Aging is when an organism, such as a cell, has accumulated an excess amount of damage over its lifetime. The
resulting damage ends up having an effect on the overall survival and status of the organism.

• As an organism ages, “degradation of their outputs leads to functional decline and death as a result of aging” .

• This degradation of outputs is related to the metabolic history of the cell which affects the cell’s function.

• The concept of aging has led to the accepted idea that aging is due to the accumulation of damage an organism as
acquired over its lifetime resulting in the inability to protect, maintain, and repair itself.

• Experiments are being conducted that are trying to determine what kind of damage, related to aging, contributes to
the loss of function for an organism.

• This is difficult as there are many factors dealing with damage that can vary across models and individuals such as
amount of damage, type of tissue, age, and simply the kind of organism in question.

• To begin, exactly what does an organism have to do in order to survive?

• First, the organism must be able to find food, shelter, and to fight off infections or predators. The organism must be
able to avoid death. Even if these factors are eliminated, death and loss of function still occur with age.

• Nonetheless, no genes have evolved to cause death. An aging organism is said to reduce the genetic contribution of
an individual for the next generation. In other words, it is disadvantageous.

• In Murphy’s paper, “Control Theory of Aging,” he states that “the genome and how it is expressed constrain
mortality and life span.” However, there is still a problem with this idea for life span varies across all organisms, even if
they are genetically similar. It is concluded that it is indeed a combination of genetic determination, variation in the
environment, and other events that occur in the organism’s life that contribute to death at the age to which the
aforementioned factors can affect.

• As a result of a lifetime of low-dose exposure to external factors such as ultraviolet radiation and gamma-ray
irradiation, the body's ability to carry out homeostatic mechanisms begins to fail and aging becomes apparent. It also
has been proposed that aging is a reflection of cellular senescence, an irreversible halt in the cell's ability to
self-replicate and grow.

• An obstacle involves mutations and environmental interventions that hinder a number of functions which proves
difficult to see what the cause for aging is.

• As mentioned before, there can be a variety of reasons why an organism may have died.

• It is important to see that one must look across all the possible biochemical and physiological entities and to observe
each one independently. This will help narrow down the cause responsible.

• Aging

Gradual change in an organism that leads to increased risk of weakness, disease, and death over the entire adult life
span of any living thing.

there is a decline in biological functions and in ability to adapt to metabolic stress.

Changes in organs include:

a. reduced immunity

b. loss of muscle strength

c. decline in memory and cognition

d. loss of colour in the hair

e. elasticity in the skin.

• Gerontology is concerned with the changes that occur between maturity and death along with factors that influence
these changes.

• Geriatrics focuses on health care of elderly people and promote health by preventing and treating diseases and
disabilities in older adults.

Factors of Aging

• Factors of Aging

1. mitochondrial damage

2. free radicals

3. telomeres
4. apoptosis and necrosis

5. diseases

1. Mitochondria

main unit of chemical power supply

During the synthesis of macroergical bio- molecules(high energy releasing potentials e.g. ATP) free radicals are being
produced as the by-product.

Free radicals released in large quantities cause intercellular oxidative stress (e.g. oxidative damage of mitochondria) >
damaging mitochondria and cause early apoptosis

2. Free radical

A molecule that contains one or more unpaired electrons and is capable of independent existence.

Harmful effect of free radicals: Because of their reactive nature, free radical can provoke inflammation or altered
cellular function through:

a. Lipid peroxidation

> React with amino acid to modify protein structure & function.

> Can crosslink lipid in cell membrane interrupting structure & fluidity.

b. Protein modification

> Proteins are major targets of free radical attack because of their high abundance & responsible for most of functional
processes.

c. DNA modification

> Free radical induced DNA damage includes – strand break. – DNA protein crosslink. – large range of base &
sugar modification.

3. Telomeres

> Repetitive DNA sequences at the ends of all human chromosomes

> aging cells have shorter telomeres

> length differs between species ]

• in humans 8-14kb long

> Telomeres are thought to be the "clock" that regulates how many times an individual cell can divide.

> Telomeric sequences shorten each time the DNA replicates. > Once the telomere shrinks to a certain level, the cell
can no longer divide. Its metabolism slows down, it ages, and dies

• Apoptosis and Necrosis

• There are two ways that a cell can die:

4. Necrosis

> occurs when a cell is damaged by an external force, such as poison, a bodily injury, an infection or getting cut off
from the blood supply (which might occur during a heart attack or stroke).

> When cells die from necrosis, it's a rather messy affair. The death causes inflammation that can cause further distress
or injury within the body.

5. Apoptosis or programmed cell death

> When a cell is compelled to commit suicide proteins called caspases go into action.

> They break down the cellular components needed for survival, > production of enzymes known as DNases, which
destroy the DNA in the nucleus of the cell.

6. Senescence – The process of aging at the cell and organismal levels.

> The process by which a cell looses its ability to divide, grow, and function. This loss of function ultimately ends in
death. A degenerative process, only and has no positive features.

7. Resveratrol, a constituent of red wine and grape skins has been found to extend life span

Postulates of free radical theory of aging

In 1956s Dr. Denham Harman studied

The rate of living theory, which holds that lifespan is an inverse function of metabolic rate which in turn is proportional
to oxygen consumption.

Rebbeca Gershman's observation that oxygen toxicity and radiation toxicity could be explained by the same underlying
phenomenon: oxygen free radicals.

On basis of these, he proposed that - Cells continuously produce free radicals by normal metabolism and oxidation of
organic compounds and these free radicals damage cellular macromolecules ( like DNA , lipid , proteins ) and due
accumulation of these damage over time period cause aging.

Mitochondrial free radical theory of aging

In 1972s, Denham Harman modified his theory-

About 95% energy of a cell is produce in mitochondria and about 97% - 99% of oxygen is use in energy formation and
1%- 3% oxygen is utilize in reactive oxygen species formation.

It states that reactive oxygen species (ROS ) are produce in mitochondria during ETC cycle and they cause damage to
macromolecules including lipid , protein, DNA of mitochondria and nucleus which lead to aging .

Wear-and-Tear Theory

The wear and tear theory of aging was first introduced by Dr. August Weismann, a German biologist, in 1882.

It is very logical because it is what happens to the nonliving things that people observe around them (i.e. components
of an aging car break due to repeated use).

Cells and tissues have vital parts that wear out resulting in aging.

Parts of the body eventually wear out from repeated use killing the parts and then the body.

Body is similar to a machine, which loses function when its parts wear out. As people age, their cells, tissues and
organs are damaged by internal or external stressors. Good health maintenance practices will reduce the rate of wear
and tear, resulting in longer and better body function.

Premature aging symptoms

Progeria [Hutchinson-gilford syndrome]

Progeria is a disease characterized by normal development in the first year of life followed by rapid aging

It is caused by genetic defect in which telomeres are considerably shorter than normal

Symptoms include dry and wrinkled skin, total baldness, and birdlike facial features.

Death usually occurs around age 13

Werner syndrome

Is a rare, inherited disease that causes a rapid acceleration of aging, usually while the person is only in his or her
twenties.

It is characterized by wrinkling of the skin, graying of the hair and baldness, cataracts, muscular atrophy, and a
tendency to develop diabetes mellitus, cancer, and cardiovascular disease.

Most afflicted individuals die before age 50.

Cockayne syndrome (CS)

also called Neill-Dingwall syndrome, is a rare and fatal autosomal recessive neurodegenerative disorder characterized
by growth failure, impaired development of the nervous system, abnormal sensitivity to sunlight (photosensitivity),
eye disorders and premature aging.